WO2014081622A1 - Amylase with maltogenic properties - Google Patents
Amylase with maltogenic properties Download PDFInfo
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- WO2014081622A1 WO2014081622A1 PCT/US2013/070237 US2013070237W WO2014081622A1 WO 2014081622 A1 WO2014081622 A1 WO 2014081622A1 US 2013070237 W US2013070237 W US 2013070237W WO 2014081622 A1 WO2014081622 A1 WO 2014081622A1
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- amylase
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- starch
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01133—Glucan 1,4-alpha-maltohydrolase (3.2.1.133), i.e. maltogenic alpha-amylase
Definitions
- the present teachings provide composition and methods relating to novel maltogen amylases.
- Starch is a mixture of amylose (15-30% w/w) and amylopectin (70-85% w/w).
- Amylose is composed of linear chains of a-1 ,4-linked glucose units having a molecular weight (MW) from about 60,000 to about 800,000.
- MW molecular weight
- Amylopectin is a branched polymer containing a-1 ,6 branch points every 24-30 glucose units; its MW may be as high as 100 million.
- Sugars from starch in the form of concentrated dextrose syrups, are currently produced by an enzyme catalyzed process involving: (1 ) liquefaction (or viscosity reduction) of solid starch with an a-amylase into dextrins having an average degree of polymerization of about 7-1 0, and (2) saccharification of the resulting liquefied starch (i.e. starch hydrolysate) with amyloglucosidase (also called glucoamylase or GA).
- the resulting syrup has a high glucose content. Much of the glucose syrup that is commercially produced is subsequently enzymatically isomerized to a dextrose/fructose mixture known as isosyrup.
- the resulting syrup also may be fermented with
- microorganisms such as yeast
- yeast to produce commercial products including ethanol, citric acid, lactic acid, succinic acid, itaconic acid, monosodium glutamate, gluconates, lysine, other organic acids, other amino acids, and other biochemicals, for example.
- Fermentation and saccharification can be conducted simultaneously (i.e., an SSF process) to achieve greater economy and efficiency.
- Alpha-amylases hydrolyze starch, glycogen, and related polysaccharides by cleaving internal a-1 ,4-glucosidic bonds at random.
- Alpha amylases particularly from Bacilli, have been used for a variety of different purposes, including starch liquefaction and saccharification, textile desizing, starch modification in the paper and pulp industry, brewing, baking, production of syrups for the food industry, production of feedstocks for fermentation processes, and in animal feed to increase digestibility. These enzymes can also be used to remove starchy soils and stains during dishwashing and laundry washing.
- Maltose a di-saccharide composed of two D-glucopyranoses joined by a ⁇ -1 ,4'-glycosidic bond
- Maltose is also a substrate for production of the non-caloric sugar sweetener, maltitol.
- High purity maltose or pure maltose is an active component of intravenous injection liquids for diabetic patients.
- maltose high conversion or low maltose syrup
- 50-55% maltose high maltose syrup
- 70-75% maltose very high maltose
- >80% maltose ultra high maltose
- the first step is a liquefaction step at high temperature, >95 °C and the second step is a saccharification step.
- the second step is called malto-saccharification and usually takes place at a temperature at or below 60°C.
- the insoluble starch granules are slurried in water, gelatinized with heat and hydrolyzed by a thermostable alpha-amylase (EC.3.2.1 .1 , a-1 ,4'- D-glucan glucanohydrolase) from Bacillus species, often in the presence of added calcium.
- a thermostable alpha-amylase EC.3.2.1 .1 , a-1 ,4'- D-glucan glucanohydrolase
- Bacterial derived thermostable alpha-amylases from Bacillus licheniformis (for example, SPEZYME ® FRED from DuPont-Genencor or Termamyl ® L-120 from Novozymes), Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus licheniformis (for example, SPEZYME ® FRED from DuPont-Genencor or Termamyl ® L-120 from Novozymes), Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus stearothermophilus (for example SPEZYME XTRA from Bacillus stearothermophilus (for example SPEZYME XTRA from
- DuPont-Genencor, Termamyl ® SC, and Termamyl ® SUPRA from Novozymes) or blends of Bacillus licheniformis and Bacillus stearothermophilus (for example ClearflowTM AA from DuPont-Genencor or Liquozyme ® Supra from Novozymes) are used to first liquefy the starch at high temperature, >95°C at pH 5.2-6.5 to a low DE (dextrose equivalent) soluble starch hydrolysate.
- maltogenic enzymes such as a fungal alpha-amylase (for example, CLARASE ® L from
- DuPont-Genencor or Fungamyl ® 800L from Novozymes a plant beta-amylase (for example, OPTIMALT ® BBA from DuPont-Genencor or Betalase 1 500L from Senson) are used at a much lower temperature to further hydrolyse the soluble starch hydrolysate.
- a debranching enzyme like pullulanase for example OPTIMAX ® L-1 000 from DuPont-Genencor, Promozyme ® D2 from Novozymes or Promozyme ® D6 from Novozymes is added during
- the present teachings provide an isolated nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1 or of a degenerate variant of SEQ I D NO: 1 .
- the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3.
- the present teachings provide an isolated nucleic acid comprising a sequence that hybridizes under stringent conditions to a hybridization probe the nucleotide sequence of which consists of SEQ ID NO: 1 , or the complement of SEQ
- the present teachings provide an isolated nucleic acid comprising a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ ID NO: 1 .
- the present teachings provide such an isolated nucleic acid wherein the nucleic acid encodes a polypeptide that has starch hydrolysis activity.
- the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ ID NO: 3, wherein the polypeptide has starch hydrolysis activity.
- the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide comprising the sequence of SEQ ID NO: 3, or SEQ I D NO: 3 with up to 50 conservative amino acid substitutions, wherein the polypeptide has starch hydrolysis activity,
- the present teachings provide a purified polypeptide, the amino acid sequence of which comprises a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ I D NO: 3.
- the present teachings provide a purified polypeptide comprising the amino acid sequence of SEQ I D NO: 3, but with 0 to 20 conservative amino acid substitutions.
- the present teachings provide an expression vector comprising the nucleic acid sequence of any of claims 1 -6 operably linked to an expression control sequence. In some embodiments, the present teachings provide a cultured cell comprising such a vector. In some embodiments, the present teachings provide cultured cell comprising any of the nucleic acids of the present teachings, operably linked to an expression control sequence. In some embodiments, the present teachings provide a cultured cell tranfected with any of the vectors provided by the present teachings, or a progeny of said cell, wherein the cell expresses the nucleic acid to form a polypeptide.
- the present teachings provide a method of producing a protein, the method comprising culturing the cells provided by the present teachings under conditions permitting expression of the polypeptide.
- the present teachings provide a method of using the polypeptide of the present teachings, the method comprising including the polypeptide in any of: starch liquefaction, starch saccharification, textile desizing, starch modification in the paper and pulp industry, brewing, baking, production of syrups for the food industry, production of feedstocks for fermentation processes, animal feed, and, removal of starchy soils and/or stains during dishwashing and/or laundry washing.
- the present teachings provide a composition comprising the polypeptide of the present teachings, and at least one accessory enzyme selected from the group consisting of phytase, protease, pullulanase, ⁇ -amylase, isoamylase, a different amylase, alpha-glucosidase, cellulase, xylanase, hemicellulase,
- beta-glucosidase transferase, pectinase, lipase, cutinase, esterase, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases,
- phospholipases phospholipases, lysophospholipases, acyltransferases, perhydrolases, arylesterases, and redox enzymes.
- FIG. 1 panels A and B depict some illustrative data according to some embodiments of the present teachings.
- FIG 2 panels A and B depict some illustrative data according to some embodiments of the present teachings.
- Figure 3 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 4 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 5 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 6 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 7 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 8 depicts some illustrative data according to some embodiments of the present teachings.
- Figure 9 depicts an illustrative cloning map according to some embodiments of the present teachings.
- SEQ ID NO: 1 sets forth the full-length nucleotide sequence for AmyMG.
- SEQ ID NO: 2 sets forth the nucleotide sequence for the native signal sequence for AmyMG.
- SEQ ID NO: 3 sets forth the full-length amino acid sequence for AmyMG.
- SEQ ID NO: 4 sets forth the amino acid sequence for the native signal peptide.
- SEQ ID NO: 5 sets forth the aprE signal nucleic acid sequence (underlined) + AGK nucleic acid sequence (italics).
- SEQ ID NO: 6 sets forth the aprE signal amino acid sequence (underlined) + AGK amino acid sequence (italics). Detailed Description
- compositions and methods relating to maltogenic amylase enzymes This enzyme was discovered and analyzed by a combination of experimental approaches, as detailed in the Examples. Exemplary applications for the variant amylase enzymes are for starch liquefaction and saccharification, for cleaning starchy stains in laundry, dishwashing, and other applications, for textile processing (e.g., desizing), in animal feed for improving digestibility, and and for baking and brewing. These and other aspects of the compositions and methods are described in detail, below.
- IPTG isopropyl ⁇ -D-thiogalactoside
- PAHBAH p-hydroxybenzoic acid hydrazide
- ppm parts per million e.g., ⁇ g protein per gram dry solid
- TrGA Trichoderma reesei glucoamylase
- maltogenic amylase refers to enzymes that are capable of producing significant amounts of maltose from starch or hydrolyzed starch. There are several enzymes falling under this definition. Some examples are:
- Fungal alpha-amylases include those obtained from filamentous fungal strains including but not limited to strains of Aspergillus (e.g., A. Niger, A. kawachi, and A. oryzae); Trichoderma sp. (e.g. Trichoderma reesie alpha-amylase, disclosed in EP 2132307), Rhisopus sp., Mucor sp., and Penicillium sp.
- Commercial fungal alpha-amylase from Aspergillus oryzae are CLARASE ® L from DuPont-Genencor and Fungamyl ® 800l_ from Novozymes.
- Beta-amylases are found in plant materials like wheat, barley, rye, shorgum, soy, sweet potato, rice and microorganisms like Bacillus cereus, Bacillus polymixa, Bacillus megaterium, Arabidopsis thaliana.
- the most common commercial beta-amylases are derived from barley and are sold under trade the names OPT I MALT ® BBA by DuPont-Genencor and Betalase 1500L by Senson.
- a commercial soy beta-amylase is -amylase#1500S from Nagase ChemteX Corporation.
- Maltogenic amylases (E.C. 3.2.1 .1 33) are produced by microorganisms Bacillus subtilis, Geobacillus stearothermophilus, Bacillus thermoalkalophilus,
- Lactobacillus gasseri Thermus sp.
- Commercial maltogenic amylases include but not limited to Maltogenase ® L from Novozymes, Veron® XTENDER from AB Enzymes and MAX-LIFETM P100 from DuPont-Danisco.
- Enzyme units refers to the amount of product formed per time under the specified conditions of the assay.
- a "glucoamylase activity unit” GAU
- a "soluble starch unit” SSU
- SSU soluble starch unit
- DP ° Diastatic Power
- This assay is based on a 30-min hydrolysis of a starch substrate at pH 4.6 and 20 °C. The reducing sugar groups produced on hydrolysis are measured in a titrimetric procedure using alkaline
- One unit of diastase activity is defined as the amount of enzyme, contained in 0.1 ml of a 5% solution of the sample enzyme preparation, that will produce sufficient reducing sugars to reduce 5 mL of Fehling's solution when the sample is incubated with 100 mL of the substrate for 1 hour at 20 °C.
- starch refers to any material comprised of the complex polysaccharide carbohydrates of plants, comprised of amylose and amylopectin with the formula (C 6 H 0 O5)x, wherein X can be any number.
- the term includes plant-based materials such as grains, cereal, grasses, tubers and roots, and more specifically materials obtained from wheat, barley, corn, rye, rice, sorghum, brans, cassava, millet, milo, potato, sweet potato, and tapioca.
- starch includes granular starch.
- granular starch refers to raw, i.e., uncooked starch, e.g., starch that has not been subject to gelatinization.
- wild-type refers to a naturally-occurring polypeptide that does not include a man-made substitution, insertion, or deletion at one or more amino acid positions.
- reference refers to a naturally-occurring polypeptide that does not include a man-made substitution, insertion, or deletion at one or more amino acid positions.
- wild-type refers to a naturally-occurring polynucleotide that does not include a man-made nucleoside change.
- a polynucleotide encoding a wild-type, parental, or reference polypeptide is not limited to a naturally-occurring polynucleotide, and encompasses any polynucleotide encoding the wild-type, parental, or reference polypeptide.
- a "mature" polypeptide or variant, thereof, is one in which a signal sequence is absent, for example, cleaved from an immature form of the polypeptide during or following expression of the polypeptide.
- variant refers to a polypeptide that differs from a specified wild-type, parental, or reference polypeptide in that it includes one or more naturally-occurring or man-made substitutions, insertions, or deletions of an amino acid.
- variant refers to a polynucleotide that differs in nucleotide sequence from a specified wild-type, parental, or reference polynucleotide. The identity of the wild-type, parental, or reference
- polypeptide or polynucleotide will be apparent from context.
- activity refers to maltogenic amylase activity, which can be measured as described herein.
- recombinant when used in reference to a subject cell, nucleic acid, protein or vector, indicates that the subject has been modified from its native state.
- recombinant cells express genes that are not found within the native (non-recombinant) form of the cell, or express native genes at different levels or under different conditions than found in nature.
- Recombinant nucleic acids differ from a native sequence by one or more nucleotides and/or are operably linked to heterologous sequences, e.g., a heterologous promoter in an expression vector.
- Recombinant proteins may differ from a native sequence by one or more amino acids and/or are fused with heterologous sequences.
- a vector comprising a nucleic acid encoding an amylase is a recombinant vector.
- isolated refers to a compound, protein (polypeptides), cell, nucleic acid, amino acid, or other specified material or component that is removed from at least one other material or component with which it is naturally associated as found in nature.
- isolated polypeptides includes, but is not limited to, a culture broth containing secreted polypeptide expressed in a heterologous host cell.
- purified refers to material (e.g., an isolated polypeptide or
- polynucleotide that is in a relatively pure state, e.g., at least about 90% pure, at least about 95% pure, at least about 98% pure, or even at least about 99% pure.
- enriched refers to material (e.g., an isolated polypeptide or
- thermostability refers to the ability of the enzyme to retain activity after exposure to an elevated temperature.
- the thermostability of an enzyme is measured by its half-life (t /2 ) given in minutes, hours, or days, during which half the enzyme activity is lost under defined conditions.
- the half-life may be calculated by measuring residual amylase activity following exposure to (i.e., challenge by) an elevated temperature.
- pH range refers to the range of pH values under which the enzyme exhibits catalytic activity.
- amino acid sequence is synonymous with the terms “polypeptide,” “protein,” and “peptide,” and are used interchangeably. Where such amino acid sequences exhibit activity, they may be referred to as an "enzyme.”
- the conventional one-letter or three-letter codes for amino acid residues are used, with amino acid sequences being presented in the standard amino-to-carboxy terminal orientation [i.e., N ⁇ C).
- nucleic acid encompasses DNA, RNA, heteroduplexes, and synthetic molecules capable of encoding a polypeptide. Nucleic acids may be single stranded or double stranded, and may be chemical modifications.
- nucleic acid and “polynucleotide” are used interchangeably.
- nucleic acid sequences that encode a particular amino acid sequence. Unless otherwise indicated, nucleic acid sequences are presented in 5'-to-3' orientation.
- Hybridization refers to the process by which one strand of nucleic acid forms a duplex with, i.e., base pairs with, a complementary strand, as occurs during blot hybridization techniques and PCR techniques.
- Hybridized, duplex nucleic acids are characterized by a melting temperature (T m ), where one half of the hybridized nucleic acids are unpaired with the complementary strand. Mismatched nucleotides within the duplex lower the T m .
- Very stringent hybridization conditions involve 68 °C and 0.1 X SSC
- a "synthetic" molecule is produced by in vitro chemical or enzymatic synthesis rather than by an organism.
- transformed means that the cell contains a non-native (e.g., heterologous) nucleic acid sequence integrated into its genome or carried as an episome that is maintained through multiple generations.
- non-native e.g., heterologous
- a “host strain” or “host cell” is an organism into which an expression vector, phage, virus, or other DNA construct, including a polynucleotide encoding a polypeptide of interest ⁇ e.g., an amylase) has been introduced.
- Exemplary host strains are microorganism cells (e.g., bacteria, filamentous fungi, and yeast) capable of expressing the polypeptide of interest and/or fermenting saccharides.
- the term “host cell” includes protoplasts created from cells.
- heterologous with reference to a polynucleotide or protein refers to a polynucleotide or protein that does not naturally occur in a host cell.
- endogenous with reference to a polynucleotide or protein refers to a polynucleotide or protein that occurs naturally in the host cell.
- expression refers to the process by which a polypeptide is produced based on a nucleic acid sequence. The process includes both transcription and translation.
- selectable marker refers to a gene capable of being expressed in a host to facilitate selection of host cells carrying the gene.
- selectable markers include but are not limited to antimicrobials (e.g., hygromycin, bleomycin, or chloramphenicol) and/or genes that confer a metabolic advantage, such as a nutritional advantage on the host cell.
- a “vector” refers to a polynucleotide sequence designed to introduce nucleic acids into one or more cell types.
- Vectors include cloning vectors, expression vectors, shuttle vectors, plasmids, phage particles, cassettes and the like.
- an "expression vector” refers to a DNA construct comprising a DNA sequence encoding a polypeptide of interest, which coding sequence is operably linked to a suitable control sequence capable of effecting expression of the DNA in a suitable host.
- control sequences may include a promoter to effect transcription, an optional operator sequence to control transcription, a sequence encoding suitable ribosome binding sites on the mRNA, enhancers and sequences which control termination of transcription and translation.
- operably linked means that specified components are in a relationship
- a regulatory sequence is operably linked to a coding sequence such that expression of the coding sequence is under control of the regulatory sequences.
- a "signal sequence” is a sequence of amino acids attached to the N-terminal portion of a protein, which facilitates the secretion of the protein outside the cell. The mature form of an extracellular protein lacks the signal sequence, which is cleaved off during the secretion process.
- Bioly active refers to a sequence having a specified biological activity, such an enzymatic activity.
- specific activity refers to the number of moles of substrate that can be converted to product by an enzyme or enzyme preparation per unit time under specific conditions. Specific activity is generally expressed as units (U)/mg of protein.
- water hardness is a measure of the minerals ⁇ e.g., calcium and magnesium) present in water.
- a “swatch” is a piece of material such as a fabric that has a stain applied thereto.
- the material can be, for example, fabrics made of cotton, polyester or mixtures of natural and synthetic fibers.
- the swatch can further be paper, such as filter paper or nitrocellulose, or a piece of a hard material such as ceramic, metal, or glass.
- the stain is starch based, but can include blood, milk, ink, grass, tea, wine, spinach, gravy, chocolate, egg, cheese, clay, pigment, oil, or mixtures of these compounds.
- a "smaller swatch” is a section of the swatch that has been cut with a single hole punch device, or has been cut with a custom manufactured 96-hole punch device, where the pattern of the multi-hole punch is matched to standard 96-well microtiter plates, or the section has been otherwise removed from the swatch.
- the swatch can be of textile, paper, metal, or other suitable material.
- the smaller swatch can have the stain affixed either before or after it is placed into the well of a 24-, 48- or 96-well microtiter plate.
- the smaller swatch can also be made by applying a stain to a small piece of material.
- the smaller swatch can be a stained piece of fabric 5/8" or 0.25" in diameter.
- the custom manufactured punch is designed in such a manner that it delivers 96 swatches simultaneously to all wells of a 96-well plate.
- the device allows delivery of more than one swatch per well by simply loading the same 96-well plate multiple times.
- Multi-hole punch devices can be conceived of to deliver simultaneously swatches to any format plate, including but not limited to 24-well, 48-well, and 96-well plates.
- the soiled test platform can be a bead made of metal, plastic, glass, ceramic, or another suitable material that is coated with the soil substrate. The one or more coated beads are then placed into wells of 96-, 48-, or 24-well plates or larger formats, containing suitable buffer and enzyme.
- a cultured cell material comprising an amylase refers to a cell lysate or supernatant (including media) that includes an amylase as a component.
- the cell material may be from a heterologous host that is grown in culture for the purpose of producing the amylase.
- percent sequence identity means that a particular sequence has at least a certain percentage of amino acid residues identical to those in a specified reference sequence, when aligned using the CLUSTAL W algorithm with default parameters. See Thompson ef a/. (1 994) Nucleic Acids Res. 22:4673-4680. Default parameters for the CLUSTAL W algorithm are:
- Gap extension penalty 0.05
- Toggle end gap separation penalty OFF Deletions are counted as non-identical residues, compared to a reference sequence. Deletions occurring at either termini are included. For example, a variant with five amino acid deletions of the C-terminus of the mature 617 residue polypeptide would have a percent sequence identity of 99% (61 2 / 617 identical residues ⁇ 1 00, rounded to the nearest whole number) relative to the mature polypeptide. Such a variant would be encompassed by a variant having "at least 99% sequence identity" to a mature polypeptide.
- Fusion polypeptide sequences are connected, i.e., operably linked, via a peptide bond between two subject polypeptide sequences.
- filamentous fungi refers to all filamentous forms of the subdivision Eumycotina, particularly Pezizomycotina species.
- degree of polymerization refers to the number (n) of
- DP1 anhydro-glucopyranose units in a given saccharide.
- DP2 the monosaccharides glucose and fructose.
- DP2 the disaccharides maltose and sucrose.
- DE or “dextrose equivalent,” is defined as the percentage of reducing sugar, i.e., D-glucose, as a fraction of total carbohydrate in a syrup.
- dry solids content refers to the total solids of a slurry in a dry weight percent basis.
- slurry refers to an aqueous mixture containing insoluble solids.
- SSF simultaneous saccharification and fermentation
- SSF includes the contemporaneous hydrolysis of starch substrates (granular, liquefied, or solubilized) to saccharides, including glucose, and the fermentation of the saccharides into alcohol or other biochemical or biomaterial in the same reactor vessel.
- an “ethanologenic microorganism” refers to a microorganism with the ability to convert a sugar or oligosaccharide to ethanol.
- the term “fermented beverage” refers to any beverage produced by a method comprising a fermentation process, such as a microbial fermentation, e.g., a bacterial and/or fungal fermentation. "Beer” is an example of such a fermented beverage, and the term “beer” is meant to comprise any fermented wort produced by
- beer is produced exclusively from malt or adjunct, or any combination of malt and adjunct.
- beers include: full malted beer, beer brewed under the "Rösgebot," ale, India pale ale, lager, pilsner, bitter, Happoshu (second beer), third beer, dry beer, near beer, light beer, low alcohol beer, low calorie beer, porter, bock, dopplebock, stout, porter, malt liquor, non-alcoholic beer, non-alcoholic malt liquor and the like, but also alternative cereal and malt beverages such as fruit flavored malt beverages, e.g., citrus flavored, such as lemon-, orange-, lime-, or berry-flavored malt beverages, liquor flavored malt beverages, e.g., vodka-, rum-, or tequila-flavored malt liquor, or coffee flavored malt beverages, such as caffeine-flavored malt liquor, and the like.
- fruit flavored malt beverages e.g., citrus flavored, such as lemon-, orange-, lime-,
- malt refers to any malted cereal grain, such as malted barley or wheat.
- adjct refers to any starch and/or sugar containing plant material that is not malt, such as barley or wheat malt.
- adjuncts include common corn grits, refined corn grits, brewer's milled yeast, rice, sorghum, refined corn starch, barley, barley starch, dehusked barley, wheat, wheat starch, torrified cereal, cereal flakes, rye, oats, potato, tapioca, cassava and syrups, such as corn syrup, sugar cane syrup, inverted sugar syrup, barley and/or wheat syrups, and the like.
- wort refers to an aqueous slurry of any starch and/or sugar containing plant material, such as grist, e.g., comprising crushed barley malt, crushed barley, and/or other adjunct or a combination thereof, mixed with water later to be separated into wort and spent grains.
- grist e.g., comprising crushed barley malt, crushed barley, and/or other adjunct or a combination thereof, mixed with water later to be separated into wort and spent grains.
- wort refers to the unfermented liquor run-off following extracting the grist during mashing.
- Iodine-positive starch or “IPS” refers to (1 ) amylose that is not hydrolyzed after liquefaction and saccharification, or (2) a retrograded starch polymer.
- saccharified starch or saccharide liquor is tested with iodine, the high DPn amylose or the retrograded starch polymer binds iodine and produces a characteristic blue color.
- the saccharide liquor is thus termed "iodine-positive saccharide,” “blue saccharide,” or “blue sac.
- starch retrogradation refers to changes that occur spontaneously in a starch paste or gel on ageing.
- the present maltogenic amylases further include one or more mutations that provide a further performance or stability benefit.
- Exemplary performance benfits include but are not limited to increased hydrolysis of a starch substrate, increased grain, cereal or other starch substrate liquifaction performance, increased cleaning performance, increased thermal stability, increased storage stability, increased solubility, an altered pH profile, decreased calcium dependence, increased specific activity, modified substrate specificity, modified substrate binding, modified pH-dependent activity, modified pH-dependent stability, increased oxidative stability, and increased expression.
- the performance benefit is realized at a relatively low temperature. In some cases, the performance benefit is realized at relatively high temperature.
- the present amylases may include any number of conservative amino acid substitutions.
- Exemplary conservative amino acid substitutions are listed in the following Table 1 .
- Table 1 Conservative amino acid substitutions
- the present maltogenic amylase may be "precursor,” “immature,” or “full-length,” in which case they include a signal sequence, or “mature,” in which case they lack a signal sequence. Mature forms of the polypeptides are generally the most useful. Unless otherwise noted, the amino acid residue numbering used herein refers to the mature forms of the respective maltogenic amylase polypeptides. The present maltogenic amylase polypeptides may also be truncated to remove the N or C-termini, so long as the resulting polypeptides retain amylase activity.
- the present maltogenic amylase may be a "chimeric" or “hybrid” polypeptide, in that it includes at least a portion of a first amylase polypeptide, and at least a portion of a second amylase polypeptide (such chimeric amylases have recently been “rediscovered” as domain-swap amylases).
- the present amylases may further include heterologous signal sequence, an epitope to allow tracking or purification, or the like.
- Exemplary heterologous signal sequences are from B. licheniformis amylase (LAT), B. subtilis (AmyE or AprE), and Streptomyces CelA.
- nucleic acids encoding a maltogenic amylase polypeptide are provided.
- the nucleic acid may encode a particular maltogenic amylase polypeptide, or a maltogenic amylase having a specified degree of amino acid sequence identity to the particular amylase.
- the nucleic acid encodes a maltogenic amylase having at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or even at least 99% identity to SEQ I D NO: 1 . It will be appreciated that due to the degeneracy of the genetic code, a plurality of nucleic acids may encode the same polypeptide.
- the nucleic acid hybridizes under stringent or very stringent conditions to a nucleic acid encoding (or complementary to a nucleic acid encoding) a maltogenic amylase having at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or even at least 99% identity to SEQ I D NO: 1 .
- Such stringent and very stringent hybridization conditions are described herein.
- Nucleic acids may encode a "full-length" (“fl” or “FL”) maltogenic amylase, which includes a signal sequence, only the mature form of a maltogenic amylase, which lacks the signal sequence, or a truncated form of a maltogenic amylase, which lacks the N or C-terminus of the mature form.
- fl full-length
- FL truncated form of a maltogenic amylase
- a nucleic acid that encodes a maltogenic amylase can be operably linked to various promoters and regulators in a vector suitable for expressing the maltogenic amylase in host cells.
- exemplary promoters are from B. licheniformis amylase (LAT), B. subtilis (AmyE or AprE), and Streptomyces CelA.
- Such a nucleic acid can also be linked to other coding sequences, e.g., to encode a chimeric polypeptide.
- the present maltogenic amylases can be produced in host cells, for example, by secretion or intracellular expression.
- a cultured cell material e.g., a whole-cell broth
- the maltogenic amylase can be isolated from the host cells, or even isolated from the cell broth, depending on the desired purity of the final maltogenic amylase.
- a gene encoding a maltogenic amylase can be cloned and expressed according to methods well known in the art.
- Suitable host cells include bacterial, fungal (including yeast and filamentous fungi), and plant cells (including algae).
- host cells include Aspergillus niger, Aspergillus oryzae or Trichoderma reesei.
- Other host cells include bacterial cells, e.g., Bacillus subtilis or B. licheniformis, as well as Streptomyces.
- the host cell further may express a nucleic acid encoding a homologous or heterologous glucoamylase, i.e., a glucoamylase that is not the same species as the host cell, or one or more other enzymes.
- the glucoamylase may be a variant glucoamylase, such as one of the glucoamylase variants disclosed in U.S. Patent No. 8,058,033 (Danisco US Inc.), for example.
- the host may express one or more accessory enzymes, proteins, peptides. These may benefit liquefaction, saccharification, fermentation, SSF, etc processes.
- the host cell may produce biochemicals in addition to enzymes used to digest the various feedstock(s). Such host cells may be useful for fermentation or simultaneous saccharification and fermentation processes to reduce or eliminate the need to add enzymes.
- Vectors in addition to enzymes used to digest the various feedstock(s). Such host cells may be useful for fermentation or simultaneous saccharification and fermentation processes to reduce or eliminate
- a DNA construct comprising a nucleic acid encoding maltogenic amylases can be constructed to be expressed in a host cell.
- Representative nucleic acids that encode maltogenic amylases include SEQ I D NO: 1 . Because of the well-known degeneracy in the genetic code, variant polynucleotides that encode an identical amino acid sequence can be designed and made with routine skill. It is also well-known in the art to optimize codon use for a particular host cell. Nucleic acids encoding maltogenic amylases can be incorporated into a vector. Vectors can be transferred to a host cell using well-known transformation techniques, such as those disclosed below.
- the vector may be any vector that can be transformed into and replicated within a host cell.
- a vector comprising a nucleic acid encoding a maltogenic amylase can be transformed and replicated in a bacterial host cell as a means of propagating and amplifying the vector.
- the vector also may be transformed into an expression host, so that the encoding nucleic acids can be expressed as a functional maltogenic amylase.
- Host cells that serve as expression hosts can include filamentous fungi, for example.
- the Fungal Genetics Stock Center (FGSC) Catalogue of Strains lists suitable vectors for expression in fungal host cells. See FGSC, Catalogue of Strains, University of Missouri, ai www.fgsc.net (last modified January 17, 2007).
- a representative vector is pJG153, a promoterless Cre expression vector that can be replicated in a bacterial host. See Harrison et al. (June 2011 ) Applied Environ. Microbiol. 77: 3916-22.
- pJG153 can be modified with routine skill to comprise and express a nucleic acid encoding a maltogenic amylase.
- a nucleic acid encoding a maltogenic amylase can be operably linked to a suitable promoter, which allows transcription in the host cell.
- the promoter may be any DNA sequence that shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell.
- Exemplary promoters for directing the transcription of the DNA sequence encoding a maltogenic amylase, especially in a bacterial host are the promoter of the lac operon of E.
- the Streptomyces coelicolor agarase gene dagA or eel A promoters the promoters of the Bacillus licheniformis ⁇ -amylase gene (amyL), the promoters of the Bacillus stearothermophilus maltogenic amylase gene (amyM), the promoters of the Bacillus amyloliquefaciens a-amylase (amyQ), the promoters of the Bacillus subtilis xylA and xylB genes etc.
- examples of useful promoters are those derived from the gene encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral a-amylase, A. niger acid stable a-amylase, A. n/ger glucoamylase, Rhizomucor miehei lipase, A. oryzae alkaline protease, A. oryzae triose phosphate isomerase, or A. nidulans acetamidase.
- TAKA amylase Rhizomucor miehei aspartic proteinase
- Aspergillus niger neutral a-amylase A. niger acid stable a-amylase
- A. n/ger glucoamylase Rhizomucor miehei lipase
- A. oryzae alkaline protease A. oryzae
- a suitable promoter can be selected, for example, from a bacteriophage promoter including a T7 promoter and a phage lambda promoter.
- suitable promoters for the expression in a yeast species include but are not limited to the Gal 1 and Gal 10 promoters of Saccharomyces cerevisiae and the Pichia pastoris AOX1 or AOX2 promoters, cbh l is an endogenous, inducible promoter from T. reesei. See Liu et al. (2008) "Improved heterologous gene expression in Trichoderma reesei by
- the coding sequence can be operably linked to a signal sequence.
- the DNA encoding the signal sequence may be the DNA sequence naturally associated with the maltogenic amylase gene to be expressed or from a different Genus or species.
- a signal sequence and a promoter sequence comprising a DNA construct or vector can be introduced into a fungal host cell and can be derived from the same source.
- the signal sequence is the cbh l signal sequence that is operably linked to a cbh l promoter.
- An expression vector may also comprise a suitable transcription terminator and, in eukaryotes, polyadenylation sequences operably linked to the DNA sequence encoding a variant amylase. Termination and polyadenylation sequences may suitably be derived from the same sources as the promoter.
- the vector may further comprise a DNA sequence enabling the vector to replicate in the host cell.
- sequences are the origins of replication of plasmids pUC19, pACYCI 77, pUB110, pE194, pAMB1 , and plJ702.
- the vector may also comprise a selectable marker, e.g., a gene the product of which complements a defect in the isolated host cell, such as the da/ genes from B. subtilis or B. licheniformis, or a gene that confers antibiotic resistance such as, e.g., ampicillin, kanamycin, chloramphenicol or tetracycline resistance.
- a selectable marker e.g., a gene the product of which complements a defect in the isolated host cell, such as the da/ genes from B. subtilis or B. licheniformis, or a gene that confers antibiotic resistance such as, e.g., ampicillin, kanamycin, chloramphenicol or tetracycline resistance.
- the vector may comprise Aspergillus selection markers such as amdS, argB, niaD and xxsC, a marker giving rise to hygromycin resistance, or the selection may be accomplished by co-transformation, such as known in the art. See
- Intracellular expression may be advantageous in some respects, e.g., when using certain bacteria or fungi as host cells to produce large amounts of maltogenic amylase for subsequent enrichment or purification.
- Extracellular secretion of amylase into the culture medium can also be used to make a cultured cell material comprising the isolated maltogenic amylase.
- the expression vector typically includes the components of a cloning vector, such as, for example, an element that permits autonomous replication of the vector in the selected host organism and one or more phenotypically detectable markers for selection purposes.
- the expression vector normally comprises control nucleotide sequences such as a promoter, operator, ribosome binding site, translation initiation signal and optionally, a repressor gene or one or more activator genes.
- the expression vector may comprise a sequence coding for an amino acid sequence capable of targeting the maltogenic amylase to a host cell organelle such as a peroxisome, or to a particular host cell compartment.
- a targeting sequence includes but is not limited to the sequence, SKL.
- the nucleic acid sequence of the amylase is operably linked to the control sequences in proper manner with respect to expression.
- An isolated cell is advantageously used as a host cell in the recombinant production of a maltogenic amylase.
- the cell may be transformed with the DNA construct encoding the enzyme, conveniently by integrating the DNA construct (in one or more copies) in the host chromosome. This integration is generally considered to be an advantage, as the DNA sequence is more likely to be stably maintained in the cell. Integration of the DNA constructs into the host chromosome may be performed according to conventional methods, e.g., by homologous or heterologous recombination. Alternatively, the cell may be transformed with an expression vector as described above in connection with the different types of host cells.
- suitable bacterial host organisms are Gram positive bacterial species such as Bacillaceae including Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Geobacillus (formerly Bacillus) stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus lautus, Bacillus megaterium, and Bacillus thuringiensis; Streptomyces species such as Streptomyces murinus; lactic acid bacterial species including Lactococcus sp. such as Lactococcus lactis; Lactobacillus sp.
- Bacillaceae including Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Geobacillus (formerly Bacillus) stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans,
- strains of a Gram negative bacterial species belonging to Enterobacteriaceae including E. coli, or to Pseudomonadaceae can be selected as the host organism.
- a suitable yeast host organism can be selected from the biotechnologically relevant yeasts species such as but not limited to yeast species such as Pichia sp., Hansenula sp., or Kluyveromyces, Yarrowinia, Schizosaccharomyces species or a species of Saccharomyces, including Saccharomyces cerevisiae or a species belonging to Schizosaccharomyces such as, for example, S. pombe species.
- a strain of the methylotrophic yeast species, Pichia pastoris can be used as the host organism.
- the host organism can be a Hansenula species.
- Suitable host organisms among filamentous fungi include species of Aspergillus, e.g., Aspergillus niger,
- strains of a Fusarium species e.g., Fusarium oxysporum or of a
- Rhizomucor species such as Rhizomucor miehei can be used as the host organism.
- Other suitable strains include Thermomyces and Mucor species.
- Trichoderma sp. can be used as a host.
- a suitable procedure for transformation of Aspergillus host cells includes, for example, that described in EP 238023.
- a maltogenic amylase expressed by a fungal host cell can be glycosylated, i.e., will comprise a glycosyl moiety.
- the glycosylation pattern can be the same or different as present in the wild-type maltogenic amylase.
- the type and/or degree of glycosylation may impart changes in enzymatic and/or biochemical properties.
- Gene inactivation may be accomplished by complete or partial deletion, by insertional inactivation or by any other means that renders a gene nonfunctional for its intended purpose, such that the gene is prevented from expression of a functional protein.
- Any gene from a Trichoderma sp. or other filamentous fungal host that has been cloned can be deleted, for example, cbh l, cbh2, egl1, and egl2 genes.
- Gene deletion may be accomplished by inserting a form of the desired gene to be inactivated into a plasmid by methods known in the art.
- Introduction of a DNA construct or vector into a host cell includes techniques such as transformation ; electroporation ; nuclear microinjection; transduction; transfection, e.g., lipofection mediated and DEAE-Dextrin mediated transfection;
- the preparation of Trichoderma sp. for transformation may involve the preparation of protoplasts from fungal mycelia. See Campbell ef al. (1 989) Curr. Genet. 1 6: 53-56.
- the mycelia can be obtained from germinated vegetative spores.
- the mycelia are treated with an enzyme that digests the cell wall, resulting in protoplasts,
- the protoplasts are protected by the presence of an osmotic stabilizer in the suspending medium.
- These stabilizers include sorbitol, mannitol, potassium chloride, magnesium sulfate, and the like. Usually the concentration of these stabilizers varies between 0.8 M and 1 .2 M, e.g., a 1 .2 M solution of sorbitol can be used in the suspension medium.
- Uptake of DNA into the host Trichoderma sp. strain depends upon the calcium ion concentration. Generally, between about 10-50 mM CaCI 2 is used in an uptake solution. Additional suitable compounds include a buffering system, such as TE buffer (10 mM Tris, pH 7.4; 1 mM EDTA) or 1 0 mM MOPS, pH 6.0 and polyethylene glycol. The polyethylene glycol is believed to fuse the cell membranes, thus permitting the contents of the medium to be delivered into the cytoplasm of the Trichoderma sp. strain. This fusion frequently leaves multiple copies of the plasmid DNA integrated into the host chromosome.
- TE buffer 10 mM Tris, pH 7.4; 1 mM EDTA
- MOPS 1 0 mM MOPS
- Trichoderma sp. uses protoplasts or cells that have been subjected to a permeability treatment, typically at a density of 10 5 to 10 7 /ml_, particularly 2x1 0 6 /mL A volume of 100 ⁇ _ of these protoplasts or cells in an appropriate solution (e.g., 1 .2 M sorbitol and 50 mM CaCI 2 ) may be mixed with the desired DNA.
- an appropriate solution e.g., 1 .2 M sorbitol and 50 mM CaCI 2
- PEG a high concentration of PEG is added to the uptake solution. From 0.1 to 1 volume of 25% PEG 4000 can be added to the protoplast suspension ; however, it is useful to add about 0.25 volumes to the protoplast suspension.
- Additives such as dimethyl sulfoxide, heparin, spermidine, potassium chloride and the like, may also be added to the uptake solution to facilitate transformation. Similar procedures are available for other fungal host cells. See, e.g., U.S. Patent No. 6,022,725.
- a method of producing a maltogenic amylase may comprise cultivating a host cell as described above under conditions conducive to the production of the enzyme and recovering the enzyme from the cells and/or culture medium.
- the medium used to cultivate the cells may be any conventional medium suitable for growing the host cell in question and obtaining expression of a maltogenic amylase. Suitable media and media components are available from commercial suppliers or may be prepared according to published recipes (e.g., as described in catalogues of the American Type Culture Collection).
- An enzyme secreted from the host cells can be used in a whole broth preparation.
- the preparation of a spent whole fermentation broth of a recombinant microorganism can be achieved using any cultivation method known in the art resulting in the expression of a maltogenic amylase. Fermentation may, therefore, be understood as comprising shake flask cultivation, small- or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermenters performed in a suitable medium and under conditions allowing the amylase to be expressed or isolated.
- the term "spent whole fermentation broth” is defined herein as unfractionated contents of fermentation material that includes culture medium, extracellular proteins (e.g., enzymes), and cellular biomass.
- spent whole fermentation broth also encompasses cellular biomass that has been lysed or permeabilized using methods well known in the art.
- An enzyme secreted from the host cells may conveniently be recovered from the culture medium by well-known procedures, including separating the cells from the medium by centrifugation or filtration, and precipitating proteinaceous components of the medium by means of a salt such as ammonium sulfate, followed by the use of chromatographic procedures such as ion exchange chromatography, affinity
- the polynucleotide encoding a maltogenic amylase in a vector can be operably linked to a control sequence that is capable of providing for the expression of the coding sequence by the host cell, i.e. the vector is an expression vector.
- the control sequences may be modified, for example by the addition of further transcriptional regulatory elements to make the level of transcription directed by the control sequences more responsive to transcriptional modulators.
- the control sequences may in particular comprise promoters.
- Host cells may be cultured under suitable conditions that allow expression of a maltogenic amylase.
- Expression of the enzymes may be constitutive such that they are continually produced, or inducible, requiring a stimulus to initiate expression.
- protein production can be initiated when required by, for example, addition of an inducer substance to the culture medium, for example dexamethasone or IPTG or Sophorose.
- Polypeptides can also be produced
- an in vitro cell-free system such as the TNTTM (Promega) rabbit reticulocyte system.
- An expression host also can be cultured in the appropriate medium for the host, under aerobic conditions. Shaking or a combination of agitation and aeration can be provided, with production occurring at the appropriate temperature for that host, e.g., from about 25 °C to about 75 °C (e.g., 30 °C to 45 °C), depending on the needs of the host and production of the desired maltogenic amylase. Culturing can occur from about 12 to about 100 hours or greater (and any hour value there between, e.g., from 24 to 72 hours). Typically, the culture broth is at a pH of about 4.0 to about 8.0, again depending on the culture conditions needed for the host relative to production of a maltogenic amylase.
- Fermentation, separation, and concentration techniques are well known in the art and conventional methods can be used in order to prepare a concentrated maltogenic amylase polypeptide-containing solution.
- a fermentation broth is obtained, the microbial cells and various suspended solids, including residual raw fermentation materials, are removed by conventional separation techniques in order to obtain a maltogenic amylase solution. Filtration, centrifugation, microfiltration, rotary vacuum drum filtration, ultrafiltration, centrifugation followed by ultra-filtration, extraction, or chromatography, or the like, are generally used.
- Exemplary methods of enrichment and purification include but are not limited to rotary vacuum filtration and/or ultrafiltration.
- the enzyme solution is concentrated into a concentrated enzyme solution until the enzyme activity of the concentrated maltogenic amylase polypeptide-containing solution is at a desired level.
- Concentration may be performed using, e.g., a precipitation agent, such as a metal halide precipitation agent.
- a precipitation agent such as a metal halide precipitation agent.
- Metal halide precipitation agents include but are not limited to alkali metal chlorides, alkali metal bromides and blends of two or more of these metal halides.
- Exemplary metal halides include sodium chloride, potassium chloride, sodium bromide, potassium bromide and blends of two or more of these metal halides.
- the metal halide precipitation agent, sodium chloride can also be used as a
- the metal halide precipitation agent is used in an amount effective to precipitate a maltogenic amylase.
- the selection of at least an effective amount and an optimum amount of metal halide effective to cause precipitation of the enzyme, as well as the conditions of the precipitation for maximum recovery including incubation time, pH, temperature and concentration of enzyme, will be readily apparent to one of ordinary skill in the art, after routine testing.
- the concentration of the metal halide precipitation agent will depend, among others, on the nature of the specific maltogenic amylase polypeptide and on its concentration in the concentrated enzyme solution.
- organic compound precipitating agents include: 4-hydroxybenzoic acid, alkali metal salts of 4-hydroxybenzoic acid, alkyl esters of 4-hydroxybenzoic acid, and blends of two or more of these organic compounds.
- the addition of the organic compound precipitation agents can take place prior to, simultaneously with or subsequent to the addition of the metal halide precipitation agent, and the addition of both precipitation agents, organic compound and metal halide, may be carried out sequentially or simultaneously.
- the organic precipitation agents are selected from the group consisting of alkali metal salts of 4-hydroxybenzoic acid, such as sodium or potassium salts, and linear or branched alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1 to 12 carbon atoms, and blends of two or more of these organic compounds.
- the organic compound precipitation agents can be, for example, linear or branched alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1 to 10 carbon atoms, and blends of two or more of these organic compounds.
- Exemplary organic compounds are linear alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1 to 6 carbon atoms, and blends of two or more of these organic compounds.
- Methyl esters of 4-hydroxybenzoic acid, propyl esters of 4-hydroxybenzoic acid, butyl ester of 4-hydroxybenzoic acid, ethyl ester of 4-hydroxybenzoic acid and blends of two or more of these organic compounds can also be used.
- Additional organic compounds also include but are not limited to 4-hydroxybenzoic acid methyl ester (named methyl PARABEN), 4-hydroxybenzoic acid propyl ester (named propyl
- PARABEN which also are both amylase preservative agents.
- U.S. Patent No. 5,281 ,526 See, e.g., U.S. Patent No. 5,281 ,526.
- Addition of the organic compound precipitation agent provides the advantage of high flexibility of the precipitation conditions with respect to pH, temperature, maltogenic amylase concentration, precipitation agent concentration, and time of incubation.
- the organic compound precipitation agent is used in an amount effective to improve precipitation of the enzyme by means of the metal halide precipitation agent.
- the selection of at least an effective amount and an optimum amount of organic compound precipitation agent, as well as the conditions of the precipitation for maximum recovery including incubation time, pH, temperature and concentration of enzyme, will be readily apparent to one of ordinary skill in the art, in light of the present disclosure, after routine testing.
- At least about 0.01 % w/v of organic compound precipitation agent is added to the concentrated enzyme solution and usually at least about 0.02% w/v.
- the concentrated polypeptide solution containing the metal halide precipitation agent, and the organic compound precipitation agent, can be adjusted to a pH, which will, of necessity, depend on the enzyme to be enriched or purified.
- the pH is adjusted at a level near the isoelectric point of the amylase.
- the pH can be adjusted at a pH in a range from about 2.5 pH units below the isoelectric point (pi) up to about 2.5 pH units above the isoelectric point.
- the incubation time necessary to obtain an enriched or purified enzyme precipitate depends on the nature of the specific enzyme, the concentration of enzyme, and the specific precipitation agent(s) and its (their) concentration. Generally, the time effective to precipitate the enzyme is between about 1 to about 30 hours; usually it does not exceed about 25 hours. In the presence of the organic compound precipitation agent, the time of incubation can still be reduced to less about 10 hours and in most cases even about 6 hours. Generally, the temperature during incubation is between about 4°C and about
- the method is carried out at a temperature between about 10 °C and about 45 °C ⁇ e.g., between about 20 °C and about 40 °C).
- the optimal temperature for inducing precipitation varies according to the solution conditions and the enzyme or precipitation agent(s) used.
- the overall recovery of enriched or purified enzyme precipitate, and the efficiency with which the process is conducted, is improved by agitating the solution comprising the enzyme, the added metal halide and the added organic compound.
- the agitation step is done both during addition of the metal halide and the organic compound, and during the subsequent incubation period. Suitable agitation methods include mechanical stirring or shaking, vigorous aeration, or any similar technique.
- the enriched or purified enzyme is then separated from the dissociated pigment and other impurities and collected by conventional separation techniques, such as filtration, centrifugation, microfiltration, rotary vacuum filtration, ultrafiltration, press filtration, cross membrane microfiltration, cross flow membrane microfiltration, or the like. Further enrichment or purification of the enzyme precipitate can be obtained by washing the precipitate with water.
- the enriched or purified enzyme precipitate is washed with water containing the metal halide precipitation agent, or with water containing the metal halide and the organic compound precipitation agents.
- a maltogenic amylase polypeptide accumulates in the culture broth.
- the culture broth is centrifuged or filtered to eliminate cells, and the resulting cell-free liquid is used for enzyme enrichment or purification.
- the cell-free broth is subjected to salting out using ammonium sulfate at about 70% saturation; the 70% saturation-precipitation fraction is then dissolved in a buffer and applied to a column such as a Sephadex G-1 00 column, and eluted to recover the enzyme-active fraction.
- a conventional procedure such as ion exchange chromatography may be used. Enriched or purified enzymes are useful for laundry and cleaning applications.
- they can be used in laundry detergents and spot removers. They can be made into a final product that is either liquid (solution, slurry) or solid (granular, powder).
- a Toyopearl HW55 column (Tosoh Bioscience, Montgomeryville, PA; Cat. No. 19812) is equilibrated with 20 mM Tris/HCI buffer (pH 7.0) containing 5 mM CaCI 2 and 1 .5 M (NH 4 ) 2 S0 4 .
- the enzyme is eluted with a linear gradient of 1 .5 to 0 M (NH 4 ) 2 S0 4 in 20 mM Tris/HCL buffer, pH 7.0 containing 5 mM CaCI 2 .
- the active fractions are collected, and the enzyme precipitated with (NH 4 ) 2 S0 4 at 80% saturation. The precipitate is recovered, re-dissolved, and dialyzed as described above.
- the dialyzed sample is then applied to a Mono Q HR5/5 column (Amersham Pharmacia; Cat. No. 17-5167-01 ) previously equilibrated with 20 mM Tris/HCI buffer (pH 7.0) containing 5 mM CaCI 2 , at a flow rate of 60 mUhour.
- the active fractions are collected and added to a 1 .5 M (NH 4 ) 2 S0 4 solution.
- the active enzyme fractions are re-chromatographed on a Toyopearl HW55 column, as before, to yield a homogeneous enzyme as determined by SDS-PAGE. See Sumitani ef al. (2000) Biochem. J. 350: 477-484, for general discussion of the method and variations thereon.
- maltogenic amylase polypeptides can be enriched or partially purified as generally described above by removing cells via flocculation with polymers.
- the enzyme can be enriched or purified by microfiltration followed by concentration by ultrafiltration using available membranes and equipment.
- the enzyme does not need to be enriched or purified, and whole broth culture can be lysed and used without further treatment.
- the enzyme can then be processed, for example, into granules.
- maltogenic amylases are useful for a variety of industrial applications.
- maltogenic amylases are useful in a starch conversion process, particularly in a saccharification process of a starch that has undergone liquefaction.
- the desired end-product may be any product that may be produced by the enzymatic conversion of the starch substrate.
- the desired product may be a syrup rich in glucose and maltose, which can be used in other processes, such as the preparation of HFCS, or which can be converted into a number of other useful products, such as ascorbic acid intermediates ⁇ e.g., gluconate;
- 2-keto-L-gulonic acid 5-keto-gluconate; and 2,5-diketogluconate) ; 1 ,3-propanediol; aromatic amino acids (e.g., tyrosine, phenylalanine and tryptophan); organic acids (e.g., lactate, pyruvate, succinate, isocitrate, and oxaloacetate); amino acids (e.g., serine and glycine) ; antibiotics; antimicrobials; enzymes; vitamins; and hormones.
- aromatic amino acids e.g., tyrosine, phenylalanine and tryptophan
- organic acids e.g., lactate, pyruvate, succinate, isocitrate, and oxaloacetate
- amino acids e.g., serine and glycine
- the starch conversion process may be a precursor to, or simultaneous with, a fermentation process designed to produce alcohol for fuel or drinking (i.e., potable alcohol).
- a fermentation process designed to produce alcohol for fuel or drinking (i.e., potable alcohol).
- One skilled in the art is aware of various fermentation conditions that may be used in the production of these end-products.
- Variant amylases are also useful in compositions and methods of food preparation. These various uses of variant amylases are described in more detail below. It will be appreciated by one of ordinary skill in the art that various accessory enzymes can be used with the maltogenic enzymes of the present teachings, as will be the case in various applications and contexts.
- maltogenic amylases can be employed in any of a variety of applications, including those described in US Provisional Application 61 /61 6,990, filed March 28, 201 2.
- a useful starch substrate may be obtained from tubers, roots, stems, legumes, cereals or whole grain. More specifically, the granular starch may be obtained from corn, cobs, wheat, barley, rye, triticale, milo, sago, millet, cassava, tapioca, sorghum, rice, peas, bean, banana, or potatoes. Corn contains about 60-68% starch; barley contains about 55-65% starch; millet contains about 75-80% starch ; wheat contains about 60-65% starch; and polished rice contains 70-72% starch.
- starch substrates are corn starch and wheat starch.
- the starch from a grain may be ground or whole and includes corn solids, such as kernels, bran and/or cobs.
- the starch may also be highly refined raw starch or feedstock from starch refinery processes.
- Various starches also are commercially available.
- corn starch is available from Cerestar, Sigma, and Katayama Chemical Industry Co. (Japan); wheat starch is available from Sigma; sweet potato starch is available from Wako Pure Chemical Industry Co. (Japan) ; and potato starch is available from Nakaari Chemical Pharmaceutical Co.
- the starch substrate can be a crude starch from milled whole grain, which contains non-starch fractions, e.g., germ residues and fibers.
- Milling may comprise either wet milling or dry milling or grinding.
- wet milling whole grain is soaked in water or dilute acid to separate the grain into its component parts, e.g., starch, protein, germ, oil, kernel fibers.
- Wet milling efficiently separates the germ and meal ⁇ i.e., starch granules and protein) and is especially suitable for production of syrups.
- whole kernels are ground into a fine powder and often processed without fractionating the grain into its component parts. In some cases, oils from the kernels are recovered.
- Dry ground grain thus will comprise significant amounts of non-starch carbohydrate compounds, in addition to starch. Dry grinding of the starch substrate can be used for production of ethanol and other biochemicals.
- the starch to be processed may be a highly refined starch quality, for example, at least 90%, at least 95%, at least 97%, or at least 99.5% pure.
- the soluble starch hydrolysate can be fermented by contacting the starch hydrolysate with a fermenting organism typically at a temperature around 32 °C, such as from 30 °C to 35 °C for alcohol-producing yeast.
- a fermenting organism typically at a temperature around 32 °C, such as from 30 °C to 35 °C for alcohol-producing yeast.
- the temperature and pH of the fermentation will depend upon the fermenting organism.
- EOF products include metabolites, such as citric acid, lactic acid, succinic acid, monosodium glutamate, gluconic acid, sodium gluconate, calcium gluconate, potassium gluconate, itaconic acid and other carboxylic acids, glucono delta-lactone, sodium erythorbate, lysine and other amino acids, omega 3 fatty acid, butanol, isoprene, 1 ,3-propanediol and other biomaterials.
- metabolites such as citric acid, lactic acid, succinic acid, monosodium glutamate, gluconic acid, sodium gluconate, calcium gluconate, potassium gluconate, itaconic acid and other carboxylic acids, glucono delta-lactone, sodium erythorbate, lysine and other amino acids, omega 3 fatty acid, butanol, isoprene, 1 ,3-propanediol and other biomaterials
- Ethanologenic microorganisms include yeast, such as Saccharomyces cerevisiae and bacteria, e.g., Zymomonas mobilis, expressing alcohol dehydrogenase and pyruvate decarboxylase.
- the ethanologenic microorganism can express xylose reductase and xylitol dehydrogenase, which convert xylose to xylulose.
- Improved strains of ethanologenic microorganisms which can withstand higher temperatures, for example, are known in the art and can be used. See Liu etal. (2011 ) Sheng Wu Gong Cheng Xue Bao 27(7): 1049-56.
- yeast Commercial sources of yeast include ETHANOL RED® (LeSaffre) ; Thermosacc® (Lallemand) ; RED STAR® (Red Star); FERMIOL® (DSM Specialties); and SUPERSTART® (Alltech).
- microorganisms that produce other metabolites, such as citric acid and lactic acid, by fermentation are also known in the art. See, e.g.,
- the broth may be distilled to produce enriched, e.g., 96% pure, solutions of ethanol.
- C0 2 generated by fermentation may be collected with a C0 2 scrubber, compressed, and marketed for other uses, e.g., carbonating beverage or dry ice production.
- fermentation process may be used as protein-rich products, e.g., livestock feed.
- an SSF process can be conducted with fungal cells that express and secrete amylase continuously throughout SSF.
- the fungal cells expressing amylase also can be the fermenting microorganism, e.g., an ethanologenic
- Ethanol production thus can be carried out using a fungal cell that expresses sufficient amylase so that less or no enzyme has to be added exogenously.
- the fungal host cell can be from an appropriately engineered fungal strain. Fungal host cells that express and secrete other enzymes, in addition to amylase, also can be used.
- Such cells may express glucoamylase and/or a pullulanase, phytase, a/pfra-glucosidase, isoamylase, beta-amylase cellulase, xylanase, other hemicellulases, protease, befa-glucosidase, pectinase, esterase, redox enzymes, transferase, or other enzyme.
- glucoamylase and/or a pullulanase phytase, a/pfra-glucosidase, isoamylase, beta-amylase cellulase, xylanase, other hemicellulases, protease, befa-glucosidase, pectinase, esterase, redox enzymes, transferase, or other enzyme.
- a variation on this process is a "fed-batch fermentation" system, where the substrate is added in increments as the fermentation progresses.
- Fed-batch systems are useful when catabolite repression may inhibit the metabolism of the cells and where it is desirable to have limited amounts of substrate in the medium.
- the actual substrate concentration in fed-batch systems is estimated by the changes of measurable factors such as pH, dissolved oxygen and the partial pressure of waste gases, such as C0 2 .
- Batch and fed-batch fermentations are common and well known in the art.
- Continuous fermentation is an open system where a defined fermentation medium is added continuously to a bioreactor, and an equal amount of conditioned medium is removed simultaneously for processing. Continuous fermentation generally maintains the cultures at a constant high density where cells are primarily in log phase growth.
- Continuous fermentation permits modulation of cell growth and/or product concentration.
- a limiting nutrient such as the carbon source or nitrogen source is maintained at a fixed rate and all other parameters are allowed to moderate. Because growth is maintained at a steady state, cell loss due to medium being drawn off should be balanced against the cell growth rate in the fermentation.
- Methods of optimizing continuous fermentation processes and maximizing the rate of product formation are well known in the art of industrial microbiology.
- compositions Comprising Maltogenic Amylases
- the maltogenic amylases of the present teachings may be combined with a glucoamylase (EC 3.2.1 .3), e.g., a Trichoderma glucoamylase or variant thereof.
- a glucoamylase EC 3.2.1 .3
- An exemplary glucoamylase is Trichoderma reesei glucoamylase (TrGA) and variants thereof that possess superior specific activity and thermal stability. See U.S. Published Applications Nos. 2006/0094080, 2007/0004018, and 2007/0015266 (Danisco US Inc.).
- Suitable variants of TrGA include those with glucoamylase activity and at least 80%, at least 90%, or at least 95% sequence identity to wild-type TrGA.
- Maltogenic amylases may advantageously increase the yield of glucose produced in a saccharification process catalyzed by TrGA.
- the glucoamylase may be another glucoamylase derived from plants (including algae), fungi, or bacteria.
- the glucoamylases may be Aspergillus niger Q ⁇ or G2 glucoamylase or its variants ⁇ e.g., Boel et al. (1984) EMBO J. 3: 1097-1 102; WO 92/00381 ; WO 00/04136 (Novo Nordisk A/S)); and A. awamori glucoamylase ⁇ e.g., WO 84/02921 (Cetus Corp.)).
- contemplated Aspergillus glucoamylase include variants with enhanced thermal stability, e.g., G137A and G1 39A (Chen etal. (1 996) Prot. Eng. 9: 499-505) ; D257E and D293E/Q (Chen et al. (1995) Prot. Eng. 8: 575-582) ; N182 (Chen et al. (1 994) Biochem. J. 301 : 275-281 ) ; A246C (Fierobe et al. (1996) Biochemistry, 35: 8698-8704); and variants with Pro residues in positions A435 and S436 (Li et al. (1 997) Protein Eng. 10: 1199-1204).
- Other contemplated glucoamylases include Talaromyces glucoamylases, in particular derived from T.
- Contemplated bacterial glucoamylases include glucoamylases from the genus Clostridium, in particular C. thermoamylolyticum ⁇ e.g., EP 135, 1 38 (CPC
- thermohydrosulfuricum ⁇ e.g., WO 86/01 831 (Michigan
- Suitable glucoamylases include the glucoamylases derived from Aspergillus oryzae, such as a glucoamylase shown in SEQ ID NO:2 in WO
- glucoamylases such as AMG 200L; AMG 300 L; SANTM SUPER and AMGTM E (Novozymes); OPTIDEX® 300 and OPTI DEX L-400 (Danisco US Inc.); AMIGASETMand AMIGASETM PLUS (DSM); G-ZYME® G900 (Enzyme Bio-Systems) ; and G-ZYME® G990 ZR ⁇ A. nigex
- glucoamylase with a low protease content.
- Still other suitable glucoamylases include Aspergillus fumigatus glucoamylase, Talaromyces glucoamylase, Thielavia
- glucoamylases typically are added in an amount of about 0.1 - 2 glucoamylase units (GAU)/g ds, e.g., about 0.1 6 GAU/g ds, 0.23 GAU/g ds, or 0.33 GAU/g ds.
- GAU glucoamylase units
- Suitable enzymes that can be used with the maltogenic amylase of the present teachings include a phytase, protease, pullulanase, ⁇ -amylase, isoamylase, a different a-amylase, alpha-glucosidase, cellulase, xylanase, other hemicellulases, beta-glucosidase, transferase, pectinase, lipase, cutinase, esterase, redox enzymes, or a combination thereof.
- a debranching enzyme such as an isoamylase (EC 3.2.1 .68), may be added in effective amounts well known to the person skilled in the art.
- a pullulanase (EC 3.2.1 .41 ), e.g., Promozyme®, is also suitable. Pullulanase typically is added at 100 U/kg ds. Further suitable enzymes include proteases, such as fungal and bacterial proteases. Fungal proteases include those obtained from
- ⁇ -Amylases (EC 3.2.1 .2) are exo-acting maltogenic amylases, which catalyze the hydrolysis of 1 ,4-a-glucosidic linkages into amylopectin and related glucose polymers, thereby releasing maltose.
- ⁇ -Amylases have been isolated from various plants and microorganisms. See Fogarty et al. (1979) in P ROGRESS IN INDUSTRIAL MICROBIOLOGY, Vol. 15, pp. 11 2-11 5.
- ⁇ -Amylases have optimum temperatures in the range from 40°C to 65°C and optimum pH in the range from about 4.5 to about 7.0.
- Contemplated ⁇ -amylases include, but are not limited to, ⁇ -amylases from barley Spezyme® BBA 1500, Spezyme® DBA, OptimaltTMME, OptimaltTMBBA (Danisco US Inc.); and NovozymTM WBA (Novozymes A/S) .
- compositions comprising the present maltogenic amylases may be aqueous or non-aqueous formulations, granules, powders, gels, slurries, pastes, etc., which may further comprise any one or more of the additional enzymes listed, herein, along with buffers, salts, preservatives, water, co-solvents, surfactants, and the like.
- Such compositions may work in combination with endogenous enzymes or other ingredients already present in a slurry, water bath, washing machine, food or drink product, etc, for example, endogenous plant (including algal) enzymes, residual enzymes from a prior processing step, and the like.
- the present teachings also relate to a "food composition,” including but not limited to a food product, animal feed and/or food/feed additives, comprising a maltogenic amylase, and methods for preparing such a food composition comprising mixing the maltogenic amylase with one or more food ingredients, or uses thereof.
- a "food composition” including but not limited to a food product, animal feed and/or food/feed additives, comprising a maltogenic amylase, and methods for preparing such a food composition comprising mixing the maltogenic amylase with one or more food ingredients, or uses thereof.
- the present teachings relate to the use of a maltogenic amylase in the preparation of a food composition, wherein the food composition is baked subsequent to the addition of the polypeptide of the present invention.
- baking composition means any composition and/or additive prepared in the process of providing a baked food product, including but not limited to bakers flour, a dough, a baking additive and/or a baked product.
- the food composition or additive may be liquid or solid.
- flour means milled or ground cereal grain.
- the term “flour” also may mean Sago or tuber products that have been ground or mashed.
- flour may also contain components in addition to the milled or mashed cereal or plant matter.
- Cereal grains include wheat, oat, rye, and barley.
- Tuber products include tapioca flour, cassava flour, and custard powder.
- the term “flour” also includes ground corn flour, maize-meal, rice flour, whole-meal flour, self-rising flour, tapioca flour, cassava flour, ground rice, enriched flower, and custard powder.
- a maltogenic amylase by itself or in combination with an a-amylase(s), may be added to the flour to augment the level of endogenous ⁇ -amylase activity in flour.
- a maltogenic amylase can further be added alone or in a combination with other amylases to prevent or retard staling, i.e., crumb firming of baked products.
- the amount of anti-staling amylase will typically be in the range of 0.01 -1 0 mg of enzyme protein per kg of flour, e.g., 0.5 mg/kg ds.
- Additional anti-staling amylases that can be used in combination with an amylase include an endo-amylase, e.g., a bacterial endo-amylase from Bacillus.
- the additional amylase can be another maltogenic a-amylase (EC 3.2.1 .133), e.g., from Bacillus.
- Novamyl® is an exemplary maltogenic a-amylase from B. stearothermophilus strain NCIB 1 1837 and is described in
- anti-staling endo-amylases include bacterial a-amylases derived from Bacillus, such as B.
- the anti-staling amylase may be an exo-amylase, such as ⁇ -amylase, e.g., from plant sources, such as soy bean, or from microbial sources, such as Bacillus.
- the baking composition comprising a maltogenic amylase further can comprise a phospholipase or enzyme with phospholipase activity.
- An enzyme with phospholipase activity has an activity that can be measured in Lipase Units (LU).
- the phospholipase may have A- ⁇ or A 2 activity to remove fatty acid from the phospholipids, forming a lysophospholipid. It may or may not have lipase activity, i.e., activity on triglyceride substrates.
- the phospholipase typically has a temperature optimum in the range of 30-90 °C, e.g., 30-70 °C.
- the added phospholipases can be of animal origin, for example, from pancreas, e.g., bovine or porcine pancreas, snake venom or bee venom.
- the phospholipase may be of microbial origin, e.g., from filamentous fungi, yeast or bacteria, for example.
- the phospholipase is added in an amount that improves the softness of the bread during the initial period after baking, particularly the first 24 hours.
- the amount of phospholipase will typically be in the range of 0.01 -1 0 mg of enzyme protein per kg of flour, e.g., 0.1 -5 mg/kg. That is, phospholipase activity generally will be in the range of 20-1000 LU/kg of flour, where a Lipase Unit is defined as the amount of enzyme required to release 1 ⁇ butyric acid per minute at 30 °C, pH 7.0, with gum arabic as emulsifier and tributyrin as substrate.
- Compositions of dough generally comprise wheat meal or wheat flour and/or other types of meal, flour or starch such as corn flour, cornstarch, rye meal, rye flour, oat flour, oatmeal, soy flour, sorghum meal, sorghum flour, potato meal, potato flour or potato starch.
- the dough may be fresh, frozen or par-baked.
- the dough can be a leavened dough or a dough to be subjected to leavening.
- the dough may be leavened in various ways, such as by adding chemical leavening agents, e.g., sodium bicarbonate or by adding a leaven, i.e., fermenting dough.
- Dough also may be leavened by adding a suitable yeast culture, such as a culture of Saccharomyces cerevisiae (baker's yeast), e.g., a commercially available strain of S. cerevisiae.
- the dough may also comprise other conventional dough ingredients, e.g., proteins, such as milk powder, gluten, and soy; eggs (e.g., whole eggs, egg yolks or egg whites); an oxidant, such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA) or ammonium persulfate; an amino acid such as L-cysteine; a sugar; or a salt, such as sodium chloride, calcium acetate, sodium sulfate or calcium sulfate.
- a suitable yeast culture such as a culture of Saccharomyces cerevisiae (baker's yeast), e.g., a commercially available strain of S. cerevisiae.
- the dough may also
- the dough further may comprise fat, e.g., triglyceride, such as granulated fat or shortening.
- the dough further may comprise an emulsifier such as mono- or diglycerides, diacetyl tartaric acid esters of mono- or diglycerides, sugar esters of fatty acids, polyglycerol esters of fatty acids, lactic acid esters of monoglycerides, acetic acid esters of monoglycerides, polyoxyethylene stearates, or lysolecithin.
- the dough can be made without addition of emulsifiers.
- the dough product may be any processed dough product, including fried, deep fried, roasted, baked, steamed and boiled doughs, such as steamed bread and rice cakes.
- the food product is a bakery product.
- Typical bakery (baked) products include bread - such as loaves, rolls, buns, bagels, pizza bases etc. pastry, pretzels, tortillas, cakes, cookies, biscuits, crackers etc.
- an additional enzyme may be used together with the anti-staling amylase and the phospholipase.
- the additional enzyme may be a second amylase, such as an amyloglucosidase, a ⁇ -amylase, a cyclodextrin glucanotransferase, or the additional enzyme may be a peptidase, in particular an exopeptidase, a transglutaminase, a lipase, a cellulase, a xylanase, a protease, a protein disulfide isomerase, e.g., a protein disulfide isomerase as disclosed in WO 95/00636, for example, a glycosyltransferase, a branching enzyme (1 ,4-a-glucan branching enzyme), a 4-a-glucanotransferase (dextrin glycosyltransferase) or an oxidoreductase
- carbohydrate oxidase may be of any origin, including mammalian and plant, and particularly of microbial (bacterial, yeast or fungal) origin and may be obtained by techniques conventionally used in the art.
- the xylanase is typically of microbial origin, e.g., derived from a bacterium or fungus, such as a strain of Aspergillus.
- Xylanases include Pentopan® and Novozym 384®, for example, which are commercially available xylanase preparations produced from Trichoderma reesei.
- the amyloglucosidase may be an A. n/ger amyloglucosidase (such as AMG®).
- Other useful amylase products include Grindamyl® A 1000 or A 5000 (Grindsted Products, Denmark) and Amylase® H or Amylase® P (DS ).
- the glucose oxidase may be a fungal glucose oxidase, in particular an Aspergillus niger glucose oxidase (such as Gluzyme®).
- An exemplary protease is Neutrase®.
- the process may be used for any kind of baked product prepared from dough, either of a soft or a crisp character, either of a white, light or dark type.
- Examples are bread, particularly white, whole-meal or rye bread, typically in the form of loaves or rolls, such as, but not limited to, French baguette-type bread, pita bread, tortillas, cakes, pancakes, biscuits, cookies, pie crusts, crisp bread, steamed bread, pizza and the like.
- a maltogenic amylase may be used in a pre-mix, comprising flour together with an anti-staling amylase, a phospholipase, and/or a phospholipid.
- the pre-mix may contain other dough-improving and/or bread-improving additives, e.g., any of the additives, including enzymes, mentioned above.
- a maltogenic amylase can be a component of an enzyme preparation comprising an anti-staling amylase and a phospholipase, for use as a baking additive.
- the enzyme preparation is optionally in the form of a granulate or agglomerated powder.
- the preparation can have a narrow particle size distribution with more than 95% (by weight) of the particles in the range from 25 to 500 ⁇ .
- Granulates and agglomerated powders may be prepared by conventional methods, e.g., by spraying an amylase onto a carrier in a fluid-bed granulator.
- the carrier may consist of particulate cores having a suitable particle size.
- the carrier may be soluble or insoluble, e.g., a salt (such as NaCI or sodium sulfate), a sugar (such as sucrose or lactose), a sugar alcohol (such as sorbitol), starch, rice, corn grits, or soy.
- Enveloped particles i.e., maltogenic amylase particles
- the enzyme is contacted with a food grade lipid in sufficient quantity to suspend all of the maltogenic amylase particles.
- Food grade lipids may be any naturally organic compound that is insoluble in water but is soluble in non-polar organic solvents such as hydrocarbon or diethyl ether. Suitable food grade lipids include, but are not limited to, triglycerides either in the form of fats or oils that are either saturated or unsaturated.
- fatty acids and combinations thereof which make up the saturated triglycerides include, but are not limited to, butyric (derived from milk fat), palmitic (derived from animal and plant fat), and/or stearic (derived from animal and plant fat).
- fatty acids and combinations thereof which make up the unsaturated triglycerides include, but are not limited to, palmitoleic (derived from animal and plant fat), oleic (derived from animal and plant fat), linoleic (derived from plant oils), and/or linolenic (derived from linseed oil).
- Other suitable food grade lipids include, but are not limited to, monoglycerides and diglycerides derived from the triglycerides discussed above, phospholipids and glycolipids.
- the food grade lipid is contacted with a powdered form of the maltogenic amylase particles in such a fashion that the lipid material covers at least a portion of the surface of at least a majority, e.g., 100% of the maltogenic amylase particles.
- each maltogenic amylase particle is individually enveloped in a lipid.
- all or substantially all of the maltogenic amylase particles are provided with a thin, continuous, enveloping film of lipid. This can be accomplished by first pouring a quantity of lipid into a container, and then slurrying the maltogenic amylase particles so that the lipid thoroughly wets the surface of each maltogenic amylase particle.
- the enveloped maltogenic amylase particles carrying a substantial amount of the lipids on their surfaces, are recovered.
- the thickness of the coating so applied to the particles of maltogenic amylase can be controlled by selection of the type of lipid used and by repeating the operation in order to build up a thicker film, when desired.
- the storing, handling and incorporation of the loaded delivery vehicle can be accomplished by means of a packaged mix.
- the packaged mix can comprise the enveloped maltogenic amylase.
- the packaged mix may further contain additional ingredients as required by the manufacturer or baker. After the enveloped maltogenic amylase has been incorporated into the dough, the baker continues through the normal production process for that product.
- the food grade lipid protects the enzyme from thermal denaturation during the baking process for those enzymes that are heat labile. Consequently, while the maltogenic amylase is stabilized and protected during the proving and baking stages, it is released from the protective coating in the final baked good product, where it hydrolyzes the glucosidic linkages in polyglucans.
- the loaded delivery vehicle also provides a sustained release of the active enzyme into the baked good. That is, following the baking process, active maltogenic amylase is continually released from the protective coating at a rate that counteracts, and therefore reduces the rate of, staling mechanisms.
- the amount of lipid applied to the maltogenic amylase particles can vary from a few percent of the total weight of the maltogenic amylase to many times that weight, depending upon the nature of the lipid, the manner in which it is applied to the maltogenic amylase particles, the composition of the dough mixture to be treated, and the severity of the dough-mixing operation involved.
- the loaded delivery vehicle i.e., the lipid-enveloped enzyme
- the baker computes the amount of enveloped maltogenic amylase, prepared as discussed above, that will be required to achieve the desired anti-staling effect.
- the amount of the enveloped maltogenic amylase required is calculated based on the concentration of enzyme enveloped and on the proportion of maltogenic amylase to flour specified. A wide range of concentrations has been found to be effective, although, as has been discussed, observable improvements in anti-staling do not correspond linearly with the maltogenic amylase concentration, but above certain minimal levels, large increases in maltogenic amylase concentration produce little additional improvement.
- the maltogenic amylase concentration actually used in a particular bakery production could be much higher than the minimum necessary to provide the baker with some insurance against inadvertent under-measurement errors by the baker.
- the lower limit of enzyme concentration is determined by the minimum anti-staling effect the baker wishes to achieve.
- a method of preparing a baked good may comprise: a) preparing lipid-coated maltogenic amylase particles, where substantially all of the maltogenic amylase particles are coated; b) mixing a dough containing flour; c) adding the lipid-coated maltogenic amylase to the dough before the mixing is complete and terminating the mixing before the lipid coating is removed from the maltogenic amylase; d) proofing the dough; and e) baking the dough to provide the baked good, where the maltogenic amylase is inactive during the mixing, proofing and baking stages and is active in the baked good.
- the enveloped maltogenic amylase can be added to the dough during the mix cycle, e.g., near the end of the mix cycle.
- the enveloped maltogenic amylase is added at a point in the mixing stage that allows sufficient distribution of the enveloped maltogenic amylase throughout the dough ; however, the mixing stage is terminated before the protective coating becomes stripped from the maltogenic amylase particle(s).
- the mixing stage is terminated before the protective coating becomes stripped from the maltogenic amylase particle(s).
- anywhere from one to six minutes or more might be required to mix the enveloped maltogenic amylase into the dough, but two to four minutes is average. Thus, several variables may determine the precise procedure.
- the quantity of enveloped maltogenic amylase should have a total volume sufficient to allow the enveloped maltogenic amylase to be spread throughout the dough mix. If the preparation of enveloped maltogenic amylase is highly concentrated, additional oil may need to be added to the pre-mix before the enveloped maltogenic amylase is added to the dough. Recipes and production processes may require specific modifications; however, good results generally can be achieved when 25% of the oil specified in a bread dough formula is held out of the dough and is used as a carrier for a concentrated enveloped a-amylase when added near the end of the mix cycle.
- an enveloped maltogenic amylase mixture of approximately 1 % of the dry flour weight is sufficient to admix the enveloped a-amylase properly with the dough.
- the range of suitable percentages is wide and depends on the formula, finished product, and production methodology requirements of the individual baker.
- the enveloped maltogenic amylase suspension should be added to the mix with sufficient time for complete mixture into the dough, but not for such a time that excessive mechanical action strips the protective lipid coating from the enveloped maltogenic amylase particles.
- the food composition is an oil, meat, lard, composition comprising a maltogenic amylase.
- oil/meat/lard composition means any composition, based on, made from and/or containing oil, meat or lard, respectively.
- Another aspect the invention relates to a method of preparing an oil or meat or lard composition and/or additive comprising a maltogenic amylase, comprising mixing the polypeptide of the invention with a oil/meat/lard composition and/or additive ingredients.
- the food composition is an animal feed composition, animal feed additive and/or pet food comprising a maltogenic amylase and variants thereof.
- the present invention further relates to a method for preparing such an animal feed composition, animal feed additive composition and/or pet food comprising mixing a maltogenic amylase and variants thereof with one or more animal feed ingredients and/or animal feed additive ingredients and/or pet food ingredients.
- the present invention relates to the use of a maltogenic amylase in the preparation of an animal feed composition and/or animal feed additive composition and/or pet food.
- the term "animal" includes all non-ruminant and ruminant animals.
- the animal is a non-ruminant animal, such as a horse and a mono-gastric animal.
- mono-gastric animals include, but are not limited to, pigs and swine, such as piglets, growing pigs, sows; poultry such as turkeys, ducks, chicken, broiler chicks, layers; fish such as salmon, trout, tilapia, catfish and carps; and crustaceans such as shrimps and prawns.
- the animal is a ruminant animal including, but not limited to, cattle, young calves, goats, sheep, giraffes, bison, moose, elk, yaks, water buffalo, deer, camels, alpacas, llamas, antelope, pronghorn and nilgai.
- pet food is understood to mean a food for a household animal such as, but not limited to dogs, cats, gerbils, hamsters, chinchillas, fancy rats, guinea pigs; avian pets, such as canaries, parakeets, and parrots; reptile pets, such as turtles, lizards and snakes; and aquatic pets, such as tropical fish and frogs.
- animal feed composition may comprise one or more feed materials selected from the group comprising a) cereals, such as small grains ⁇ e.g., wheat, barley, rye, oats and combinations thereof) and/or large grains such as maize or sorghum ; b) by products from cereals, such as corn gluten meal, Distillers Dried Grain Solubles (DDGS) (particularly corn based Distillers Dried Grain Solubles (cDDGS), wheat bran, wheat middlings, wheat shorts, rice bran, rice hulls, oat hulls, palm kernel, and citrus pulp; c) protein obtained from sources such as soya, sunflower, peanut, lupin, peas, fava beans, cotton, canola, fish meal, dried plasma protein, meat and bone meal, potato protein, whey, copra, sesame; d) oils and fats obtained from vegetable and animal sources; e) minerals
- compositions and methods of treating fabrics ⁇ e.g., to desize a textile) using a maltogenic amylase.
- Fabric-treating methods are well known in the art ⁇ see, e.g., U.S. Patent No. 6,077,316).
- the feel and appearance of a fabric can be improved by a method comprising contacting the fabric with a maltogenic amylase in a solution.
- the fabric can be treated with the solution under pressure.
- a maltogenic amylase can be applied during or after the weaving of a textile, or during the desizing stage, or one or more additional fabric processing steps. During the weaving of textiles, the threads are exposed to considerable mechanical strain. Prior to weaving on mechanical looms, warp yarns are often coated with sizing starch or starch derivatives to increase their tensile strength and to prevent breaking. A maltogenic amylase can be applied during or after the weaving to remove these sizing starch or starch derivatives. After weaving, a maltogenic amylase can be used to remove the size coating before further processing the fabric to ensure a homogeneous and wash-proof result.
- a maltogenic amylase can be used alone or with other desizing chemical reagents and/or desizing enzymes to desize fabrics, including cotton-containing fabrics, as detergent additives, e.g., in aqueous compositions.
- a maltogenic amylase also can be used in compositions and methods for producing a stonewashed look on indigo-dyed denim fabric and garments.
- the fabric can be cut and sewn into clothes or garments, which are afterwards finished.
- different enzymatic finishing methods have been developed.
- the finishing of denim garment normally is initiated with an enzymatic desizing step, during which garments are subjected to the action of amylolytic enzymes to provide softness to the fabric and make the cotton more accessible to the subsequent enzymatic finishing steps.
- a maltogenic amylase can be used in methods of finishing denim garments ⁇ e.g., a "bio-stoning process"), enzymatic desizing and providing softness to fabrics, and/or finishing process.
- An aspect of the present compositions and methods is a cleaning composition that includes a maltogenic amylase as a component.
- a maltogenic amylase polypeptide can be used as a component in detergent compositions for hand washing, laundry washing, dishwashing, and other hard-surface cleaning.
- a maltogenic amylase is incorporated into detergents at or near a concentration conventionally used for amylase in detergents.
- a maltogenic amylase polypeptide may be added in amount corresponding to 0.00001 - 1 mg (calculated as pure enzyme protein) of maltogenic amylase per liter of
- wash/dishwash liquor exemplary formulations are provided herein, as exemplified by the following:
- a maltogenic amylase polypeptide may be a component of a detergent composition, as the only enzyme or with other enzymes including other amylolytic enzymes. As such, it may be included in the detergent composition in the form of a non-dusting granulate, a stabilized liquid, or a protected enzyme. Non-dusting granulates may be produced, e.g., as disclosed in U.S. Patent Nos. 4,1 06,991 and 4,661 ,452 and may optionally be coated by methods known in the art.
- waxy coating materials are poly(ethylene oxide) products (polyethyleneglycol, PEG) with mean molar weights of 1 ,000 to 20,000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids.
- film-forming coating materials suitable for application by fluid bed techniques are given in, for example, GB 1483591 .
- Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other enzyme stabilizers are known in the art.
- Protected enzymes may be prepared according to the method disclosed in for example EP 238 21 6. Polyols have long been recognized as stabilizers of proteins, as well as improving protein solubility.
- the detergent composition may be in any useful form, e.g., as powders, granules, pastes, or liquid.
- a liquid detergent may be aqueous, typically containing up to about 70% of water and 0% to about 30% of organic solvent. It may also be in the form of a compact gel type containing only about 30% water.
- the detergent composition comprises one or more surfactants, each of which may be anionic, nonionic, cationic, or zwitterionic.
- the detergent will usually contain 0% to about 50% of anionic surfactant, such as linear alkylbenzenesulfonate (LAS) ;
- AOS alkyl sulfate (fatty alcohol sulfate)
- AS alkyl sulfate
- AEOS or AES alcohol ethoxysulfate
- SAS secondary alkanesulfonates
- a-sulfo fatty acid methyl esters alkyl- or alkenylsuccinic acid; or soap.
- the composition may also contain 0% to about 40% of nonionic surfactant such as alcohol ethoxylate (AEO or AE), carboxylated alcohol ethoxylates, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (as described for example in WO 92/061 54).
- nonionic surfactant such as alcohol ethoxylate (AEO or AE), carboxylated alcohol ethoxylates, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (as described for example in WO 92/061 54).
- the detergent composition may additionally comprise one or more other enzymes, such as proteases, another amylolytic enzyme, cutinase, lipase, cellulase, pectate lyase, perhydrolase, xylanase, peroxidase, and/or laccase in any combination.
- enzymes such as proteases, another amylolytic enzyme, cutinase, lipase, cellulase, pectate lyase, perhydrolase, xylanase, peroxidase, and/or laccase in any combination.
- the detergent may contain about 1 % to about 65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA),
- a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA),
- the detergent may also be unbuilt, i.e. essentially free of detergent builder.
- the enzymes can be used in any composition compatible with the stability of the enzyme. Enzymes generally can be protected against deleterious components by known forms of encapsulation, for example, by granulation or sequestration in hydro gels. Enzymes, and specifically maltoogenic amylases, either with or without starch binding domains, can be used in a variety of compositions including laundry and dishwashing applications, surface cleaners, as well as in compositions for ethanol production from starch or biomass.
- the detergent may comprise one or more polymers. Examples include carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (PVP), polyethyleneglycol (PEG), polyvinyl alcohol) (PVA), polycarboxylates such as polyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid copolymers.
- CMC carboxymethylcellulose
- PVP poly(vinylpyrrolidone)
- PEG polyethyleneglycol
- PVA polyvinyl alcohol
- polycarboxylates such as polyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid copolymers.
- the detergent may contain a bleaching system, which may comprise a H 2 0 2 source such as perborate or percarbonate, which may be combined with a
- peracid-forming bleach activator such as tetraacetylethylenediamine (TAED) or nonanoyloxybenzenesulfonate (NOBS).
- TAED tetraacetylethylenediamine
- NOBS nonanoyloxybenzenesulfonate
- the bleaching system may comprise peroxyacids (e.g., the amide, imide, or sulfone type peroxyacids).
- the bleaching system can also be an enzymatic bleaching system, for example, perhydrolase, such as that described in International PCT Application WO 2005/056783.
- the enzymes of the detergent composition may be stabilized using conventional stabilizing agents, e.g., a polyol such as propylene glycol or glycerol; a sugar or sugar alcohol; lactic acid; boric acid or a boric acid derivative such as, e.g., an aromatic borate ester; and the composition may be formulated as described in, e.g., WO 92/19709 and WO 92/19708.
- stabilizing agents e.g., a polyol such as propylene glycol or glycerol
- a sugar or sugar alcohol lactic acid
- boric acid or a boric acid derivative such as, e.g., an aromatic borate ester
- the composition may be formulated as described in, e.g., WO 92/19709 and WO 92/19708.
- the detergent may also contain other conventional detergent ingredients such as e.g., fabric conditioners including clays, foam boosters, suds suppressors, anti-corrosion agents, soil-suspending agents, anti-soil redeposition agents, dyes, bactericides, tarnish inhibiters, optical brighteners, or perfumes.
- fabric conditioners including clays, foam boosters, suds suppressors, anti-corrosion agents, soil-suspending agents, anti-soil redeposition agents, dyes, bactericides, tarnish inhibiters, optical brighteners, or perfumes.
- the pH (measured in aqueous solution at use concentration) is usually neutral or alkaline, e.g., pH about 7.0 to about 1 1 .0.
- Exemplary HDL laundry detergent compositions includes a detersive surfactant (10%-40% wt/wt), including an anionic detersive surfactant (selected from a group of linear or branched or random chain, substituted or unsubstituted alkyl sulphates, alkyl sulphonates, alkyl alkoxylated sulphate, alkyl phosphates, alkyl phosphonates, alkyl carboxylates, and/or mixtures thereof), and optionally non-ionic surfactant (selected from a group of linear or branched or random chain, substituted or unsubstituted alkyl alkoxylated alcohol, for example a Cs-Ci 8 alkyl ethoxylated alcohol and/or C 6 -Ci 2 alkyl phenol alkoxylates), wherein the weight ratio of anionic detersive surfactant (with a hydrophilic index (Hlc) of from 6.0 to 9) to non-ionic detersive
- Suitable detersive surfactants also include cationic detersive surfactants (selected from a group of alkyl pyridinium compounds, alkyl quarternary ammonium compounds, alkyl quarternary phosphonium compounds, alkyl ternary sulphonium compounds, and/or mixtures thereof); zwitterionic and/or amphoteric detersive surfactants (selected from a group of alkanolamine sulpho-betaines) ; ampholytic surfactants; semi-polar non-ionic surfactants and mixtures thereof.
- cationic detersive surfactants selected from a group of alkyl pyridinium compounds, alkyl quarternary ammonium compounds, alkyl quarternary phosphonium compounds, alkyl ternary sulphonium compounds, and/or mixtures thereof
- zwitterionic and/or amphoteric detersive surfactants selected from a group of alkanolamine sulpho-betaines
- the composition may optionally include, a surfactancy boosting polymer consisting of amphiphilic alkoxylated grease cleaning polymers (selected from a group of alkoxylated polymers having branched hydrophilic and hydrophobic properties, such as alkoxylated polyalkylenimines in the range of 0.05wt%-10wt%) and/or random graft polymers (typically comprising of hydrophilic backbone comprising monomers selected from the group consisting of: unsaturated C-
- the composition may include additional polymers such as soil release polymers (include anionically end-capped polyesters, for example SRP1 , polymers comprising at least one monomer unit selected from saccharide, dicarboxylic acid, polyol and combinations thereof, in random or block configuration, ethylene terephthalate-based polymers and co-polymers thereof in random or block configuration, for example Repel-o-tex SF, SF-2 and SRP6, Texcare SRA1 00, SRA300, SRN100, SRN170, SRN240, SRN300 and SRN325, Marloquest SL), anti-redeposition polymers (0.1 wt% to 10wt%, include carboxylate polymers, such as polymers comprising at least one monomer selected from acrylic acid, maleic acid (or maleic anhydride), fumaric acid, itaconic acid, aconitic acid, mesaconic acid, citraconic acid, methylenemalonic acid, and any mixture thereof, vinylpyrrolidone homopol
- the composition may further include saturated or unsaturated fatty acid, preferably saturated or unsaturated Ci 2 -C 2 4 fatty acid (0 wt% to 10 wt%); deposition aids (examples for which include polysaccharides, preferably cellulosic polymers, poly diallyl dimethyl ammonium halides (DADMAC), and co-polymers of DAD MAC with vinyl pyrrolidone, acrylamides, imidazoles, imidazolinium halides, and mixtures thereof, in random or block configuration, cationic guar gum, cationic cellulose such as cationic hydoxyethyl cellulose, cationic starch, cationic polyacylamides, and mixtures thereof.
- deposition aids include polysaccharides, preferably cellulosic polymers, poly diallyl dimethyl ammonium halides (DADMAC), and co-polymers of DAD MAC with vinyl pyrrolidone, acrylamides, imi
- the composition may further include dye transfer inhibiting agents, examples of which include manganese phthalocyanine, peroxidases, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles and/or mixtures thereof; chelating agents, examples of which include ethylene-diamine-tetraacetic acid (EDTA), diethylene triamine penta methylene phosphonic acid (DTPMP), hydroxy-ethane diphosphonic acid (HEDP), ethylenediamine ⁇ , ⁇ '-disuccinic acid (EDDS), methyl glycine diacetic acid (MGDA), diethylene triamine penta acetic acid (DTPA), propylene diamine tetracetic acid (PDT A), 2-hydroxypyridine-N-oxide (HPNO), or methyl glycine diacetic acid (MGDA), glut
- TTHA triethylenetetraaminehexaacetic acid
- HEIDA N-hydroxyethyliminodiacetic acid
- DHEG dihydroxyethylglycine
- EDTP ethylenediaminetetrapropionic acid
- the composition preferably includes enzymes (generally about 0.01 wt% active enzyme to 0.03wt% active enzyme) selected from proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferases,
- enzymes generally about 0.01 wt% active enzyme to 0.03wt% active enzyme selected from proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferases,
- the composition may include an enzyme stabilizer (examples of which include polyols such as propylene glycol or glycerol, sugar or sugar alcohol, lactic acid, reversible protease inhibitor, boric acid, or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid).
- an enzyme stabilizer examples of which include polyols such as propylene glycol or glycerol, sugar or sugar alcohol, lactic acid, reversible protease inhibitor, boric acid, or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid).
- composition optionally includes silicone or fatty-acid based suds
- hueing dyes 0.001 wt% to about 4.0wt%), and/or structurant/thickener (0.01 wt% to 5wt%, selected from the group consisting of diglycerides and triglycerides, ethylene glycol distearate, microcrystalline cellulose, cellulose based materials, microfiber cellulose, biopolymers, xanthan gum, gellan gum, and mixtures thereof).
- the composition can be any liquid form, for example a liquid or gel form, or any combination thereof.
- the composition may be in any unit dose form, for example a pouch.
- Exemplary HDD laundry detergent compositions includes a detersive surfactant, including anionic detersive surfactants (e.g., linear or branched or random chain, substituted or unsubstituted alkyi sulphates, alkyi sulphonates, alkyi alkoxylated sulphate, alkyi phosphates, alkyi phosphonates, alkyi carboxylates and/or mixtures thereof), non-ionic detersive surfactant (e.g., linear or branched or random chain, substituted or unsubstituted C 8 -C 8 alkyi ethoxylates, and/or C 6 -C 2 alkyi phenol alkoxylates), cationic detersive surfactants (e.g., alkyi pyridinium compounds, alkyi quaternary ammonium compounds, alkyi quaternary phosphonium compounds, alkyi ternary sulphonium compounds, and mixtures thereof), zwitterionic
- carbonate salt e.g., sodium carbonate and/or sodium bicarbonate in the range of 0 wt% to less than 80 wt%)
- bleaching agents including photobleaches (e.g., sulfonated zinc phthalocyanines, sulfonated aluminum phthalocyanines, xanthenes dyes, and mixtures thereof) hydrophobic or hydrophilic bleach activators (e.g., dodecanoyl oxybenzene sulfonate, decanoyl oxybenzene sulfonate, decanoyl oxybenzoic acid or salts thereof, 3,5,5-trimethy hexanoyl oxybenzene sulfonate, tetraacetyl ethylene diamine-TAED, nonanoyloxybenzene sulfonate-NOBS, nitrile quats, and mixtures thereof), sources of hydrogen peroxide (e.g., inorganic perhydrate salts examples of
- the composition preferably includes enzymes, e.g., proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases,
- enzymes e.g., proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases,
- mannanases cutinases, laccases, phospholipases, lysophospholipases, acyltransferase, perhydrolase, arylesterase, and any mixture thereof.
- composition may optionally include additional detergent ingredients including perfume microcapsules, starch encapsulated perfume accord, hueing agents, additional polymers, including fabric integrity and cationic polymers, dye-lock ingredients, fabric-softening agents, brighteners (for example C.I. Fluorescent brighteners), flocculating agents, chelating agents, alkoxylated polyamines, fabric deposition aids, and/or cyclodextrin.
- additional detergent ingredients including perfume microcapsules, starch encapsulated perfume accord, hueing agents, additional polymers, including fabric integrity and cationic polymers, dye-lock ingredients, fabric-softening agents, brighteners (for example C.I. Fluorescent brighteners), flocculating agents, chelating agents, alkoxylated polyamines, fabric deposition aids, and/or cyclodextrin.
- ADW detergent composition includes non-ionic surfactants, including ethoxylated non-ionic surfactants, alcohol alkoxylated surfactants, epoxy-capped poly(oxyalkylated) alcohols, or amine oxide surfactants present in amounts from 0 to 1 0% by weight; builders in the range of 5-60% including phosphate builders ⁇ e.g.,
- phosphate-free builders e.g., amino acid-based compounds including methyl-glycine-diacetic acid (MGDA) and salts and derivatives thereof, glutamic-N,N-diacetic acid (GLDA) and salts and derivatives thereof,
- iminodisuccinic acid and salts and derivatives thereof, carboxy methyl inulin and salts and derivatives thereof, nitrilotriacetic acid (NTA), diethylene triamine penta acetic acid (DTPA), B-alaninediacetic acid (B-ADA) and their salts, homopolymers and copolymers of poly-carboxylic acids and their partially or completely neutralized salts, monomeric polycarboxylic acids and hydroxycarboxylic acids and their salts in the range of 0.5% to 50% by weight; sulfonated/carboxylated polymers in the range of about 0.1 % to about 50% by weight to provide dimensional stability; drying aids in the range of about 0.1 % to about 10% by weight ⁇ e.g., polyesters, especially anionic polyesters, optionally together with further monomers with 3 to 6 functionalities - typically acid, alcohol or ester functionalities which are conducive to polycondensation, polycarbonate-, polyurethane- and
- the present maltogenic amylase polypeptide may be incorporated at a concentration conventionally employed in detergents. It is at present contemplated that, in the detergent composition, the enzyme may be added in an amount corresponding to 0.00001 -1 .0 mg (calculated as pure enzyme protein) of amylase polypeptide per liter of wash liquor.
- the detergent composition may also contain other conventional detergent ingredients, e.g., deflocculant material, filler material, foam depressors, anti-corrosion agents, soil-suspending agents, sequestering agents, anti-soil redeposition agents, dehydrating agents, dyes, bactericides, fluorescers, thickeners, and perfumes.
- the detergent composition may be formulated as a hand (manual) or machine (automatic) laundry detergent composition, including a laundry additive composition suitable for pre-treatment of stained fabrics and a rinse added fabric softener composition, or be formulated as a detergent composition for use in general household hard surface cleaning operations, or be formulated for manual or automatic dishwashing operations.
- any of the cleaning compositions described, herein, may include any number of additional enzymes.
- the enzyme(s) should be compatible with the selected detergent, ⁇ e.g., with respect to pH-optimum, compatibility with other enzymatic and non-enzymatic ingredients, and the like), and the enzyme(s) should be present in effective amounts.
- the following enzymes are provided as examples.
- proteases include those of animal, vegetable or microbial origin. Chemically modified or protein engineered mutants are included, as well as naturally processed proteins.
- the protease may be a serine protease or a metalloprotease, an alkaline microbial protease, a trypsin-like protease, or a
- subtilisin-like protease examples include subtilisins, especially those derived from Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin 309, subtilisin 147, and subtilisin 168 ⁇ see, e.g., WO 89/06279).
- trypsin-like proteases are trypsin (e.g., of porcine or bovine origin), and Fusarium proteases ⁇ see, e.g., WO 89/06270 and WO 94/25583).
- proteases also include but are not limited to the variants described in WO 92/1 9729, WO 98/2011 5, WO 98/20116, and WO 98/34946.
- Commercially available protease enzymes include but are not limited to: ALCALASE®, SAVINASE®, PRIMASETM, DURALASETM, ESPERASE®, KANNASETM, and BLAZETM (Novo Nordisk A/S and Novozymes A/S); MAXATASE®, MAXACALTM, MAXAPEMTM, PROPERASE®, PURAFECT®, PURAFECT OXPTM, FN2TM, and FN3TM (Danisco US Inc.).
- Other exemplary proteases include NprE from Bacillus amyloliquifaciens and ASP from Cellulomonas sp. strain 69B4.
- Suitable lipases include those of bacterial or fungal origin. Chemically modified, proteolytically modified, or protein engineered mutants are included. Examples of useful lipases include but are not limited to lipases from Humicola (synonym Thermomyces), e.g., from H. lanuginosa ( T. lanuginosus) ⁇ see e.g., EP 258068 and EP 305216), from H. insolens ⁇ see e.g., WO 96/13580); a Pseudomonas lipase ⁇ e.g., from P. alcaligenes or P. pseudoalcaligenes; see, e.g., EP 218 272), P.
- cepacia see e.g., EP 331 376
- P. stutzeri see e.g., GB 1 ,372,034
- P. fluorescens Pseudomonas sp. strain SD 705 ⁇ see e.g., WO 95/06720 and WO 96/27002
- P. wisconsinensis ⁇ see e.g., WO 96/12012
- Bacillus lipase ⁇ e.g., from B. subtilis; see e.g., Dartois et al. Biochemica et Biophysica Acta, 11 31 : 253-360 (1993)
- B. stearothermophilus see e.g., JP
- Additional lipase variants contemplated for use in the formulations include those described for example in: WO 92/05249, WO 94/01541 , WO 95/35381 , WO 96/00292, WO 95/30744, WO 94/25578, WO 95/14783, WO 95/2261 5, WO 97/04079, WO 97/07202, EP 407225, and EP 260105.
- Some commercially available lipase enzymes include LIPOLASE® and LI POLASE ULTRATM (Novo Nordisk A/S and Novozymes A/S).
- Poly esterases Suitable polyesterases can be included in the composition, such as those described in, for example, WO 01 /34899, WO 01 /14629, and US6933140.
- Amylases The compositions can be combined with other amylases, such as non-production enhanced amylase. These can include commercially available amylases, such as but not limited to STAINZYME®, NATALASE®, DURAMYL®, TERMAMYL®, FUNGAMYL® and BANTM (Novo Nordisk A/S and Novozymes A/S) ; RAPIDASE®, POWERASE®, and PURASTAR® (from Danisco US Inc.).
- amylases such as but not limited to STAINZYME®, NATALASE®, DURAMYL®, TERMAMYL®, FUNGAMYL® and BANTM (Novo Nordisk A/S and Novozymes A/S) ; RAPIDASE®, POWERASE®, and PURASTAR® (from Danisco US Inc.).
- Cellulases can be added to the compositions. Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed for example in U.S. Patent Nos. 4,435,307; 5,648,263; 5,691 ,178; 5,776,757; and WO 89/09259.
- Exemplary cellulases contemplated for use are those having color care benefit for the textile.
- Examples of such cellulases are cellulases described in for example EP 0495257, EP 0531372, WO 96/11262, WO 96/29397, and WO 98/08940.
- Other examples are cellulase variants, such as those described in WO 94/07998; WO 98/1 2307; WO 95/24471 ; PCT/DK98/00299; EP
- cellulases include CELLUZYME® and CAR EZYME® (Novo Nordisk A/S and Novozymes A/S); CLAZINASE® and PURADAX HA® (Danisco US Inc.); and
- Peroxidases/Oxidases Suitable peroxidases/oxidases contemplated for use in the compositions include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinus, e.g., from C. cinereus, and variants thereof as those described in WO 93/24618, WO 95/1 0602, and WO 98/1 5257. Commercially available peroxidases include for example GUARDZYMETM(Novo Nordisk A/S and Novozymes A/S).
- the detergent composition can also comprise 2,6-p-D-fructan hydrolase, which is effective for removal/cleaning of biofilm present on household and/or industrial textile/laundry.
- the detergent enzyme(s) may be included in a detergent composition by adding separate additives containing one or more enzymes, or by adding a combined additive comprising all of these enzymes.
- a detergent additive i.e. a separate additive or a combined additive, can be formulated e.g., as a granulate, a liquid, a slurry, and the like.
- Exemplary detergent additive formulations include but are not limited to granulates, in particular non-dusting granulates, liquids, in particular stabilized liquids or slurries.
- Non-dusting granulates may be produced, e.g., as disclosed in U.S. Patent Nos. 4,106,991 and 4,661 ,452 and may optionally be coated by methods known in the art.
- waxy coating materials are poly(ethylene oxide) products ⁇ e.g.,
- polyethyleneglycol, PEG polyethyleneglycol, PEG with mean molar weights of 1 ,000 to 20,000; ethoxylated nonylphenols having from 1 6 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids.
- film-forming coating materials suitable for application by fluid bed techniques are given in, for example, GB 1483591 .
- Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods.
- Protected enzymes may be prepared according to the method disclosed in EP 238,216.
- the detergent composition may be in any convenient form, e.g., a bar, a tablet, a powder, a granule, a paste, or a liquid.
- a liquid detergent may be aqueous, typically containing up to about 70% water, and 0% to about 30% organic solvent.
- Compact detergent gels containing about 30% or less water are also contemplated.
- the detergent composition can optionally comprise one or more surfactants, which may be non-ionic, including semi-polar and/or anionic and/or cationic and/or zwitterionic.
- the surfactants can be present in a wide range, from about 0.1 % to about 60% by weight.
- the detergent When included therein the detergent will typically contain from about 1 % to about 40% of an anionic surfactant, such as linear alkylbenzenesulfonate,
- alkyl sulfate fatty alcohol sulfate
- alcohol ethoxysulfate secondary alkanesulfonate
- a-sulfo fatty acid methyl ester alkyl- or alkenylsuccinic acid, or soap.
- the detergent When included therein, the detergent will usually contain from about 0.2% to about 40% of a non-ionic surfactant such as alcohol ethoxylate, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, polyhydroxy alkyl fatty acid amide, or N-acyl-N-alkyl derivatives of glucosamine (“glucamides").
- a non-ionic surfactant such as alcohol ethoxylate, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, polyhydroxy alkyl fatty acid amide, or N-acyl-N-alkyl derivatives of glucosamine (“glucamides”).
- glucamides N-acyl-N-alkyl derivatives of glucosamine
- the detergent may contain 0% to about 65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, carbonate, citrate, nitrilotriacetic acid, ethylenediaminetetraacetic acid (EDTA),
- a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, carbonate, citrate, nitrilotriacetic acid, ethylenediaminetetraacetic acid (EDTA),
- diethylenetriaminepentaacetic acid alkyl- or alkenylsuccinic acid, soluble silicates or layered silicates (e.g.,SKS-6 from Hoechst).
- the detergent may comprise one or more polymers.
- Exemplary polymers include carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (PVP), poly(ethylene glycol) (PEG), polyvinyl alcohol) (PVA), poly(vinylpyridine-N-oxide), poly(vinylimidazole), polycarboxylates e.g., polyacrylates, maleic/acrylic acid copolymers), and lauryl methacrylate/acrylic acid copolymers.
- the enzyme(s) of the detergent composition may be stabilized using conventional stabilizing agents, e.g., as polyol [e.g., propylene glycol or glycerol), a sugar or sugar alcohol, lactic acid, boric acid, or a boric acid derivative ⁇ e.g., an aromatic borate ester), or a phenyl boronic acid derivative (e.g., 4-formylphenyl boronic acid).
- the composition may be formulated as described in WO 92/19709 and WO 92/19708.
- the maltogenic amylase of the present teachings may be added in an amount corresponding to about 0.01 to about 100 mg of enzyme protein per liter of wash liquor (e.g., about 0.05 to about 5.0 mg of enzyme protein per liter of wash liquor or 0.1 to about 1 .0 mg of enzyme protein per liter of wash liquor).
- a-amylase cleaning assays are known in the art, including swatch and micro-swatch assays.
- the present maltogenic amylase may be a component of a brewing composition used in a process of brewing, i.e., making a fermented malt beverage.
- Non-fermentable carbohydrates form the majority of the dissolved solids in the final beer. This residue remains because of the inability of malt amylases to hydrolyze the alpha-1 ,6-linkages of the starch.
- the non-fermentable carbohydrates contribute about 50 calories per 12 ounces of beer.
- adjuncts such as common corn grits, refined corn grits, brewer's milled yeast, rice, sorghum, refined corn starch, barley, barley starch, dehusked barley, wheat, wheat starch, torrified cereal, cereal flakes, rye, oats, potato, tapioca, and syrups, such as corn syrup, sugar cane syrup, inverted sugar syrup, barley and/or wheat syrups, and the like may be used as a source of starch.
- adjuncts such as common corn grits, refined corn grits, brewer's milled yeast, rice, sorghum, refined corn starch, barley, barley starch, dehusked barley, wheat, wheat starch, torrified cereal, cereal flakes, rye, oats, potato, tapioca, and syrups, such as corn syrup, sugar cane syrup, inverted sugar syrup, barley and/or wheat syrups, and the like may be used as a source
- the malt which is produced principally from selected varieties of barley, has the greatest effect on the overall character and quality of the beer.
- the malt is the primary flavoring agent in beer.
- the malt provides the major portion of the fermentable sugar.
- the malt provides the proteins, which will contribute to the body and foam character of the beer.
- the malt provides the necessary enzymatic activity during mashing.
- Hops also contribute significantly to beer quality, including flavoring.
- hops or hops constituents
- the hops act as protein precipitants, establish preservative agents and aid in foam formation and stabilization.
- Grains such as barley, oats, wheat, as well as plant components, such as corn, hops, and rice, also are used for brewing, both in industry and for home brewing.
- the components used in brewing may be unmalted or may be malted, i.e., partially germinated, resulting in an increase in the levels of enzymes, including a-amylase.
- a-amylase enzymes
- An amylase by itself or in combination with another a-amylase(s), accordingly may be added to the components used for brewing.
- the term "stock” means grains and plant components that are crushed or broken.
- barley used in beer production is a grain that has been coarsely ground or crushed to yield a consistency appropriate for producing a mash for fermentation.
- the term "stock” includes any of the aforementioned types of plants and grains in crushed or coarsely ground forms. The methods described herein may be used to determine maltogenic amylase activity levels in both flours and stock. Processes for making beer are well known in the art. See, e.g., Wolfgang
- the process involves: (a) preparing a mash, (b) filtering the mash to prepare a wort, and (c) fermenting the wort to obtain a fermented beverage, such as beer.
- a fermented beverage such as beer.
- milled or crushed malt is mixed with water and held for a period of time under controlled temperatures to permit the enzymes present in the malt to convert the starch present in the malt into fermentable sugars.
- the mash is then transferred to a mash filter where the liquid is separated from the grain residue.
- This sweet liquid is called "wort," and the left over grain residue is called “spent grain.”
- the mash is typically subjected to an extraction, which involves adding water to the mash in order to recover the residual soluble extract from the spent grain.
- the wort is then boiled vigorously to sterilize the wort and help develop the color, flavor and odor. Hops are added at some point during the boiling.
- the wort is cooled and transferred to a fermentor.
- the wort is then contacted in a fermentor with yeast.
- the fermentor may be chilled to stop fermentation.
- the beer is cooled and stored for a period of time, during which the beer clarifies and its flavor develops, and any material that might impair the appearance, flavor and shelf life of the beer settles out.
- the beer usually contains from about 2% to about 10% v/v alcohol, although beer with a higher alcohol content, e.g., 18% v/v, may be obtained.
- the beer Prior to packaging, the beer is carbonated and, optionally, filtered and pasteurized.
- the brewing composition comprising a maltogenic amylase, in combination with a glucoamylase and optionally a pullulanase and/or isoamylase, may be added to the mash of step (a) above, i.e., during the preparation of the mash.
- the brewing composition may be added to the mash of step (b) above, i.e., during the filtration of the mash.
- the brewing composition may be added to the wort of step (c) above, i.e., during the fermenting of the wort.
- a fermented beverage such as a beer
- the fermented beverage can be a beer, such as full malted beer, beer brewed under the "Rösgebot,” ale, IPA, lager, bitter, Happoshu (second beer), third beer, dry beer, near beer, light beer, low alcohol beer, low calorie beer, porter, bock beer, stout, malt liquor, non-alcoholic beer, non-alcoholic malt liquor and the like, but also alternative cereal and malt beverages such as fruit flavored malt beverages, e.g., citrus flavored, such as lemon-, orange-, lime-, or berry-flavored malt beverages, liquor flavored malt beverages, e.g., vodka-, rum-, or tequila-flavored malt liquor, or coffee flavored malt beverages, such as caffeine-flavored malt liquor, and the like.
- fruit flavored malt beverages e.g., citrus flavored, such as lemon-, orange-, lime-, or berry-flavored malt beverages
- the maltogenic amylases of the present teachings may reduce the
- I PS iodine-positive starch
- amylose escapes hydrolysis and/or from retrograded starch polymer.
- Starch retrogradation occurs spontaneously in a starch paste, or gel on aging, because of the tendency of starch molecules to bind to one another followed by an increase in crystallinity. Solutions of low concentration become increasingly cloudy due to the progressive association of starch molecules into larger articles. Spontaneous precipitation takes place and the precipitated starch appears to be reverting to its original condition of cold-water insolubility.
- Pastes of higher concentration on cooling set to a gel, which on aging becomes steadily firmer due to the increasing association of the starch molecules. This arises because of the strong tendency for hydrogen bond formation between hydroxy groups on adjacent starch molecules. See J. A. Radley, ed., STARCH AND ITS DERIVATIVES 1 94-201 (Chapman and Hall, London (1968)).
- IPS in saccharide liquor negatively affects final product quality and represents a major issue with downstream processing.
- the amount of IPS can be reduced by isolating the saccharification tank and blending the contents back. IPS nevertheless will accumulate in carbon columns and filter systems, among other things.
- the use of the maltogenic amylases of the present teachings is expected to improve overall process performance by reducing the amount of IPS.
- a putative novel amylase was identified from a metagenomic library constructed by conventional cloning techniques. Protein sequence analysis indicates that the amylase belongs to the glycosyl hydrolase family 13 (GH13), and shows less than 66% sequence identity to known proteins in the public NCBI database. Following identification of the putative amylase in silico, the gene was cloned using conventional molecular biology PCR techniques and expressed in Bacillus subtilis. The plasmid cloning map is shown in Figure 9. Cloning and expression of AmyMG
- the full gene sequence is SEQ ID NO:1 .
- the full protein sequence is SEQ ID NO:3.
- the native signal peptide is SEQ I D NO: 4.
- the mature gene sequence was identified by conventional procedures, PCR amplified, digested by BssHii/Xhol, and ligated into the p2JM plasmid digested with same restriction enzymes.
- the AmyMG gene was put under the control of the aprE promoter.
- the aprE signal sequence was used to direct protein secretion.
- nucleotides coding for three additional amino acids (AGK) were placed between the aprE signal sequence and the mature AmyMG gene to facilitate secretion of the target protein.
- the resulting plasmid was used to transform competent Bacillus subtilis cells. Expression evaluation of AmyMG showed that the enzyme expresses very well in both strains.
- the aprE signal nucleic acid sequence (underlined) + AGK nucleic acid sequence (italics) is SEQ ID NO: 5.
- the aprE signal amino acid sequence (underlined) + AGK amino acid sequence (italics) is SEQ ID NO: 6.
- Figure 1 shows the typical chromatograms of oligosaccharide product profile analysis of A) AmyMG (10 ppm) incubated with Maltodextrin (DE1 0) (0.5%, w/v) under pH 5.3 at 50°C for 2 h and B) the mixture of standard compounds of DP1 to DP 7 (0.01 25%, w/v).
- HPLC separation was done using an Agilent 1200 series HPLC system with an Aminex HPX-42A column (300 mm x 7.8 mm) at 85 °C.
- the sample (10 ⁇ _) was subjected to the HPLC column and separated with an isocratic gradient of Milli-Q water as the mobile phase at a flow rate of 0.6 mL/min.
- the oligosaccharide products were detected using a refractive index detector.
- the sample (25 ⁇ _) was subjected to the column and separated with the gradient: 0-1 0 min, 50mM NaOH; 10-15 min, 50-1 OOmM NaOH ; 15-35min, 1 00mM NaOH, 0-200mM NaAc; 35-45min, 50mM NaOH, at a flow rate of 0.5 mL/min.
- oligosaccharide products were detected using a pulsed amperometric detector.
- AmyMG was tested for alpha-amylase characterization (Amylopectin/PAHBAH method), including dose dependent assay ( Figure 5, a dose-dependant assay for AmyMG using amylopectin as the substrate), pH ( Figure 6, showing a normalized pH profile), temperature profile (Figure 7, showing a normalized temperature profile), and, a thermostability assay ( Figure 8, showing the thermostability of AmyMG).
- dose dependent assay Figure 5
- pH Figure 6, showing a normalized pH profile
- temperature profile Figure 7, showing a normalized temperature profile
- thermostability assay Figure 8, showing the thermostability of AmyMG.
- the specific activity of this enzyme towards amylopectin is 218.4 U/mg, with optimum pH at 7 and optimum temperature at 63°C.
- the present teachings provide a polypeptide with maltogenic acitivity wherein at least 70%, at least 75%, at least 80%, at least 85%, at least 90%., or at least 95% of the maximum enzyme activity is present in the pH range of 6-8, as assessed by DP ° Units. Additionally, in some embodiments, the present teachings provide a polypeptide with maximum activity, as assessed by DP ° Units, at 60°C-70°C, or 62°C-64°C.
- the present teachings provide a polypeptide that produces at least 80%, at least 85%, at least 90%, or at least 95% DP2 of the DP1 to DP7 products resulting from a hydrolysis reaction performed according to the procedures depicted in Figure 3.
- polypeptides of the present teachings can be engineered to provide various improved properties. Such engineering efforts can be guided by the atomic coordinate structural information provided in Table 3.
- ATOM 14 O ALA A 9 Ill . , 263 81 , .146 16. .368 1. , 00 16. .65 0
- ATOM 15 N ASP A 10 109. , 823 82 , , 651 15. , 494 1. , 00 13. ,87 N
- ATOM 21 C ASP A 10 108. , 159 80 , , 918 14. , 856 1. , 00 12. ,27 c
- ATOM 22 O ASP A 10 107. , 934 81 , .294 13. .725 1. , 00 12. .47 O
- ATOM 24 CA TYR A 11 107. , 418 78 , , 635 14. ,367 1. , 00 10. ,00 C
- ATOM 36 CA TYR A 12 103. , 913 77 , ,379 15. , 195 1. , 00 10. ,87 C
- ATOM 110 CA GLU A 21 93 , , 772 69 , , 145 24. , 120 1. , 00 10. ,20 C
- ATOM 118 N ALA A 22 93 , , 096 66 , .848 23. , 730 1. , 00 8. , 78 N
- ATOM 119 CA ALA A 22 92 , .593 65 , .712 22. .987 1. , 00 8. .73 c
- ATOM 122 O ALA A 22 91 , , 118 65, .264 24. , 825 1. , 00 8. ,96 O
- ATOM 160 N MET A 27 78. , 173 60 , .214 23. .856 1. , 00 11. .76 N
- ATOM 162 CB MET A 27 75. , 931 59 , .590 23. .020 1. , 00 10. .86 C
- ATOM 166 C MET A 27 76. , 471 60 , .071 25. .434 1. , 00 9. .42 C
- ATOM 181 C ASP A 29 73 , .468 57. , 249 27. .485 1. , 00 7. .72 C
- ATOM 212 N ASN A 33 75 , .576 54. , 595 33. .054 1. , 00 8. .65 N
- ATOM 213 CA ASN A 33 74 , .448 54. , 927 33. .919 1. , 00 8. .72 c
- ATOM 214 CB ASN A 33 73 , .284 53. , 940 33. .699 1. , 00 9. .37 C
- ATOM 230 C ASP A 35 71 , , 232 56 , , 072 38. ,323 1. , 00 10. ,65 C
- ATOM 232 N PRO A 36 70 , , 756 54 , , 890 38. , 748 1. , 00 12. , 71 N
- ATOM 238 O PRO A 36 67, , 231 55 , , 293 38. , 015 1. , 00 11. ,01 O
- ATOM 241 C GLY A 37 66 , , 935 58 , , 041 39. , 421 1. , 00 10. ,34 C
- ATOM 244 CA ASN A 38 67, , 296 59 , , 500 37. ,503 1. , 00 9. ,39 C
- ATOM 245 CB ASN A 38 68 , ,383 60 , , 581 37. ,382 1. , 00 8. ,58 C
- ATOM 249 C ASN A 38 66 , , 887 58 , , 982 36. , 090 1. , 00 10. , 78 C
- ATOM 251 N ASP A 39 66 , , 742 57 , , 667 35. ,940 1. , 00 9. ,62 N
- ATOM 257 C ASP A 39 65, , 060 57 , , 445 34. , 072 1. , 00 9. ,97 C
- ATOM 268 O HIS A 40 61 , , 626 57 , , 254 32. ,609 1. , 00 8. ,28 O ATOM 269 N
- ATOM 278 C ARG A 41 60. , 477 54. , 561 32. .763 1. , 00 11. .82 C
- ATOM 280 N ASP A 42 59. , 542 55. , 067 33. .555 1. , 00 11. .44 N
- ATOM 282 CB ASP A 42 57. , 222 54. , 616 34. .279 1. , 00 17. .56 C
- ATOM 286 C ASP A 42 57. , 673 56. , 351 32. .473 1. , 00 11. .61 C
- ATOM 302 CA GLY A 45 53. , 802 55. , 703 29. .382 1. , 00 13. .62 C
- ATOM 306 CA ALA A 46 52. , 977 54. , 192 25. .952 1. , 00 11. .74 C
- ATOM 308 C ALA A 46 53. , 770 55. , 097 24. .988 1. , 00 11. .26 C
- ATOM 340 N ASP A 51 64. , 188 53. ,345 26. .794 1. , 00 11. .49 N
- ATOM 342 CB ASP A 51 64. , 505 50. , 962 27. .220 1. , 00 10. .45 C
- ATOM 346 C ASP A 51 63. , 502 51. , 637 25. .083 1. , 00 9. .86 C
- ATOM 349 CA ILE A 52 62. , 968 51. , 829 22. .697 1. , 00 8. .39 c
- ATOM 354 C ILE A 52 63. , 794 51. , 030 21. .687 1. , 00 9. .20 C
- ATOM 356 N PRO A 53 63. , 784 49. , 695 21. .798 1. , 00 10. .75 N
- ATOM 372 CA PRO A 55 64. , 404 45 , .388 15. .169 1. , 00 14. .57 C
- ATOM 376 C PRO A 55 65. , 885 45 , .260 14. .757 1. , 00 14. .76 C
- ATOM 378 N PRO A 56 66. , 517 44 , .102 15. .003 1. , 00 15. .11 N
- ATOM 379 CA PRO A 56 67. , 954 44 , .066 14. .773 1. , 00 16. .52 C
- ATOM 382 CD PRO A 56 66. , 007 42 , .830 15. .546 1. , 00 16. .27 c
- ATOM 400 CA GLY A 59 72. , 592 40 , .921 14. .213 1. , 00 16. .76 C
- ATOM 402 O GLY A 59 72. , 911 41 , .331 16. .529 1. , 00 15. .63 O
- ATOM 422 CB ASN A 63 68. , 261 51. , 617 21. .847 1. , 00 9. .76 C
- ATOM 426 C ASN A 63 67. , 569 53. ,377 20. .166 1. , 00 9. .97 c
- ATOM 434 C ILE A 64 65. , 391 55. , 904 21. .267 1. , 00 9. .04 c
- ATOM 435 O ILE A 64 64. , 819 56. , 989 21. .184 1. , 00 10. .11 O
- ATOM 436 N GLY A 65 65. , 573 55. , 292 22. .432 1. , 00 8. .54 N
- ATOM 437 CA GLY A 65 65. , 170 55. , 915 23. .699 1. , 00 7. .46 C
- ATOM 438 C GLY A 65 66. , 325 56. , 136 24. .643 1. , 00 7. .13 c
- ATOM 441 CA TYR A 66 67. , 043 56. , 821 26. .835 1. , 00 7. .42 C
- ATOM 448 CE2 TYR A 66 67. , 223 61. , 456 28. .041 1. , 00 7. .41 C
- ATOM 449 CD2 TYR A 66 67., 368 60 ,, 086 28., 234 1., 00 6., 55 C
- ATOM 453 CA LEU A 67 69. , 190 54 , , 027 28. , 261 1. , 00 8. ,10 C
- ATOM 456 CD1 LEU A 67 70. , 429 51 , .912 25. .188 1. , 00 7. .21 c
- ATOM 462 C GLY A 68 71. , 595 56 , , 416 31. , 776 1. , 00 7. , 86 C
- ATOM 468 N ASP A 70 74. , 560 58 , .951 32. .381 1. , 00 8. .04 N
- ATOM 474 C ASP A 70 75. , 608 60 , , 877 33. , 573 1. , 00 8. ,59 C
- ATOM 508 N ALA A 75 81 , ,315 61 , , 047 35. , 501 1. , 00 8. ,68 N
- ATOM 511 C ALA A 75 83 , , 005 60 , ,398 37. , 138 1. , 00 10. , 79 C
- ATOM 512 O ALA A 75 84, , 224 60 , , 477 37. , 292 1. , 00 12. , 06 O
- ATOM 530 O HIS A 77 86 , , 426 57 , .730 33. , 199 1. , 00 12. , 44 O
- ATOM 545 C ASP A 79 90 , ,340 57 , ,361 34. , 192 1. , 00 10. ,54 C
- ATOM 548 CA TYR A 80 89 , , 452 55 , , 668 32. ,692 1. , 00 11. ,17 C
- ATOM 560 CA ILE A 81 89 , , 204 58 , , 453 30. , 060 1. , 00 9. ,52 C
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Abstract
The present teachings provide an amylase with maltogenic properties. Nucleic acids encoding the maltogenic amylase and variants thereof, expression vectors, formulations, and host cells are also provided. Additional embodiments of the present teachings provide various methods of use and methods of manufacturing.
Description
Amylase with Maltogenic Properties
Cross-Reference to Related Applications
This application claims benefit to international patent application no.
PCT/CN2012/084883, filed on November 20, 2012, the contents of which are hereby incorporated by reference in its entirety.
Field of the Invention
The present teachings provide composition and methods relating to novel maltogen amylases.
Background
Starch is a mixture of amylose (15-30% w/w) and amylopectin (70-85% w/w). Amylose is composed of linear chains of a-1 ,4-linked glucose units having a molecular weight (MW) from about 60,000 to about 800,000. Amylopectin is a branched polymer containing a-1 ,6 branch points every 24-30 glucose units; its MW may be as high as 100 million.
Sugars from starch, in the form of concentrated dextrose syrups, are currently produced by an enzyme catalyzed process involving: (1 ) liquefaction (or viscosity reduction) of solid starch with an a-amylase into dextrins having an average degree of polymerization of about 7-1 0, and (2) saccharification of the resulting liquefied starch (i.e. starch hydrolysate) with amyloglucosidase (also called glucoamylase or GA). The resulting syrup has a high glucose content. Much of the glucose syrup that is commercially produced is subsequently enzymatically isomerized to a dextrose/fructose mixture known as isosyrup. The resulting syrup also may be fermented with
microorganisms, such as yeast, to produce commercial products including ethanol, citric acid, lactic acid, succinic acid, itaconic acid, monosodium glutamate, gluconates, lysine, other organic acids, other amino acids, and other biochemicals, for example.
Fermentation and saccharification can be conducted simultaneously (i.e., an SSF process) to achieve greater economy and efficiency.
Alpha-amylases hydrolyze starch, glycogen, and related polysaccharides by cleaving internal a-1 ,4-glucosidic bonds at random. Alpha amylases, particularly from Bacilli, have been used for a variety of different purposes, including starch liquefaction and saccharification, textile desizing, starch modification in the paper and pulp industry, brewing, baking, production of syrups for the food industry, production of feedstocks for fermentation processes, and in animal feed to increase digestibility. These enzymes can also be used to remove starchy soils and stains during dishwashing and laundry washing.
Maltose, a di-saccharide composed of two D-glucopyranoses joined by a β-1 ,4'-glycosidic bond, has high commercial value in applications for the food/frozen foods, baking, brewing and beverage industries. Maltose is also a substrate for production of the non-caloric sugar sweetener, maltitol. High purity maltose or pure maltose is an active component of intravenous injection liquids for diabetic patients. Commercial processes for the production of syrup containing different levels of maltose content, i.e. <50% maltose (high conversion or low maltose syrup), 50-55% maltose (high maltose syrup), 70-75% maltose (very high maltose) and >80% maltose (ultra high maltose) have been established depending on the applications. A common factor for these processes is that they involve a dual enzyme process with two different steps, i.e. liquefaction and saccharification.
Historically, two enzyme steps are involved in the hydrolysis of starch to produce glucose or maltose syrups. The first step is a liquefaction step at high temperature, >95 °C and the second step is a saccharification step. In maltose production the second step is called malto-saccharification and usually takes place at a temperature at or below 60°C. In the liquefaction step, the insoluble starch granules are slurried in water, gelatinized with heat and hydrolyzed by a thermostable alpha-amylase (EC.3.2.1 .1 , a-1 ,4'- D-glucan glucanohydrolase) from Bacillus species, often in the presence of added calcium. Bacterial derived thermostable alpha-amylases from Bacillus licheniformis (for example, SPEZYME® FRED from DuPont-Genencor or Termamyl® L-120 from
Novozymes), Bacillus stearothermophilus (for example SPEZYME XTRA from
DuPont-Genencor, Termamyl® SC, and Termamyl® SUPRA from Novozymes) or blends of Bacillus licheniformis and Bacillus stearothermophilus (for example Clearflow™ AA from DuPont-Genencor or Liquozyme® Supra from Novozymes) are used to first liquefy the starch at high temperature, >95°C at pH 5.2-6.5 to a low DE (dextrose equivalent) soluble starch hydrolysate. In the malto-saccharification step, generally maltogenic enzymes such as a fungal alpha-amylase (for example, CLARASE® L from
DuPont-Genencor or Fungamyl® 800L from Novozymes), a plant beta-amylase (for example, OPTIMALT® BBA from DuPont-Genencor or Betalase 1 500L from Senson) are used at a much lower temperature to further hydrolyse the soluble starch hydrolysate. For maltose syrup containing greater than 60% maltose, a debranching enzyme like pullulanase (for example OPTIMAX® L-1 000 from DuPont-Genencor, Promozyme® D2 from Novozymes or Promozyme® D6 from Novozymes) is added during
malto-saccharification of liquefied starch.
Summary
In some embodiments, the present teachings provide an isolated nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1 or of a degenerate variant of SEQ I D NO: 1 .
In some embodiments, the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3.
In some embodiments, the present teachings provide an isolated nucleic acid comprising a sequence that hybridizes under stringent conditions to a hybridization probe the nucleotide sequence of which consists of SEQ ID NO: 1 , or the complement of SEQ
I D NO: 1 .
In some embodiments, the present teachings provide an isolated nucleic acid comprising a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ ID NO: 1 . In some embodiments, the present teachings provide such an isolated nucleic acid wherein the nucleic acid encodes a
polypeptide that has starch hydrolysis activity.
In some embodiments, the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ ID NO: 3, wherein the polypeptide has starch hydrolysis activity.
In some embodiments, the present teachings provide an isolated nucleic acid comprising a sequence that encodes a polypeptide comprising the sequence of SEQ ID NO: 3, or SEQ I D NO: 3 with up to 50 conservative amino acid substitutions, wherein the polypeptide has starch hydrolysis activity,
In some embodiments, the present teachings provide a purified polypeptide, the amino acid sequence of which comprises a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ I D NO: 3.
In some embodiments, the present teachings provide a purified polypeptide comprising the amino acid sequence of SEQ I D NO: 3, but with 0 to 20 conservative amino acid substitutions.
In some embodiments, the present teachings provide an expression vector comprising the nucleic acid sequence of any of claims 1 -6 operably linked to an expression control sequence. In some embodiments, the present teachings provide a cultured cell comprising such a vector. In some embodiments, the present teachings provide cultured cell comprising any of the nucleic acids of the present teachings, operably linked to an expression control sequence. In some embodiments, the present teachings provide a cultured cell tranfected with any of the vectors provided by the present teachings, or a progeny of said cell, wherein the cell expresses the nucleic acid to form a polypeptide.
In some embodiments, the present teachings provide a method of producing a protein, the method comprising culturing the cells provided by the present teachings under conditions permitting expression of the polypeptide.
In some embodiments, the present teachings provide a method of using the polypeptide of the present teachings, the method comprising including the polypeptide in any of: starch liquefaction, starch saccharification, textile desizing, starch modification in
the paper and pulp industry, brewing, baking, production of syrups for the food industry, production of feedstocks for fermentation processes, animal feed, and, removal of starchy soils and/or stains during dishwashing and/or laundry washing.
In some embodiments, the present teachings provide a composition comprising the polypeptide of the present teachings, and at least one accessory enzyme selected from the group consisting of phytase, protease, pullulanase, β-amylase, isoamylase, a different amylase, alpha-glucosidase, cellulase, xylanase, hemicellulase,
beta-glucosidase, transferase, pectinase, lipase, cutinase, esterase, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases,
phospholipases, lysophospholipases, acyltransferases, perhydrolases, arylesterases, and redox enzymes.
These and other aspects and embodiments of the compositions and methods of the present teachings will be apparent from the present description and drawings.
Brief Description of the Drawings
Figure 1 , panels A and B depict some illustrative data according to some embodiments of the present teachings.
Figure 2, panels A and B depict some illustrative data according to some embodiments of the present teachings. Figure 3 depicts some illustrative data according to some embodiments of the present teachings.
Figure 4 depicts some illustrative data according to some embodiments of the present teachings.
Figure 5 depicts some illustrative data according to some embodiments of the present teachings.
Figure 6 depicts some illustrative data according to some embodiments of the present teachings.
Figure 7 depicts some illustrative data according to some embodiments of the present teachings.
Figure 8 depicts some illustrative data according to some embodiments of the present teachings. Figure 9 depicts an illustrative cloning map according to some embodiments of the present teachings.
Brief Description of the Sequences
SEQ ID NO: 1 sets forth the full-length nucleotide sequence for AmyMG. SEQ ID NO: 2 sets forth the nucleotide sequence for the native signal sequence for AmyMG.
SEQ ID NO: 3 sets forth the full-length amino acid sequence for AmyMG.
SEQ ID NO: 4 sets forth the amino acid sequence for the native signal peptide.
SEQ ID NO: 5 sets forth the aprE signal nucleic acid sequence (underlined) + AGK nucleic acid sequence (italics).
SEQ ID NO: 6 sets forth the aprE signal amino acid sequence (underlined) + AGK amino acid sequence (italics). Detailed Description
Described are compositions and methods relating to maltogenic amylase enzymes. This enzyme was discovered and analyzed by a combination of experimental approaches, as detailed in the Examples. Exemplary applications for the variant amylase enzymes are for starch liquefaction and saccharification, for cleaning starchy stains in laundry, dishwashing, and other applications, for textile processing (e.g., desizing), in animal feed for improving digestibility, and and for baking and brewing. These and other aspects of the compositions and methods are described in detail, below.
Prior to describing the various aspects and embodiments of the present compositions and methods, the following definitions and abbreviations are described.
Definitions and Abbreviations
In accordance with this detailed description, the following abbreviations and definitions apply. Note that the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an enzyme" includes a plurality of such enzymes, and reference to "the dosage" includes reference to one or more dosages and equivalents thereof known to those skilled in the art, and so forth.
The present document is organized into a number of sections for ease of reading ; however, the reader will appreciate that statements made in one section may apply to other sections. In this manner, the headings used for different sections of the disclosure should not be construed as limiting.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The following terms are provided below.
Abbreviations and Acronyms
The following abbreviations/acronyms, when and if present, have the following meanings unless otherwise specified:
ABTS 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid
AE or AEO alcohol ethoxylate
AES or AEOS alcohol ethoxysulfate
AkAA Aspergillus kawachii a-amylase
AnGA Aspergillus n/ger glucoamylase
AOS a-olefinsulfonate
AS alkyl sulfate
cDNA complementary DNA
CMC carboxymethylcellulose
DE dextrose equivalent
DNA deoxyribonucleic acid
DPn degree of saccharide polymerization having n subunits ds or DS dry solids
DTMPA diethylenetriaminepentaacetic acid
EC Enzyme Commission
EDTA ethylenediaminetetraacetic acid
EO ethylene oxide (polymer fragment)
EOF End of Fermentation
GA glucoamylase
GAU/g ds glucoamylase activity unit/gram dry solids
HFCS high fructose corn syrup
HgGA Humicola grisea glucoamylase
IPTG isopropyl β-D-thiogalactoside
IRS insoluble residual starch
kDa kiloDalton
LAS linear alkylbenzenesulfonate
LAT, BLA B. licheniformis amylase
MW molecular weight
MWU modified Wohlgemuth unit; 1 .6x10"5 mg/MWU = unit of activity
NCBI National Center for Biotechnology Information
NOBS nonanoyloxybenzenesulfonate
NTA nitriloacetic acid
OxAm Purastar HPAM 5000L (Danisco US Inc.)
PAHBAH p-hydroxybenzoic acid hydrazide
PEG polyethyleneglycol
pi isoelectric point
PI performance index
ppm parts per million, e.g., μg protein per gram dry solid
PVA polyvinyl alcohol)
PVP poly(vinylpyrrolidone)
RCF relative centrifugal/centripetal force (i.e., x gravity)
RNA ribonucleic acid
SAS alkanesulfonate
SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis SSF simultaneous saccharification and fermentation
SSU/g solid soluble starch unit/gram dry solids
sp. species
TAED tetraacetylethylenediamine
Tm melting temperature
TrGA Trichoderma reesei glucoamylase
w/v weight/volume
w/w weight/weight
v/v volume/volume
wt% weight percent
°C degrees Centigrade
H2O water
dH2O or Dl deionized water
dlH2O deionized water, Milli-Q filtration
g or gm grams
μg micrograms
mg milligrams
kg kilograms
μΙ_ and μΙ microliters
ml_ and ml milliliters
mm millimeters
μιη micrometer
M molar
mM millimolar
μΜ micromolar
U units
sec seconds
min(s) minute/minutes
hr(s) hour/hours
DO dissolved oxygen
Ncm Newton centimeter
ETOH ethanol
eq. equivalents
N normal
uPWA variant a-amylase derived from Pyrococcus woesei
PWA a-amylase from Pyrococcus woesei
MWCO molecular weight cut-off
SSRL Stanford Synchrotron Radiation Lightsource
PDB Protein Database
CAZy Carbohydrate-Active Enzymes database
Tris-HCI tris(hydroxymethyl)aminomethane hydrochloride
HEPES 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid
Definitions
The term "maltogenic amylase" refers to enzymes that are capable of producing significant amounts of maltose from starch or hydrolyzed starch. There are several enzymes falling under this definition. Some examples are:
1 ) Fungal alpha-amylases include those obtained from filamentous fungal strains including but not limited to strains of Aspergillus (e.g., A. Niger, A. kawachi, and A. oryzae); Trichoderma sp. (e.g. Trichoderma reesie alpha-amylase, disclosed in EP 2132307), Rhisopus sp., Mucor sp., and Penicillium sp. Commercial fungal alpha-amylase from Aspergillus oryzae are CLARASE® L from DuPont-Genencor and Fungamyl® 800l_ from Novozymes.
2) Acid stable fungal amylase from Aspergillus niger {For example, from Shin Nihon Chemicals).
3) Beta-amylases are found in plant materials like wheat, barley, rye, shorgum, soy, sweet potato, rice and microorganisms like Bacillus cereus, Bacillus polymixa, Bacillus megaterium, Arabidopsis thaliana. The most common commercial beta-amylases are derived from barley and are sold under trade the names OPT I MALT® BBA by DuPont-Genencor and Betalase 1500L by Senson. A commercial soy beta-amylase is -amylase#1500S from Nagase ChemteX Corporation.
4) Maltogenic amylases (E.C. 3.2.1 .1 33) are produced by microorganisms Bacillus subtilis, Geobacillus stearothermophilus, Bacillus thermoalkalophilus,
Lactobacillus gasseri, Thermus sp. Commercial maltogenic amylases include but not limited to Maltogenase® L from Novozymes, Veron® XTENDER from AB Enzymes and MAX-LIFE™ P100 from DuPont-Danisco.
5) Maltogenic amylases provided by the present teachings, as illustrated for
example by SEQ I D NO: 1 and SEQ I D NO: 3, and variants thereof taught herein.
As used herein, the term "Enzyme units" refers to the amount of product formed per time under the specified conditions of the assay. For example, a "glucoamylase activity unit" (GAU) is defined as the amount of enzyme that produces 1 g of glucose per hour from soluble starch substrate (4% DS) at 60 °C, pH 4.2. A "soluble starch unit" (SSU) is the amount of enzyme that produces 1 mg of glucose per minute from soluble starch substrate (4% DS) at pH 4.5, 50 °C. As another example, maltogenic amylase activity can be measured in degrees Diastatic Power (DP °) Units. This assay is based on a 30-min hydrolysis of a starch substrate at pH 4.6 and 20 °C. The reducing sugar groups produced on hydrolysis are measured in a titrimetric procedure using alkaline
ferricyanide. One unit of diastase activity, expressed as degrees DP (DP 0), is defined as the amount of enzyme, contained in 0.1 ml of a 5% solution of the sample enzyme preparation, that will produce sufficient reducing sugars to reduce 5 mL of Fehling's solution when the sample is incubated with 100 mL of the substrate for 1 hour at 20 °C.
The term "starch" refers to any material comprised of the complex polysaccharide carbohydrates of plants, comprised of amylose and amylopectin with the formula (C6H 0O5)x, wherein X can be any number. The term includes plant-based materials
such as grains, cereal, grasses, tubers and roots, and more specifically materials obtained from wheat, barley, corn, rye, rice, sorghum, brans, cassava, millet, milo, potato, sweet potato, and tapioca. The term "starch" includes granular starch. The term "granular starch" refers to raw, i.e., uncooked starch, e.g., starch that has not been subject to gelatinization.
The terms, "wild-type," "parental," or "reference," with respect to a polypeptide, refer to a naturally-occurring polypeptide that does not include a man-made substitution, insertion, or deletion at one or more amino acid positions. Similarly, the terms
"wild-type," "parental," or "reference," with respect to a polynucleotide, refer to a naturally-occurring polynucleotide that does not include a man-made nucleoside change. However, note that a polynucleotide encoding a wild-type, parental, or reference polypeptide is not limited to a naturally-occurring polynucleotide, and encompasses any polynucleotide encoding the wild-type, parental, or reference polypeptide.
Reference to the wild-type polypeptide is understood to include the mature form of the polypeptide. A "mature" polypeptide or variant, thereof, is one in which a signal sequence is absent, for example, cleaved from an immature form of the polypeptide during or following expression of the polypeptide.
The term "variant," with respect to a polypeptide, refers to a polypeptide that differs from a specified wild-type, parental, or reference polypeptide in that it includes one or more naturally-occurring or man-made substitutions, insertions, or deletions of an amino acid. Similarly, the term "variant," with respect to a polynucleotide, refers to a polynucleotide that differs in nucleotide sequence from a specified wild-type, parental, or reference polynucleotide. The identity of the wild-type, parental, or reference
polypeptide or polynucleotide will be apparent from context. In the case of the present maltogenic amylases, "activity" refers to maltogenic amylase activity, which can be measured as described herein.
The term "recombinant," when used in reference to a subject cell, nucleic acid, protein or vector, indicates that the subject has been modified from its native state. Thus, for example, recombinant cells express genes that are not found within the native
(non-recombinant) form of the cell, or express native genes at different levels or under different conditions than found in nature. Recombinant nucleic acids differ from a native sequence by one or more nucleotides and/or are operably linked to heterologous sequences, e.g., a heterologous promoter in an expression vector. Recombinant proteins may differ from a native sequence by one or more amino acids and/or are fused with heterologous sequences. A vector comprising a nucleic acid encoding an amylase is a recombinant vector.
The terms "recovered," "isolated," and "separated," refer to a compound, protein (polypeptides), cell, nucleic acid, amino acid, or other specified material or component that is removed from at least one other material or component with which it is naturally associated as found in nature. An "isolated" polypeptides, thereof, includes, but is not limited to, a culture broth containing secreted polypeptide expressed in a heterologous host cell.
The term "purified" refers to material (e.g., an isolated polypeptide or
polynucleotide) that is in a relatively pure state, e.g., at least about 90% pure, at least about 95% pure, at least about 98% pure, or even at least about 99% pure.
The term "enriched" refers to material (e.g., an isolated polypeptide or
polynucleotide) that is in about 50% pure, at least about 60% pure, at least about 70% pure, or even at least about 70% pure. The terms "thermostable" and "thermostability," with reference to an enzyme, refer to the ability of the enzyme to retain activity after exposure to an elevated temperature. The thermostability of an enzyme, such as a maltogenic amylase enzyme, is measured by its half-life (t /2) given in minutes, hours, or days, during which half the enzyme activity is lost under defined conditions. The half-life may be calculated by measuring residual amylase activity following exposure to (i.e., challenge by) an elevated temperature.
A "pH range," with reference to an enzyme, refers to the range of pH values under which the enzyme exhibits catalytic activity.
The terms "pH stable" and "pH stability," with reference to an enzyme, relate to the ability of the enzyme to retain activity over a wide range of pH values for a predetermined period of time (e.g., 1 5 min., 30 min., 1 hour).
The term "amino acid sequence" is synonymous with the terms "polypeptide," "protein," and "peptide," and are used interchangeably. Where such amino acid sequences exhibit activity, they may be referred to as an "enzyme." The conventional one-letter or three-letter codes for amino acid residues are used, with amino acid sequences being presented in the standard amino-to-carboxy terminal orientation [i.e., N→C). The term "nucleic acid" encompasses DNA, RNA, heteroduplexes, and synthetic molecules capable of encoding a polypeptide. Nucleic acids may be single stranded or double stranded, and may be chemical modifications. The terms "nucleic acid" and "polynucleotide" are used interchangeably. Because the genetic code is degenerate, more than one codon may be used to encode a particular amino acid, and the present compositions and methods encompass nucleotide sequences that encode a particular amino acid sequence. Unless otherwise indicated, nucleic acid sequences are presented in 5'-to-3' orientation.
"Hybridization" refers to the process by which one strand of nucleic acid forms a duplex with, i.e., base pairs with, a complementary strand, as occurs during blot hybridization techniques and PCR techniques. Stringent hybridization conditions are exemplified by hybridization under the following conditions: 65 °C and 0.1 X SSC (where 1 X SSC = 0.15 M NaCI, 0.015 M Na3 citrate, pH 7.0). Hybridized, duplex nucleic acids are characterized by a melting temperature (Tm), where one half of the hybridized nucleic acids are unpaired with the complementary strand. Mismatched nucleotides within the duplex lower the Tm. Very stringent hybridization conditions involve 68 °C and 0.1 X SSC
A "synthetic" molecule is produced by in vitro chemical or enzymatic synthesis rather than by an organism.
The terms "transformed," "stably transformed," and "transgenic," used with reference to a cell means that the cell contains a non-native (e.g., heterologous) nucleic
acid sequence integrated into its genome or carried as an episome that is maintained through multiple generations.
The term "introduced" in the context of inserting a nucleic acid sequence into a cell, means "transfection", "transformation" or "transduction," as known in the art. A "host strain" or "host cell" is an organism into which an expression vector, phage, virus, or other DNA construct, including a polynucleotide encoding a polypeptide of interest {e.g., an amylase) has been introduced. Exemplary host strains are microorganism cells (e.g., bacteria, filamentous fungi, and yeast) capable of expressing the polypeptide of interest and/or fermenting saccharides. The term "host cell" includes protoplasts created from cells.
The term "heterologous" with reference to a polynucleotide or protein refers to a polynucleotide or protein that does not naturally occur in a host cell.
The term "endogenous" with reference to a polynucleotide or protein refers to a polynucleotide or protein that occurs naturally in the host cell. The term "expression" refers to the process by which a polypeptide is produced based on a nucleic acid sequence. The process includes both transcription and translation.
A "selective marker" or "selectable marker" refers to a gene capable of being expressed in a host to facilitate selection of host cells carrying the gene. Examples of selectable markers include but are not limited to antimicrobials (e.g., hygromycin, bleomycin, or chloramphenicol) and/or genes that confer a metabolic advantage, such as a nutritional advantage on the host cell.
A "vector" refers to a polynucleotide sequence designed to introduce nucleic acids into one or more cell types. Vectors include cloning vectors, expression vectors, shuttle vectors, plasmids, phage particles, cassettes and the like.
An "expression vector" refers to a DNA construct comprising a DNA sequence encoding a polypeptide of interest, which coding sequence is operably linked to a suitable control sequence capable of effecting expression of the DNA in a suitable host.
Such control sequences may include a promoter to effect transcription, an optional operator sequence to control transcription, a sequence encoding suitable ribosome binding sites on the mRNA, enhancers and sequences which control termination of transcription and translation. The term "operably linked" means that specified components are in a relationship
(including but not limited to juxtaposition) permitting them to function in an intended manner. For example, a regulatory sequence is operably linked to a coding sequence such that expression of the coding sequence is under control of the regulatory sequences. A "signal sequence" is a sequence of amino acids attached to the N-terminal portion of a protein, which facilitates the secretion of the protein outside the cell. The mature form of an extracellular protein lacks the signal sequence, which is cleaved off during the secretion process.
"Biologically active" refers to a sequence having a specified biological activity, such an enzymatic activity.
The term "specific activity" refers to the number of moles of substrate that can be converted to product by an enzyme or enzyme preparation per unit time under specific conditions. Specific activity is generally expressed as units (U)/mg of protein.
As used herein, "water hardness" is a measure of the minerals {e.g., calcium and magnesium) present in water.
A "swatch" is a piece of material such as a fabric that has a stain applied thereto. The material can be, for example, fabrics made of cotton, polyester or mixtures of natural and synthetic fibers. The swatch can further be paper, such as filter paper or nitrocellulose, or a piece of a hard material such as ceramic, metal, or glass. For amylases, the stain is starch based, but can include blood, milk, ink, grass, tea, wine, spinach, gravy, chocolate, egg, cheese, clay, pigment, oil, or mixtures of these compounds.
A "smaller swatch" is a section of the swatch that has been cut with a single hole punch device, or has been cut with a custom manufactured 96-hole punch device, where
the pattern of the multi-hole punch is matched to standard 96-well microtiter plates, or the section has been otherwise removed from the swatch. The swatch can be of textile, paper, metal, or other suitable material. The smaller swatch can have the stain affixed either before or after it is placed into the well of a 24-, 48- or 96-well microtiter plate. The smaller swatch can also be made by applying a stain to a small piece of material. For example, the smaller swatch can be a stained piece of fabric 5/8" or 0.25" in diameter. The custom manufactured punch is designed in such a manner that it delivers 96 swatches simultaneously to all wells of a 96-well plate. The device allows delivery of more than one swatch per well by simply loading the same 96-well plate multiple times. Multi-hole punch devices can be conceived of to deliver simultaneously swatches to any format plate, including but not limited to 24-well, 48-well, and 96-well plates. In another conceivable method, the soiled test platform can be a bead made of metal, plastic, glass, ceramic, or another suitable material that is coated with the soil substrate. The one or more coated beads are then placed into wells of 96-, 48-, or 24-well plates or larger formats, containing suitable buffer and enzyme.
As used herein, "a cultured cell material comprising an amylase" or similar language, refers to a cell lysate or supernatant (including media) that includes an amylase as a component. The cell material may be from a heterologous host that is grown in culture for the purpose of producing the amylase.
As used herein, "percent sequence identity" means that a particular sequence has at least a certain percentage of amino acid residues identical to those in a specified reference sequence, when aligned using the CLUSTAL W algorithm with default parameters. See Thompson ef a/. (1 994) Nucleic Acids Res. 22:4673-4680. Default parameters for the CLUSTAL W algorithm are:
Gap opening penalty: 10.0
Gap extension penalty: 0.05
Protein weight matrix: BLOSUM series
DNA weight matrix: I UB
Delay divergent sequences %: 40
Gap separation distance: 8
DNA transitions weight: 0.50
List hydrophilic residues: GPSNDQEKR
Use negative matrix: OFF
Toggle Residue specific penalties: ON
Toggle hydrophilic penalties: ON
Toggle end gap separation penalty OFF. Deletions are counted as non-identical residues, compared to a reference sequence. Deletions occurring at either termini are included. For example, a variant with five amino acid deletions of the C-terminus of the mature 617 residue polypeptide would have a percent sequence identity of 99% (61 2 / 617 identical residues χ 1 00, rounded to the nearest whole number) relative to the mature polypeptide. Such a variant would be encompassed by a variant having "at least 99% sequence identity" to a mature polypeptide.
"Fused" polypeptide sequences are connected, i.e., operably linked, via a peptide bond between two subject polypeptide sequences.
The term "filamentous fungi" refers to all filamentous forms of the subdivision Eumycotina, particularly Pezizomycotina species.
The term "degree of polymerization" (DP) refers to the number (n) of
anhydro-glucopyranose units in a given saccharide. Examples of DP1 are the monosaccharides glucose and fructose. Examples of DP2 are the disaccharides maltose and sucrose. The term "DE," or "dextrose equivalent," is defined as the percentage of reducing sugar, i.e., D-glucose, as a fraction of total carbohydrate in a syrup.
The term "dry solids content" (ds) refers to the total solids of a slurry in a dry weight percent basis. The term "slurry" refers to an aqueous mixture containing insoluble solids. The phrase "simultaneous saccharification and fermentation (SSF)" refers to a process in the production of biochemicals in which a microbial organism, such as an ethanologenic microorganism, and at least one enzyme, such as an amylase, are present during the same process step. SSF includes the contemporaneous hydrolysis of starch substrates (granular, liquefied, or solubilized) to saccharides, including glucose,
and the fermentation of the saccharides into alcohol or other biochemical or biomaterial in the same reactor vessel.
An "ethanologenic microorganism" refers to a microorganism with the ability to convert a sugar or oligosaccharide to ethanol. The term "fermented beverage" refers to any beverage produced by a method comprising a fermentation process, such as a microbial fermentation, e.g., a bacterial and/or fungal fermentation. "Beer" is an example of such a fermented beverage, and the term "beer" is meant to comprise any fermented wort produced by
fermentation/brewing of a starch-containing plant material. Often, beer is produced exclusively from malt or adjunct, or any combination of malt and adjunct. Examples of beers include: full malted beer, beer brewed under the "Reinheitsgebot," ale, India pale ale, lager, pilsner, bitter, Happoshu (second beer), third beer, dry beer, near beer, light beer, low alcohol beer, low calorie beer, porter, bock, dopplebock, stout, porter, malt liquor, non-alcoholic beer, non-alcoholic malt liquor and the like, but also alternative cereal and malt beverages such as fruit flavored malt beverages, e.g., citrus flavored, such as lemon-, orange-, lime-, or berry-flavored malt beverages, liquor flavored malt beverages, e.g., vodka-, rum-, or tequila-flavored malt liquor, or coffee flavored malt beverages, such as caffeine-flavored malt liquor, and the like.
The term "malt" refers to any malted cereal grain, such as malted barley or wheat. The term "adjunct" refers to any starch and/or sugar containing plant material that is not malt, such as barley or wheat malt. Examples of adjuncts include common corn grits, refined corn grits, brewer's milled yeast, rice, sorghum, refined corn starch, barley, barley starch, dehusked barley, wheat, wheat starch, torrified cereal, cereal flakes, rye, oats, potato, tapioca, cassava and syrups, such as corn syrup, sugar cane syrup, inverted sugar syrup, barley and/or wheat syrups, and the like.
The term "mash" refers to an aqueous slurry of any starch and/or sugar containing plant material, such as grist, e.g., comprising crushed barley malt, crushed barley, and/or other adjunct or a combination thereof, mixed with water later to be separated into wort and spent grains.
The term "wort" refers to the unfermented liquor run-off following extracting the grist during mashing.
"Iodine-positive starch" or "IPS" refers to (1 ) amylose that is not hydrolyzed after liquefaction and saccharification, or (2) a retrograded starch polymer. When
saccharified starch or saccharide liquor is tested with iodine, the high DPn amylose or the retrograded starch polymer binds iodine and produces a characteristic blue color. The saccharide liquor is thus termed "iodine-positive saccharide," "blue saccharide," or "blue sac.
The terms "retrograded starch" or "starch retrogradation" refer to changes that occur spontaneously in a starch paste or gel on ageing.
The term "about" refers to ± 5% to the referenced value.
Additional mutations
In some embodiments, the present maltogenic amylases further include one or more mutations that provide a further performance or stability benefit. Exemplary performance benfits include but are not limited to increased hydrolysis of a starch substrate, increased grain, cereal or other starch substrate liquifaction performance, increased cleaning performance, increased thermal stability, increased storage stability, increased solubility, an altered pH profile, decreased calcium dependence, increased specific activity, modified substrate specificity, modified substrate binding, modified pH-dependent activity, modified pH-dependent stability, increased oxidative stability, and increased expression. In some cases, the performance benefit is realized at a relatively low temperature. In some cases, the performance benefit is realized at relatively high temperature.
Furthermore, the present amylases may include any number of conservative amino acid substitutions. Exemplary conservative amino acid substitutions are listed in the following Table 1 .
Table 1 . Conservative amino acid substitutions
The reader will appreciate that some of the above mentioned conservative mutations can be produced by genetic manpulation, while others are produced by introducing synthetic amino acids into a polypeptide by genetic or other means.
The present maltogenic amylase may be "precursor," "immature," or "full-length," in which case they include a signal sequence, or "mature," in which case they lack a signal sequence. Mature forms of the polypeptides are generally the most useful. Unless otherwise noted, the amino acid residue numbering used herein refers to the mature forms of the respective maltogenic amylase polypeptides. The present
maltogenic amylase polypeptides may also be truncated to remove the N or C-termini, so long as the resulting polypeptides retain amylase activity.
The present maltogenic amylase may be a "chimeric" or "hybrid" polypeptide, in that it includes at least a portion of a first amylase polypeptide, and at least a portion of a second amylase polypeptide (such chimeric amylases have recently been "rediscovered" as domain-swap amylases). The present amylases may further include heterologous signal sequence, an epitope to allow tracking or purification, or the like. Exemplary heterologous signal sequences are from B. licheniformis amylase (LAT), B. subtilis (AmyE or AprE), and Streptomyces CelA.
Nucleotides encoding maltogenic amylase polypeptides
In another aspect, nucleic acids encoding a maltogenic amylase polypeptide are provided. The nucleic acid may encode a particular maltogenic amylase polypeptide, or a maltogenic amylase having a specified degree of amino acid sequence identity to the particular amylase.
In one example, the nucleic acid encodes a maltogenic amylase having at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or even at least 99% identity to SEQ I D NO: 1 . It will be appreciated that due to the degeneracy of the genetic code, a plurality of nucleic acids may encode the same polypeptide.
In another example, the nucleic acid hybridizes under stringent or very stringent conditions to a nucleic acid encoding (or complementary to a nucleic acid encoding) a maltogenic amylase having at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or
even at least 99% identity to SEQ I D NO: 1 . Such stringent and very stringent hybridization conditions are described herein.
Nucleic acids may encode a "full-length" ("fl" or "FL") maltogenic amylase, which includes a signal sequence, only the mature form of a maltogenic amylase, which lacks the signal sequence, or a truncated form of a maltogenic amylase, which lacks the N or C-terminus of the mature form.
A nucleic acid that encodes a maltogenic amylase can be operably linked to various promoters and regulators in a vector suitable for expressing the maltogenic amylase in host cells. Exemplary promoters are from B. licheniformis amylase (LAT), B. subtilis (AmyE or AprE), and Streptomyces CelA. Such a nucleic acid can also be linked to other coding sequences, e.g., to encode a chimeric polypeptide.
Production of Variant Amylases
The present maltogenic amylases can be produced in host cells, for example, by secretion or intracellular expression. A cultured cell material (e.g., a whole-cell broth) comprising a maltogenic amylase can be obtained following secretion of the maltogenic amylase into the cell medium. Optionally, the maltogenic amylase can be isolated from the host cells, or even isolated from the cell broth, depending on the desired purity of the final maltogenic amylase. A gene encoding a maltogenic amylase can be cloned and expressed according to methods well known in the art. Suitable host cells include bacterial, fungal (including yeast and filamentous fungi), and plant cells (including algae). Particularly useful host cells include Aspergillus niger, Aspergillus oryzae or Trichoderma reesei. Other host cells include bacterial cells, e.g., Bacillus subtilis or B. licheniformis, as well as Streptomyces.
The host cell further may express a nucleic acid encoding a homologous or heterologous glucoamylase, i.e., a glucoamylase that is not the same species as the host cell, or one or more other enzymes. The glucoamylase may be a variant glucoamylase, such as one of the glucoamylase variants disclosed in U.S. Patent No. 8,058,033 (Danisco US Inc.), for example. Additionally, the host may express one or more accessory enzymes, proteins, peptides. These may benefit liquefaction,
saccharification, fermentation, SSF, etc processes. Furthermore, the host cell may produce biochemicals in addition to enzymes used to digest the various feedstock(s). Such host cells may be useful for fermentation or simultaneous saccharification and fermentation processes to reduce or eliminate the need to add enzymes. Vectors
A DNA construct comprising a nucleic acid encoding maltogenic amylases can be constructed to be expressed in a host cell. Representative nucleic acids that encode maltogenic amylases include SEQ I D NO: 1 . Because of the well-known degeneracy in the genetic code, variant polynucleotides that encode an identical amino acid sequence can be designed and made with routine skill. It is also well-known in the art to optimize codon use for a particular host cell. Nucleic acids encoding maltogenic amylases can be incorporated into a vector. Vectors can be transferred to a host cell using well-known transformation techniques, such as those disclosed below.
The vector may be any vector that can be transformed into and replicated within a host cell. For example, a vector comprising a nucleic acid encoding a maltogenic amylase can be transformed and replicated in a bacterial host cell as a means of propagating and amplifying the vector. The vector also may be transformed into an expression host, so that the encoding nucleic acids can be expressed as a functional maltogenic amylase. Host cells that serve as expression hosts can include filamentous fungi, for example. The Fungal Genetics Stock Center (FGSC) Catalogue of Strains lists suitable vectors for expression in fungal host cells. See FGSC, Catalogue of Strains, University of Missouri, ai www.fgsc.net (last modified January 17, 2007). A representative vector is pJG153, a promoterless Cre expression vector that can be replicated in a bacterial host. See Harrison et al. (June 2011 ) Applied Environ. Microbiol. 77: 3916-22. pJG153can be modified with routine skill to comprise and express a nucleic acid encoding a maltogenic amylase.
A nucleic acid encoding a maltogenic amylase can be operably linked to a suitable promoter, which allows transcription in the host cell. The promoter may be any DNA sequence that shows transcriptional activity in the host cell of choice and may be
derived from genes encoding proteins either homologous or heterologous to the host cell. Exemplary promoters for directing the transcription of the DNA sequence encoding a maltogenic amylase, especially in a bacterial host, are the promoter of the lac operon of E. coli, the Streptomyces coelicolor agarase gene dagA or eel A promoters, the promoters of the Bacillus licheniformis α-amylase gene (amyL), the promoters of the Bacillus stearothermophilus maltogenic amylase gene (amyM), the promoters of the Bacillus amyloliquefaciens a-amylase (amyQ), the promoters of the Bacillus subtilis xylA and xylB genes etc. For transcription in a fungal host, examples of useful promoters are those derived from the gene encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral a-amylase, A. niger acid stable a-amylase, A. n/ger glucoamylase, Rhizomucor miehei lipase, A. oryzae alkaline protease, A. oryzae triose phosphate isomerase, or A. nidulans acetamidase. When a gene encoding a maltogenic amylase is expressed in a bacterial species such as E. coli, a suitable promoter can be selected, for example, from a bacteriophage promoter including a T7 promoter and a phage lambda promoter. Examples of suitable promoters for the expression in a yeast species include but are not limited to the Gal 1 and Gal 10 promoters of Saccharomyces cerevisiae and the Pichia pastoris AOX1 or AOX2 promoters, cbh l is an endogenous, inducible promoter from T. reesei. See Liu et al. (2008) "Improved heterologous gene expression in Trichoderma reesei by
cellobiohydrolase I gene (cbhl ) promoter optimization," Acta Biochim. Biophys. Sin (Shanghai) 40(2): 1 58-65.
The coding sequence can be operably linked to a signal sequence. The DNA encoding the signal sequence may be the DNA sequence naturally associated with the maltogenic amylase gene to be expressed or from a different Genus or species. A signal sequence and a promoter sequence comprising a DNA construct or vector can be introduced into a fungal host cell and can be derived from the same source. For example, the signal sequence is the cbh l signal sequence that is operably linked to a cbh l promoter.
An expression vector may also comprise a suitable transcription terminator and, in eukaryotes, polyadenylation sequences operably linked to the DNA sequence
encoding a variant amylase. Termination and polyadenylation sequences may suitably be derived from the same sources as the promoter.
The vector may further comprise a DNA sequence enabling the vector to replicate in the host cell. Examples of such sequences are the origins of replication of plasmids pUC19, pACYCI 77, pUB110, pE194, pAMB1 , and plJ702.
The vector may also comprise a selectable marker, e.g., a gene the product of which complements a defect in the isolated host cell, such as the da/ genes from B. subtilis or B. licheniformis, or a gene that confers antibiotic resistance such as, e.g., ampicillin, kanamycin, chloramphenicol or tetracycline resistance. Furthermore, the vector may comprise Aspergillus selection markers such as amdS, argB, niaD and xxsC, a marker giving rise to hygromycin resistance, or the selection may be accomplished by co-transformation, such as known in the art. See e.g., International PCT Application WO 91 /17243.
Intracellular expression may be advantageous in some respects, e.g., when using certain bacteria or fungi as host cells to produce large amounts of maltogenic amylase for subsequent enrichment or purification. Extracellular secretion of amylase into the culture medium can also be used to make a cultured cell material comprising the isolated maltogenic amylase.
The expression vector typically includes the components of a cloning vector, such as, for example, an element that permits autonomous replication of the vector in the selected host organism and one or more phenotypically detectable markers for selection purposes. The expression vector normally comprises control nucleotide sequences such as a promoter, operator, ribosome binding site, translation initiation signal and optionally, a repressor gene or one or more activator genes. Additionally, the expression vector may comprise a sequence coding for an amino acid sequence capable of targeting the maltogenic amylase to a host cell organelle such as a peroxisome, or to a particular host cell compartment. Such a targeting sequence includes but is not limited to the sequence, SKL. For expression under the direction of control sequences, the nucleic
acid sequence of the amylase is operably linked to the control sequences in proper manner with respect to expression.
The procedures used to ligate the DNA construct encoding a maltogenic amylase, the promoter, terminator and other elements, respectively, and to insert them into suitable vectors containing the information necessary for replication, are well known to persons skilled in the art {see, e.g., Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2nd ed., Cold Spring Harbor, 1 989, and 3rd ed., 2001 ).
Transformation and Culture of Host Cells
An isolated cell, either comprising a DNA construct or an expression vector, is advantageously used as a host cell in the recombinant production of a maltogenic amylase. The cell may be transformed with the DNA construct encoding the enzyme, conveniently by integrating the DNA construct (in one or more copies) in the host chromosome. This integration is generally considered to be an advantage, as the DNA sequence is more likely to be stably maintained in the cell. Integration of the DNA constructs into the host chromosome may be performed according to conventional methods, e.g., by homologous or heterologous recombination. Alternatively, the cell may be transformed with an expression vector as described above in connection with the different types of host cells.
Examples of suitable bacterial host organisms are Gram positive bacterial species such as Bacillaceae including Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Geobacillus (formerly Bacillus) stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus lautus, Bacillus megaterium, and Bacillus thuringiensis; Streptomyces species such as Streptomyces murinus; lactic acid bacterial species including Lactococcus sp. such as Lactococcus lactis; Lactobacillus sp. including Lactobacillus reuteri; Leuconostoc sp. ; Pediococcus sp. ; and Streptococcus sp. Alternatively, strains of a Gram negative bacterial species belonging to Enterobacteriaceae including E. coli, or to Pseudomonadaceae can be selected as the host organism.
A suitable yeast host organism can be selected from the biotechnologically relevant yeasts species such as but not limited to yeast species such as Pichia sp., Hansenula sp., or Kluyveromyces, Yarrowinia, Schizosaccharomyces species or a species of Saccharomyces, including Saccharomyces cerevisiae or a species belonging to Schizosaccharomyces such as, for example, S. pombe species. A strain of the methylotrophic yeast species, Pichia pastoris, can be used as the host organism.
Alternatively, the host organism can be a Hansenula species. Suitable host organisms among filamentous fungi include species of Aspergillus, e.g., Aspergillus niger,
Aspergillus oryzae, Aspergillus tubigensis, Aspergillus awamori, or Aspergillus nidulans. Alternatively, strains of a Fusarium species, e.g., Fusarium oxysporum or of a
Rhizomucor species such as Rhizomucor miehei can be used as the host organism. Other suitable strains include Thermomyces and Mucor species. In addition,
Trichoderma sp. can be used as a host. A suitable procedure for transformation of Aspergillus host cells includes, for example, that described in EP 238023. A maltogenic amylase expressed by a fungal host cell can be glycosylated, i.e., will comprise a glycosyl moiety. The glycosylation pattern can be the same or different as present in the wild-type maltogenic amylase. The type and/or degree of glycosylation may impart changes in enzymatic and/or biochemical properties.
It is advantageous to delete genes from expression hosts, where the gene deficiency can be cured by the transformed expression vector. Known methods may be used to obtain a fungal host cell having one or more inactivated genes. Gene inactivation may be accomplished by complete or partial deletion, by insertional inactivation or by any other means that renders a gene nonfunctional for its intended purpose, such that the gene is prevented from expression of a functional protein. Any gene from a Trichoderma sp. or other filamentous fungal host that has been cloned can be deleted, for example, cbh l, cbh2, egl1, and egl2 genes. Gene deletion may be accomplished by inserting a form of the desired gene to be inactivated into a plasmid by methods known in the art.
Introduction of a DNA construct or vector into a host cell includes techniques such as transformation ; electroporation ; nuclear microinjection; transduction;
transfection, e.g., lipofection mediated and DEAE-Dextrin mediated transfection;
incubation with calcium phosphate DNA precipitate; high velocity bombardment with DNA-coated microprojectiles; and protoplast fusion. General transformation techniques are known in the art. See, e.g., Sambrook et al. (2001 ), supra. The expression of heterologous protein in Trichoderma is described, for example, in U.S. Patent No.
6,022,725. Reference is also made to Cao ef al. (2000) Science 9:991 -1001 for transformation of Aspergillus strains. Genetically stable transformants can be constructed with vector systems whereby the nucleic acid encoding a maltogenic amylase is stably integrated into a host cell chromosome. Transformants are then selected and purified by known techniques.
The preparation of Trichoderma sp. for transformation, for example, may involve the preparation of protoplasts from fungal mycelia. See Campbell ef al. (1 989) Curr. Genet. 1 6: 53-56. The mycelia can be obtained from germinated vegetative spores. The mycelia are treated with an enzyme that digests the cell wall, resulting in protoplasts, The protoplasts are protected by the presence of an osmotic stabilizer in the suspending medium. These stabilizers include sorbitol, mannitol, potassium chloride, magnesium sulfate, and the like. Usually the concentration of these stabilizers varies between 0.8 M and 1 .2 M, e.g., a 1 .2 M solution of sorbitol can be used in the suspension medium.
Uptake of DNA into the host Trichoderma sp. strain depends upon the calcium ion concentration. Generally, between about 10-50 mM CaCI2 is used in an uptake solution. Additional suitable compounds include a buffering system, such as TE buffer (10 mM Tris, pH 7.4; 1 mM EDTA) or 1 0 mM MOPS, pH 6.0 and polyethylene glycol. The polyethylene glycol is believed to fuse the cell membranes, thus permitting the contents of the medium to be delivered into the cytoplasm of the Trichoderma sp. strain. This fusion frequently leaves multiple copies of the plasmid DNA integrated into the host chromosome.
Usually transformation of Trichoderma sp. uses protoplasts or cells that have been subjected to a permeability treatment, typically at a density of 105 to 107/ml_, particularly 2x1 06/mL A volume of 100 μΙ_ of these protoplasts or cells in an appropriate
solution (e.g., 1 .2 M sorbitol and 50 mM CaCI2) may be mixed with the desired DNA. Generally, a high concentration of PEG is added to the uptake solution. From 0.1 to 1 volume of 25% PEG 4000 can be added to the protoplast suspension ; however, it is useful to add about 0.25 volumes to the protoplast suspension. Additives, such as dimethyl sulfoxide, heparin, spermidine, potassium chloride and the like, may also be added to the uptake solution to facilitate transformation. Similar procedures are available for other fungal host cells. See, e.g., U.S. Patent No. 6,022,725.
Expression A method of producing a maltogenic amylase may comprise cultivating a host cell as described above under conditions conducive to the production of the enzyme and recovering the enzyme from the cells and/or culture medium.
The medium used to cultivate the cells may be any conventional medium suitable for growing the host cell in question and obtaining expression of a maltogenic amylase. Suitable media and media components are available from commercial suppliers or may be prepared according to published recipes (e.g., as described in catalogues of the American Type Culture Collection).
An enzyme secreted from the host cells can be used in a whole broth preparation. In the present methods, the preparation of a spent whole fermentation broth of a recombinant microorganism can be achieved using any cultivation method known in the art resulting in the expression of a maltogenic amylase. Fermentation may, therefore, be understood as comprising shake flask cultivation, small- or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermenters performed in a suitable medium and under conditions allowing the amylase to be expressed or isolated. The term "spent whole fermentation broth" is defined herein as unfractionated contents of fermentation material that includes culture medium, extracellular proteins (e.g., enzymes), and cellular biomass. It is understood that the term "spent whole fermentation broth" also encompasses cellular biomass that has been lysed or permeabilized using methods well known in the art.
An enzyme secreted from the host cells may conveniently be recovered from the culture medium by well-known procedures, including separating the cells from the medium by centrifugation or filtration, and precipitating proteinaceous components of the medium by means of a salt such as ammonium sulfate, followed by the use of chromatographic procedures such as ion exchange chromatography, affinity
chromatography, or the like.
The polynucleotide encoding a maltogenic amylase in a vector can be operably linked to a control sequence that is capable of providing for the expression of the coding sequence by the host cell, i.e. the vector is an expression vector. The control sequences may be modified, for example by the addition of further transcriptional regulatory elements to make the level of transcription directed by the control sequences more responsive to transcriptional modulators. The control sequences may in particular comprise promoters.
Host cells may be cultured under suitable conditions that allow expression of a maltogenic amylase. Expression of the enzymes may be constitutive such that they are continually produced, or inducible, requiring a stimulus to initiate expression. In the case of inducible expression, protein production can be initiated when required by, for example, addition of an inducer substance to the culture medium, for example dexamethasone or IPTG or Sophorose. Polypeptides can also be produced
recombinantly in an in vitro cell-free system, such as the TNT™ (Promega) rabbit reticulocyte system.
An expression host also can be cultured in the appropriate medium for the host, under aerobic conditions. Shaking or a combination of agitation and aeration can be provided, with production occurring at the appropriate temperature for that host, e.g., from about 25 °C to about 75 °C (e.g., 30 °C to 45 °C), depending on the needs of the host and production of the desired maltogenic amylase. Culturing can occur from about 12 to about 100 hours or greater (and any hour value there between, e.g., from 24 to 72 hours). Typically, the culture broth is at a pH of about 4.0 to about 8.0, again depending on the culture conditions needed for the host relative to production of a maltogenic amylase.
Methods for Enriching and Purifying Maltogenic Amylases
Fermentation, separation, and concentration techniques are well known in the art and conventional methods can be used in order to prepare a concentrated maltogenic amylase polypeptide-containing solution.
After fermentation, a fermentation broth is obtained, the microbial cells and various suspended solids, including residual raw fermentation materials, are removed by conventional separation techniques in order to obtain a maltogenic amylase solution. Filtration, centrifugation, microfiltration, rotary vacuum drum filtration, ultrafiltration, centrifugation followed by ultra-filtration, extraction, or chromatography, or the like, are generally used.
It is desirable to concentrate a maltogenic amylase polypeptide-containing solution in order to optimize recovery. Use of unconcentrated solutions requires increased incubation time in order to collect the enriched or purified enzyme precipitate. The enzyme containing solution is concentrated using conventional concentration techniques until the desired enzyme level is obtained. Concentration of the enzyme containing solution may be achieved by any of the techniques discussed herein.
Exemplary methods of enrichment and purification include but are not limited to rotary vacuum filtration and/or ultrafiltration. The enzyme solution is concentrated into a concentrated enzyme solution until the enzyme activity of the concentrated maltogenic amylase polypeptide-containing solution is at a desired level.
Concentration may be performed using, e.g., a precipitation agent, such as a metal halide precipitation agent. Metal halide precipitation agents include but are not limited to alkali metal chlorides, alkali metal bromides and blends of two or more of these metal halides. Exemplary metal halides include sodium chloride, potassium chloride, sodium bromide, potassium bromide and blends of two or more of these metal halides.
The metal halide precipitation agent, sodium chloride, can also be used as a
preservative.
The metal halide precipitation agent is used in an amount effective to precipitate a maltogenic amylase. The selection of at least an effective amount and an optimum amount of metal halide effective to cause precipitation of the enzyme, as well as the conditions of the precipitation for maximum recovery including incubation time, pH, temperature and concentration of enzyme, will be readily apparent to one of ordinary skill in the art, after routine testing.
Generally, at least about 5% w/v (weight/volume) to about 25% w/v of metal halide is added to the concentrated enzyme solution, and usually at least 8% w/v. Generally, no more than about 25% w/v of metal halide is added to the concentrated enzyme solution and usually no more than about 20% w/v. The optimal concentration of the metal halide precipitation agent will depend, among others, on the nature of the specific maltogenic amylase polypeptide and on its concentration in the concentrated enzyme solution.
Another alternative way to precipitate the enzyme is to use organic compounds. Exemplary organic compound precipitating agents include: 4-hydroxybenzoic acid, alkali metal salts of 4-hydroxybenzoic acid, alkyl esters of 4-hydroxybenzoic acid, and blends of two or more of these organic compounds. The addition of the organic compound precipitation agents can take place prior to, simultaneously with or subsequent to the addition of the metal halide precipitation agent, and the addition of both precipitation agents, organic compound and metal halide, may be carried out sequentially or simultaneously.
Generally, the organic precipitation agents are selected from the group consisting of alkali metal salts of 4-hydroxybenzoic acid, such as sodium or potassium salts, and linear or branched alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1 to 12 carbon atoms, and blends of two or more of these organic compounds. The organic compound precipitation agents can be, for example, linear or branched alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1
to 10 carbon atoms, and blends of two or more of these organic compounds. Exemplary organic compounds are linear alkyl esters of 4-hydroxybenzoic acid, wherein the alkyl group contains from 1 to 6 carbon atoms, and blends of two or more of these organic compounds. Methyl esters of 4-hydroxybenzoic acid, propyl esters of 4-hydroxybenzoic acid, butyl ester of 4-hydroxybenzoic acid, ethyl ester of 4-hydroxybenzoic acid and blends of two or more of these organic compounds can also be used. Additional organic compounds also include but are not limited to 4-hydroxybenzoic acid methyl ester (named methyl PARABEN), 4-hydroxybenzoic acid propyl ester (named propyl
PARABEN), which also are both amylase preservative agents. For further descriptions, see, e.g., U.S. Patent No. 5,281 ,526.
Addition of the organic compound precipitation agent provides the advantage of high flexibility of the precipitation conditions with respect to pH, temperature, maltogenic amylase concentration, precipitation agent concentration, and time of incubation.
The organic compound precipitation agent is used in an amount effective to improve precipitation of the enzyme by means of the metal halide precipitation agent. The selection of at least an effective amount and an optimum amount of organic compound precipitation agent, as well as the conditions of the precipitation for maximum recovery including incubation time, pH, temperature and concentration of enzyme, will be readily apparent to one of ordinary skill in the art, in light of the present disclosure, after routine testing.
Generally, at least about 0.01 % w/v of organic compound precipitation agent is added to the concentrated enzyme solution and usually at least about 0.02% w/v.
Generally, no more than about 0.3% w/v of organic compound precipitation agent is added to the concentrated enzyme solution and usually no more than about 0.2% w/v. The concentrated polypeptide solution, containing the metal halide precipitation agent, and the organic compound precipitation agent, can be adjusted to a pH, which will, of necessity, depend on the enzyme to be enriched or purified. Generally, the pH is adjusted at a level near the isoelectric point of the amylase. The pH can be adjusted at
a pH in a range from about 2.5 pH units below the isoelectric point (pi) up to about 2.5 pH units above the isoelectric point.
The incubation time necessary to obtain an enriched or purified enzyme precipitate depends on the nature of the specific enzyme, the concentration of enzyme, and the specific precipitation agent(s) and its (their) concentration. Generally, the time effective to precipitate the enzyme is between about 1 to about 30 hours; usually it does not exceed about 25 hours. In the presence of the organic compound precipitation agent, the time of incubation can still be reduced to less about 10 hours and in most cases even about 6 hours. Generally, the temperature during incubation is between about 4°C and about
50 °C. Usually, the method is carried out at a temperature between about 10 °C and about 45 °C {e.g., between about 20 °C and about 40 °C). The optimal temperature for inducing precipitation varies according to the solution conditions and the enzyme or precipitation agent(s) used. The overall recovery of enriched or purified enzyme precipitate, and the efficiency with which the process is conducted, is improved by agitating the solution comprising the enzyme, the added metal halide and the added organic compound. The agitation step is done both during addition of the metal halide and the organic compound, and during the subsequent incubation period. Suitable agitation methods include mechanical stirring or shaking, vigorous aeration, or any similar technique.
After the incubation period, the enriched or purified enzyme is then separated from the dissociated pigment and other impurities and collected by conventional separation techniques, such as filtration, centrifugation, microfiltration, rotary vacuum filtration, ultrafiltration, press filtration, cross membrane microfiltration, cross flow membrane microfiltration, or the like. Further enrichment or purification of the enzyme precipitate can be obtained by washing the precipitate with water. For example, the enriched or purified enzyme precipitate is washed with water containing the metal halide precipitation agent, or with water containing the metal halide and the organic compound precipitation agents.
During fermentation, a maltogenic amylase polypeptide accumulates in the culture broth. For the isolation, enrichment, or purification of the desired maltogenic amylase, the culture broth is centrifuged or filtered to eliminate cells, and the resulting cell-free liquid is used for enzyme enrichment or purification. In one embodiment, the cell-free broth is subjected to salting out using ammonium sulfate at about 70% saturation; the 70% saturation-precipitation fraction is then dissolved in a buffer and applied to a column such as a Sephadex G-1 00 column, and eluted to recover the enzyme-active fraction. For further enrichment or purification, a conventional procedure such as ion exchange chromatography may be used. Enriched or purified enzymes are useful for laundry and cleaning applications.
For example, they can be used in laundry detergents and spot removers. They can be made into a final product that is either liquid (solution, slurry) or solid (granular, powder).
A more specific example of enrichment or purification, is described in Sumitani et al. (2000) "New type of starch-binding domain : the direct repeat motif in the C-terminal region of Bacillus sp. 195 a-amylase contributes to starch binding and raw starch degrading," Biochem. J. 350: 477-484, and is briefly summarized here. The enzyme obtained from 4 liters of a Streptomyces lividans TK24 culture supernatant is treated with (NH4)2S04 at 80% saturation. The precipitate is recovered by centrifugation at 10,000 x g (20 min. and 4°C) and re-dissolved in 20 mM Tris/HCI buffer (pH 7.0) containing 5 mM CaCI2. The solubilized precipitate is then dialyzed against the same buffer. The dialyzed sample is then applied to a Sephacryl S-200 column, which had previously been equilibrated with 20 mM Tris/HCI buffer, (pH 7.0), 5 mM CaCI2, and eluted at a linear flow rate of 7 mL/hr with the same buffer. Fractions from the column are collected and assessed for activity as judged by enzyme assay and SDS-PAGE. The protein is further purified as follows. A Toyopearl HW55 column (Tosoh Bioscience, Montgomeryville, PA; Cat. No. 19812) is equilibrated with 20 mM Tris/HCI buffer (pH 7.0) containing 5 mM CaCI2 and 1 .5 M (NH4)2S04. The enzyme is eluted with a linear gradient of 1 .5 to 0 M (NH4)2S04 in 20 mM Tris/HCL buffer, pH 7.0 containing 5 mM CaCI2. The active fractions are collected, and the enzyme precipitated with (NH4)2S04 at 80% saturation. The precipitate is recovered, re-dissolved, and dialyzed as described above. The
dialyzed sample is then applied to a Mono Q HR5/5 column (Amersham Pharmacia; Cat. No. 17-5167-01 ) previously equilibrated with 20 mM Tris/HCI buffer (pH 7.0) containing 5 mM CaCI2, at a flow rate of 60 mUhour. The active fractions are collected and added to a 1 .5 M (NH4)2S04 solution. The active enzyme fractions are re-chromatographed on a Toyopearl HW55 column, as before, to yield a homogeneous enzyme as determined by SDS-PAGE. See Sumitani ef al. (2000) Biochem. J. 350: 477-484, for general discussion of the method and variations thereon.
For production scale recovery, maltogenic amylase polypeptides can be enriched or partially purified as generally described above by removing cells via flocculation with polymers. Alternatively, the enzyme can be enriched or purified by microfiltration followed by concentration by ultrafiltration using available membranes and equipment. However, for some applications, the enzyme does not need to be enriched or purified, and whole broth culture can be lysed and used without further treatment. The enzyme can then be processed, for example, into granules. Compositions and Uses of Maltogenic Amylases
The maltogenic amylases provided by the present teachings are useful for a variety of industrial applications. For example, maltogenic amylases are useful in a starch conversion process, particularly in a saccharification process of a starch that has undergone liquefaction. The desired end-product may be any product that may be produced by the enzymatic conversion of the starch substrate. For example, the desired product may be a syrup rich in glucose and maltose, which can be used in other processes, such as the preparation of HFCS, or which can be converted into a number of other useful products, such as ascorbic acid intermediates {e.g., gluconate;
2-keto-L-gulonic acid; 5-keto-gluconate; and 2,5-diketogluconate) ; 1 ,3-propanediol; aromatic amino acids (e.g., tyrosine, phenylalanine and tryptophan); organic acids (e.g., lactate, pyruvate, succinate, isocitrate, and oxaloacetate); amino acids (e.g., serine and glycine) ; antibiotics; antimicrobials; enzymes; vitamins; and hormones.
The starch conversion process may be a precursor to, or simultaneous with, a fermentation process designed to produce alcohol for fuel or drinking (i.e., potable
alcohol). One skilled in the art is aware of various fermentation conditions that may be used in the production of these end-products. Variant amylases are also useful in compositions and methods of food preparation. These various uses of variant amylases are described in more detail below. It will be appreciated by one of ordinary skill in the art that various accessory enzymes can be used with the maltogenic enzymes of the present teachings, as will be the case in various applications and contexts.
In the field of grain processing to produce maltose syrups, the maltogenic amylases can be employed in any of a variety of applications, including those described in US Provisional Application 61 /61 6,990, filed March 28, 201 2.
Preparation of Starch Substrates from Plants
Those of general skill in the art are well aware of available methods that may be used to prepare starch substrates for use in the processes disclosed herein. For example, a useful starch substrate may be obtained from tubers, roots, stems, legumes, cereals or whole grain. More specifically, the granular starch may be obtained from corn, cobs, wheat, barley, rye, triticale, milo, sago, millet, cassava, tapioca, sorghum, rice, peas, bean, banana, or potatoes. Corn contains about 60-68% starch; barley contains about 55-65% starch; millet contains about 75-80% starch ; wheat contains about 60-65% starch; and polished rice contains 70-72% starch. Specifically contemplated starch substrates are corn starch and wheat starch. The starch from a grain may be ground or whole and includes corn solids, such as kernels, bran and/or cobs. The starch may also be highly refined raw starch or feedstock from starch refinery processes. Various starches also are commercially available. For example, corn starch is available from Cerestar, Sigma, and Katayama Chemical Industry Co. (Japan); wheat starch is available from Sigma; sweet potato starch is available from Wako Pure Chemical Industry Co. (Japan) ; and potato starch is available from Nakaari Chemical Pharmaceutical Co.
(Japan).
The starch substrate can be a crude starch from milled whole grain, which contains non-starch fractions, e.g., germ residues and fibers. Milling may comprise either wet milling or dry milling or grinding. In wet milling, whole grain is soaked in water or dilute acid to separate the grain into its component parts, e.g., starch, protein, germ, oil, kernel fibers. Wet milling efficiently separates the germ and meal {i.e., starch granules and protein) and is especially suitable for production of syrups. In dry milling or grinding, whole kernels are ground into a fine powder and often processed without fractionating the grain into its component parts. In some cases, oils from the kernels are recovered. Dry ground grain thus will comprise significant amounts of non-starch carbohydrate compounds, in addition to starch. Dry grinding of the starch substrate can be used for production of ethanol and other biochemicals. The starch to be processed may be a highly refined starch quality, for example, at least 90%, at least 95%, at least 97%, or at least 99.5% pure.
All references cited herein are herein incorporated by reference in their entirety for all purposes. In order to further illustrate the compositions and methods, and
advantages thereof, the following specific examples are given with the understanding that they are illustrative rather than limiting.
Fermentation The soluble starch hydrolysate, particularly a glucose rich syrup, can be fermented by contacting the starch hydrolysate with a fermenting organism typically at a temperature around 32 °C, such as from 30 °C to 35 °C for alcohol-producing yeast. The temperature and pH of the fermentation will depend upon the fermenting organism. EOF products include metabolites, such as citric acid, lactic acid, succinic acid, monosodium glutamate, gluconic acid, sodium gluconate, calcium gluconate, potassium gluconate, itaconic acid and other carboxylic acids, glucono delta-lactone, sodium erythorbate, lysine and other amino acids, omega 3 fatty acid, butanol, isoprene, 1 ,3-propanediol and other biomaterials.
Ethanologenic microorganisms include yeast, such as Saccharomyces cerevisiae and bacteria, e.g., Zymomonas mobilis, expressing alcohol dehydrogenase and pyruvate decarboxylase. The ethanologenic microorganism can express xylose reductase and xylitol dehydrogenase, which convert xylose to xylulose. Improved strains of ethanologenic microorganisms, which can withstand higher temperatures, for example, are known in the art and can be used. See Liu etal. (2011 ) Sheng Wu Gong Cheng Xue Bao 27(7): 1049-56. Commercial sources of yeast include ETHANOL RED® (LeSaffre) ; Thermosacc® (Lallemand) ; RED STAR® (Red Star); FERMIOL® (DSM Specialties); and SUPERSTART® (Alltech). Microorganisms that produce other metabolites, such as citric acid and lactic acid, by fermentation are also known in the art. See, e.g.,
Papagianni (2007) "Advances in citric acid fermentation by Aspergillus niger: biochemical aspects, membrane transport and modeling," Biotechnol. Adv. 25(3): 244-63; John et al. (2009) "Direct lactic acid fermentation : focus on simultaneous saccharification and lactic acid production," Biotechnol. Adv. 27(2): 145-52. The saccharification and fermentation processes may be carried out as an SSF process. Fermentation may comprise subsequent enrichment, purification, and recovery of ethanol, for example. During the fermentation, the ethanol content of the broth or "beer" may reach about 8-18% v/v, e.g., 14-15% v/v. The broth may be distilled to produce enriched, e.g., 96% pure, solutions of ethanol. Further, C02 generated by fermentation may be collected with a C02 scrubber, compressed, and marketed for other uses, e.g., carbonating beverage or dry ice production. Solid waste from the
fermentation process may be used as protein-rich products, e.g., livestock feed.
As mentioned above, an SSF process can be conducted with fungal cells that express and secrete amylase continuously throughout SSF. The fungal cells expressing amylase also can be the fermenting microorganism, e.g., an ethanologenic
microorganism. Ethanol production thus can be carried out using a fungal cell that expresses sufficient amylase so that less or no enzyme has to be added exogenously. The fungal host cell can be from an appropriately engineered fungal strain. Fungal host cells that express and secrete other enzymes, in addition to amylase, also can be used. Such cells may express glucoamylase and/or a pullulanase, phytase, a/pfra-glucosidase,
isoamylase, beta-amylase cellulase, xylanase, other hemicellulases, protease, befa-glucosidase, pectinase, esterase, redox enzymes, transferase, or other enzyme.
A variation on this process is a "fed-batch fermentation" system, where the substrate is added in increments as the fermentation progresses. Fed-batch systems are useful when catabolite repression may inhibit the metabolism of the cells and where it is desirable to have limited amounts of substrate in the medium. The actual substrate concentration in fed-batch systems is estimated by the changes of measurable factors such as pH, dissolved oxygen and the partial pressure of waste gases, such as C02. Batch and fed-batch fermentations are common and well known in the art. Continuous fermentation is an open system where a defined fermentation medium is added continuously to a bioreactor, and an equal amount of conditioned medium is removed simultaneously for processing. Continuous fermentation generally maintains the cultures at a constant high density where cells are primarily in log phase growth. Continuous fermentation permits modulation of cell growth and/or product concentration. For example, a limiting nutrient such as the carbon source or nitrogen source is maintained at a fixed rate and all other parameters are allowed to moderate. Because growth is maintained at a steady state, cell loss due to medium being drawn off should be balanced against the cell growth rate in the fermentation. Methods of optimizing continuous fermentation processes and maximizing the rate of product formation are well known in the art of industrial microbiology.
Compositions Comprising Maltogenic Amylases
The maltogenic amylases of the present teachings may be combined with a glucoamylase (EC 3.2.1 .3), e.g., a Trichoderma glucoamylase or variant thereof. An exemplary glucoamylase is Trichoderma reesei glucoamylase (TrGA) and variants thereof that possess superior specific activity and thermal stability. See U.S. Published Applications Nos. 2006/0094080, 2007/0004018, and 2007/0015266 (Danisco US Inc.). Suitable variants of TrGA include those with glucoamylase activity and at least 80%, at least 90%, or at least 95% sequence identity to wild-type TrGA. Maltogenic amylases
may advantageously increase the yield of glucose produced in a saccharification process catalyzed by TrGA.
Alternatively, the glucoamylase may be another glucoamylase derived from plants (including algae), fungi, or bacteria. For example, the glucoamylases may be Aspergillus niger Q† or G2 glucoamylase or its variants {e.g., Boel et al. (1984) EMBO J. 3: 1097-1 102; WO 92/00381 ; WO 00/04136 (Novo Nordisk A/S)); and A. awamori glucoamylase {e.g., WO 84/02921 (Cetus Corp.)). Other contemplated Aspergillus glucoamylase include variants with enhanced thermal stability, e.g., G137A and G1 39A (Chen etal. (1 996) Prot. Eng. 9: 499-505) ; D257E and D293E/Q (Chen et al. (1995) Prot. Eng. 8: 575-582) ; N182 (Chen et al. (1 994) Biochem. J. 301 : 275-281 ) ; A246C (Fierobe et al. (1996) Biochemistry, 35: 8698-8704); and variants with Pro residues in positions A435 and S436 (Li et al. (1 997) Protein Eng. 10: 1199-1204). Other contemplated glucoamylases include Talaromyces glucoamylases, in particular derived from T.
emersonii {e.g., WO 99/28448 (Novo Nordisk A/S), T. leycettanus {e.g., U.S. Patent No. RE 32, 153 (CPC International, Inc.)), T. duponti, or T. thermophilus {e.g., U.S. Patent No. 4,587,21 5). Contemplated bacterial glucoamylases include glucoamylases from the genus Clostridium, in particular C. thermoamylolyticum {e.g., EP 135, 1 38 (CPC
International, Inc.) and C. thermohydrosulfuricum {e.g., WO 86/01 831 (Michigan
Biotechnology Institute)). Suitable glucoamylases include the glucoamylases derived from Aspergillus oryzae, such as a glucoamylase shown in SEQ ID NO:2 in WO
00/041 36 (Novo Nordisk A/S). Also suitable are commercial glucoamylases, such as AMG 200L; AMG 300 L; SAN™ SUPER and AMG™ E (Novozymes); OPTIDEX® 300 and OPTI DEX L-400 (Danisco US Inc.); AMIGASE™and AMIGASE™ PLUS (DSM); G-ZYME® G900 (Enzyme Bio-Systems) ; and G-ZYME® G990 ZR {A. nigex
glucoamylase with a low protease content). Still other suitable glucoamylases include Aspergillus fumigatus glucoamylase, Talaromyces glucoamylase, Thielavia
glucoamylase, Trametes glucoamylase, Thermomyces glucoamylase, Athelia glucoamylase, or Humicola glucoamylase {e.g., HgGA). Glucoamylases typically are added in an amount of about 0.1 - 2 glucoamylase units (GAU)/g ds, e.g., about 0.1 6 GAU/g ds, 0.23 GAU/g ds, or 0.33 GAU/g ds.
Other suitable enzymes that can be used with the maltogenic amylase of the present teachings include a phytase, protease, pullulanase, β-amylase, isoamylase, a different a-amylase, alpha-glucosidase, cellulase, xylanase, other hemicellulases, beta-glucosidase, transferase, pectinase, lipase, cutinase, esterase, redox enzymes, or a combination thereof. For example, a debranching enzyme, such as an isoamylase (EC 3.2.1 .68), may be added in effective amounts well known to the person skilled in the art. A pullulanase (EC 3.2.1 .41 ), e.g., Promozyme®, is also suitable. Pullulanase typically is added at 100 U/kg ds. Further suitable enzymes include proteases, such as fungal and bacterial proteases. Fungal proteases include those obtained from
Aspergillus, such as A. niger, A. awamori, A. oryzae; Mucor (e.g., M. miehei); Rhizopus; and Trichoderma. β-Amylases (EC 3.2.1 .2) are exo-acting maltogenic amylases, which catalyze the hydrolysis of 1 ,4-a-glucosidic linkages into amylopectin and related glucose polymers, thereby releasing maltose. β-Amylases have been isolated from various plants and microorganisms. See Fogarty et al. (1979) in P ROGRESS IN INDUSTRIAL MICROBIOLOGY, Vol. 15, pp. 11 2-11 5. These β-Amylases have optimum temperatures in the range from 40°C to 65°C and optimum pH in the range from about 4.5 to about 7.0. Contemplated β-amylases include, but are not limited to, β-amylases from barley Spezyme® BBA 1500, Spezyme® DBA, Optimalt™ME, Optimalt™BBA (Danisco US Inc.); and Novozym™ WBA (Novozymes A/S) .
Compositions comprising the present maltogenic amylases may be aqueous or non-aqueous formulations, granules, powders, gels, slurries, pastes, etc., which may further comprise any one or more of the additional enzymes listed, herein, along with buffers, salts, preservatives, water, co-solvents, surfactants, and the like. Such compositions may work in combination with endogenous enzymes or other ingredients already present in a slurry, water bath, washing machine, food or drink product, etc, for example, endogenous plant (including algal) enzymes, residual enzymes from a prior processing step, and the like.
Compositions and Methods for Baking and Food Preparation
The present teachings also relate to a "food composition," including but not limited to a food product, animal feed and/or food/feed additives, comprising a maltogenic amylase, and methods for preparing such a food composition comprising mixing the maltogenic amylase with one or more food ingredients, or uses thereof.
Furthermore, the present teachings relate to the use of a maltogenic amylase in the preparation of a food composition, wherein the food composition is baked subsequent to the addition of the polypeptide of the present invention. As used herein the term "baking composition" means any composition and/or additive prepared in the process of providing a baked food product, including but not limited to bakers flour, a dough, a baking additive and/or a baked product. The food composition or additive may be liquid or solid.
As used herein, the term "flour" means milled or ground cereal grain. The term "flour" also may mean Sago or tuber products that have been ground or mashed. In some embodiments, flour may also contain components in addition to the milled or mashed cereal or plant matter. An example of an additional component, although not intended to be limiting, is a leavening agent. Cereal grains include wheat, oat, rye, and barley. Tuber products include tapioca flour, cassava flour, and custard powder. The term "flour" also includes ground corn flour, maize-meal, rice flour, whole-meal flour, self-rising flour, tapioca flour, cassava flour, ground rice, enriched flower, and custard powder.
For the commercial and home use of flour for baking and food production, it is important to maintain an appropriate level of a-amylase activity in the flour. A level of activity that is too high may result in a product that is sticky and/or doughy and therefore unmarketable. Flour with insufficient a-amylase activity may not contain enough sugar for proper yeast function, resulting in dry, crumbly bread, or baked products.
Accordingly, a maltogenic amylase, by itself or in combination with an a-amylase(s), may be added to the flour to augment the level of endogenous α-amylase activity in flour.
A maltogenic amylase can further be added alone or in a combination with other amylases to prevent or retard staling, i.e., crumb firming of baked products. The amount of anti-staling amylase will typically be in the range of 0.01 -1 0 mg of enzyme protein per kg of flour, e.g., 0.5 mg/kg ds. Additional anti-staling amylases that can be used in combination with an amylase include an endo-amylase, e.g., a bacterial endo-amylase from Bacillus. The additional amylase can be another maltogenic a-amylase (EC 3.2.1 .133), e.g., from Bacillus. Novamyl® is an exemplary maltogenic a-amylase from B. stearothermophilus strain NCIB 1 1837 and is described in
Christophersen et al. (1997) Starch 50: 39-45. Other examples of anti-staling endo-amylases include bacterial a-amylases derived from Bacillus, such as B.
licheniformis or B. amyloliquefaciens. The anti-staling amylase may be an exo-amylase, such as β-amylase, e.g., from plant sources, such as soy bean, or from microbial sources, such as Bacillus.
The baking composition comprising a maltogenic amylase further can comprise a phospholipase or enzyme with phospholipase activity. An enzyme with phospholipase activity has an activity that can be measured in Lipase Units (LU). The phospholipase may have A-ι or A2 activity to remove fatty acid from the phospholipids, forming a lysophospholipid. It may or may not have lipase activity, i.e., activity on triglyceride substrates. The phospholipase typically has a temperature optimum in the range of 30-90 °C, e.g., 30-70 °C. The added phospholipases can be of animal origin, for example, from pancreas, e.g., bovine or porcine pancreas, snake venom or bee venom. Alternatively, the phospholipase may be of microbial origin, e.g., from filamentous fungi, yeast or bacteria, for example.
The phospholipase is added in an amount that improves the softness of the bread during the initial period after baking, particularly the first 24 hours. The amount of phospholipase will typically be in the range of 0.01 -1 0 mg of enzyme protein per kg of flour, e.g., 0.1 -5 mg/kg. That is, phospholipase activity generally will be in the range of 20-1000 LU/kg of flour, where a Lipase Unit is defined as the amount of enzyme required to release 1 μιηοΙ butyric acid per minute at 30 °C, pH 7.0, with gum arabic as emulsifier and tributyrin as substrate.
Compositions of dough generally comprise wheat meal or wheat flour and/or other types of meal, flour or starch such as corn flour, cornstarch, rye meal, rye flour, oat flour, oatmeal, soy flour, sorghum meal, sorghum flour, potato meal, potato flour or potato starch. The dough may be fresh, frozen or par-baked. The dough can be a leavened dough or a dough to be subjected to leavening. The dough may be leavened in various ways, such as by adding chemical leavening agents, e.g., sodium bicarbonate or by adding a leaven, i.e., fermenting dough. Dough also may be leavened by adding a suitable yeast culture, such as a culture of Saccharomyces cerevisiae (baker's yeast), e.g., a commercially available strain of S. cerevisiae. The dough may also comprise other conventional dough ingredients, e.g., proteins, such as milk powder, gluten, and soy; eggs (e.g., whole eggs, egg yolks or egg whites); an oxidant, such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA) or ammonium persulfate; an amino acid such as L-cysteine; a sugar; or a salt, such as sodium chloride, calcium acetate, sodium sulfate or calcium sulfate. The dough further may comprise fat, e.g., triglyceride, such as granulated fat or shortening. The dough further may comprise an emulsifier such as mono- or diglycerides, diacetyl tartaric acid esters of mono- or diglycerides, sugar esters of fatty acids, polyglycerol esters of fatty acids, lactic acid esters of monoglycerides, acetic acid esters of monoglycerides, polyoxyethylene stearates, or lysolecithin. In particular, the dough can be made without addition of emulsifiers.
The dough product may be any processed dough product, including fried, deep fried, roasted, baked, steamed and boiled doughs, such as steamed bread and rice cakes. In one embodiment, the food product is a bakery product. Typical bakery (baked) products include bread - such as loaves, rolls, buns, bagels, pizza bases etc. pastry, pretzels, tortillas, cakes, cookies, biscuits, crackers etc.
Optionally, an additional enzyme may be used together with the anti-staling amylase and the phospholipase. The additional enzyme may be a second amylase, such as an amyloglucosidase, a β-amylase, a cyclodextrin glucanotransferase, or the additional enzyme may be a peptidase, in particular an exopeptidase, a transglutaminase,
a lipase, a cellulase, a xylanase, a protease, a protein disulfide isomerase, e.g., a protein disulfide isomerase as disclosed in WO 95/00636, for example, a glycosyltransferase, a branching enzyme (1 ,4-a-glucan branching enzyme), a 4-a-glucanotransferase (dextrin glycosyltransferase) or an oxidoreductase, e.g., a peroxidase, a laccase, a glucose oxidase, a pyranose oxidase, a lipooxygenase, an L-amino acid oxidase or a
carbohydrate oxidase. The additional enzyme(s) may be of any origin, including mammalian and plant, and particularly of microbial (bacterial, yeast or fungal) origin and may be obtained by techniques conventionally used in the art.
The xylanase is typically of microbial origin, e.g., derived from a bacterium or fungus, such as a strain of Aspergillus. Xylanases include Pentopan® and Novozym 384®, for example, which are commercially available xylanase preparations produced from Trichoderma reesei. The amyloglucosidase may be an A. n/ger amyloglucosidase (such as AMG®). Other useful amylase products include Grindamyl® A 1000 or A 5000 (Grindsted Products, Denmark) and Amylase® H or Amylase® P (DS ). The glucose oxidase may be a fungal glucose oxidase, in particular an Aspergillus niger glucose oxidase (such as Gluzyme®). An exemplary protease is Neutrase®.
The process may be used for any kind of baked product prepared from dough, either of a soft or a crisp character, either of a white, light or dark type. Examples are bread, particularly white, whole-meal or rye bread, typically in the form of loaves or rolls, such as, but not limited to, French baguette-type bread, pita bread, tortillas, cakes, pancakes, biscuits, cookies, pie crusts, crisp bread, steamed bread, pizza and the like.
A maltogenic amylase may be used in a pre-mix, comprising flour together with an anti-staling amylase, a phospholipase, and/or a phospholipid. The pre-mix may contain other dough-improving and/or bread-improving additives, e.g., any of the additives, including enzymes, mentioned above. A maltogenic amylase can be a component of an enzyme preparation comprising an anti-staling amylase and a phospholipase, for use as a baking additive.
The enzyme preparation is optionally in the form of a granulate or agglomerated powder. The preparation can have a narrow particle size distribution with
more than 95% (by weight) of the particles in the range from 25 to 500 μιη. Granulates and agglomerated powders may be prepared by conventional methods, e.g., by spraying an amylase onto a carrier in a fluid-bed granulator. The carrier may consist of particulate cores having a suitable particle size. The carrier may be soluble or insoluble, e.g., a salt (such as NaCI or sodium sulfate), a sugar (such as sucrose or lactose), a sugar alcohol (such as sorbitol), starch, rice, corn grits, or soy.
Enveloped particles, i.e., maltogenic amylase particles, can comprise a maltogenic amylase. To prepare enveloped maltogenic amylase particles, the enzyme is contacted with a food grade lipid in sufficient quantity to suspend all of the maltogenic amylase particles. Food grade lipids, as used herein, may be any naturally organic compound that is insoluble in water but is soluble in non-polar organic solvents such as hydrocarbon or diethyl ether. Suitable food grade lipids include, but are not limited to, triglycerides either in the form of fats or oils that are either saturated or unsaturated. Examples of fatty acids and combinations thereof which make up the saturated triglycerides include, but are not limited to, butyric (derived from milk fat), palmitic (derived from animal and plant fat), and/or stearic (derived from animal and plant fat). Examples of fatty acids and combinations thereof which make up the unsaturated triglycerides include, but are not limited to, palmitoleic (derived from animal and plant fat), oleic (derived from animal and plant fat), linoleic (derived from plant oils), and/or linolenic (derived from linseed oil). Other suitable food grade lipids include, but are not limited to, monoglycerides and diglycerides derived from the triglycerides discussed above, phospholipids and glycolipids.
The food grade lipid, particularly in the liquid form, is contacted with a powdered form of the maltogenic amylase particles in such a fashion that the lipid material covers at least a portion of the surface of at least a majority, e.g., 100% of the maltogenic amylase particles. Thus, each maltogenic amylase particle is individually enveloped in a lipid. For example, all or substantially all of the maltogenic amylase particles are provided with a thin, continuous, enveloping film of lipid. This can be accomplished by first pouring a quantity of lipid into a container, and then slurrying the maltogenic amylase particles so that the lipid thoroughly wets the surface of each maltogenic amylase particle. After a
short period of stirring, the enveloped maltogenic amylase particles, carrying a substantial amount of the lipids on their surfaces, are recovered. The thickness of the coating so applied to the particles of maltogenic amylase can be controlled by selection of the type of lipid used and by repeating the operation in order to build up a thicker film, when desired.
The storing, handling and incorporation of the loaded delivery vehicle can be accomplished by means of a packaged mix. The packaged mix can comprise the enveloped maltogenic amylase. However, the packaged mix may further contain additional ingredients as required by the manufacturer or baker. After the enveloped maltogenic amylase has been incorporated into the dough, the baker continues through the normal production process for that product.
The advantages of enveloping the maltogenic amylase particles are two-fold. First, the food grade lipid protects the enzyme from thermal denaturation during the baking process for those enzymes that are heat labile. Consequently, while the maltogenic amylase is stabilized and protected during the proving and baking stages, it is released from the protective coating in the final baked good product, where it hydrolyzes the glucosidic linkages in polyglucans. The loaded delivery vehicle also provides a sustained release of the active enzyme into the baked good. That is, following the baking process, active maltogenic amylase is continually released from the protective coating at a rate that counteracts, and therefore reduces the rate of, staling mechanisms.
In general, the amount of lipid applied to the maltogenic amylase particles can vary from a few percent of the total weight of the maltogenic amylase to many times that weight, depending upon the nature of the lipid, the manner in which it is applied to the maltogenic amylase particles, the composition of the dough mixture to be treated, and the severity of the dough-mixing operation involved.
The loaded delivery vehicle, i.e., the lipid-enveloped enzyme, is added to the ingredients used to prepare a baked good in an effective amount to extend the shelf-life of the baked good. The baker computes the amount of enveloped maltogenic amylase, prepared as discussed above, that will be required to achieve the desired anti-staling
effect. The amount of the enveloped maltogenic amylase required is calculated based on the concentration of enzyme enveloped and on the proportion of maltogenic amylase to flour specified. A wide range of concentrations has been found to be effective, although, as has been discussed, observable improvements in anti-staling do not correspond linearly with the maltogenic amylase concentration, but above certain minimal levels, large increases in maltogenic amylase concentration produce little additional improvement. The maltogenic amylase concentration actually used in a particular bakery production could be much higher than the minimum necessary to provide the baker with some insurance against inadvertent under-measurement errors by the baker. The lower limit of enzyme concentration is determined by the minimum anti-staling effect the baker wishes to achieve.
A method of preparing a baked good may comprise: a) preparing lipid-coated maltogenic amylase particles, where substantially all of the maltogenic amylase particles are coated; b) mixing a dough containing flour; c) adding the lipid-coated maltogenic amylase to the dough before the mixing is complete and terminating the mixing before the lipid coating is removed from the maltogenic amylase; d) proofing the dough; and e) baking the dough to provide the baked good, where the maltogenic amylase is inactive during the mixing, proofing and baking stages and is active in the baked good.
The enveloped maltogenic amylase can be added to the dough during the mix cycle, e.g., near the end of the mix cycle. The enveloped maltogenic amylase is added at a point in the mixing stage that allows sufficient distribution of the enveloped maltogenic amylase throughout the dough ; however, the mixing stage is terminated before the protective coating becomes stripped from the maltogenic amylase particle(s). Depending on the type and volume of dough, and mixer action and speed, anywhere from one to six minutes or more might be required to mix the enveloped maltogenic amylase into the dough, but two to four minutes is average. Thus, several variables may determine the precise procedure. First, the quantity of enveloped maltogenic amylase should have a total volume sufficient to allow the enveloped maltogenic amylase to be spread throughout the dough mix. If the preparation of enveloped maltogenic amylase is highly concentrated, additional oil may need to be added to the pre-mix before the
enveloped maltogenic amylase is added to the dough. Recipes and production processes may require specific modifications; however, good results generally can be achieved when 25% of the oil specified in a bread dough formula is held out of the dough and is used as a carrier for a concentrated enveloped a-amylase when added near the end of the mix cycle. In bread or other baked goods, particularly those having a low fat content, e.g., French-style breads, an enveloped maltogenic amylase mixture of approximately 1 % of the dry flour weight is sufficient to admix the enveloped a-amylase properly with the dough. The range of suitable percentages is wide and depends on the formula, finished product, and production methodology requirements of the individual baker. Second, the enveloped maltogenic amylase suspension should be added to the mix with sufficient time for complete mixture into the dough, but not for such a time that excessive mechanical action strips the protective lipid coating from the enveloped maltogenic amylase particles.
In a further aspect of the invention, the food composition is an oil, meat, lard, composition comprising a maltogenic amylase. In this context the term "oil/meat/lard" composition" means any composition, based on, made from and/or containing oil, meat or lard, respectively. Another aspect the invention relates to a method of preparing an oil or meat or lard composition and/or additive comprising a maltogenic amylase, comprising mixing the polypeptide of the invention with a oil/meat/lard composition and/or additive ingredients.
In a further aspect of the invention, the food composition is an animal feed composition, animal feed additive and/or pet food comprising a maltogenic amylase and variants thereof. The present invention further relates to a method for preparing such an animal feed composition, animal feed additive composition and/or pet food comprising mixing a maltogenic amylase and variants thereof with one or more animal feed ingredients and/or animal feed additive ingredients and/or pet food ingredients.
Furthermore, the present invention relates to the use of a maltogenic amylase in the preparation of an animal feed composition and/or animal feed additive composition and/or pet food.
The term "animal" includes all non-ruminant and ruminant animals. In a particular embodiment, the animal is a non-ruminant animal, such as a horse and a mono-gastric animal. Examples of mono-gastric animals include, but are not limited to, pigs and swine, such as piglets, growing pigs, sows; poultry such as turkeys, ducks, chicken, broiler chicks, layers; fish such as salmon, trout, tilapia, catfish and carps; and crustaceans such as shrimps and prawns. In a further embodiment the animal is a ruminant animal including, but not limited to, cattle, young calves, goats, sheep, giraffes, bison, moose, elk, yaks, water buffalo, deer, camels, alpacas, llamas, antelope, pronghorn and nilgai.
In the present context, it is intended that the term "pet food" is understood to mean a food for a household animal such as, but not limited to dogs, cats, gerbils, hamsters, chinchillas, fancy rats, guinea pigs; avian pets, such as canaries, parakeets, and parrots; reptile pets, such as turtles, lizards and snakes; and aquatic pets, such as tropical fish and frogs.
The terms "animal feed composition," "feedstuff" and "fodder" are used interchangeably and may comprise one or more feed materials selected from the group comprising a) cereals, such as small grains {e.g., wheat, barley, rye, oats and combinations thereof) and/or large grains such as maize or sorghum ; b) by products from cereals, such as corn gluten meal, Distillers Dried Grain Solubles (DDGS) (particularly corn based Distillers Dried Grain Solubles (cDDGS), wheat bran, wheat middlings, wheat shorts, rice bran, rice hulls, oat hulls, palm kernel, and citrus pulp; c) protein obtained from sources such as soya, sunflower, peanut, lupin, peas, fava beans, cotton, canola, fish meal, dried plasma protein, meat and bone meal, potato protein, whey, copra, sesame; d) oils and fats obtained from vegetable and animal sources; e) minerals and vitamins. Textile Desizing Compositions and Use
Also contemplated are compositions and methods of treating fabrics {e.g., to desize a textile) using a maltogenic amylase. Fabric-treating methods are well known in the art {see, e.g., U.S. Patent No. 6,077,316). For example, the feel and appearance of
a fabric can be improved by a method comprising contacting the fabric with a maltogenic amylase in a solution. The fabric can be treated with the solution under pressure.
A maltogenic amylase can be applied during or after the weaving of a textile, or during the desizing stage, or one or more additional fabric processing steps. During the weaving of textiles, the threads are exposed to considerable mechanical strain. Prior to weaving on mechanical looms, warp yarns are often coated with sizing starch or starch derivatives to increase their tensile strength and to prevent breaking. A maltogenic amylase can be applied during or after the weaving to remove these sizing starch or starch derivatives. After weaving, a maltogenic amylase can be used to remove the size coating before further processing the fabric to ensure a homogeneous and wash-proof result.
A maltogenic amylase can be used alone or with other desizing chemical reagents and/or desizing enzymes to desize fabrics, including cotton-containing fabrics, as detergent additives, e.g., in aqueous compositions. A maltogenic amylase also can be used in compositions and methods for producing a stonewashed look on indigo-dyed denim fabric and garments. For the manufacture of clothes, the fabric can be cut and sewn into clothes or garments, which are afterwards finished. In particular, for the manufacture of denim jeans, different enzymatic finishing methods have been developed. The finishing of denim garment normally is initiated with an enzymatic desizing step, during which garments are subjected to the action of amylolytic enzymes to provide softness to the fabric and make the cotton more accessible to the subsequent enzymatic finishing steps. A maltogenic amylase can be used in methods of finishing denim garments {e.g., a "bio-stoning process"), enzymatic desizing and providing softness to fabrics, and/or finishing process. Cleaning Compositions
An aspect of the present compositions and methods is a cleaning composition that includes a maltogenic amylase as a component. A maltogenic amylase polypeptide can be used as a component in detergent compositions for hand washing, laundry washing, dishwashing, and other hard-surface cleaning.
Overview of Cleaning Compositions
Preferably, a maltogenic amylase is incorporated into detergents at or near a concentration conventionally used for amylase in detergents. For example, a maltogenic amylase polypeptide may be added in amount corresponding to 0.00001 - 1 mg (calculated as pure enzyme protein) of maltogenic amylase per liter of
wash/dishwash liquor. Exemplary formulations are provided herein, as exemplified by the following:
A maltogenic amylase polypeptide may be a component of a detergent composition, as the only enzyme or with other enzymes including other amylolytic enzymes. As such, it may be included in the detergent composition in the form of a non-dusting granulate, a stabilized liquid, or a protected enzyme. Non-dusting granulates may be produced, e.g., as disclosed in U.S. Patent Nos. 4,1 06,991 and 4,661 ,452 and may optionally be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxide) products (polyethyleneglycol, PEG) with mean molar weights of 1 ,000 to 20,000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in, for example, GB 1483591 . Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other enzyme stabilizers are known in the art.
Protected enzymes may be prepared according to the method disclosed in for example EP 238 21 6. Polyols have long been recognized as stabilizers of proteins, as well as improving protein solubility.
The detergent composition may be in any useful form, e.g., as powders, granules, pastes, or liquid. A liquid detergent may be aqueous, typically containing up to about 70% of water and 0% to about 30% of organic solvent. It may also be in the form of a compact gel type containing only about 30% water.
The detergent composition comprises one or more surfactants, each of which may be anionic, nonionic, cationic, or zwitterionic. The detergent will usually contain 0% to about 50% of anionic surfactant, such as linear alkylbenzenesulfonate (LAS) ;
a-olefinsulfonate (AOS); alkyl sulfate (fatty alcohol sulfate) (AS); alcohol ethoxysulfate (AEOS or AES); secondary alkanesulfonates (SAS); a-sulfo fatty acid methyl esters; alkyl- or alkenylsuccinic acid; or soap. The composition may also contain 0% to about 40% of nonionic surfactant such as alcohol ethoxylate (AEO or AE), carboxylated alcohol ethoxylates, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (as described for example in WO 92/061 54).
The detergent composition may additionally comprise one or more other enzymes, such as proteases, another amylolytic enzyme, cutinase, lipase, cellulase, pectate lyase, perhydrolase, xylanase, peroxidase, and/or laccase in any combination.
The detergent may contain about 1 % to about 65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA),
diethylenetriaminepentaacetic acid (DT PA), alkyl- or alkenylsuccinic acid, soluble silicates or layered silicates {e.g., SKS-6 from Hoechst). The detergent may also be unbuilt, i.e. essentially free of detergent builder. The enzymes can be used in any composition compatible with the stability of the enzyme. Enzymes generally can be protected against deleterious components by known forms of encapsulation, for example, by granulation or sequestration in hydro gels. Enzymes, and specifically maltoogenic amylases, either with or without starch binding domains, can be used in a variety of compositions including laundry and dishwashing applications, surface cleaners, as well as in compositions for ethanol production from starch or biomass.
The detergent may comprise one or more polymers. Examples include carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (PVP), polyethyleneglycol (PEG), polyvinyl alcohol) (PVA), polycarboxylates such as polyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid copolymers.
The detergent may contain a bleaching system, which may comprise a H202 source such as perborate or percarbonate, which may be combined with a
peracid-forming bleach activator such as tetraacetylethylenediamine (TAED) or nonanoyloxybenzenesulfonate (NOBS). Alternatively, the bleaching system may comprise peroxyacids (e.g., the amide, imide, or sulfone type peroxyacids). The bleaching system can also be an enzymatic bleaching system, for example, perhydrolase, such as that described in International PCT Application WO 2005/056783.
The enzymes of the detergent composition may be stabilized using conventional stabilizing agents, e.g., a polyol such as propylene glycol or glycerol; a sugar or sugar alcohol; lactic acid; boric acid or a boric acid derivative such as, e.g., an aromatic borate ester; and the composition may be formulated as described in, e.g., WO 92/19709 and WO 92/19708.
The detergent may also contain other conventional detergent ingredients such as e.g., fabric conditioners including clays, foam boosters, suds suppressors, anti-corrosion agents, soil-suspending agents, anti-soil redeposition agents, dyes, bactericides, tarnish inhibiters, optical brighteners, or perfumes.
The pH (measured in aqueous solution at use concentration) is usually neutral or alkaline, e.g., pH about 7.0 to about 1 1 .0.
Particular forms of detergent compositions for inclusion of the present a-amylase are described, below.
Heavy Duty Liquid (HDL) laundry detergent composition
Exemplary HDL laundry detergent compositions includes a detersive surfactant (10%-40% wt/wt), including an anionic detersive surfactant (selected from a group of linear or branched or random chain, substituted or unsubstituted alkyl sulphates, alkyl sulphonates, alkyl alkoxylated sulphate, alkyl phosphates, alkyl phosphonates, alkyl carboxylates, and/or mixtures thereof), and optionally non-ionic surfactant (selected from a group of linear or branched or random chain, substituted or unsubstituted alkyl alkoxylated alcohol, for example a Cs-Ci8 alkyl ethoxylated alcohol and/or C6-Ci2 alkyl phenol alkoxylates), wherein the weight ratio of anionic detersive surfactant (with a
hydrophilic index (Hlc) of from 6.0 to 9) to non-ionic detersive surfactant is greater than 1 : 1 . Suitable detersive surfactants also include cationic detersive surfactants (selected from a group of alkyl pyridinium compounds, alkyl quarternary ammonium compounds, alkyl quarternary phosphonium compounds, alkyl ternary sulphonium compounds, and/or mixtures thereof); zwitterionic and/or amphoteric detersive surfactants (selected from a group of alkanolamine sulpho-betaines) ; ampholytic surfactants; semi-polar non-ionic surfactants and mixtures thereof.
The composition may optionally include, a surfactancy boosting polymer consisting of amphiphilic alkoxylated grease cleaning polymers (selected from a group of alkoxylated polymers having branched hydrophilic and hydrophobic properties, such as alkoxylated polyalkylenimines in the range of 0.05wt%-10wt%) and/or random graft polymers (typically comprising of hydrophilic backbone comprising monomers selected from the group consisting of: unsaturated C-|-C6 carboxylic acids, ethers, alcohols, aldehydes, ketones, esters, sugar units, alkoxy units, maleic anhydride, saturated polyalcohols such as glycerol, and mixtures thereof; and hydrophobic side chain(s) selected from the group consisting of: C -C25 alkyl group, polypropylene, polybutylene, vinyl ester of a saturated CrC6 mono-carboxylic acid, CrC6 alkyl ester of acrylic or methacrylic acid, and mixtures thereof.
The composition may include additional polymers such as soil release polymers (include anionically end-capped polyesters, for example SRP1 , polymers comprising at least one monomer unit selected from saccharide, dicarboxylic acid, polyol and combinations thereof, in random or block configuration, ethylene terephthalate-based polymers and co-polymers thereof in random or block configuration, for example Repel-o-tex SF, SF-2 and SRP6, Texcare SRA1 00, SRA300, SRN100, SRN170, SRN240, SRN300 and SRN325, Marloquest SL), anti-redeposition polymers (0.1 wt% to 10wt%, include carboxylate polymers, such as polymers comprising at least one monomer selected from acrylic acid, maleic acid (or maleic anhydride), fumaric acid, itaconic acid, aconitic acid, mesaconic acid, citraconic acid, methylenemalonic acid, and any mixture thereof, vinylpyrrolidone homopolymer, and/or polyethylene glycol, molecular weight in the range of from 500 to 100,000 Da); cellulosic polymer (including
those selected from alkyl cellulose, alkyl alkoxyalkyl cellulose, carboxyalkyl cellulose, alkyl carboxyalkyl cellulose examples of which include carboxymethyl cellulose, methyl cellulose, methyl hydroxyethyl cellulose, methyl carboxymethyl cellulose, and mixures thereof) and polymeric carboxylate (such as maleate/acrylate random copolymer or polyacrylate homopolymer).
The composition may further include saturated or unsaturated fatty acid, preferably saturated or unsaturated Ci2-C24 fatty acid (0 wt% to 10 wt%); deposition aids (examples for which include polysaccharides, preferably cellulosic polymers, poly diallyl dimethyl ammonium halides (DADMAC), and co-polymers of DAD MAC with vinyl pyrrolidone, acrylamides, imidazoles, imidazolinium halides, and mixtures thereof, in random or block configuration, cationic guar gum, cationic cellulose such as cationic hydoxyethyl cellulose, cationic starch, cationic polyacylamides, and mixtures thereof.
The composition may further include dye transfer inhibiting agents, examples of which include manganese phthalocyanine, peroxidases, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles and/or mixtures thereof; chelating agents, examples of which include ethylene-diamine-tetraacetic acid (EDTA), diethylene triamine penta methylene phosphonic acid (DTPMP), hydroxy-ethane diphosphonic acid (HEDP), ethylenediamine Ν,Ν'-disuccinic acid (EDDS), methyl glycine diacetic acid (MGDA), diethylene triamine penta acetic acid (DTPA), propylene diamine tetracetic acid (PDT A), 2-hydroxypyridine-N-oxide (HPNO), or methyl glycine diacetic acid (MGDA), glutamic acid Ν,Ν-diacetic acid (Ν,Ν-dicarboxymethyl glutamic acid tetrasodium salt (GLDA), nitrilotriacetic acid (NTA), 4,5-dihydroxy-m-benzenedisulfonic acid, citric acid and any salts thereof, N-hydroxyethylethylenediaminetri-acetic acid (HEDTA),
triethylenetetraaminehexaacetic acid (TTHA), N-hydroxyethyliminodiacetic acid (HEIDA), dihydroxyethylglycine (DHEG), ethylenediaminetetrapropionic acid (EDTP), and derivatives thereof.
The composition preferably includes enzymes (generally about 0.01 wt% active enzyme to 0.03wt% active enzyme) selected from proteases, amylases, lipases,
cellulases, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferases,
perhydrolases, arylesterases, and any mixture thereof. The composition may include an enzyme stabilizer (examples of which include polyols such as propylene glycol or glycerol, sugar or sugar alcohol, lactic acid, reversible protease inhibitor, boric acid, or a boric acid derivative, e.g., an aromatic borate ester, or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid).
The composition optionally includes silicone or fatty-acid based suds
suppressors; hueing dyes, calcium and magnesium cations, visual signaling ingredients, anti-foam (0.001 wt% to about 4.0wt%), and/or structurant/thickener (0.01 wt% to 5wt%, selected from the group consisting of diglycerides and triglycerides, ethylene glycol distearate, microcrystalline cellulose, cellulose based materials, microfiber cellulose, biopolymers, xanthan gum, gellan gum, and mixtures thereof).
The composition can be any liquid form, for example a liquid or gel form, or any combination thereof. The composition may be in any unit dose form, for example a pouch.
Heavy Duty Dry/Solid (HDD) laundry detergent composition
Exemplary HDD laundry detergent compositions includes a detersive surfactant, including anionic detersive surfactants (e.g., linear or branched or random chain, substituted or unsubstituted alkyi sulphates, alkyi sulphonates, alkyi alkoxylated sulphate, alkyi phosphates, alkyi phosphonates, alkyi carboxylates and/or mixtures thereof), non-ionic detersive surfactant (e.g., linear or branched or random chain, substituted or unsubstituted C8-C 8 alkyi ethoxylates, and/or C6-C 2 alkyi phenol alkoxylates), cationic detersive surfactants (e.g., alkyi pyridinium compounds, alkyi quaternary ammonium compounds, alkyi quaternary phosphonium compounds, alkyi ternary sulphonium compounds, and mixtures thereof), zwitterionic and/or amphoteric detersive surfactants (e.g., alkanolamine sulpho-betaines), ampholytic surfactants, semi-polar non-ionic surfactants, and mixtures thereof ; builders including phosphate free builders (for example zeolite builders examples which include zeolite A, zeolite X, zeolite P and
zeolite MAP in the range of 0wt% to less than 1 0wt%), phosphate builders (for example sodium tri-polyphosphate in the range of 0wt% to less than 1 0wt%), citric acid, citrate salts and nitrilotriacetic acid, silicate salt (e.g., sodium or potassium silicate or sodium meta-silicate in the range of 0wt% to less than 1 0wt%, or layered silicate (SKS-6)) ;
carbonate salt (e.g., sodium carbonate and/or sodium bicarbonate in the range of 0 wt% to less than 80 wt%) ; and bleaching agents including photobleaches (e.g., sulfonated zinc phthalocyanines, sulfonated aluminum phthalocyanines, xanthenes dyes, and mixtures thereof) hydrophobic or hydrophilic bleach activators (e.g., dodecanoyl oxybenzene sulfonate, decanoyl oxybenzene sulfonate, decanoyl oxybenzoic acid or salts thereof, 3,5,5-trimethy hexanoyl oxybenzene sulfonate, tetraacetyl ethylene diamine-TAED, nonanoyloxybenzene sulfonate-NOBS, nitrile quats, and mixtures thereof), sources of hydrogen peroxide (e.g., inorganic perhydrate salts examples of which include mono or tetra hydrate sodium salt of perborate, percarbonate, persulfate, perphosphate, or persilicate), preformed hydrophilic and/or hydrophobic peracids (e.g., percarboxylic acids and salts, percarbonic acids and salts, perimidic acids and salts, peroxymonosulfuric acids and salts, and mixtures thereof), and/or bleach catalysts (e.g., imine bleach boosters (examples of which include iminium cations and polyions), iminium zwitterions, modified amines, modified amine oxides, N-sulphonyl imines, N-phosphonyl imines, N-acyl imines, thiadiazole dioxides, perfluoroimines, cyclic sugar ketones, and mixtures thereof, and metal-containing bleach catalysts (e.g., copper, iron, titanium, ruthenium, tungsten, molybdenum, or manganese cations along with an auxiliary metal cations such as zinc or aluminum and a sequestrate such as ethylenediaminetetraacetic acid, ethylenediaminetetra(methylenephosphonic acid), and water-soluble salts thereof).
The composition preferably includes enzymes, e.g., proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases,
mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferase, perhydrolase, arylesterase, and any mixture thereof.
The composition may optionally include additional detergent ingredients including perfume microcapsules, starch encapsulated perfume accord, hueing agents, additional polymers, including fabric integrity and cationic polymers, dye-lock ingredients,
fabric-softening agents, brighteners (for example C.I. Fluorescent brighteners), flocculating agents, chelating agents, alkoxylated polyamines, fabric deposition aids, and/or cyclodextrin.
Automatic dishwashing (ADW) detergent composition Exemplary ADW detergent composition includes non-ionic surfactants, including ethoxylated non-ionic surfactants, alcohol alkoxylated surfactants, epoxy-capped poly(oxyalkylated) alcohols, or amine oxide surfactants present in amounts from 0 to 1 0% by weight; builders in the range of 5-60% including phosphate builders {e.g.,
mono-phosphates, di-phosphates, tri-polyphosphates, other oligomeric-poylphosphates, sodium tripolyphosphate-STPP) and phosphate-free builders (e.g., amino acid-based compounds including methyl-glycine-diacetic acid (MGDA) and salts and derivatives thereof, glutamic-N,N-diacetic acid (GLDA) and salts and derivatives thereof,
iminodisuccinic acid (IDS) and salts and derivatives thereof, carboxy methyl inulin and salts and derivatives thereof, nitrilotriacetic acid (NTA), diethylene triamine penta acetic acid (DTPA), B-alaninediacetic acid (B-ADA) and their salts, homopolymers and copolymers of poly-carboxylic acids and their partially or completely neutralized salts, monomeric polycarboxylic acids and hydroxycarboxylic acids and their salts in the range of 0.5% to 50% by weight; sulfonated/carboxylated polymers in the range of about 0.1 % to about 50% by weight to provide dimensional stability; drying aids in the range of about 0.1 % to about 10% by weight {e.g., polyesters, especially anionic polyesters, optionally together with further monomers with 3 to 6 functionalities - typically acid, alcohol or ester functionalities which are conducive to polycondensation, polycarbonate-, polyurethane- and/or polyurea-polyorganosiloxane compounds or precursor compounds, thereof, particularly of the reactive cyclic carbonate and urea type); silicates in the range from about 1 % to about 20% by weight (including sodium or potassium silicates for example sodium disilicate, sodium meta-silicate and crystalline phyllosilicates) ; inorganic bleach (e.g., perhydrate salts such as perborate, percarbonate, perphosphate, persulfate and persilicate salts) and organic bleach (e.g., organic peroxyacids, including diacyl and tetraacylperoxides, especially diperoxydodecanedioc acid, diperoxytetradecanedioc acid, and diperoxyhexadecanedioc acid) ; bleach activators (i.e., organic peracid precursors in
the range from about 0.1 % to about 10% by weight); bleach catalysts {e.g., manganese triazacyclononane and related complexes, Co, Cu, Mn, and Fe bispyridylamine and related complexes, and pentamine acetate cobalt(l l l) and related complexes); metal care agents in the range from about 0.1 % to 5% by weight {e.g., benzatriazoles, metal salts and complexes, and/or silicates); enzymes in the range from about 0.01 to 5.0 mg of active enzyme per gram of automatic dishwashing detergent composition {e.g., proteases, amylases, lipases, cellulases, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferase, perhydrolase, arylesterase, and mixtures thereof); and enzyme stabilizer components (e.g., oligosaccharides, polysaccharides, and inorganic divalent metal salts).
The present maltogenic amylase polypeptide may be incorporated at a concentration conventionally employed in detergents. It is at present contemplated that, in the detergent composition, the enzyme may be added in an amount corresponding to 0.00001 -1 .0 mg (calculated as pure enzyme protein) of amylase polypeptide per liter of wash liquor.
The detergent composition may also contain other conventional detergent ingredients, e.g., deflocculant material, filler material, foam depressors, anti-corrosion agents, soil-suspending agents, sequestering agents, anti-soil redeposition agents, dehydrating agents, dyes, bactericides, fluorescers, thickeners, and perfumes.
The detergent composition may be formulated as a hand (manual) or machine (automatic) laundry detergent composition, including a laundry additive composition suitable for pre-treatment of stained fabrics and a rinse added fabric softener composition, or be formulated as a detergent composition for use in general household hard surface cleaning operations, or be formulated for manual or automatic dishwashing operations.
Any of the cleaning compositions described, herein, may include any number of additional enzymes. In general the enzyme(s) should be compatible with the selected detergent, {e.g., with respect to pH-optimum, compatibility with other enzymatic and
non-enzymatic ingredients, and the like), and the enzyme(s) should be present in effective amounts. The following enzymes are provided as examples.
Proteases: Suitable proteases include those of animal, vegetable or microbial origin. Chemically modified or protein engineered mutants are included, as well as naturally processed proteins. The protease may be a serine protease or a metalloprotease, an alkaline microbial protease, a trypsin-like protease, or a
chymotrypsin-like protease. Examples of alkaline proteases are subtilisins, especially those derived from Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin 309, subtilisin 147, and subtilisin 168 {see, e.g., WO 89/06279). Examples of trypsin-like proteases are trypsin (e.g., of porcine or bovine origin), and Fusarium proteases {see, e.g., WO 89/06270 and WO 94/25583). Examples of useful proteases also include but are not limited to the variants described in WO 92/1 9729, WO 98/2011 5, WO 98/20116, and WO 98/34946. Commercially available protease enzymes include but are not limited to: ALCALASE®, SAVINASE®, PRIMASE™, DURALASE™, ESPERASE®, KANNASE™, and BLAZE™ (Novo Nordisk A/S and Novozymes A/S); MAXATASE®, MAXACAL™, MAXAPEM™, PROPERASE®, PURAFECT®, PURAFECT OXP™, FN2™, and FN3™ (Danisco US Inc.). Other exemplary proteases include NprE from Bacillus amyloliquifaciens and ASP from Cellulomonas sp. strain 69B4.
Lipases: Suitable lipases include those of bacterial or fungal origin. Chemically modified, proteolytically modified, or protein engineered mutants are included. Examples of useful lipases include but are not limited to lipases from Humicola (synonym Thermomyces), e.g., from H. lanuginosa ( T. lanuginosus) {see e.g., EP 258068 and EP 305216), from H. insolens {see e.g., WO 96/13580); a Pseudomonas lipase {e.g., from P. alcaligenes or P. pseudoalcaligenes; see, e.g., EP 218 272), P. cepacia {see e.g., EP 331 376), P. stutzeri {see e.g., GB 1 ,372,034), P. fluorescens, Pseudomonas sp. strain SD 705 {see e.g., WO 95/06720 and WO 96/27002), P. wisconsinensis {see e.g., WO 96/12012); a Bacillus lipase {e.g., from B. subtilis; see e.g., Dartois et al. Biochemica et Biophysica Acta, 11 31 : 253-360 (1993)), B. stearothermophilus {see e.g., JP
64/744992), or B. pumilus {see e.g., WO 91/16422). Additional lipase variants contemplated for use in the formulations include those described for example in: WO
92/05249, WO 94/01541 , WO 95/35381 , WO 96/00292, WO 95/30744, WO 94/25578, WO 95/14783, WO 95/2261 5, WO 97/04079, WO 97/07202, EP 407225, and EP 260105. Some commercially available lipase enzymes include LIPOLASE® and LI POLASE ULTRA™ (Novo Nordisk A/S and Novozymes A/S). Poly esterases: Suitable polyesterases can be included in the composition, such as those described in, for example, WO 01 /34899, WO 01 /14629, and US6933140.
Amylases: The compositions can be combined with other amylases, such as non-production enhanced amylase. These can include commercially available amylases, such as but not limited to STAINZYME®, NATALASE®, DURAMYL®, TERMAMYL®, FUNGAMYL® and BAN™ (Novo Nordisk A/S and Novozymes A/S) ; RAPIDASE®, POWERASE®, and PURASTAR® (from Danisco US Inc.).
Cellulases: Cellulases can be added to the compositions. Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed for example in U.S. Patent Nos. 4,435,307; 5,648,263; 5,691 ,178; 5,776,757; and WO 89/09259. Exemplary cellulases contemplated for use are those having color care benefit for the textile. Examples of such cellulases are cellulases described in for example EP 0495257, EP 0531372, WO 96/11262, WO 96/29397, and WO 98/08940. Other examples are cellulase variants, such as those described in WO 94/07998; WO 98/1 2307; WO 95/24471 ; PCT/DK98/00299; EP
531315; U.S. Patent Nos. 5,457,046; 5,686,593; and 5,763,254. Commercially available cellulases include CELLUZYME® and CAR EZYME® (Novo Nordisk A/S and Novozymes A/S); CLAZINASE® and PURADAX HA® (Danisco US Inc.); and
KAC-500(B)™(Kao Corporation).
Peroxidases/Oxidases: Suitable peroxidases/oxidases contemplated for use in the compositions include those of plant, bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful peroxidases
include peroxidases from Coprinus, e.g., from C. cinereus, and variants thereof as those described in WO 93/24618, WO 95/1 0602, and WO 98/1 5257. Commercially available peroxidases include for example GUARDZYME™(Novo Nordisk A/S and Novozymes A/S). The detergent composition can also comprise 2,6-p-D-fructan hydrolase, which is effective for removal/cleaning of biofilm present on household and/or industrial textile/laundry.
The detergent enzyme(s) may be included in a detergent composition by adding separate additives containing one or more enzymes, or by adding a combined additive comprising all of these enzymes. A detergent additive, i.e. a separate additive or a combined additive, can be formulated e.g., as a granulate, a liquid, a slurry, and the like. Exemplary detergent additive formulations include but are not limited to granulates, in particular non-dusting granulates, liquids, in particular stabilized liquids or slurries.
Non-dusting granulates may be produced, e.g., as disclosed in U.S. Patent Nos. 4,106,991 and 4,661 ,452 and may optionally be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxide) products {e.g.,
polyethyleneglycol, PEG) with mean molar weights of 1 ,000 to 20,000; ethoxylated nonylphenols having from 1 6 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in, for example, GB 1483591 . Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Protected enzymes may be prepared according to the method disclosed in EP 238,216.
The detergent composition may be in any convenient form, e.g., a bar, a tablet, a powder, a granule, a paste, or a liquid. A liquid detergent may be aqueous, typically containing up to about 70% water, and 0% to about 30% organic solvent. Compact detergent gels containing about 30% or less water are also contemplated. The
detergent composition can optionally comprise one or more surfactants, which may be non-ionic, including semi-polar and/or anionic and/or cationic and/or zwitterionic. The surfactants can be present in a wide range, from about 0.1 % to about 60% by weight.
When included therein the detergent will typically contain from about 1 % to about 40% of an anionic surfactant, such as linear alkylbenzenesulfonate,
a-olefinsulfonate, alkyl sulfate (fatty alcohol sulfate), alcohol ethoxysulfate, secondary alkanesulfonate, a-sulfo fatty acid methyl ester, alkyl- or alkenylsuccinic acid, or soap.
When included therein, the detergent will usually contain from about 0.2% to about 40% of a non-ionic surfactant such as alcohol ethoxylate, nonylphenol ethoxylate, alkylpolyglycoside, alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, polyhydroxy alkyl fatty acid amide, or N-acyl-N-alkyl derivatives of glucosamine ("glucamides").
The detergent may contain 0% to about 65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, carbonate, citrate, nitrilotriacetic acid, ethylenediaminetetraacetic acid (EDTA),
diethylenetriaminepentaacetic acid, alkyl- or alkenylsuccinic acid, soluble silicates or layered silicates (e.g.,SKS-6 from Hoechst).
The detergent may comprise one or more polymers. Exemplary polymers include carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (PVP), poly(ethylene glycol) (PEG), polyvinyl alcohol) (PVA), poly(vinylpyridine-N-oxide), poly(vinylimidazole), polycarboxylates e.g., polyacrylates, maleic/acrylic acid copolymers), and lauryl methacrylate/acrylic acid copolymers.
The enzyme(s) of the detergent composition may be stabilized using conventional stabilizing agents, e.g., as polyol [e.g., propylene glycol or glycerol), a sugar or sugar alcohol, lactic acid, boric acid, or a boric acid derivative {e.g., an aromatic borate ester), or a phenyl boronic acid derivative (e.g., 4-formylphenyl boronic acid). The composition may be formulated as described in WO 92/19709 and WO 92/19708.
It is contemplated that in the detergent compositions, in particular the maltogenic amylase of the present teachings, may be added in an amount corresponding
to about 0.01 to about 100 mg of enzyme protein per liter of wash liquor (e.g., about 0.05 to about 5.0 mg of enzyme protein per liter of wash liquor or 0.1 to about 1 .0 mg of enzyme protein per liter of wash liquor).
Methods of Assessing Amylase Activity in Detergent Compositions
Numerous a-amylase cleaning assays are known in the art, including swatch and micro-swatch assays.
Brewing Compositions
The present maltogenic amylase may be a component of a brewing composition used in a process of brewing, i.e., making a fermented malt beverage. Non-fermentable carbohydrates form the majority of the dissolved solids in the final beer. This residue remains because of the inability of malt amylases to hydrolyze the alpha-1 ,6-linkages of the starch. The non-fermentable carbohydrates contribute about 50 calories per 12 ounces of beer. An amylase, in combination with a glucoamylase and optionally a pullulanase and/or isoamylase, assist in converting the starch into dextrins and fermentable sugars, lowering the residual non-fermentable carbohydrates in the final beer.
The principal raw materials used in making these beverages are water, hops and malt. In addition, adjuncts such as common corn grits, refined corn grits, brewer's milled yeast, rice, sorghum, refined corn starch, barley, barley starch, dehusked barley, wheat, wheat starch, torrified cereal, cereal flakes, rye, oats, potato, tapioca, and syrups, such as corn syrup, sugar cane syrup, inverted sugar syrup, barley and/or wheat syrups, and the like may be used as a source of starch.
For a number of reasons, the malt, which is produced principally from selected varieties of barley, has the greatest effect on the overall character and quality of the beer. First, the malt is the primary flavoring agent in beer. Second, the malt provides the major portion of the fermentable sugar. Third, the malt provides the proteins, which will contribute to the body and foam character of the beer. Fourth, the malt provides the
necessary enzymatic activity during mashing. Hops also contribute significantly to beer quality, including flavoring. In particular, hops (or hops constituents) add desirable bittering substances to the beer. In addition, the hops act as protein precipitants, establish preservative agents and aid in foam formation and stabilization. Grains, such as barley, oats, wheat, as well as plant components, such as corn, hops, and rice, also are used for brewing, both in industry and for home brewing. The components used in brewing may be unmalted or may be malted, i.e., partially germinated, resulting in an increase in the levels of enzymes, including a-amylase. For successful brewing, adequate levels of a-amylase enzyme activity are necessary to ensure the appropriate levels of sugars for fermentation. An amylase, by itself or in combination with another a-amylase(s), accordingly may be added to the components used for brewing.
As used herein, the term "stock" means grains and plant components that are crushed or broken. For example, barley used in beer production is a grain that has been coarsely ground or crushed to yield a consistency appropriate for producing a mash for fermentation. As used herein, the term "stock" includes any of the aforementioned types of plants and grains in crushed or coarsely ground forms. The methods described herein may be used to determine maltogenic amylase activity levels in both flours and stock. Processes for making beer are well known in the art. See, e.g., Wolfgang
Kunze (2004) "Technology Brewing and Malting," Research and Teaching Institute of Brewing, Berlin (VLB), 3rd edition. Briefly, the process involves: (a) preparing a mash, (b) filtering the mash to prepare a wort, and (c) fermenting the wort to obtain a fermented beverage, such as beer. Typically, milled or crushed malt is mixed with water and held for a period of time under controlled temperatures to permit the enzymes present in the malt to convert the starch present in the malt into fermentable sugars. The mash is then transferred to a mash filter where the liquid is separated from the grain residue. This sweet liquid is called "wort," and the left over grain residue is called "spent grain." The mash is typically subjected to an extraction, which involves adding water to the mash in
order to recover the residual soluble extract from the spent grain. The wort is then boiled vigorously to sterilize the wort and help develop the color, flavor and odor. Hops are added at some point during the boiling. The wort is cooled and transferred to a fermentor. The wort is then contacted in a fermentor with yeast. The fermentor may be chilled to stop fermentation. The yeast flocculates and is removed. Finally, the beer is cooled and stored for a period of time, during which the beer clarifies and its flavor develops, and any material that might impair the appearance, flavor and shelf life of the beer settles out. The beer usually contains from about 2% to about 10% v/v alcohol, although beer with a higher alcohol content, e.g., 18% v/v, may be obtained. Prior to packaging, the beer is carbonated and, optionally, filtered and pasteurized.
The brewing composition comprising a maltogenic amylase, in combination with a glucoamylase and optionally a pullulanase and/or isoamylase, may be added to the mash of step (a) above, i.e., during the preparation of the mash. Alternatively, or in addition, the brewing composition may be added to the mash of step (b) above, i.e., during the filtration of the mash. Alternatively, or in addition, the brewing composition may be added to the wort of step (c) above, i.e., during the fermenting of the wort.
A fermented beverage, such as a beer, can be produced by one of the methods above. The fermented beverage can be a beer, such as full malted beer, beer brewed under the "Reinheitsgebot," ale, IPA, lager, bitter, Happoshu (second beer), third beer, dry beer, near beer, light beer, low alcohol beer, low calorie beer, porter, bock beer, stout, malt liquor, non-alcoholic beer, non-alcoholic malt liquor and the like, but also alternative cereal and malt beverages such as fruit flavored malt beverages, e.g., citrus flavored, such as lemon-, orange-, lime-, or berry-flavored malt beverages, liquor flavored malt beverages, e.g., vodka-, rum-, or tequila-flavored malt liquor, or coffee flavored malt beverages, such as caffeine-flavored malt liquor, and the like.
Reduction of Iodine-Positive Starch
The maltogenic amylases of the present teachings may reduce the
iodine-positive starch (I PS), when used in a method of liquefaction and/or
saccharification. One source of I PS is from amylose that escapes hydrolysis and/or from retrograded starch polymer. Starch retrogradation occurs spontaneously in a starch paste, or gel on aging, because of the tendency of starch molecules to bind to one another followed by an increase in crystallinity. Solutions of low concentration become increasingly cloudy due to the progressive association of starch molecules into larger articles. Spontaneous precipitation takes place and the precipitated starch appears to be reverting to its original condition of cold-water insolubility. Pastes of higher concentration on cooling set to a gel, which on aging becomes steadily firmer due to the increasing association of the starch molecules. This arises because of the strong tendency for hydrogen bond formation between hydroxy groups on adjacent starch molecules. See J. A. Radley, ed., STARCH AND ITS DERIVATIVES 1 94-201 (Chapman and Hall, London (1968)).
The presence of IPS in saccharide liquor negatively affects final product quality and represents a major issue with downstream processing. IPS plugs or slows filtration system, and fouls the carbon columns used for purification. When IPS reaches sufficiently high levels, it may leak through the carbon columns and decrease production efficiency. Additionally, it may results in hazy final product upon storage, which is unacceptable for final product quality. The amount of IPS can be reduced by isolating the saccharification tank and blending the contents back. IPS nevertheless will accumulate in carbon columns and filter systems, among other things. The use of the maltogenic amylases of the present teachings is expected to improve overall process performance by reducing the amount of IPS.
Examples
A putative novel amylase was identified from a metagenomic library constructed by conventional cloning techniques. Protein sequence analysis indicates that the amylase belongs to the glycosyl hydrolase family 13 (GH13), and shows less than 66% sequence identity to known proteins in the public NCBI database.
Following identification of the putative amylase in silico, the gene was cloned using conventional molecular biology PCR techniques and expressed in Bacillus subtilis. The plasmid cloning map is shown in Figure 9. Cloning and expression of AmyMG
The full gene sequence is SEQ ID NO:1 .
CACACCCCGACGACCCGGCAGGCCGATTACTACGGCACGCTGGAGCCGTTTGCG CGTGAAGCGGTGTACTTCGTGATGACCGATCGCTTCGTCAACGGCGACCCCGGCA ACGACCACCGCGACCAAGGCGGCGCCCTGGGCACGTTCGACATCCCGCTGCCGC CATGCAATGGCGTGTCCGGCAACATCGGCTACCTGGGTGGCGACTTCAAGGGCCT GGCCGATCATCTGGATTACATCCGCGAAATGGGCTTCACCGCGGTGTGGATCACG CCGATCGTGGACAATCCGGACCAGCGCTTCACTGGCGGCAGCGCACCAACCTGC GGCGGCATTCTGGCTGACCAGGGCAAAGCCGGCTATCACGGCTACTGGGGCGTG AATTTCTACCAAGTGGACGAGCACCTGCCCAGCCCAGGCATGGACTTCCGCGACC TGGCGGCGGCGATGCATCGCAAGGGCATGAAGCTGGTGCTGGACATCGTGGGGA ACCACGGCTCGCCGGCCTGGGGCATGGCCTTCGACCAGCCCAAGTTCGGCAAGA TCTACGACAAGGACGGCACGCTGATTGCCGATCACCAGAACCTGCCGCCGCAGCA GCTGGATCCCGAGCACAACCCGCTGCACCGCTTCTACAACACGGTCGGCCCGGT GGACGGGGCCAAGGGATCGATCTTCGACGGCAATCTGGCCCAGCTGTCGGATCT CAATGAACGCAACCCGGACGTGCTGGACTATCTGGTCGGGGCCTATCTGCAATGG ATCGACCAGGGTGCCGATGCGTTTCGCATCGACACCATCGCCTGGATGCCGGACA GCTTCTGGCAGGCCTTCACCACCCGCATCCGGGCAAAGCACCCCGGCTTTTTCAT GTTCGGCGAGGCCTTCGACTACGACGCCGCCAGGATTGCCACCCACACCCTGCC CGGCCACGGCGAAACCAGCGTGCTGGACTTCCCGATGAAACAGGCGATGGAAGA GGTCTTCGGGCGCAAGCAGGCCGGTTTTGAACGGATGATACCGGCGCTGCATCTG ACTGGCGGCCCGTATGCCAACCCCTACGAGCTGGCCACCTTCTACGACAATCACG ACATGCCGCGGCTGGATGCCAGCGATGAAGGCTTCATTGATGCACACAACTGGCT GTTCACCGCGCGTGGCATTCCGGTGGTCTATTACGGCTCGGAAATGGGCTTCATG CGCGGCCGACCCGAGCACGGCGGCAACCGCAACTACTTCGGCACCGAAGGCATT GCCGCCGCCAAGGCAAGCCCGATCCGGGCAGCGCTGACCCGCATTGCGCAGGT GCGTGCCGCTTCACCAGCGCTGCAGCgCGGGCTGCAACTCAATCTGGAGCTGCA AGGCAACCGCGCCGCGTTCTATCGGGTGTACCAGCACGACGGTGTGCACCAGATC GCGCTGGTCCTGCTCAACAAGGGCGACGCCCCGGAACACTTTGCCGTCCAGACG ATGCTGCAACCCGGCcGCTGGCATGACGCGATTGGCGGTGAGACGCTGACCATAC AGGCTGGCGAGGCGCTGCACGCCGAGGTTCCGGCGCATGGCGTGCGGGTCTTT CTGCTCGACGCCCAGGTGACTGAGCCGACGCTGGCGGCTGCGCTGGATGCCGC CATGGCCGATGCACGCCGGTCACGGTAA
The nucleic acid fragment encoding the native signal peptide is SEQ ID NO: 2.
ATGATCACCATGCCCCTGCGTTCTGCCCGCCTCGGCCTGAGTTTGCTTTGCGCGC TTGCCTCGACGGCCTGTGCA
The full protein sequence is SEQ ID NO:3.
HTPTTRQADYYGTLEPFAREAVYFVMTDRFVNGDPGNDHRDQGGALGTFDI PLPPCN GVSGNIGYLGGDFKGLADHLDYIREMGFTAVWITPIVDNPDQRFTGGSAPTCGGILAD QGKAGYHGYWGVNFYQVDEHLPSPGMDFRDLAAAMHRKGMKLVLDIVGNHGSPAW GMAFDQPKFGKIYDKDGTLIADHQNLPPQQLDPEHNPLHRFYNTVGPVDGAKGSIFD GNLAQLSDLNERNPDVLDYLVGAYLQWI DQGADAFRI DTIAWMPDSFWQAFTTRIRAK HPGFFMFGEAFDYDAARIATHTLPGHGETSVLDFPMKQAMEEVFGRKQAGFERMIPA LHLTGGPYANPYELATFYDNHDMPRLDASDEGFIDAHNWLFTARGIPVVYYGSEMGF MRGRPEHGGNRNYFGTEGIAAAKASPI RAALTRIAQVRAASPALQRGLQLNLELQGNR AAFYRVYQHDGVHQIALVLLNKGDAPEHFAVQTMLQPGRWHDAIGGETLTIQAGEALH AEVPAHGVRVFLLDAQVTEPTLAAALDAAMADARRSR
The native signal peptide is SEQ I D NO: 4.
MITMPLRSARLGLSLLCALASTACA
The mature gene sequence was identified by conventional procedures, PCR amplified, digested by BssHii/Xhol, and ligated into the p2JM plasmid digested with same restriction enzymes. As shown in the plasmid map of Figure 9, the AmyMG gene was put under the control of the aprE promoter. The aprE signal sequence was used to direct protein secretion. In addition, nucleotides coding for three additional amino acids (AGK) were placed between the aprE signal sequence and the mature AmyMG gene to facilitate secretion of the target protein. The resulting plasmid was used to transform competent Bacillus subtilis cells. Expression evaluation of AmyMG showed that the enzyme expresses very well in both strains.
The aprE signal nucleic acid sequence (underlined) + AGK nucleic acid sequence (italics) is SEQ ID NO: 5. qtqaqaaqcaaaaaattqtqqatcaqcttqttqtttqcqttaacqttaatctttacqatqqcqttcaqcaacatqaqcqcqca ggcagctggtaaa
The aprE signal amino acid sequence (underlined) + AGK amino acid sequence (italics) is SEQ ID NO: 6. MRSKKLWISLLFALALIFTMAFGSTSSAQA/4GK'
Product profile analysis and biochemical characterization of AmyMG
Preliminary product profile analysis showed that AmyMG exhibited DP2 as its major product, with the composition (%) higher than 90% (if one only considered the product profile from DP1 to DP7) (Figure 1 and Table 1 ).
Figure 1 shows the typical chromatograms of oligosaccharide product profile analysis of A) AmyMG (10 ppm) incubated with Maltodextrin (DE1 0) (0.5%, w/v) under pH 5.3 at 50°C for 2 h and B) the mixture of standard compounds of DP1 to DP 7 (0.01 25%, w/v). HPLC separation was done using an Agilent 1200 series HPLC system with an Aminex HPX-42A column (300 mm x 7.8 mm) at 85 °C. The sample (10 μΙ_) was subjected to the HPLC column and separated with an isocratic gradient of Milli-Q water as the mobile phase at a flow rate of 0.6 mL/min. The oligosaccharide products were detected using a refractive index detector.
Table 2. The oligosaccharide product compositions (%) of AmyMG
Ion exchange chromatography results confirmed that AmyMG showed maltogenic activity with maltose as its major product (Figure 2). The typical chromatograms of oligosaccharide product profile analysis of AmyMG (1 0 ppm) incubated with Maltodextrin (DE1 0) (1 5%, w/v) under pH 5.3 at 50°C for 24h. The separation was done using a Dionex ICS-5000 ion exchange chromatography with a CarboPac PA 200 column at 30 °C. The sample (25 μΙ_) was subjected to the column and separated with the gradient: 0-1 0 min, 50mM NaOH; 10-15 min, 50-1 OOmM NaOH ; 15-35min, 1 00mM NaOH, 0-200mM NaAc; 35-45min, 50mM NaOH, at a flow rate of 0.5 mL/min. The
oligosaccharide products were detected using a pulsed amperometric detector.
More detailed product profile analysis (Figure 3) with different incubation time points showed the peak area of DP2 increased while the peak area of DP10+ decreased with the extension of incubation time from 0 h up to 48 h, indicating that maltose is the major product of AmyMG by hydrolyzing Maltrin040. As shown, the typical
chromatograms of oligosaccharide product profile analysis of AmyMG (25 ppm) incubated with Maltrin040 (30%, w/v) under pH 5.3 at 50°C. HPLC separation was done using an Agilent 1 200 series HPLC system with an Aminex HPX-42A column (300 mm x 7.8 mm) at 85°C. The sample (10 μί) was subjected to the HPLC column and separated with an isocratic gradient of Milli-Q water as the mobile phase at a flow rate of 0.6 mL/min. The oligosaccharide products were detected using a refractive index detector.
In order to figure out the exact anomeric form of maltose produced from maltodextrin catalyzed by AmyMG, the real time NMR assay was done by analyzing the enzyme product at different reaction times (Figure 4). The results suggested that the product is alpha-maltose and AmyMG is a maltogenic alpha-amylase. As shown, 1 H-NMR of the product from Maltrin040 incubated with AmyMG. The reaction was done by incubating the mixture of 10 mg Maltrin040 that was dissolved into 1 mL of D20 and 33 ppm of AmyMG at 25 °C. The NMR assay was done by a Bruker NMR spectrometer that operated at 500MHz in D20.
AmyMG was tested for alpha-amylase characterization (Amylopectin/PAHBAH method), including dose dependent assay (Figure 5, a dose-dependant assay for
AmyMG using amylopectin as the substrate), pH (Figure 6, showing a normalized pH profile), temperature profile (Figure 7, showing a normalized temperature profile), and, a thermostability assay (Figure 8, showing the thermostability of AmyMG). The specific activity of this enzyme towards amylopectin is 218.4 U/mg, with optimum pH at 7 and optimum temperature at 63°C.
Thus, in some embodiments, the present teachings provide a polypeptide with maltogenic acitivity wherein at least 70%, at least 75%, at least 80%, at least 85%, at least 90%., or at least 95% of the maximum enzyme activity is present in the pH range of 6-8, as assessed by DP ° Units. Additionally, in some embodiments, the present teachings provide a polypeptide with maximum activity, as assessed by DP ° Units, at 60°C-70°C, or 62°C-64°C.
Additionally, in some embodiments, the present teachings provide a polypeptide that produces at least 80%, at least 85%, at least 90%, or at least 95% DP2 of the DP1 to DP7 products resulting from a hydrolysis reaction performed according to the procedures depicted in Figure 3.
Finally, in some embodiments, the polypeptides of the present teachings can be engineered to provide various improved properties. Such engineering efforts can be guided by the atomic coordinate structural information provided in Table 3.
Table 3
ATOM 1 N GLN A 8 114. , 737 81 , , 675 18. ,562 1. , 00 37. , 71 N
ATOM 2 CA GLN A 8 114. .425 83 , .092 18. .184 1. , 00 34. .91 C
ATOM 3 CB GLN A 8 115. , 667 83 , , 800 17. ,610 1. , 00 42. , 85 C
ATOM 4 CG GLN A 8 115. , 634 85, ,314 17. , 761 1. , 00 49. ,20 C
ATOM 5 CD GLN A 8 115. , 695 85, , 734 19. ,218 1. , 00 59. ,92 C
ATOM 6 OE1 GLN A 8 114. , 734 85 , .547 19. .968 1. , 00 58. .08 0
ATOM 7 NE2 GLN A 8 116. , 832 86 , , 289 19. ,633 1. , 00 55. ,51 N
ATOM 8 C GLN A 8 113. , 216 83 , , 263 17. ,240 1. , 00 26. , 15 C
ATOM 9 O GLN A 8 112. , 182 83 , , 767 17. ,675 1. , 00 24. ,80 O
ATOM 10 N ALA A 9 113. ,343 82 , .897 15. .960 1. , 00 21. .01 N
ATOM 11 CA ALA A 9 112. , 213 82 , , 903 15. , 027 1. , 00 18. ,94 C
ATOM 12 CB ALA A 9 112. , 613 82 , , 251 13. , 712 1. , 00 19. , 85 C
ATOM 13 C ALA A 9 Ill . , 041 82 , , 146 15. ,671 1. , 00 15. ,96 C
ATOM 14 O ALA A 9 Ill . , 263 81 , .146 16. .368 1. , 00 16. .65 0
ATOM 15 N ASP A 10 109. , 823 82 , , 651 15. , 494 1. , 00 13. ,87 N
ATOM 16 CA ASP A 10 108. , 612 81 , , 920 15. ,913 1. , 00 15. ,61 C
ATOM 17 CB ASP A 10 107. , 434 82 , , 870 16. , 119 1. , 00 19. , 46 C
ATOM 18 CG ASP A 10 107. , 469 83 , .592 17. .448 1. , 00 23. .75 c
ATOM 19 OD1 ASP A 10 107. , 906 83 , , 023 18. , 466 1. , 00 20. , 82 O
ATOM 20 OD2 ASP A 10 107. , 009 84, , 743 17. , 468 1. , 00 32. , 09 O
ATOM 21 C ASP A 10 108. , 159 80 , , 918 14. , 856 1. , 00 12. ,27 c
ATOM 22 O ASP A 10 107. , 934 81 , .294 13. .725 1. , 00 12. .47 O
ATOM 23 N TYR A 11 107. , 976 79 , , 664 15. ,244 1. , 00 11. , 78 N
ATOM 24 CA TYR A 11 107. , 418 78 , , 635 14. ,367 1. , 00 10. ,00 C
ATOM 25 CB TYR A 11 108. .508 77 , .669 13. .941 1. , 00 10. .62 c
ATOM 26 CG TYR A 11 109. , 637 78 , , 276 13. , 132 1. , 00 10. ,60 C
ATOM 27 CD1 TYR A 11 109. ,376 79 , , 006 11. ,986 1. , 00 12. ,14 C
ATOM 28 CE1 TYR A 11 110. ,393 79 , , 548 11. , 225 1. , 00 11. , 89 C
ATOM 29 CZ TYR A 11 Ill . , 713 79 , .360 11. .601 1. , 00 13. .20 c
ATOM 30 OH TYR A 11 112. , 721 79 , , 895 10. , 822 1. , 00 13. ,22 O
ATOM 31 CE2 TYR A 11 112. , 008 78 , , 630 12. , 736 1. , 00 12. , 02 C
ATOM 32 CD2 TYR A 11 110. , 962 78 , , 079 13. , 485 1. , 00 12. ,17 C
ATOM 33 C TYR A 11 106. , 294 77 , .907 15. .116 1. , 00 10. .63 c
ATOM 34 O TYR A 11 106. , 515 77 , , 293 16. , 167 1. , 00 10. , 75 O
ATOM 35 N TYR A 12 105. , 067 78 , , 040 14. ,612 1. , 00 10. , 78 N
ATOM 36 CA TYR A 12 103. , 913 77 , ,379 15. , 195 1. , 00 10. ,87 C
ATOM 37 CB TYR A 12 103. , 411 78 , .087 16. .449 1. , 00 11. .39 c
ATOM 38 CG TYR A 12 103. , 189 79 , , 570 16. , 322 1. , 00 12. , 09 C
ATOM 39 CD1 TYR A 12 102. , 050 80 , , 066 15. , 729 1. , 00 13. ,04 C
ATOM 40 CE1 TYR A 12 101. , 834 81 , , 424 15. ,603 1. , 00 13. ,98 C
ATOM 41 CZ TYR A 12 102. , 771 82 , .298 16. .120 1. , 00 14. .07 c
ATOM 42 OH TYR A 12 102. , 570 83 , , 635 16. , 006 1. , 00 19. ,59 O
ATOM 43 CE2 TYR A 12 103. , 906 81 , , 846 16. , 726 1. , 00 13. , 46 C
ATOM 44 CD2 TYR A 12 104., 110 80 ,.476 16..841 1., 00 14..72 C
ATOM 45 C TYR A 12 102. , 783 77 , .285 14. .178 1. , 00 10. .60 C
ATOM 46 O TYR A 12 102. , 615 78 , .167 13. .319 1. , 00 8. .70 O
ATOM 47 N GLY A 13 102. .040 76 , .190 14. .257 1. , 00 9. .04 N
ATOM 48 CA GLY A 13 100. , 830 76 , .045 13. .454 1. , 00 9. .97 C
ATOM 49 C GLY A 13 99. , 734 76 , .942 13. .991 1. , 00 9. .94 C
ATOM 50 O GLY A 13 99. , 001 77 , .567 13. .229 1. , 00 9. .84 O
ATOM 51 N THR A 14 99. , 632 76 , .993 15. .312 1. , 00 9. .70 N
ATOM 52 CA THR A 14 98. , 663 77 , .856 15. .980 1. , 00 10. .20 C
ATOM 53 CB THR A 14 97. , 245 77 , .241 15. .994 1. , 00 10. .01 C
ATOM 54 OG1 THR A 14 96. ,360 78 , .172 16. .611 1. , 00 10. .36 O
ATOM 55 CG2 THR A 14 97. , 189 75 , .925 16. .776 1. , 00 8. .48 c
ATOM 56 C THR A 14 99. , 067 78 , .090 17. .419 1. , 00 10. .24 C
ATOM 57 O THR A 14 99. , 709 77 , .220 18. .020 1. , 00 9. .49 O
ATOM 58 N LEU A 15 98. , 661 79 , .244 17. .964 1. , 00 10. .52 N
ATOM 59 CA LEU A 15 98. , 798 79 , .553 19. .390 1. , 00 11. .07 c
ATOM 60 CB LEU A 15 99. ,342 80 , .990 19. .605 1. , 00 12. .17 C
ATOM 61 CG LEU A 15 100. , 793 81 , .221 19. .135 1. , 00 14. .65 C
ATOM 62 CD1 LEU A 15 101. , 260 82 , .655 19. .445 1. , 00 13. .18 c
ATOM 63 CD2 LEU A 15 101. , 724 80 , .178 19. .758 1. , 00 13. .48 c
ATOM 64 C LEU A 15 97. , 459 79 , .397 20. .128 1. , 00 12. .49 C
ATOM 65 O LEU A 15 97. ,373 79 , .717 21. .299 1. , 00 12. .03 O
ATOM 66 N GLU A 16 96. , 411 78 , .891 19. .462 1. , 00 12. .24 N
ATOM 67 CA GLU A 16 95. , 139 78 , .627 20. .163 1. , 00 11. .45 c
ATOM 68 CB GLU A 16 94. , 117 78 , .040 19. .189 1. , 00 12. .06 C
ATOM 69 CG GLU A 16 92. , 920 77 , .366 19. .815 1. , 00 13. .55 C
ATOM 70 CD GLU A 16 92. , 156 78 , .277 20. .763 1. , 00 15. .28 c
ATOM 71 OE1 GLU A 16 91. , 845 79 , .402 20. .326 1. , 00 14. .11 O
ATOM 72 OE2 GLU A 16 91. , 851 77 , .844 21. .918 1. , 00 12. .98 O
ATOM 73 C GLU A 16 95. ,392 77 , .679 21. .343 1. , 00 10. .66 C
ATOM 74 O GLU A 16 95. , 931 76 , .586 21. .153 1. , 00 10. .99 O
ATOM 75 N PRO A 17 95. , 031 78 , .099 22. .573 1. , 00 12. .38 N
ATOM 76 CA PRO A 17 95. , 229 77 , .237 23. .730 1. , 00 11. .70 C
ATOM 77 CB PRO A 17 94. , 522 78 , .016 24. .857 1. , 00 15. .92 C
ATOM 78 CG PRO A 17 94. , 750 79 , .456 24. .485 1. , 00 15. .36 c
ATOM 79 CD PRO A 17 94. , 605 79 , .467 22. .979 1. , 00 13. .18 C
ATOM 80 C PRO A 17 94. , 617 75 , .842 23. .584 1. , 00 12. .59 C
ATOM 81 O PRO A 17 95. , 176 74 , .874 24. .084 1. , 00 10. .90 O
ATOM 82 N PHE A 18 93. , 473 75 , .723 22. .911 1. , 00 10. .98 N
ATOM 83 CA PHE A 18 92. , 771 74 , .431 22. .893 1. , 00 10. .65 C
ATOM 84 CB PHE A 18 91. ,300 74 , .592 22. .466 1. , 00 11. .32 C
ATOM 85 CG PHE A 18 90. ,399 73 , .608 23. .124 1. , 00 11. .42 c
ATOM 86 CD1 PHE A 18 89. , 862 73 , .884 24. .366 1. , 00 12. .54 c
ATOM 87 CE1 PHE A 18 89. , 031 72 , .960 24. .998 1. , 00 14. .32 C
ATOM 88 CZ PHE A 18 88. , 755 71 , .758 24. .381 1. , 00 12. .10 C
ATOM 89 CE2 PHE A 18 89 ,, 291 71 ,, 476 23., 144 1., 00 12.,87 C
ATOM 90 CD2 PHE A 18 90 , , 118 72 , ,399 22. , 524 1. , 00 11. ,38 C
ATOM 91 C PHE A 18 93 , , 467 73 , ,352 22. , 046 1. , 00 10. , 19 C
ATOM 92 O PHE A 18 93. .154 72 , .163 22. .153 1. , 00 9. .58 0
ATOM 93 N ALA A 19 94 , , 467 73 , , 744 21. , 266 1. , 00 11. ,62 N
ATOM 94 CA ALA A 19 95, , 189 72 , , 795 20. , 451 1. , 00 12. ,01 C
ATOM 95 CB ALA A 19 96 , , 140 73 , , 525 19. , 489 1. , 00 12. , 82 C
ATOM 96 C ALA A 19 95 , .939 71 , .753 21. .275 1. , 00 11. .44 c
ATOM 97 O ALA A 19 96 , , 264 70 , , 661 20. , 767 1. , 00 12. ,23 O
ATOM 98 N ARG A 20 96 , , 239 72 , , 063 22. , 524 1. , 00 10. ,20 N
ATOM 99 CA ARG A 20 96 , , 929 71 , , 113 23. ,389 1. , 00 12. , 13 C
ATOM 100 CB ARG A 20 97 , .459 71 , .771 24. .681 1. , 00 14. .90 c
ATOM 101 CG ARG A 20 96 , ,379 72 , , 227 25. ,683 1. , 00 22. ,99 C
ATOM 102 CD ARG A 20 96 , , 968 72 , , 828 26. ,976 1. , 00 26. ,81 C
ATOM 103 NE ARG A 20 97, , 421 71 , , 776 27. , 886 1. , 00 36. ,93 N
ATOM 104 CZ ARG A 20 96 , .644 71 , .115 28. .753 1. , 00 43. .46 c
ATOM 105 NH1 ARG A 20 95, ,347 71 , ,380 28. , 870 1. , 00 39. , 73 N
ATOM 106 NH2 ARG A 20 97, , 172 70 , , 172 29. , 519 1. , 00 44. ,22 N
ATOM 107 C ARG A 20 96 , , 045 69 , , 954 23. , 780 1. , 00 11. , 42 C
ATOM 108 O ARG A 20 96 , .552 68 , .960 24. .302 1. , 00 9. .84 O
ATOM 109 N GLU A 21 94 , , 730 70 , , 098 23. , 574 1. , 00 10. , 02 N
ATOM 110 CA GLU A 21 93 , , 772 69 , , 145 24. , 120 1. , 00 10. ,20 C
ATOM 111 CB GLU A 21 92 , , 403 69 , , 807 24. ,361 1. , 00 10. ,61 c
ATOM 112 CG GLU A 21 92 , .428 71 , .104 25. .167 1. , 00 11. .29 c
ATOM 113 CD GLU A 21 92 , , 854 70 , .952 26. ,607 1. , 00 13. ,95 C
ATOM 114 OE1 GLU A 21 92 , , 766 69 , .823 27. , 154 1. , 00 16. ,27 O
ATOM 115 OE2 GLU A 21 93 , .226 71 , .998 27. .215 1. , 00 14. .14 0
ATOM 116 C GLU A 21 93 , , 579 67 , .955 23. , 172 1. , 00 9. , 85 C
ATOM 117 O GLU A 21 93 , , 870 68 , .044 21. ,988 1. , 00 9. , 19 O
ATOM 118 N ALA A 22 93 , , 096 66 , .848 23. , 730 1. , 00 8. , 78 N
ATOM 119 CA ALA A 22 92 , .593 65 , .712 22. .987 1. , 00 8. .73 c
ATOM 120 CB ALA A 22 93 , , 543 64 , .520 23. , 099 1. , 00 7. , 76 C
ATOM 121 C ALA A 22 91 , , 244 65, .378 23. , 595 1. , 00 9. ,11 C
ATOM 122 O ALA A 22 91 , , 118 65, .264 24. , 825 1. , 00 8. ,96 O
ATOM 123 N VAL A 23 90 , .250 65 , .196 22. .726 1. , 00 9. .91 N
ATOM 124 CA VAL A 23 88 , , 816 65, .108 23. , 110 1. , 00 9. , 02 C
ATOM 125 CB VAL A 23 87, , 955 66 , .035 22. ,243 1. , 00 7. ,58 C
ATOM 126 CGI VAL A 23 86 , , 471 65, .954 22. , 657 1. , 00 8. ,23 C
ATOM 127 CG2 VAL A 23 88 , .484 67 , .474 22. .301 1. , 00 7. .05 c
ATOM 128 C VAL A 23 88 , , 236 63 , .700 22. ,987 1. , 00 9. ,21 C
ATOM 129 O VAL A 23 88 , ,382 63 , .056 21. ,958 1. , 00 9. ,50 O
ATOM 130 N TYR A 24 87, , 580 63 , .238 24. , 052 1. , 00 9. , 44 N
ATOM 131 CA TYR A 24 86 , .818 61 , .989 24. .058 1. , 00 8. .12 c
ATOM 132 CB TYR A 24 87, , 082 61 , .245 25. ,356 1. , 00 7. ,57 C
ATOM 133 CG TYR A 24 86 , , 537 59 , .849 25. , 446 1. , 00 8. ,57 C
ATOM 134 CD1 TYR A 24 87., 149 58 ,.797 24..783 1., 00 8..16 C
ATOM 135 CE1 TYR A 24 86. , 678 57 , .505 24. .909 1. , 00 8. .46 C
ATOM 136 CZ TYR A 24 85. , 583 57 , .244 25. .703 1. , 00 9. .22 C
ATOM 137 OH TYR A 24 85. .107 55 , .949 25. .829 1. , 00 9. .23 0
ATOM 138 CE2 TYR A 24 84. , 939 58 , .275 26. .352 1. , 00 8. .75 C
ATOM 139 CD2 TYR A 24 85. , 420 59 , .569 26. .231 1. , 00 7. .98 C
ATOM 140 C TYR A 24 85. ,353 62 , .400 23. .990 1. , 00 7. .92 C
ATOM 141 O TYR A 24 84. , 898 63 , .193 24. .803 1. , 00 8. .03 0
ATOM 142 N PHE A 25 84. , 641 61 , .900 22. .991 1. , 00 8. .21 N
ATOM 143 CA PHE A 25 83. , 280 62 , .320 22. .721 1. , 00 8. .18 C
ATOM 144 CB PHE A 25 83. , 144 62 , .689 21. .242 1. , 00 8. .40 C
ATOM 145 CG PHE A 25 81. , 709 62 , .743 20. .739 1. , 00 8. .66 c
ATOM 146 CD1 PHE A 25 80. , 696 63 , .328 21. .500 1. , 00 9. .58 C
ATOM 147 CE1 PHE A 25 79. ,383 63 , .358 21. .027 1. , 00 9. .05 C
ATOM 148 CZ PHE A 25 79. , 088 62 , .816 19. .801 1. , 00 8. .26 C
ATOM 149 CE2 PHE A 25 80. , 084 62 , .252 19. .045 1. , 00 8. .37 c
ATOM 150 CD2 PHE A 25 81. ,381 62 , .208 19. .518 1. , 00 8. .39 C
ATOM 151 C PHE A 25 82. ,311 61 , .213 23. .151 1. , 00 8. .63 C
ATOM 152 O PHE A 25 82. , 355 60 , .090 22. .636 1. , 00 8. .21 0
ATOM 153 N VAL A 26 81. , 459 61 , .555 24. .113 1. , 00 9. .31 N
ATOM 154 CA VAL A 26 80. , 425 60 , .653 24. .615 1. , 00 9. .72 C
ATOM 155 CB VAL A 26 80. , 466 60 , .446 26. .154 1. , 00 8. .23 C
ATOM 156 CGI VAL A 26 81. , 757 59 , .776 26. .598 1. , 00 7. .98 c
ATOM 157 CG2 VAL A 26 80. , 294 61 , .762 26. .914 1. , 00 7. .37 c
ATOM 158 C VAL A 26 79. , 036 61 , .152 24. .255 1. , 00 11. .35 C
ATOM 159 O VAL A 26 78. , 736 62 , .349 24. .348 1. , 00 10. .46 0
ATOM 160 N MET A 27 78. , 173 60 , .214 23. .856 1. , 00 11. .76 N
ATOM 161 CA MET A 27 76. , 753 60 , .442 23. .995 1. , 00 10. .64 C
ATOM 162 CB MET A 27 75. , 931 59 , .590 23. .020 1. , 00 10. .86 C
ATOM 163 CG MET A 27 76. , 252 59 , .893 21. .569 1. , 00 11. .29 C
ATOM 164 SD MET A 27 74. , 897 59 , .499 20. .447 1. , 00 15. .44 s
ATOM 165 CE MET A 27 73. , 641 60 , .680 20. .974 1. , 00 12. .17 C
ATOM 166 C MET A 27 76. , 471 60 , .071 25. .434 1. , 00 9. .42 C
ATOM 167 O MET A 27 76. , 507 58 , .884 25. .808 1. , 00 9. .40 0
ATOM 168 N THR A 28 76. , 180 61 , .084 26. .247 1. , 00 8. .45 N
ATOM 169 CA THR A 28 76. , 110 60 , .889 27. .690 1. , 00 7. .76 C
ATOM 170 CB THR A 28 75. , 717 62 , .188 28. .370 1. , 00 6. .89 C
ATOM 171 OG1 THR A 28 76. , 648 63 , .190 27. .968 1. , 00 7. .28 0
ATOM 172 CG2 THR A 28 75. , 745 62 , .030 29. .895 1. , 00 6. .66 c
ATOM 173 C THR A 28 75. , 150 59 , .759 28. .107 1. , 00 7. .56 C
ATOM 174 O THR A 28 75. , 475 58 , .929 28. .976 1. , 00 7. .08 O
ATOM 175 N ASP A 29 74. , 007 59 , .685 27. .433 1. , 00 7. .84 N
ATOM 176 CA ASP A 29 72. , 986 58 , .674 27. .754 1. , 00 7. .69 c
ATOM 177 CB ASP A 29 71. , 700 58 , .919 26. .974 1. , 00 8. .29 C
ATOM 178 CG ASP A 29 70. , 632 57 , .883 27. .301 1. , 00 10. .64 C
ATOM 179 OD1 ASP A 29 70 ,.103 57., 872 28..458 1., 00 12..48 O
ATOM 180 OD2 ASP A 29 70 , .353 57. , 039 26. .419 1. , 00 11. .07 O
ATOM 181 C ASP A 29 73 , .468 57. , 249 27. .485 1. , 00 7. .72 C
ATOM 182 O ASP A 29 72. .907 56. .278 28. .012 1. .00 7. .59 O
ATOM 183 N ARG A 30 74 , .480 57. , 125 26. .641 1. , 00 7. .20 N
ATOM 184 CA ARG A 30 74 , .952 55. , 837 26. .162 1. , 00 7. .50 C
ATOM 185 CB ARG A 30 74 , .920 55. , 807 24. .630 1. , 00 7. .56 C
ATOM 186 CG ARG A 30 73 , .550 56. , 253 24. .070 1. , 00 9. .17 c
ATOM 187 CD ARG A 30 72 , .466 55. , 295 24. .557 1. , 00 8. .86 C
ATOM 188 NE ARG A 30 71 , .119 55. , 514 24. .018 1. , 00 8. .92 N
ATOM 189 CZ ARG A 30 70 , .680 55. , 058 22. .842 1. , 00 9. .72 C
ATOM 190 NH1 ARG A 30 71 , .499 54. , 428 21. .991 1. , 00 9. .20 N
ATOM 191 NH2 ARG A 30 69 , .425 55. , 273 22. .493 1. , 00 9. .24 N
ATOM 192 C ARG A 30 76 , .346 55. , 501 26. .675 1. , 00 7. .67 C
ATOM 193 O ARG A 30 76 , .925 54. , 501 26. .243 1. , 00 8. .07 O
ATOM 194 N PHE A 31 76 , .872 56. , 288 27. .611 1. , 00 7. .18 N
ATOM 195 CA PHE A 31 78 , .207 55. , 982 28. .186 1. , 00 7. .41 C
ATOM 196 CB PHE A 31 79 , .034 57. , 257 28. .374 1. , 00 7. .88 C
ATOM 197 CG PHE A 31 80 , .440 57. , 012 28. .842 1. , 00 6. .80 c
ATOM 198 CD1 PHE A 31 81. .313 56. , 267 28. .073 1. , 00 6. .81 c
ATOM 199 CE1 PHE A 31 82 , .598 56. , 001 28. .500 1. , 00 7. .26 C
ATOM 200 CZ PHE A 31 83 , .041 56. , 504 29. .708 1. , 00 8. .10 C
ATOM 201 CE2 PHE A 31 82 , .179 57. , 260 30. .494 1. , 00 8. .20 c
ATOM 202 CD2 PHE A 31 80. .881 57. , 524 30. .041 1. , 00 6. .85 c
ATOM 203 C PHE A 31 78 , .088 55. , 141 29. .467 1. , 00 7. .98 C
ATOM 204 O PHE A 31 78 , .222 53. , 923 29. .398 1. , 00 8. .68 O
ATOM 205 N VAL A 32 77. .808 55. .739 30. .626 1. .00 8. .35 N
ATOM 206 CA VAL A 32 77 , .642 54. , 953 31. .861 1. , 00 7. .55 C
ATOM 207 CB VAL A 32 78 , .937 54. , 908 32. .728 1. , 00 7. .27 C
ATOM 208 CGI VAL A 32 78 , .729 54. , 076 34. .014 1. , 00 6. .36 C
ATOM 209 CG2 VAL A 32 80 , .103 54. , 351 31. .905 1. , 00 7. .01 c
ATOM 210 C VAL A 32 76 , .499 55. , 491 32. .714 1. , 00 8. .33 C
ATOM 211 O VAL A 32 76 , .424 56. , 699 33. .016 1. , 00 7. .21 O
ATOM 212 N ASN A 33 75 , .576 54. , 595 33. .054 1. , 00 8. .65 N
ATOM 213 CA ASN A 33 74 , .448 54. , 927 33. .919 1. , 00 8. .72 c
ATOM 214 CB ASN A 33 73 , .284 53. , 940 33. .699 1. , 00 9. .37 C
ATOM 215 CG ASN A 33 72 , .068 54. , 236 34. .577 1. , 00 10. .57 C
ATOM 216 OD1 ASN A 33 72 , .071 55. , 169 35. .402 1. , 00 11. .90 O
ATOM 217 ND2 ASN A 33 71 , .015 53. , 471 34. .380 1. , 00 8. .33 N
ATOM 218 C ASN A 33 74 , .994 54. , 804 35. .329 1. , 00 9. .69 C
ATOM 219 O ASN A 33 75 , .061 53. , 712 35. .860 1. , 00 7. .47 O
ATOM 220 N GLY A 34 75 , .395 55. , 918 35. .931 1. , 00 9. .95 N
ATOM 221 CA GLY A 34 75 , .913 55. , 884 37. .292 1. , 00 12. .12 c
ATOM 222 C GLY A 34 74 , .819 55. , 969 38. .343 1. , 00 12. .68 C
ATOM 223 O GLY A 34 75 , .054 55. , 654 39. .478 1. , 00 10. .41 O
ATOM 224 N ASP A 35 73 ,, 620 56 ,,380 37., 960 1., 00 11., 76 N
ATOM 225 CA ASP A 35 72 , , 556 56 , , 583 38. ,916 1. , 00 10. ,54 C
ATOM 226 CB ASP A 35 72 , , 474 58 , , 078 39. , 257 1. , 00 11. ,10 C
ATOM 227 CG ASP A 35 71. .409 58 , .396 40. .311 1. .00 14. .76 c
ATOM 228 OD1 ASP A 35 70 , , 843 57 , , 458 40. , 896 1. , 00 13. ,11 0
ATOM 229 OD2 ASP A 35 71 , , 154 59 , , 593 40. , 547 1. , 00 15. ,92 O
ATOM 230 C ASP A 35 71 , , 232 56 , , 072 38. ,323 1. , 00 10. ,65 C
ATOM 231 O ASP A 35 70 , .640 56 , .733 37. .500 1. , 00 8. .08 0
ATOM 232 N PRO A 36 70 , , 756 54 , , 890 38. , 748 1. , 00 12. , 71 N
ATOM 233 CA PRO A 36 69 , , 449 54 , , 454 38. ,202 1. , 00 12. ,84 C
ATOM 234 CB PRO A 36 69 , , 282 53 , , 022 38. , 723 1. , 00 16. , 77 C
ATOM 235 CG PRO A 36 70 , .570 52 , .639 39. .374 1. , 00 17. .55 c
ATOM 236 CD PRO A 36 71 , , 444 53 , , 846 39. ,530 1. , 00 14. , 77 C
ATOM 237 C PRO A 36 68 , , 282 55 , ,318 38. ,641 1. , 00 10. ,21 C
ATOM 238 O PRO A 36 67, , 231 55 , , 293 38. , 015 1. , 00 11. ,01 O
ATOM 239 N GLY A 37 68 , .469 56 , .107 39. .684 1. , 00 9. .71 N
ATOM 240 CA GLY A 37 67, ,371 56 , , 865 40. ,274 1. , 00 9. , 82 C
ATOM 241 C GLY A 37 66 , , 935 58 , , 041 39. , 421 1. , 00 10. ,34 C
ATOM 242 O GLY A 37 65, , 861 58 , , 577 39. ,623 1. , 00 9. ,92 0
ATOM 243 N ASN A 38 67 , .738 58 , .444 38. .436 1. , 00 10. .06 N
ATOM 244 CA ASN A 38 67, , 296 59 , , 500 37. ,503 1. , 00 9. ,39 C
ATOM 245 CB ASN A 38 68 , ,383 60 , , 581 37. ,382 1. , 00 8. ,58 C
ATOM 246 CG ASN A 38 69 , , 672 60 , , 042 36. , 769 1. , 00 8. , 41 c
ATOM 247 OD1 ASN A 38 69 , .678 58 , .989 36. .141 1. , 00 8. .21 0
ATOM 248 ND2 ASN A 38 70 , , 773 60 , , 744 36. ,986 1. , 00 8. ,14 N
ATOM 249 C ASN A 38 66 , , 887 58 , , 982 36. , 090 1. , 00 10. , 78 C
ATOM 250 O ASN A 38 66 , .684 59 , .784 35. .143 1. .00 9. .29 0
ATOM 251 N ASP A 39 66 , , 742 57 , , 667 35. ,940 1. , 00 9. ,62 N
ATOM 252 CA ASP A 39 66 , , 450 57 , , 084 34. ,619 1. , 00 9. ,27 C
ATOM 253 CB ASP A 39 66 , , 578 55 , , 568 34. ,681 1. , 00 10. , 46 C
ATOM 254 CG ASP A 39 68 , .017 55 , .098 34. .786 1. , 00 10. .85 c
ATOM 255 OD1 ASP A 39 68 , , 998 55 , , 898 34. , 764 1. , 00 10. ,52 O
ATOM 256 OD2 ASP A 39 68 , , 157 53 , , 880 34. , 883 1. , 00 12. ,39 O
ATOM 257 C ASP A 39 65, , 060 57 , , 445 34. , 072 1. , 00 9. ,97 C
ATOM 258 O ASP A 39 64 , .848 57 , .441 32. .873 1. , 00 8. .89 0
ATOM 259 N HIS A 40 64 , , 102 57 , , 714 34. ,963 1. , 00 11. ,07 N
ATOM 260 CA HIS A 40 62 , , 726 58 , , 040 34. ,569 1. , 00 9. , 48 C
ATOM 261 CB HIS A 40 62 , , 674 59 , , 424 33. ,915 1. , 00 9. ,93 C
ATOM 262 CG HIS A 40 62 , .912 60 , .562 34. .887 1. , 00 10. .71 c
ATOM 263 ND1 HIS A 40 64 , , 149 60 , , 898 35. ,329 1. , 00 10. ,36 N
ATOM 264 CE1 HIS A 40 64, , 046 61 , , 924 36. ,211 1. , 00 9. , 83 C
ATOM 265 NE2 HIS A 40 62 , , 757 62 , , 248 36. , 321 1. , 00 11. ,22 N
ATOM 266 CD2 HIS A 40 62 , .023 61 , .440 35. .508 1. , 00 10. .32 C
ATOM 267 C HIS A 40 62 , , 167 56 , , 971 33. ,674 1. , 00 10. ,33 C
ATOM 268 O HIS A 40 61 , , 626 57 , , 254 32. ,609 1. , 00 8. ,28 O
ATOM 269 N ARG A 41 62., 288 55., 710 34..093 1., 00 10..34 N
ATOM 270 CA ARG A 41 61. , 934 54. , 575 33. .212 1. , 00 12. .39 C
ATOM 271 CB ARG A 41 62. ,300 53. , 217 33. .861 1. , 00 16. .40 C
ATOM 272 CG ARG A 41 63. .811 53. , 020 33. .956 1. , 00 23. .60 c
ATOM 273 CD ARG A 41 64. , 263 51. , 753 34. .702 1. , 00 25. .64 C
ATOM 274 NE ARG A 41 65. , 733 51. , 754 34. .724 1. , 00 30. .83 N
ATOM 275 CZ ARG A 41 66. , 530 50. , 916 34. .052 1. , 00 39. .84 C
ATOM 276 NH1 ARG A 41 66. , 027 49. , 918 33. .323 1. , 00 35. .55 N
ATOM 277 NH2 ARG A 41 67. , 858 51. , 068 34. .119 1. , 00 31. .93 N
ATOM 278 C ARG A 41 60. , 477 54. , 561 32. .763 1. , 00 11. .82 C
ATOM 279 O ARG A 41 60. , 201 54. , 121 31. .680 1. , 00 13. .51 O
ATOM 280 N ASP A 42 59. , 542 55. , 067 33. .555 1. , 00 11. .44 N
ATOM 281 CA ASP A 42 58. , 147 55. , 034 33. .123 1. , 00 13. .15 C
ATOM 282 CB ASP A 42 57. , 222 54. , 616 34. .279 1. , 00 17. .56 C
ATOM 283 CG ASP A 42 57. , 116 55. , 666 35. .367 1. , 00 24. .61 C
ATOM 284 OD1 ASP A 42 57. , 542 56. , 815 35. .146 1. , 00 34. .56 0
ATOM 285 OD2 ASP A 42 56. , 594 55. , 351 36. .461 1. , 00 34. .16 O
ATOM 286 C ASP A 42 57. , 673 56. , 351 32. .473 1. , 00 11. .61 C
ATOM 287 O ASP A 42 56. , 514 56. , 487 32. .134 1. , 00 10. .98 O
ATOM 288 N GLN A 43 58. , 574 57. ,301 32. .280 1. , 00 11. .71 N
ATOM 289 CA GLN A 43 58. , 196 58. , 624 31. .763 1. , 00 11. .62 C
ATOM 290 CB GLN A 43 59. , 440 59. , 508 31. .675 1. , 00 12. .35 C
ATOM 291 CG GLN A 43 59. , 251 60. , 838 30. .952 1. , 00 11. .65 c
ATOM 292 CD GLN A 43 60. , 564 61. , 586 30. .802 1. , 00 8. .67 c
ATOM 293 OE1 GLN A 43 61. ,348 61. ,308 29. .880 1. , 00 8. .21 O
ATOM 294 NE2 GLN A 43 60. , 802 62. , 545 31. .689 1. , 00 6. .92 N
ATOM 295 C GLN A 43 57. , 516 58. , 486 30. .397 1. , 00 12. .63 c
ATOM 296 O GLN A 43 58. , 061 57. , 837 29. .496 1. , 00 10. .97 O
ATOM 297 N GLY A 44 56. ,318 59. , 075 30. .268 1. , 00 12. .17 N
ATOM 298 CA GLY A 44 55. , 587 59. , 090 29. .005 1. , 00 11. .64 C
ATOM 299 C GLY A 44 54. , 684 57. , 890 28. .717 1. , 00 12. .56 c
ATOM 300 O GLY A 44 53. , 997 57. , 886 27. .708 1. , 00 11. .13 O
ATOM 301 N GLY A 45 54. , 715 56. , 854 29. .551 1. , 00 12. .20 N
ATOM 302 CA GLY A 45 53. , 802 55. , 703 29. .382 1. , 00 13. .62 C
ATOM 303 C GLY A 45 54. , 015 54. , 996 28. .038 1. , 00 14. .01 c
ATOM 304 O GLY A 45 55. , 151 54. , 639 27. .681 1. , 00 12. .57 O
ATOM 305 N ALA A 46 52. , 926 54. , 811 27. .288 1. , 00 11. .00 N
ATOM 306 CA ALA A 46 52. , 977 54. , 192 25. .952 1. , 00 11. .74 C
ATOM 307 CB ALA A 46 51. , 547 53. , 981 25. .439 1. , 00 10. .90 c
ATOM 308 C ALA A 46 53. , 770 55. , 097 24. .988 1. , 00 11. .26 C
ATOM 309 O ALA A 46 54. ,331 54. , 640 24. .005 1. , 00 11. .81 O
ATOM 310 N LEU A 47 53. , 824 56. ,392 25. .272 1. , 00 10. .12 N
ATOM 311 CA LEU A 47 54. , 630 57. ,313 24. .452 1. , 00 12. .21 c
ATOM 312 CB LEU A 47 53. , 886 58. , 641 24. .261 1. , 00 12. .67 C
ATOM 313 CG LEU A 47 52. , 528 58. , 507 23. .558 1. , 00 13. .74 C
ATOM 314 CD1 LEU A 47 51., 905 59., 883 23..411 1., 00 13..95 C
ATOM 315 CD2 LEU A 47 52. , 682 57. , 807 22. .212 1. , 00 12. .69 C
ATOM 316 C LEU A 47 55. , 969 57. , 543 25. .174 1. , 00 10. .46 C
ATOM 317 O LEU A 47 56. .521 58. , 632 25. .164 1. , 00 11. .30 0
ATOM 318 N GLY A 48 56. , 486 56. , 483 25. .768 1. , 00 9. .33 N
ATOM 319 CA GLY A 48 57. , 547 56. , 588 26. .747 1. , 00 11. .14 C
ATOM 320 C GLY A 48 58. , 881 56. , 922 26. .102 1. , 00 10. .67 c
ATOM 321 O GLY A 48 59. ,114 56. , 625 24. .932 1. , 00 11. .69 0
ATOM 322 N THR A 49 59. , 762 57. , 496 26. .897 1. , 00 10. .50 N
ATOM 323 CA THR A 49 61. , 084 57. , 895 26. .435 1. , 00 9. .90 C
ATOM 324 CB THR A 49 61. , 455 59. , 229 27. .095 1. , 00 10. .00 c
ATOM 325 OG1 THR A 49 61. , 221 59. , 107 28. .511 1. , 00 8. .33 0
ATOM 326 CG2 THR A 49 60. , 615 60. ,392 26. .512 1. , 00 8. .41 c
ATOM 327 C THR A 49 62. , 195 56. , 905 26. .755 1. , 00 9. .60 C
ATOM 328 O THR A 49 63. ,308 57. , 005 26. .175 1. , 00 8. .74 O
ATOM 329 N PHE A 50 61. , 928 55. , 943 27. .648 1. , 00 10. .00 N
ATOM 330 CA PHE A 50 62. , 978 55. , 023 28. .067 1. , 00 9. .04 C
ATOM 331 CB PHE A 50 62. , 885 54. , 707 29. .539 1. , 00 9. .49 C
ATOM 332 CG PHE A 50 64. , 168 54. , 168 30. .125 1. , 00 9. .26 c
ATOM 333 CD1 PHE A 50 65. ,170 55. , 033 30. .544 1. , 00 9. .95 c
ATOM 334 CE1 PHE A 50 66. ,358 54. , 539 31. .079 1. , 00 11. .12 C
ATOM 335 CZ PHE A 50 66. , 560 53. , 171 31. .188 1. , 00 11. .10 C
ATOM 336 CE2 PHE A 50 65. , 555 52. , 291 30. .788 1. , 00 10. .93 c
ATOM 337 CD2 PHE A 50 64. , 365 52. , 799 30. .262 1. , 00 9. .67 c
ATOM 338 C PHE A 50 62. , 988 53. , 733 27. .267 1. , 00 10. .78 C
ATOM 339 O PHE A 50 61. , 946 53. , 116 27. .105 1. , 00 11. .80 O
ATOM 340 N ASP A 51 64. , 188 53. ,345 26. .794 1. , 00 11. .49 N
ATOM 341 CA ASP A 51 64. , 482 52. , 029 26. .166 1. , 00 9. .54 c
ATOM 342 CB ASP A 51 64. , 505 50. , 962 27. .220 1. , 00 10. .45 C
ATOM 343 CG ASP A 51 65. , 172 49. , 689 26. .753 1. , 00 11. .12 c
ATOM 344 OD1 ASP A 51 65. , 426 49. , 481 25. .545 1. , 00 9. .80 0
ATOM 345 OD2 ASP A 51 65. , 442 48. , 873 27. .633 1. , 00 15. .31 O
ATOM 346 C ASP A 51 63. , 502 51. , 637 25. .083 1. , 00 9. .86 C
ATOM 347 O ASP A 51 62. , 473 50. , 997 25. .354 1. , 00 8. .71 O
ATOM 348 N ILE A 52 63. , 813 52. , 053 23. .857 1. , 00 9. .39 N
ATOM 349 CA ILE A 52 62. , 968 51. , 829 22. .697 1. , 00 8. .39 c
ATOM 350 CB ILE A 52 62. , 484 53. , 158 22. .073 1. , 00 8. .12 C
ATOM 351 CGI ILE A 52 61. , 810 54. , 041 23. .133 1. , 00 7. .93 c
ATOM 352 CD1 ILE A 52 61. , 567 55. , 486 22. .695 1. , 00 7. .27 c
ATOM 353 CG2 ILE A 52 61. , 508 52. , 888 20. .905 1. , 00 6. .95 C
ATOM 354 C ILE A 52 63. , 794 51. , 030 21. .687 1. , 00 9. .20 C
ATOM 355 O ILE A 52 64. , 460 51. , 606 20. .843 1. , 00 9. .82 0
ATOM 356 N PRO A 53 63. , 784 49. , 695 21. .798 1. , 00 10. .75 N
ATOM 357 CA PRO A 53 64. , 610 48. , 880 20. .916 1. , 00 10. .80 C
ATOM 358 CB PRO A 53 64. , 512 47. , 459 21. .507 1. , 00 12. .29 C
ATOM 359 CG PRO A 53 63., 856 47 ,.608 22..821 1., 00 13..76 C
ATOM 360 CD PRO A 53 63. , 093 48 , .894 22. .826 1. , 00 12. .07 C
ATOM 361 C PRO A 53 64. , 082 48 , .862 19. .505 1. , 00 11. .93 C
ATOM 362 O PRO A 53 62. .878 48 , .866 19. .303 1. , 00 11. .09 O
ATOM 363 N LEU A 54 64. , 994 48 , .848 18. .547 1. , 00 12. .47 N
ATOM 364 CA LEU A 54 64. , 671 48 , .619 17. .170 1. , 00 12. .45 C
ATOM 365 CB LEU A 54 65. , 573 49 , .462 16. .269 1. , 00 12. .54 C
ATOM 366 CG LEU A 54 65. , 460 50 , .972 16. .440 1. , 00 13. .96 c
ATOM 367 CD1 LEU A 54 66. , 534 51 , .671 15. .601 1. , 00 14. .06 C
ATOM 368 CD2 LEU A 54 64. , 060 51 , .454 16. .088 1. , 00 13. .08 C
ATOM 369 C LEU A 54 64. , 845 47 , .136 16. .800 1. , 00 12. .65 C
ATOM 370 O LEU A 54 65. ,644 46 , .391 17. .425 1. , 00 11. .14 0
ATOM 371 N PRO A 55 64. , 127 46 , .705 15. .759 1. , 00 12. .63 N
ATOM 372 CA PRO A 55 64. , 404 45 , .388 15. .169 1. , 00 14. .57 C
ATOM 373 CB PRO A 55 63. , 486 45 , .354 13. .942 1. , 00 15. .45 C
ATOM 374 CG PRO A 55 62. ,385 46 , .338 14. .259 1. , 00 14. .72 c
ATOM 375 CD PRO A 55 63. , 024 47 , .410 15. .082 1. , 00 12. .19 C
ATOM 376 C PRO A 55 65. , 885 45 , .260 14. .757 1. , 00 14. .76 C
ATOM 377 O PRO A 55 66. , 444 46 , .208 14. .243 1. , 00 12. .76 O
ATOM 378 N PRO A 56 66. , 517 44 , .102 15. .003 1. , 00 15. .11 N
ATOM 379 CA PRO A 56 67. , 954 44 , .066 14. .773 1. , 00 16. .52 C
ATOM 380 CB PRO A 56 68. ,372 42 , .661 15. .226 1. , 00 17. .32 C
ATOM 381 CG PRO A 56 67. , 112 41 , .852 15. .270 1. , 00 18. .80 c
ATOM 382 CD PRO A 56 66. , 007 42 , .830 15. .546 1. , 00 16. .27 c
ATOM 383 C PRO A 56 68. ,319 44 , .262 13. .325 1. , 00 17. .80 C
ATOM 384 O PRO A 56 67. , 532 43 , .962 12. .439 1. , 00 14. .03 O
ATOM 385 N CYS A 57 69. , 523 44 , .766 13. .115 1. , 00 19. .16 N
ATOM 386 CA CYS A 57 70. , 044 45 , .023 11. .791 1. , 00 22. .01 C
ATOM 387 CB CYS A 57 70. , 167 46 , .535 11. .580 1. , 00 34. .43 C
ATOM 388 SG CYS A 57 71. , 206 46 , .936 10. .171 1. , 00 49. .84 S
ATOM 389 C CYS A 57 71. , 403 44 , .343 11. .723 1. , 00 17. .97 c
ATOM 390 O CYS A 57 72. , 285 44 , .641 12. .518 1. , 00 13. .58 O
ATOM 391 N ASN A 58 71. , 550 43 , .373 10. .827 1. , 00 17. .43 N
ATOM 392 CA ASN A 58 72. , 755 42 , .529 10. .790 1. , 00 18. .31 C
ATOM 393 CB ASN A 58 73. , 898 43 , .266 10. .086 1. , 00 22. .78 c
ATOM 394 CG ASN A 58 73. , 557 43 , .599 8. .649 1. , 00 25. .97 C
ATOM 395 OD1 ASN A 58 73. , 669 44 , .746 8. .215 1. , 00 30. .21 O
ATOM 396 ND2 ASN A 58 73. , 099 42 , .603 7. .918 1. , 00 26. .44 N
ATOM 397 C ASN A 58 73. , 215 42 , .063 12. .158 1. , 00 16. .62 c
ATOM 398 O ASN A 58 74. ,391 42 , .180 12. .515 1. , 00 13. .20 O
ATOM 399 N GLY A 59 72. , 272 41 , .559 12. .939 1. , 00 16. .73 N
ATOM 400 CA GLY A 59 72. , 592 40 , .921 14. .213 1. , 00 16. .76 C
ATOM 401 C GLY A 59 72. , 719 41 , .825 15. .429 1. , 00 15. .94 c
ATOM 402 O GLY A 59 72. , 911 41 , .331 16. .529 1. , 00 15. .63 O
ATOM 403 N VAL A 60 72. , 580 43 , .136 15. .258 1. , 00 15. .07 N
ATOM 404 CA VAL A 60 72., 674 44., 055 16..381 1., 00 14..25 C
ATOM 405 CB VAL A 60 73. , 967 44. , 916 16. .310 1. , 00 16. .22 C
ATOM 406 CGI VAL A 60 73. , 908 46. , 018 17. .356 1. , 00 14. .41 C
ATOM 407 CG2 VAL A 60 75. .210 44. , 056 16. .532 1. , 00 13. .03 c
ATOM 408 C VAL A 60 71. , 445 44. , 963 16. .403 1. , 00 14. .55 C
ATOM 409 O VAL A 60 71. , 011 45. , 468 15. .372 1. , 00 15. .71 O
ATOM 410 N SER A 61 70. , 875 45. , 149 17. .580 1. , 00 13. .04 N
ATOM 411 CA SER A 61 69. , 716 46. , 022 17. .760 1. , 00 13. .92 c
ATOM 412 CB SER A 61 68. , 715 45. ,358 18. .735 1. , 00 16. .79 C
ATOM 413 OG SER A 61 67. , 721 46. , 284 19. .166 1. , 00 22. .16 O
ATOM 414 C SER A 61 70. , 151 47. ,382 18. .292 1. , 00 12. .07 C
ATOM 415 O SER A 61 70. , 854 47. , 471 19. .298 1. , 00 13. .14 0
ATOM 416 N GLY A 62 69. , 776 48. , 442 17. .593 1. , 00 11. .47 N
ATOM 417 CA GLY A 62 69. , 943 49. , 791 18. .112 1. , 00 11. .67 C
ATOM 418 C GLY A 62 68. , 742 50. , 194 18. .946 1. , 00 10. .60 C
ATOM 419 O GLY A 62 67. ,814 49. , 418 19. .111 1. , 00 9. .27 0
ATOM 420 N ASN A 63 68. , 758 51. , 426 19. .444 1. , 00 9. .89 N
ATOM 421 CA ASN A 63 67. , 763 51. , 889 20. .416 1. , 00 10. .25 C
ATOM 422 CB ASN A 63 68. , 261 51. , 617 21. .847 1. , 00 9. .76 C
ATOM 423 CG ASN A 63 67. , 137 51. , 549 22. .862 1. , 00 9. .92 c
ATOM 424 OD1 ASN A 63 66. , 575 52. , 579 23. .252 1. , 00 8. .35 O
ATOM 425 ND2 ASN A 63 66. , 820 50. ,336 23. .321 1. , 00 9. .25 N
ATOM 426 C ASN A 63 67. , 569 53. ,377 20. .166 1. , 00 9. .97 c
ATOM 427 O ASN A 63 68. , 551 54. , 092 19. .875 1. , 00 10. .00 O
ATOM 428 N ILE A 64 66. ,319 53. , 827 20. .236 1. , 00 8. .58 N
ATOM 429 CA ILE A 64 65. , 974 55. , 225 20. .019 1. , 00 8. .85 C
ATOM 430 CB ILE A 64 65. , 096 55. , 422 18. .747 1. , 00 9. .33 c
ATOM 431 CGI ILE A 64 63. , 729 54. , 746 18. .825 1. , 00 10. .39 C
ATOM 432 CD1 ILE A 64 62. , 865 54. , 975 17. .570 1. , 00 9. .04 C
ATOM 433 CG2 ILE A 64 65. , 862 54. , 919 17. .522 1. , 00 8. .98 C
ATOM 434 C ILE A 64 65. , 391 55. , 904 21. .267 1. , 00 9. .04 c
ATOM 435 O ILE A 64 64. , 819 56. , 989 21. .184 1. , 00 10. .11 O
ATOM 436 N GLY A 65 65. , 573 55. , 292 22. .432 1. , 00 8. .54 N
ATOM 437 CA GLY A 65 65. , 170 55. , 915 23. .699 1. , 00 7. .46 C
ATOM 438 C GLY A 65 66. , 325 56. , 136 24. .643 1. , 00 7. .13 c
ATOM 439 O GLY A 65 67. , 495 55. , 830 24. .319 1. , 00 7. .21 O
ATOM 440 N TYR A 66 66. , 021 56. , 671 25. .821 1. , 00 6. .49 N
ATOM 441 CA TYR A 66 67. , 043 56. , 821 26. .835 1. , 00 7. .42 C
ATOM 442 CB TYR A 66 66. , 581 57. , 694 28. .007 1. , 00 7. .97 c
ATOM 443 CG TYR A 66 66. , 436 59. , 182 27. .744 1. , 00 6. .94 C
ATOM 444 CD1 TYR A 66 65. ,335 59. , 679 27. .102 1. , 00 6. .70 C
ATOM 445 CE1 TYR A 66 65. , 178 61. , 059 26. .898 1. , 00 7. .52 C
ATOM 446 CZ TYR A 66 66. , 122 61. , 950 27. .381 1. , 00 6. .30 c
ATOM 447 OH TYR A 66 65. , 950 63. , 298 27. .236 1. , 00 6. .38 O
ATOM 448 CE2 TYR A 66 67. , 223 61. , 456 28. .041 1. , 00 7. .41 C
ATOM 449 CD2 TYR A 66 67., 368 60 ,, 086 28., 234 1., 00 6., 55 C
ATOM 450 C TYR A 66 67. ,399 55 , , 426 27. ,379 1. , 00 8. ,28 C
ATOM 451 O TYR A 66 66. , 519 54 , , 538 27. , 553 1. , 00 7. , 15 O
ATOM 452 N LEU A 67 68. .694 55 , .243 27. .633 1. , 00 8. .66 N
ATOM 453 CA LEU A 67 69. , 190 54 , , 027 28. , 261 1. , 00 8. ,10 C
ATOM 454 CB LEU A 67 70. , 163 53 , ,308 27. ,319 1. , 00 8. , 12 C
ATOM 455 CG LEU A 67 69. , 433 52 , , 587 26. , 158 1. , 00 7. , 40 C
ATOM 456 CD1 LEU A 67 70. , 429 51 , .912 25. .188 1. , 00 7. .21 c
ATOM 457 CD2 LEU A 67 68. , 402 51 , , 600 26. , 727 1. , 00 7. , 16 C
ATOM 458 C LEU A 67 69. , 798 54 , , 262 29. ,648 1. , 00 8. ,35 C
ATOM 459 O LEU A 67 70. , 173 53 , ,306 30. ,298 1. , 00 7. ,99 O
ATOM 460 N GLY A 68 69. , 866 55 , .523 30. .100 1. , 00 8. .46 N
ATOM 461 CA GLY A 68 70. , 217 55 , , 844 31. , 489 1. , 00 7. ,95 C
ATOM 462 C GLY A 68 71. , 595 56 , , 416 31. , 776 1. , 00 7. , 86 C
ATOM 463 O GLY A 68 71. , 880 56 , , 765 32. ,907 1. , 00 8. ,36 O
ATOM 464 N GLY A 69 72. , 449 56 , .556 30. .762 1. , 00 8. .50 N
ATOM 465 CA GLY A 69 73. , 783 57 , , 136 30. , 956 1. , 00 7. ,95 C
ATOM 466 C GLY A 69 73. , 696 58 , , 565 31. , 448 1. , 00 7. , 55 C
ATOM 467 O GLY A 69 72. , 831 59 , ,314 31. , 024 1. , 00 6. , 85 O
ATOM 468 N ASP A 70 74. , 560 58 , .951 32. .381 1. , 00 8. .04 N
ATOM 469 CA ASP A 70 74. ,328 60 , , 212 33. , 078 1. , 00 8. , 48 C
ATOM 470 CB ASP A 70 73. , 298 59 , , 999 34. , 195 1. , 00 8. ,88 C
ATOM 471 CG ASP A 70 73. , 687 58 , , 869 35. , 150 1. , 00 9. ,84 c
ATOM 472 OD1 ASP A 70 74. ,914 58 , .609 35. .274 1. , 00 10. .67 O
ATOM 473 OD2 ASP A 70 72. , 767 58 , , 255 35. , 746 1. , 00 10. ,62 O
ATOM 474 C ASP A 70 75. , 608 60 , , 877 33. , 573 1. , 00 8. ,59 C
ATOM 475 O ASP A 70 76. , 697 60 , .358 33. .358 1. , 00 9. .98 0
ATOM 476 N PHE A 71 75. , 468 62 , , 048 34. , 196 1. , 00 9. ,01 N
ATOM 477 CA PHE A 71 76. , 618 62 , , 807 34. ,697 1. , 00 8. ,64 C
ATOM 478 CB PHE A 71 76. , 196 64 , , 118 35. ,348 1. , 00 9. , 16 C
ATOM 479 CG PHE A 71 75. , 640 65 , .159 34. .390 1. , 00 9. .25 c
ATOM 480 CD1 PHE A 71 75. , 812 65, , 069 33. , 015 1. , 00 9. , 79 C
ATOM 481 CE1 PHE A 71 75. ,301 66 , , 058 32. , 177 1. , 00 9. , 12 C
ATOM 482 CZ PHE A 71 74. , 639 67, , 157 32. , 703 1. , 00 8. , 06 C
ATOM 483 CE2 PHE A 71 74. , 461 67 , .253 34. .052 1. , 00 9. .63 c
ATOM 484 CD2 PHE A 71 74. , 976 66 , , 272 34. , 892 1. , 00 9. , 71 C
ATOM 485 C PHE A 71 77. ,398 62 , , 007 35. , 712 1. , 00 9. ,92 C
ATOM 486 O PHE A 71 78. , 632 61 , , 988 35. ,653 1. , 00 10. ,26 O
ATOM 487 N LYS A 72 76. , 689 61 , .323 36. .626 1. , 00 9. .44 N
ATOM 488 CA LYS A 72 77. ,370 60 , , 524 37. ,645 1. , 00 10. ,90 C
ATOM 489 CB LYS A 72 76. ,330 59 , , 961 38. ,599 1. , 00 15. ,21 C
ATOM 490 CG LYS A 72 76. , 803 58 , , 855 39. , 519 1. , 00 20. ,58 C
ATOM 491 CD LYS A 72 77. ,341 59 , .372 40. .807 1. , 00 24. .80 c
ATOM 492 CE LYS A 72 77. , 427 58 , , 245 41. , 827 1. , 00 27. , 45 C
ATOM 493 NZ LYS A 72 78. , 820 58 , .284 42. , 294 1. , 00 25. ,67 N
ATOM 494 C LYS A 72 78 ,, 238 59 ,, 404 37., 059 1., 00 10., 76 C
ATOM 495 O LYS A 72 79 , ,369 59 , , 207 37. , 494 1. , 00 9. ,54 O
ATOM 496 N GLY A 73 77 , , 691 58 , , 657 36. , 106 1. , 00 9. , 89 N
ATOM 497 CA GLY A 73 78. .440 57 , .596 35. .443 1. .00 9. .50 C
ATOM 498 C GLY A 73 79 , , 681 58 , , 114 34. , 737 1. , 00 9. , 49 C
ATOM 499 O GLY A 73 80 , , 728 57 , .472 34. , 757 1. , 00 8. , 42 O
ATOM 500 N LEU A 74 79 , , 564 59 , , 289 34. , 124 1. , 00 10. ,36 N
ATOM 501 CA LEU A 74 80 , .691 59 , .910 33. .426 1. , 00 9. .14 c
ATOM 502 CB LEU A 74 80 , , 226 61 , , 090 32. , 554 1. , 00 8. ,52 C
ATOM 503 CG LEU A 74 81 , ,304 61 , , 914 31. , 834 1. , 00 8. ,84 C
ATOM 504 CD1 LEU A 74 82 , , 273 61 , , 059 30. ,977 1. , 00 10. ,65 C
ATOM 505 CD2 LEU A 74 80 , .677 63 , .028 31. .000 1. , 00 7. .62 c
ATOM 506 C LEU A 74 81 , , 753 60 , ,353 34. , 452 1. , 00 9. ,81 C
ATOM 507 O LEU A 74 82 , , 950 60 , , 059 34. ,283 1. , 00 8. ,81 O
ATOM 508 N ALA A 75 81 , ,315 61 , , 047 35. , 501 1. , 00 8. ,68 N
ATOM 509 CA ALA A 75 82 , .230 61 , .568 36. .539 1. , 00 9. .78 c
ATOM 510 CB ALA A 75 81 , , 462 62 , , 294 37. , 655 1. , 00 8. ,66 C
ATOM 511 C ALA A 75 83 , , 005 60 , ,398 37. , 138 1. , 00 10. , 79 C
ATOM 512 O ALA A 75 84, , 224 60 , , 477 37. , 292 1. , 00 12. , 06 O
ATOM 513 N ASP A 76 82. .295 59 , .319 37. .460 1. , 00 9. .31 N
ATOM 514 CA ASP A 76 82 , , 917 58 , .134 38. , 053 1. , 00 11. , 46 C
ATOM 515 CB ASP A 76 81 , , 867 57 , .057 38. ,344 1. , 00 12. ,69 C
ATOM 516 CG ASP A 76 80 , , 948 57 , .397 39. ,502 1. , 00 15. ,11 c
ATOM 517 OD1 ASP A 76 81. .246 58 , .296 40. .316 1. , 00 15. .40 O
ATOM 518 OD2 ASP A 76 79 , , 902 56 , .730 39. , 565 1. , 00 19. ,57 O
ATOM 519 C ASP A 76 83 , , 932 57 , .471 37. , 139 1. , 00 10. ,81 C
ATOM 520 O ASP A 76 84. .696 56 , .641 37. .581 1. .00 10. .99 O
ATOM 521 N HIS A 77 83 , , 950 57 , .785 35. , 852 1. , 00 10. , 46 N
ATOM 522 CA HIS A 77 84, , 919 57 , . Ill 34. ,953 1. , 00 10. ,51 C
ATOM 523 CB HIS A 77 84, , 161 56 , .175 34. , 016 1. , 00 10. ,91 C
ATOM 524 CG HIS A 77 83 , .519 55 , .052 34. .756 1. , 00 12. .17 c
ATOM 525 ND1 HIS A 77 82 , , 228 55 , .079 35. , 129 1. , 00 13. , 06 N
ATOM 526 CE1 HIS A 77 81 , , 952 53 , .975 35. , 854 1. , 00 14. , 09 C
ATOM 527 NE2 HIS A 77 83 , , 088 53 , .264 35. ,982 1. , 00 15. ,67 N
ATOM 528 CD2 HIS A 77 84 , .074 53 , .902 35. .333 1. , 00 12. .48 C
ATOM 529 C HIS A 77 85, , 838 58 , .059 34. , 234 1. , 00 11. ,61 C
ATOM 530 O HIS A 77 86 , , 426 57 , .730 33. , 199 1. , 00 12. , 44 O
ATOM 531 N LEU A 78 86 , , 016 59 , .246 34. , 799 1. , 00 10. ,69 N
ATOM 532 CA LEU A 78 86 , .935 60 , .197 34. .188 1. , 00 9. .80 c
ATOM 533 CB LEU A 78 86 , , 851 61 , .569 34. , 859 1. , 00 9. , 78 C
ATOM 534 CG LEU A 78 85, , 520 62 , .289 34. , 728 1. , 00 10. ,17 C
ATOM 535 CD1 LEU A 78 85, , 455 63 , .491 35. ,658 1. , 00 9. ,98 C
ATOM 536 CD2 LEU A 78 85 , .365 62 , .704 33. .278 1. , 00 8. .35 c
ATOM 537 C LEU A 78 88 , ,364 59 , .670 34. ,247 1. , 00 10. ,80 C
ATOM 538 O LEU A 78 89 , , 158 59 , .972 33. ,350 1. , 00 10. ,68 O
ATOM 539 N ASP A 79 88 ,, 713 58 ,, 887 35.,270 1., 00 10.,17 N
ATOM 540 CA ASP A 79 90 , , 067 58 , ,320 35. ,338 1. , 00 12. ,18 C
ATOM 541 CB ASP A 79 90 , ,345 57 , , 628 36. ,685 1. , 00 14. , 77 C
ATOM 542 CG ASP A 79 90. .595 58 , .635 37. .808 1. .00 22. .62 c
ATOM 543 OD1 ASP A 79 90 , , 902 59 , , 821 37. , 522 1. , 00 28. ,51 0
ATOM 544 OD2 ASP A 79 90 , , 482 58 , , 245 38. ,983 1. , 00 32. , 76 0
ATOM 545 C ASP A 79 90 , ,340 57 , ,361 34. , 192 1. , 00 10. ,54 C
ATOM 546 O ASP A 79 91 , .427 57 , .349 33. .605 1. , 00 10. .10 0
ATOM 547 N TYR A 80 89 , ,347 56 , , 575 33. , 855 1. , 00 10. ,50 N
ATOM 548 CA TYR A 80 89 , , 452 55 , , 668 32. ,692 1. , 00 11. ,17 C
ATOM 549 CB TYR A 80 88 , , 166 54 , , 852 32. , 567 1. , 00 10. , 47 C
ATOM 550 CG TYR A 80 87 , .945 54 , .222 31. .208 1. , 00 11. .09 c
ATOM 551 CD1 TYR A 80 88 , , 549 53 , , 031 30. , 883 1. , 00 11. ,31 C
ATOM 552 CE1 TYR A 80 88 , ,364 52 , .447 29. ,656 1. , 00 12. , 06 C
ATOM 553 CZ TYR A 80 87, , 562 53 , , 035 28. , 731 1. , 00 11. ,32 C
ATOM 554 OH TYR A 80 87 , .380 52 , .400 27. .529 1. , 00 11. .71 0
ATOM 555 CE2 TYR A 80 86 , , 925 54 , , 228 29. , 017 1. , 00 10. , 47 C
ATOM 556 CD2 TYR A 80 87, , 129 54 , , 818 30. ,258 1. , 00 11. ,11 C
ATOM 557 C TYR A 80 89 , , 710 56 , , 437 31. ,378 1. , 00 11. , 71 C
ATOM 558 O TYR A 80 90 , .493 55 , .992 30. .521 1. , 00 10. .23 0
ATOM 559 N ILE A 81 89 , , 050 57 , , 591 31. ,239 1. , 00 10. ,10 N
ATOM 560 CA ILE A 81 89 , , 204 58 , , 453 30. , 060 1. , 00 9. ,52 C
ATOM 561 CB ILE A 81 88 , , 086 59 , , 520 30. , 024 1. , 00 9. ,50 C
ATOM 562 CGI ILE A 81 86 , .723 58 , .866 29. .806 1. , 00 9. .90 C
ATOM 563 CD1 ILE A 81 85, , 527 59 , , 795 29. ,987 1. , 00 9. ,62 C
ATOM 564 CG2 ILE A 81 88 , ,340 60 , , 546 28. ,933 1. , 00 8. , 47 C
ATOM 565 C ILE A 81 90 , .590 59 , .136 30. .063 1. .00 10. .89 c
ATOM 566 O ILE A 81 91 , , 333 59 , , 084 29. , 088 1. , 00 10. ,33 0
ATOM 567 N ARG A 82 90 , , 929 59 , , 776 31. , 173 1. , 00 11. ,96 N
ATOM 568 CA ARG A 82 92 , , 170 60 , , 502 31. ,289 1. , 00 13. , 12 C
ATOM 569 CB ARG A 82 92 , .257 61 , .192 32. .643 1. , 00 17. .16 c
ATOM 570 CG ARG A 82 93 , , 471 62 , , 089 32. , 809 1. , 00 24. ,97 C
ATOM 571 CD ARG A 82 93 , , 534 62 , , 787 34. , 154 1. , 00 33. , 12 C
ATOM 572 NE ARG A 82 93 , , 284 61 , , 889 35. , 292 1. , 00 40. ,22 N
ATOM 573 CZ ARG A 82 93 , .544 62 , .195 36. .564 1. , 00 40. .67 c
ATOM 574 NH1 ARG A 82 94 , , 097 63 , ,360 36. , 887 1. , 00 36. ,36 N
ATOM 575 NH2 ARG A 82 93 , , 271 61 , ,314 37. , 516 1. , 00 41. , 45 N
ATOM 576 C ARG A 82 93 , ,379 59 , , 596 31. , 105 1. , 00 13. ,92 C
ATOM 577 O ARG A 82 94 , .382 60 , .037 30. .557 1. , 00 11. .59 0
ATOM 578 N GLU A 83 93 , ,318 58 , ,352 31. , 576 1. , 00 13. , 09 N
ATOM 579 CA GLU A 83 94, , 507 57 , , 495 31. , 470 1. , 00 16. ,54 C
ATOM 580 CB GLU A 83 94 , , 453 56 , , 324 32. , 449 1. , 00 21. ,64 C
ATOM 581 CG GLU A 83 93 , .624 55 , .146 32. .035 1. , 00 28. .98 c
ATOM 582 CD GLU A 83 93 , , 406 54 , , 136 33. , 176 1. , 00 34. ,23 C
ATOM 583 OE1 GLU A 83 93 , , 801 54 , ,398 34. ,331 1. , 00 36. ,99 0
ATOM 584 OE2 GLU A 83 92., 799 53., 078 32..914 1., 00 31..15 O
ATOM 585 C GLU A 83 94. , 801 57. , 059 30. .028 1. , 00 14. .04 C
ATOM 586 O GLU A 83 95. , 934 56. , 677 29. .720 1. , 00 11. .54 O
ATOM 587 N MET A 84 93. .812 57. , 172 29. .129 1. , 00 11. .61 N
ATOM 588 CA MET A 84 94. , 061 56. , 954 27. .693 1. , 00 12. .16 C
ATOM 589 CB MET A 84 92. , 760 56. , 693 26. .932 1. , 00 10. .60 C
ATOM 590 CG MET A 84 92. , 128 55. ,398 27. .359 1. , 00 10. .96 C
ATOM 591 SD MET A 84 90. , 704 54. , 942 26. .397 1. , 00 11. .99 s
ATOM 592 CE MET A 84 89. , 468 55. , 997 27. .179 1. , 00 10. .41 C
ATOM 593 C MET A 84 94. , 789 58. , 118 27. .030 1. , 00 11. .00 C
ATOM 594 O MET A 84 95. , 168 58. , 008 25. .879 1. , 00 13. .79 O
ATOM 595 N GLY A 85 94. , 904 59. , 242 27. .730 1. , 00 11. .45 N
ATOM 596 CA GLY A 85 95. , 580 60. , 440 27. .215 1. , 00 11. .01 C
ATOM 597 C GLY A 85 94. , 653 61. , 562 26. .751 1. , 00 10. .97 C
ATOM 598 O GLY A 85 95. , 111 62. , 560 26. .179 1. , 00 11. .10 O
ATOM 599 N PHE A 86 93. , 355 61. , 435 27. .030 1. , 00 10. .66 N
ATOM 600 CA PHE A 86 92. , 405 62. , 508 26. .710 1. , 00 9. .23 C
ATOM 601 CB PHE A 86 90. , 986 61. , 931 26. .577 1. , 00 8. .82 C
ATOM 602 CG PHE A 86 90. , 857 60. , 984 25. .427 1. , 00 9. .20 c
ATOM 603 CD1 PHE A 86 90. , 776 61. , 471 24. .121 1. , 00 9. .07 c
ATOM 604 CE1 PHE A 86 90. , 694 60. , 598 23. .042 1. , 00 8. .92 C
ATOM 605 CZ PHE A 86 90. , 703 59. , 220 23. .257 1. , 00 9. .67 C
ATOM 606 CE2 PHE A 86 90. , 795 58. , 730 24. .557 1. , 00 9. .45 c
ATOM 607 CD2 PHE A 86 90. ,877 59. ,611 25. .623 1. , 00 9. .13 c
ATOM 608 C PHE A 86 92. , 467 63. , 572 27. .788 1. , 00 9. .38 C
ATOM 609 O PHE A 86 92. , 593 63. , 243 28. .951 1. , 00 9. .10 O
ATOM 610 N THR A 87 92. .330 64. , 838 27. .403 1. , 00 9. .49 N
ATOM 611 CA THR A 87 92. , 333 65. , 935 28. .353 1. , 00 10. .37 C
ATOM 612 CB THR A 87 93. , 479 66. , 888 28. .012 1. , 00 13. .23 C
ATOM 613 OG1 THR A 87 93. , 245 67. ,376 26. .696 1. , 00 13. .80 O
ATOM 614 CG2 THR A 87 94. , 867 66. , 153 28. .076 1. , 00 12. .69 c
ATOM 615 C THR A 87 91. , 037 66. , 734 28. .362 1. , 00 10. .11 C
ATOM 616 O THR A 87 90. , 935 67. , 713 29. .096 1. , 00 11. .86 O
ATOM 617 N ALA A 88 90. , 056 66. ,326 27. .547 1. , 00 9. .26 N
ATOM 618 CA ALA A 88 88. , 782 67. , 001 27. .412 1. , 00 8. .97 c
ATOM 619 CB ALA A 88 88. , 832 68. , 081 26. .331 1. , 00 7. .85 C
ATOM 620 C ALA A 88 87. , 713 65. , 981 27. .064 1. , 00 8. .38 C
ATOM 621 O ALA A 88 87. , 999 65. , 001 26. .402 1. , 00 7. .89 O
ATOM 622 N VAL A 89 86. , 484 66. , 250 27. .488 1. , 00 7. .77 N
ATOM 623 CA VAL A 89 85. ,333 65. , 430 27. .150 1. , 00 7. .88 C
ATOM 624 CB VAL A 89 84. , 742 64. , 750 28. .369 1. , 00 7. .81 C
ATOM 625 CGI VAL A 89 83. , 621 63. , 809 27. .937 1. , 00 7. .23 c
ATOM 626 CG2 VAL A 89 85. , 835 63. , 992 29. .131 1. , 00 8. .30 c
ATOM 627 C VAL A 89 84. , 237 66. , 271 26. .505 1. , 00 8. .68 C
ATOM 628 O VAL A 89 83. , 821 67. , 297 27. .047 1. , 00 10. .69 O
ATOM 629 N TRP A 90 83 ,.795 65., 827 25..345 1., 00 8..80 N
ATOM 630 CA TRP A 90 82 , .647 66. , 432 24. .645 1. , 00 9. .05 C
ATOM 631 CB TRP A 90 82 , .965 66. , 553 23. .141 1. , 00 8. .66 C
ATOM 632 CG TRP A 90 81. .820 66. .563 22. .143 1. , 00 8. .98 c
ATOM 633 CD1 TRP A 90 80 , .475 66. , 749 22. .391 1. , 00 8. .53 C
ATOM 634 NE1 TRP A 90 79 , .755 66. , 666 21. .233 1. , 00 10. .03 N
ATOM 635 CE2 TRP A 90 80 , .572 66. , 463 20. .168 1. , 00 9. .10 C
ATOM 636 CD2 TRP A 90 81 , .926 66. , 417 20. .676 1. , 00 8. .23 c
ATOM 637 CE3 TRP A 90 82 , .984 66. , 178 19. .776 1. , 00 8. .44 C
ATOM 638 CZ3 TRP A 90 82 , .699 66. , 058 18. .416 1. , 00 8. .17 C
ATOM 639 CH2 TRP A 90 81 , .373 66. , 109 17. .943 1. , 00 9. .05 C
ATOM 640 CZ2 TRP A 90 80 , .286 66. , 285 18. .819 1. , 00 9. .11 c
ATOM 641 C TRP A 90 81 , .440 65. , 590 25. .015 1. , 00 7. .90 C
ATOM 642 O TRP A 90 81 , .422 64. ,348 24. .841 1. , 00 8. .27 O
ATOM 643 N ILE A 91 80 , .485 66. , 256 25. .649 1. , 00 7. .66 N
ATOM 644 CA ILE A 91 79 , .228 65. , 659 26. .041 1. , 00 7. .40 c
ATOM 645 CB ILE A 91 78 , .918 65. , 864 27. .546 1. , 00 7. .67 C
ATOM 646 CGI ILE A 91 78 , .493 67. ,323 27. .834 1. , 00 7. .56 C
ATOM 647 CD1 ILE A 91 78 , .135 67. , 600 29. .300 1. , 00 6. .81 c
ATOM 648 CG2 ILE A 91 80. .108 65. , 406 28. .419 1. , 00 7. .40 c
ATOM 649 C ILE A 91 78 , .099 66. , 224 25. .165 1. , 00 7. .63 C
ATOM 650 O ILE A 91 78 , .171 67. ,372 24. .682 1. , 00 6. .31 O
ATOM 651 N THR A 92 77 , .074 65. ,394 24. .950 1. , 00 7. .17 N
ATOM 652 CA THR A 92 75. .894 65. , 784 24. .184 1. , 00 7. .86 c
ATOM 653 CB THR A 92 75 , .000 64. , 561 23. .863 1. , 00 7. .77 C
ATOM 654 OG1 THR A 92 74 , .892 63. , 711 25. .021 1. , 00 6. .92 O
ATOM 655 CG2 THR A 92 75. .618 63. , 748 22. .716 1. , 00 7. .73 c
ATOM 656 C THR A 92 75 , .120 66. , 869 24. .940 1. , 00 8. .10 C
ATOM 657 O THR A 92 75 , .376 67. , 110 26. .127 1. , 00 6. .99 O
ATOM 658 N PRO A 93 74 , .248 67. , 603 24. .231 1. , 00 8. .13 N
ATOM 659 CA PRO A 93 73 , .575 68. , 734 24. .878 1. , 00 7. .93 c
ATOM 660 CB PRO A 93 72 , .618 69. , 240 23. .800 1. , 00 7. .98 C
ATOM 661 CG PRO A 93 73 , .285 68. , 843 22. .527 1. , 00 8. .35 C
ATOM 662 CD PRO A 93 73 , .847 67. , 480 22. .822 1. , 00 8. .00 C
ATOM 663 C PRO A 93 72 , .821 68. ,314 26. .125 1. , 00 8. .83 c
ATOM 664 O PRO A 93 72 , .265 67. , 198 26. .188 1. , 00 9. .93 O
ATOM 665 N ILE A 94 72 , .835 69. , 206 27. .102 1. , 00 8. .49 N
ATOM 666 CA ILE A 94 72 , .417 68. , 917 28. .456 1. , 00 8. .19 C
ATOM 667 CB ILE A 94 73 , .391 69. , 548 29. .495 1. , 00 7. .74 c
ATOM 668 CGI ILE A 94 73 , .431 71. , 092 29. .388 1. , 00 7. .40 C
ATOM 669 CD1 ILE A 94 74 , .265 71. , 742 30. .478 1. , 00 7. .41 C
ATOM 670 CG2 ILE A 94 74 , .802 68. , 970 29. .327 1. , 00 6. .92 c
ATOM 671 C ILE A 94 71 , .037 69. , 454 28. .758 1. , 00 8. .68 c
ATOM 672 O ILE A 94 70 , .513 69. , 175 29. .849 1. , 00 7. .38 O
ATOM 673 N VAL A 95 70 , .452 70. , 222 27. .825 1. , 00 8. .70 N
ATOM 674 CA VAL A 95 69 ,, 220 70 ,.974 28.. Ill 1., 00 8..89 C
ATOM 675 CB VAL A 95 69 , , 027 72 , .148 27. . 136 1. , 00 9. .88 C
ATOM 676 CGI VAL A 95 70 , , 243 73 , .077 27. . 203 1. , 00 9. .93 C
ATOM 677 CG2 VAL A 95 68. .785 71 , .625 25. .724 1. .00 8. .69 c
ATOM 678 C VAL A 95 67 , , 984 70 , .088 28. .087 1. , 00 8. . 23 C
ATOM 679 O VAL A 95 67, , 997 69 , .019 27. .487 1. , 00 9. .61 O
ATOM 680 N ASP A 96 66 , , 913 70 , .508 28. .749 1. , 00 8. .61 N
ATOM 681 CA ASP A 96 65 , .703 69 , .664 28. .823 1. , 00 7. .56 c
ATOM 682 CB ASP A 96 64 , , 590 70 , . 332 29. .648 1. , 00 7. .93 C
ATOM 683 CG ASP A 96 63 , , 355 69 , .443 29. .808 1. , 00 8. . 23 C
ATOM 684 OD1 ASP A 96 63 , , 489 68 , .190 29. .980 1. , 00 8. .77 O
ATOM 685 OD2 ASP A 96 62 , .228 69 , .985 29. .796 1. , 00 7. .69 0
ATOM 686 C ASP A 96 65, , 194 69 , .336 27. .428 1. , 00 7. .56 C
ATOM 687 O ASP A 96 64, , 994 70 , . 233 26. .624 1. , 00 7. . 31 O
ATOM 688 N ASN A 97 65, , 018 68 , .041 27. .144 1. , 00 8. .12 N
ATOM 689 CA ASN A 97 64 , .407 67 , .568 25. .884 1. , 00 7. .65 c
ATOM 690 CB ASN A 97 65, , 234 66 , .442 25. .249 1. , 00 7. .08 C
ATOM 691 CG ASN A 97 66 , , 444 66 , .945 24. .432 1. , 00 7. .51 C
ATOM 692 OD1 ASN A 97 66 , , 673 66 , .451 23. .339 1. , 00 6. .84 O
ATOM 693 ND2 ASN A 97 67 , .212 67 , .899 24. .963 1. , 00 6. .08 N
ATOM 694 C ASN A 97 63 , , 015 67, .059 26. .258 1. , 00 8. .12 C
ATOM 695 O ASN A 97 62 , , 697 66 , .915 27. .454 1. , 00 8. .47 O
ATOM 696 N PRO A 98 62 , , 157 66 , .810 25. .265 1. , 00 8. .50 N
ATOM 697 CA PRO A 98 60 , .791 66 , .405 25. .614 1. , 00 8. .16 C
ATOM 698 CB PRO A 98 60 , , 109 66 , .292 24. .252 1. , 00 8. .27 C
ATOM 699 CG PRO A 98 60 , , 910 67, .217 23. .355 1. , 00 8. .40 C
ATOM 700 CD PRO A 98 62 , .317 66 , .944 23 . .802 1. .00 8. .79 c
ATOM 701 C PRO A 98 60 , , 665 65 , .088 26. .404 1. , 00 8. .40 C
ATOM 702 O PRO A 98 61 , , 570 64 , .262 26. .398 1. , 00 8. .90 O
ATOM 703 N ASP A 99 59 , , 536 64 , .937 27. .085 1. , 00 8. .91 N
ATOM 704 CA ASP A 99 59 , .307 63 , .858 28. .053 1. , 00 10. .21 c
ATOM 705 CB ASP A 99 58 , .765 64 , .455 29. .352 1. , 00 11. .40 C
ATOM 706 CG ASP A 99 59 , .751 65, .439 30. .001 1. , 00 11. .08 C
ATOM 707 OD1 ASP A 99 60 , .934 65, .470 29. .627 1. , 00 9. .70 O
ATOM 708 OD2 ASP A 99 59 , .343 66 , .204 30. .876 1. , 00 11. .83 0
ATOM 709 C ASP A 99 58 , .357 62 , .800 27. .489 1. , 00 9. .78 C
ATOM 710 O ASP A 99 57 , .805 62 , .000 28. .230 1. , 00 9. .41 O
ATOM 711 N GLN A 100 58 , .196 62 , .814 26. .168 1. , 00 9. .73 N
ATOM 712 CA GLN A 100 57 , .585 61 , .738 25. .386 1. , 00 10. .34 c
ATOM 713 CB GLN A 100 56 , .166 62 , . Ill 24. .915 1. , 00 9. .23 C
ATOM 714 CG GLN A 100 55 , .161 62 , .353 26. .049 1. , 00 9. .49 C
ATOM 715 CD GLN A 100 53 , .773 62 , .763 25. .568 1. , 00 10. .49 C
ATOM 716 OE1 GLN A 100 53 , .595 63 , .210 24. .417 1. , 00 10. .91 0
ATOM 717 NE2 GLN A 100 52 , .763 62 , .627 26. .467 1. , 00 8. .16 N
ATOM 718 C GLN A 100 58 , .443 61 , .468 24. .161 1. , 00 10. .06 C
ATOM 719 O GLN A 100 59., 230 62.319 23.759 1.00 11.00 O
ATOM 720 N ARG A 101 5 i 3.265 6C 1.300 23 ; .562 l . O C s 1.65
ATOM 721 CA ARG A 101 58. , 781 60. 044 22. 213 1 .00 9. 72 C
ATOM 722 CB ARG A 101 58. .788 58. 557 21. 859 1 .00 9. 97 C
ATOM 723 CG ARG A 101 57. , 405 57. 950 21. 685 1 .00 10. 82 c
ATOM 724 CD ARG A 101 57. , 491 56. 459 21. 389 1 .00 11. 39 C
ATOM 725 NE ARG A 101 57. , 797 55. 699 22. 601 1 .00 12. 66 N
ATOM 726 CZ ARG A 101 57. , 859 54. 369 22. 674 1 .00 15. 60 c
ATOM 727 NH1 ARG A 101 57. , 615 53. 615 21. 611 1 .00 17. 16 N
ATOM 728 NH2 ARG A 101 58. , 166 53. 777 23. 821 1 .00 13. 77 N
ATOM 729 C ARG A 101 57. , 948 60. 787 21. 169 1 .00 9. 87 C
ATOM 730 O ARG A 101 56. , 778 61. 115 21. 408 1 .00 10. 29 0
ATOM 731 N PHE A 102 58. , 576 61. 041 20. 014 1 .00 8. 92 N
ATOM 732 CA PHE A 102 57. , 954 61. 798 18. 916 1 .00 9. 48 C
ATOM 733 CB PHE A 102 59. , 055 62. 345 18. 010 1 .00 9. 36 c
ATOM 734 CG PHE A 102 58. , 565 63. 258 16. 899 1 .00 8. 41 c
ATOM 735 CD1 PHE A 102 57. , 745 64. 339 17. 165 1 .00 8. 84 C
ATOM 736 CE1 PHE A 102 57. ,339 65. 193 16. 148 1 .00 8. 66 C
ATOM 737 CZ PHE A 102 57. , 798 64. 988 14. 858 1 .00 8. 96 c
ATOM 738 CE2 PHE A 102 58. , 622 63. 941 14. 595 1 .00 9. 20 c
ATOM 739 CD2 PHE A 102 59. , 015 63. 081 15. 625 1 .00 9. 33 C
ATOM 740 C PHE A 102 56. , 959 60. 956 18. 094 1 .00 10. 34 C
ATOM 741 O PHE A 102 57. ,301 59. 854 17. 632 1 .00 10. 41 O
ATOM 742 N THR A 103 55. , 727 61. 452 17. 915 1 .00 10. 57 N
ATOM 743 CA THR A 103 54. , 721 60. 690 17. 136 1 .00 9. 76 C
ATOM 744 CB THR A 103 53. ,315 60. 712 17. 789 1 .00 10. 24 C
ATOM 745 OG1 THR A 103 52. , 801 62. 051 17. 784 1 .00 10. 00 O
ATOM 746 CG2 THR A 103 53. ,383 60. 193 19. 221 1 .00 8. 11 c
ATOM 747 C THR A 103 54. , 624 61. 152 15. 681 1 .00 11. 29 C
ATOM 748 O THR A 103 53. , 801 60. 660 14. 944 1 .00 10. 52 O
ATOM 749 N GLY A 104 55. , 499 62. 054 15. 241 1 .00 10. 63 N
ATOM 750 CA GLY A 104 55. , 557 62. 418 13. 818 1 .00 11. 41 c
ATOM 751 C GLY A 104 56. , 723 61. 734 13. 121 1 .00 11. 50 C
ATOM 752 O GLY A 104 57. , 230 60. 713 13. 597 1 .00 12. 94 O
ATOM 753 N GLY A 105 57. , 153 62. 310 12. 005 1 .00 11. 93 N
ATOM 754 CA GLY A 105 58. , 451 62. 001 11. 422 1 .00 12. 45 C
ATOM 755 C GLY A 105 58. , 443 60. 770 10. 535 1 .00 11. 99 C
ATOM 756 O GLY A 105 57. , 410 60. 211 10. 236 1 .00 12. 76 O
ATOM 757 N SER A 106 59. , 615 60. 370 10. 090 1 .00 13. 86 N
ATOM 758 CA SER A 106 59. , 743 59. 201 9. 243 1 .00 15. 64 C
ATOM 759 CB SER A 106 60. , 402 59. 601 7. 932 1 .00 21. 06 C
ATOM 760 OG SER A 106 61. , 592 60. 283 8. 227 1 .00 33. 58 O
ATOM 761 C SER A 106 60. , 514 58. 092 9. 959 1 .00 13. 96 c
ATOM 762 O SER A 106 61. , 283 58. 335 10. 894 1 .00 12. 62 O
ATOM 763 N ALA A 107 60. , 265 56. 860 9. 526 1 .00 16. 61 N
ATOM 764 CA ALA A 107 60 ,, 715 55 ,, 655 10., 221 1., 00 16.,25 C
ATOM 765 CB ALA A 107 59 , , 991 54 , , 423 9. ,678 1. , 00 17. , 15 C
ATOM 766 C ALA A 107 62 , , 225 55 , , 466 10. , 090 1. , 00 17. , 43 C
ATOM 767 O ALA A 107 62. .806 55. , 670 9. .026 1. , 00 15. .32 O
ATOM 768 N PRO A 108 62 , , 874 55 , , 096 11. , 192 1. , 00 18. , 09 N
ATOM 769 CA PRO A 108 64, ,308 54 , , 883 11. , 165 1. , 00 15. ,95 C
ATOM 770 CB PRO A 108 64 , , 656 54 , , 723 12. , 651 1. , 00 17. ,38 C
ATOM 771 CG PRO A 108 63 , .410 54. ,141 13. .254 1. , 00 18. .61 c
ATOM 772 CD PRO A 108 62 , , 294 54 , , 849 12. , 527 1. , 00 18. ,07 C
ATOM 773 C PRO A 108 64, , 646 53 , , 635 10. ,376 1. , 00 18. ,29 C
ATOM 774 O PRO A 108 63 , , 933 52 , , 655 10. , 436 1. , 00 16. , 49 O
ATOM 775 N THR A 109 65 , .697 53. , 686 9. .574 1. , 00 21. .18 N
ATOM 776 CA THR A 109 66 , , 147 52 , , 510 8. , 872 1. , 00 21. ,58 C
ATOM 777 CB THR A 109 65, , 984 52 , , 635 7. ,340 1. , 00 26. , 72 C
ATOM 778 OG1 THR A 109 66 , , 829 53 , , 675 6. , 855 1. , 00 29. , 49 O
ATOM 779 CG2 THR A 109 64 , .545 52. , 945 6. .942 1. , 00 33. .30 c
ATOM 780 C THR A 109 67, , 628 52 , ,364 9. ,210 1. , 00 24. ,94 C
ATOM 781 O THR A 109 68 , , 252 53 , , 288 9. , 757 1. , 00 25. ,33 O
ATOM 782 N CYS A 110 68 , , 196 51 , , 223 8. , 856 1. , 00 24. ,91 N
ATOM 783 CA CYS A 110 69 , .579 50. ,941 9. .206 1. , 00 39. .83 c
ATOM 784 CB CYS A 110 69 , , 771 49 , , 443 9. ,309 1. , 00 49. , 43 C
ATOM 785 SG CYS A 110 71 , ,304 48 , , 984 10. , 127 1. , 00 57. ,18 s
ATOM 786 C CYS A 110 70 , , 480 51 , , 540 8. , 140 1. , 00 36. ,26 c
ATOM 787 O CYS A 110 70 , .895 50. , 878 7. .222 1. , 00 43. .68 O
ATOM 788 N GLY A 111 70 , , 780 52 , , 815 8. ,268 1. , 00 45. ,28 N
ATOM 789 CA GLY A 111 71 , , 282 53 , , 574 7. , 132 1. , 00 45. , 13 C
ATOM 790 C GLY A 111 70 , .829 54. , 982 7. .369 1. , 00 51. .84 c
ATOM 791 O GLY A 111 69 , , 719 55 , , 207 7. , 847 1. , 00 68. , 71 O
ATOM 792 N GLY A 112 71 , , 678 55 , , 934 7. , 023 1. , 00 61. ,20 N
ATOM 793 CA GLY A 112 71 , , 602 57 , , 273 7. ,603 1. , 00 62. , 75 C
ATOM 794 C GLY A 112 70 , .648 58. , 259 6. .955 1. , 00 60. .42 c
ATOM 795 O GLY A 112 71 , , 066 59 , ,353 6. , 559 1. , 00 79. ,36 O
ATOM 796 N ILE A 113 69 , ,372 57 , , 900 6. , 834 1. , 00 49. ,88 N
ATOM 797 CA ILE A 113 68 , ,366 58 , , 943 6. ,669 1. , 00 47. ,96 C
ATOM 798 CB ILE A 113 66 , .986 58. , 484 6. .122 1. , 00 59. .41 c
ATOM 799 CGI ILE A 113 67, , 064 57 , , 179 5. ,306 1. , 00 64. ,07 C
ATOM 800 CD1 ILE A 113 65, , 701 56 , , 600 4. , 957 1. , 00 62. ,95 C
ATOM 801 CG2 ILE A 113 66 , ,372 59 , , 608 5. ,288 1. , 00 41. , 47 C
ATOM 802 C ILE A 113 68 , .203 59. , 451 8. .100 1. , 00 39. .00 c
ATOM 803 O ILE A 113 68 , , 182 58 , , 647 9. , 053 1. , 00 39. ,36 O
ATOM 804 N LEU A 114 68 , , 138 60 , , 764 8. ,260 1. , 00 29. ,33 N
ATOM 805 CA LEU A 114 67, , 740 61 , ,371 9. ,528 1. , 00 24. ,96 c
ATOM 806 CB LEU A 114 67 , .936 62. , 884 9. .458 1. , 00 27. .04 c
ATOM 807 CG LEU A 114 67, , 777 63 , , 738 10. , 711 1. , 00 32. ,37 C
ATOM 808 CD1 LEU A 114 68 , ,312 65, , 144 10. , 463 1. , 00 27. ,25 C
ATOM 809 CD2 LEU A 114 66.,316 63 ,.798 11..126 1., 00 31..93 C
ATOM 810 C LEU A 114 66. , 266 60 , .986 9. .738 1. , 00 30. .91 C
ATOM 811 O LEU A 114 65. , 444 61 , .280 8. .882 1. , 00 32. .11 O
ATOM 812 N ALA A 115 65. .938 60 , .317 10. .854 1. , 00 22. .31 N
ATOM 813 CA ALA A 115 64. , 601 59 , .687 11. .022 1. , 00 17. .88 C
ATOM 814 CB ALA A 115 64. , 625 58 , .229 10. .534 1. , 00 16. .41 C
ATOM 815 C ALA A 115 64. , 188 59 , .757 12. .472 1. , 00 15. .02 C
ATOM 816 O ALA A 115 64. , 820 59 , .135 13. .334 1. , 00 15. .07 0
ATOM 817 N ASP A 116 63. , 163 60 , .559 12. .752 1. , 00 13. .37 N
ATOM 818 CA ASP A 116 62. , 755 60 , .875 14. .143 1. , 00 11. .94 C
ATOM 819 CB ASP A 116 62. , 602 62 , .399 14. .288 1. , 00 13. .11 C
ATOM 820 CG ASP A 116 63. , 939 63 , .128 14. .215 1. , 00 17. .42 c
ATOM 821 OD1 ASP A 116 64. , 960 62 , .454 14. .445 1. , 00 18. .27 0
ATOM 822 OD2 ASP A 116 63. , 968 64, .362 13. .963 1. , 00 18. .41 O
ATOM 823 C ASP A 116 61. , 482 60 , .178 14. .645 1. , 00 9. .91 C
ATOM 824 O ASP A 116 61. , 099 60 , .351 15. .794 1. , 00 11. .02 0
ATOM 825 N GLN A 117 60. , 825 59 , .394 13. .806 1. , 00 10. .99 N
ATOM 826 CA GLN A 117 59. , 582 58 , .729 14. .191 1. , 00 11. .26 C
ATOM 827 CB GLN A 117 59. , 030 57 , .961 12. .988 1. , 00 12. .70 c
ATOM 828 CG GLN A 117 57. , 681 57 , .320 13. .209 1. , 00 12. .29 c
ATOM 829 CD GLN A 117 57. , 202 56 , .602 11. .973 1. , 00 13. .35 C
ATOM 830 OE1 GLN A 117 56. , 837 57 , .232 10. .974 1. , 00 19. .23 O
ATOM 831 NE2 GLN A 117 57. , 181 55 , .308 12. .029 1. , 00 10. .56 N
ATOM 832 C GLN A 117 59. , 790 57 , .773 15. .366 1. , 00 11. .00 c
ATOM 833 O GLN A 117 60. , 649 56 , .884 15. .314 1. , 00 9. .15 O
ATOM 834 N GLY A 118 59. , 005 57 , .966 16. .430 1. , 00 11. .48 N
ATOM 835 CA GLY A 118 59. .034 57 , .065 17. .608 1. , 00 9. .34 c
ATOM 836 C GLY A 118 60. , 262 57 , .255 18. .488 1. , 00 8. .98 C
ATOM 837 O GLY A 118 60. , 474 56 , .490 19. .421 1. , 00 9. .55 O
ATOM 838 N LYS A 119 61. , 025 58 , .319 18. .248 1. , 00 8. .15 N
ATOM 839 CA LYS A 119 62. , 330 58 , .538 18. .888 1. , 00 7. .91 c
ATOM 840 CB LYS A 119 63. ,339 59 , .042 17. .843 1. , 00 8. .40 C
ATOM 841 CG LYS A 119 64. , 707 59 , .476 18. .364 1. , 00 9. .03 C
ATOM 842 CD LYS A 119 65. , 685 59 , .851 17. .232 1. , 00 8. .89 C
ATOM 843 CE LYS A 119 66. , 210 58 , .623 16. .473 1. , 00 9. .64 c
ATOM 844 NZ LYS A 119 66. , 918 58 , .935 15. .184 1. , 00 9. .41 N
ATOM 845 C LYS A 119 62. , 194 59 , .513 20. .045 1. , 00 8. .61 C
ATOM 846 O LYS A 119 61. ,330 60 , .404 20. .022 1. , 00 10. .22 O
ATOM 847 N ALA A 120 63. , 012 59 , .308 21. .077 1. , 00 7. .51 N
ATOM 848 CA ALA A 120 63. , 098 60 , .222 22. .209 1. , 00 7. .13 C
ATOM 849 CB ALA A 120 63. , 038 59 , .429 23. .513 1. , 00 6. .28 C
ATOM 850 C ALA A 120 64. ,395 61 , .036 22. .153 1. , 00 7. .14 c
ATOM 851 O ALA A 120 65. , 351 60 , .647 21. .470 1. , 00 6. .72 0
ATOM 852 N GLY A 121 64. , 425 62 , .135 22. .901 1. , 00 6. .81 N
ATOM 853 CA GLY A 121 65. , 573 63 , .034 22. .957 1. , 00 8. .27 C
ATOM 854 C GLY A 121 66 ,, 733 62 ,, 599 23., 829 1., 00 8.,37 C
ATOM 855 O GLY A 121 67, ,340 63 , , 440 24. , 512 1. , 00 8. , 71 O
ATOM 856 N TYR A 122 67, , 052 61 , , 293 23. , 813 1. , 00 7. ,94 N
ATOM 857 CA TYR A 122 68 , .169 60 , .752 24. .598 1. , 00 7. .85 C
ATOM 858 CB TYR A 122 68 , , 257 59 , , 243 24. , 429 1. , 00 7. ,36 C
ATOM 859 CG TYR A 122 68 , , 793 58 , , 827 23. , 065 1. , 00 8. ,14 C
ATOM 860 CD1 TYR A 122 70 , , 154 58 , , 724 22. , 841 1. , 00 9. , 19 C
ATOM 861 CE1 TYR A 122 70 , .661 58 , .394 21. .585 1. , 00 9. .17 c
ATOM 862 CZ TYR A 122 69 , , 802 58 , , 127 20. , 549 1. , 00 8. ,00 C
ATOM 863 OH TYR A 122 70 , ,303 57 , , 803 19. ,310 1. , 00 7. ,95 O
ATOM 864 CE2 TYR A 122 68 , , 437 58 , , 237 20. , 747 1. , 00 8. , 48 C
ATOM 865 CD2 TYR A 122 67 , .942 58 , .592 22. .003 1. , 00 8. .41 c
ATOM 866 C TYR A 122 69 , , 509 61 , , 407 24. ,209 1. , 00 8. , 49 C
ATOM 867 O TYR A 122 70 , , 467 61 , , 414 24. ,990 1. , 00 9. ,08 O
ATOM 868 N HIS A 123 69 , , 541 61 , , 905 22. ,976 1. , 00 8. , 09 N
ATOM 869 CA HIS A 123 70 , .703 62 , .505 22. .340 1. , 00 8. .06 c
ATOM 870 CB HIS A 123 70 , , 578 62 , , 357 20. , 829 1. , 00 7. , 45 C
ATOM 871 CG HIS A 123 69 , , 227 62 , , 750 20. ,311 1. , 00 8. ,31 C
ATOM 872 ND1 HIS A 123 68 , , 819 64 , , 030 20. ,303 1. , 00 7. , 16 N
ATOM 873 CE1 HIS A 123 67 , .558 64 , .081 19. .841 1. , 00 8. .00 C
ATOM 874 NE2 HIS A 123 67, , 144 62 , , 824 19. , 576 1. , 00 8. , 19 N
ATOM 875 CD2 HIS A 123 68 , , 147 61 , , 979 19. , 859 1. , 00 8. ,31 C
ATOM 876 C HIS A 123 70 , , 913 63 , , 946 22. , 723 1. , 00 8. , 09 c
ATOM 877 O HIS A 123 72 , .015 64 , .469 22. .542 1. , 00 8. .47 O
ATOM 878 N GLY A 124 69 , , 866 64 , , 582 23. , 252 1. , 00 8. , 41 N
ATOM 879 CA GLY A 124 69 , , 897 65, , 966 23. , 755 1. , 00 8. ,27 C
ATOM 880 C GLY A 124 69 , .664 67 , .107 22. .760 1. , 00 8. .38 c
ATOM 881 O GLY A 124 69 , , 557 68 , , 254 23. , 174 1. , 00 10. , 78 O
ATOM 882 N TYR A 125 69 , , 553 66 , , 802 21. , 472 1. , 00 8. ,01 N
ATOM 883 CA TYR A 125 69 , , 474 67, , 838 20. , 411 1. , 00 8. ,90 C
ATOM 884 CB TYR A 125 69 , .973 67 , .287 19. .078 1. , 00 9. .02 c
ATOM 885 CG TYR A 125 71 , , 420 66 , , 886 19. , 093 1. , 00 9. , 71 C
ATOM 886 CD1 TYR A 125 72 , , 439 67, , 857 19. , 184 1. , 00 11. , 89 C
ATOM 887 CE1 TYR A 125 73 , , 765 67, , 500 19. ,209 1. , 00 11. ,14 C
ATOM 888 CZ TYR A 125 74 , .099 66 , .139 19. .154 1. , 00 11. .90 c
ATOM 889 OH TYR A 125 75 , ,397 65, , 737 19. , 181 1. , 00 11. ,84 O
ATOM 890 CE2 TYR A 125 73 , , 109 65, , 171 19. , 076 1. , 00 11. ,88 C
ATOM 891 CD2 TYR A 125 71 , , 785 65, , 550 19. , 043 1. , 00 9. ,99 C
ATOM 892 C TYR A 125 68 , .098 68 , .475 20. .199 1. , 00 9. .13 c
ATOM 893 O TYR A 125 67, , 929 69 , ,328 19. ,305 1. , 00 9. ,20 O
ATOM 894 N TRP A 126 67, , 134 68 , , 092 21. , 033 1. , 00 8. , 70 N
ATOM 895 CA TRP A 126 65, , 784 68 , , 631 20. ,948 1. , 00 8. , 45 C
ATOM 896 CB TRP A 126 64 , .786 67 , .497 20. .828 1. , 00 8. .19 c
ATOM 897 CG TRP A 126 64 , , 948 66 , , 595 19. ,647 1. , 00 7. ,80 C
ATOM 898 CD1 TRP A 126 65, , 703 66 , , 793 18. , 501 1. , 00 7. ,99 C
ATOM 899 NE1 TRP A 126 65., 559 65., 735 17..643 1., 00 8..77 N
ATOM 900 CE2 TRP A 126 64. , 714 64. , 818 18. .163 1. , 00 8. .26 C
ATOM 901 CD2 TRP A 126 64. , 280 65. ,316 19. .450 1. , 00 7. .54 C
ATOM 902 CE3 TRP A 126 63. .409 64. , 562 20. .198 1. , 00 8. .14 c
ATOM 903 CZ3 TRP A 126 62. , 983 63. ,341 19. .699 1. , 00 8. .06 C
ATOM 904 CH2 TRP A 126 63. ,393 62. , 880 18. .459 1. , 00 8. .34 C
ATOM 905 CZ2 TRP A 126 64. , 285 63. , 598 17. .672 1. , 00 8. .33 C
ATOM 906 C TRP A 126 65. , 433 69. , 473 22. .149 1. , 00 8. .53 c
ATOM 907 O TRP A 126 64. , 646 69. , 066 23. .004 1. , 00 8. .34 O
ATOM 908 N GLY A 127 66. , 007 70. , 670 22. .222 1. , 00 8. .52 N
ATOM 909 CA GLY A 127 65. , 856 71. , 519 23. .371 1. , 00 8. .25 C
ATOM 910 C GLY A 127 64. , 478 72. , 126 23. .556 1. , 00 8. .41 c
ATOM 911 O GLY A 127 63. , 875 72. , 583 22. .599 1. , 00 8. .48 O
ATOM 912 N VAL A 128 64. , 006 72. , 132 24. .810 1. , 00 8. .05 N
ATOM 913 CA . AVAL A 128 62. , 735 72. , 722 25. .228 0. , 50 7. .57 C
ATOM 914 CA BVAL A 128 62. , 752 72. , 796 25. .150 0. , 50 8. .37 c
ATOM 915 CB . AVAL A 128 61. , 847 71. , 645 25. .883 0. , 50 7. .06 C
ATOM 916 CB BVAL A 128 61. , 597 71. , 801 25. .476 0. , 50 9. .15 C
ATOM 917 CG1AVAL A 128 60. , 541 72. , 240 26. .427 0. , 50 6. .29 C
ATOM 918 CG1BVAL A 128 61. , 962 70. , 400 25. .015 0. , 50 9. .16 C
ATOM 919 CG2AVAL A 128 61. , 545 70. , 549 24. .870 0. , 50 6. .90 C
ATOM 920 CG2BVAL A 128 61. , 169 71. , 823 26. .941 0. , 50 9. .05 C
ATOM 921 C VAL A 128 62. , 981 73. , 865 26. .209 1. , 00 8. .10 C
ATOM 922 O VAL A 128 62. , 403 74. , 939 26. .113 1. , 00 9. .16 O
ATOM 923 N ASN A 129 63. , 859 73. , 627 27. .171 1. , 00 8. .52 N
ATOM 924 CA ASN A 129 64. , 257 74. , 687 28. .090 1. , 00 9. .76 C
ATOM 925 CB ASN A 129 63. .715 74. , 475 29. .504 1. , 00 9. .54 c
ATOM 926 CG ASN A 129 63. , 887 75. , 714 30. .370 1. , 00 11. .86 C
ATOM 927 OD1 ASN A 129 64. , 876 76. , 456 30. .223 1. , 00 11. .47 O
ATOM 928 ND2 ASN A 129 62. , 928 75. , 953 31. .282 1. , 00 10. .82 N
ATOM 929 C ASN A 129 65. , 764 74. , 750 28. .122 1. , 00 9. .14 c
ATOM 930 O ASN A 129 66. , 407 73. , 896 28. .725 1. , 00 8. .31 O
ATOM 931 N PHE A 130 66. , 324 75. , 752 27. .459 1. , 00 8. .93 N
ATOM 932 CA PHE A 130 67. , 783 75. , 830 27. .286 1. , 00 9. .13 C
ATOM 933 CB PHE A 130 68. , 126 76. , 736 26. .089 1. , 00 9. .44 c
ATOM 934 CG PHE A 130 67. , 823 76. , 108 24. .739 1. , 00 9. .95 C
ATOM 935 CD1 PHE A 130 66. , 509 75. , 854 24. .328 1. , 00 8. .88 C
ATOM 936 CE1 PHE A 130 66. , 244 75. , 287 23. .100 1. , 00 10. .21 C
ATOM 937 CZ PHE A 130 67. , 284 74. , 958 22. .239 1. , 00 10. .77 c
ATOM 938 CE2 PHE A 130 68. , 591 75. , 198 22. .633 1. , 00 11. .17 C
ATOM 939 CD2 PHE A 130 68. , 851 75. , 777 23. .877 1. , 00 10. .30 C
ATOM 940 C PHE A 130 68. , 473 76. ,312 28. .572 1. , 00 8. .31 C
ATOM 941 O PHE A 130 69. , 695 76. ,311 28. .655 1. , 00 8. .16 0
ATOM 942 N TYR A 131 67. , 682 76. , 706 29. .579 1. , 00 8. .67 N
ATOM 943 CA TYR A 131 68. , 204 77. , 137 30. .871 1. , 00 9. .14 C
ATOM 944 CB TYR A 131 67., 446 78 ,.364 31..385 1., 00 9..45 C
ATOM 945 CG TYR A 131 67. , 771 79 , .608 30. .586 1. , 00 10. .55 C
ATOM 946 CD1 TYR A 131 67. , 201 79 , .817 29. .350 1. , 00 10. .18 C
ATOM 947 CE1 TYR A 131 67. .483 80 , .947 28. .610 1. , 00 11. .25 c
ATOM 948 CZ TYR A 131 68. , 363 81 , .875 29. .088 1. , 00 11. .51 C
ATOM 949 OH TYR A 131 68. , 660 82 , .957 28. .302 1. , 00 12. .41 O
ATOM 950 CE2 TYR A 131 68. , 962 81 , .701 30. .314 1. , 00 12. .46 C
ATOM 951 CD2 TYR A 131 68. , 672 80 , .555 31. .058 1. , 00 12. .49 c
ATOM 952 C TYR A 131 68. , 165 76 , .046 31. .942 1. , 00 10. .15 C
ATOM 953 O TYR A 131 68. , 588 76 , .290 33. .034 1. , 00 9. .65 O
ATOM 954 N GLN A 132 67. , 650 74 , .863 31. .633 1. , 00 10. .60 N
ATOM 955 CA GLN A 132 67. , 530 73 , .796 32. .635 1. , 00 10. .64 c
ATOM 956 CB GLN A 132 66. , 059 73 , .529 32. .986 1. , 00 12. .22 C
ATOM 957 CG GLN A 132 65. ,381 74 , .646 33. .750 1. , 00 15. .14 C
ATOM 958 CD GLN A 132 63. , 929 74 , .334 34. .139 1. , 00 15. .81 C
ATOM 959 OE1 GLN A 132 63. , 416 73 , .222 33. .943 1. , 00 19. .68 0
ATOM 960 NE2 GLN A 132 63. , 275 75 , .319 34. .685 1. , 00 16. .95 N
ATOM 961 C GLN A 132 68. , 137 72 , .521 32. .084 1. , 00 9. .69 C
ATOM 962 O GLN A 132 67. , 989 72 , .206 30. .904 1. , 00 10. .13 O
ATOM 963 N VAL A 133 68. , 795 71 , .787 32. .960 1. , 00 10. .21 N
ATOM 964 CA VAL A 133 69. ,376 70 , .502 32. .632 1. , 00 10. .16 C
ATOM 965 CB VAL A 133 70. , 437 70 , .114 33. .675 1. , 00 11. .32 C
ATOM 966 CGI VAL A 133 70. , 946 68 , .707 33. .419 1. , 00 12. .83 c
ATOM 967 CG2 VAL A 133 71. , 586 71 , .103 33. .599 1. , 00 13. .12 c
ATOM 968 C VAL A 133 68. , 282 69 , .460 32. .527 1. , 00 8. .89 C
ATOM 969 O VAL A 133 67. ,335 69 , .468 33. .281 1. , 00 7. .09 O
ATOM 970 N ASP A 134 68. ,370 68 , .593 31. .536 1. , 00 8. .88 N
ATOM 971 CA ASP A 134 67. , 397 67 , .527 31. .412 1. , 00 9. .39 C
ATOM 972 CB ASP A 134 67. , 660 66 , .665 30. .179 1. , 00 9. .63 C
ATOM 973 CG ASP A 134 66. , 513 65, .686 29. .932 1. , 00 9. .77 C
ATOM 974 OD1 ASP A 134 66. , 530 64 , .554 30. .510 1. , 00 9. .09 0
ATOM 975 OD2 ASP A 134 65. , 579 66 , .102 29. .227 1. , 00 8. .12 O
ATOM 976 C ASP A 134 67. , 412 66 , .628 32. .657 1. , 00 9. .22 C
ATOM 977 O ASP A 134 68. , 477 66 , .309 33. .179 1. , 00 10. .21 O
ATOM 978 N GLU A 135 66. , 224 66 , .240 33. .118 1. , 00 9. .96 N
ATOM 979 CA GLU A 135 66. , 047 65, .459 34. .343 1. , 00 9. .72 C
ATOM 980 CB GLU A 135 64. , 532 65, .295 34. .660 1. , 00 12. .37 C
ATOM 981 CG GLU A 135 63. , 748 64 , .378 33. .718 1. , 00 11. .48 C
ATOM 982 CD GLU A 135 63. , 500 64 , .952 32. .336 1. , 00 12. .01 c
ATOM 983 OE1 GLU A 135 63. , 634 66 , .184 32. .127 1. , 00 12. .59 O
ATOM 984 OE2 GLU A 135 63. , 190 64, .152 31. .433 1. , 00 9. .85 O
ATOM 985 C GLU A 135 66. , 732 64 , .091 34. .379 1. , 00 8. .97 c
ATOM 986 O GLU A 135 66. , 926 63 , .511 35. .456 1. , 00 10. .04 0
ATOM 987 N HIS A 136 67. , 071 63 , .542 33. .221 1. , 00 8. .92 N
ATOM 988 CA HIS A 136 67. , 820 62 , .289 33. .178 1. , 00 8. .25 C
ATOM 989 CB HIS A 136 67 ,, 767 61., 695 31., 801 1., 00 8., 45 C
ATOM 990 CG HIS A 136 66 , ,390 61. , 264 31. ,372 1. , 00 8. ,53 C
ATOM 991 ND1 HIS A 136 65, , 501 62. , 107 30. , 821 1. , 00 8. ,93 N
ATOM 992 CE1 HIS A 136 64. .387 61. , 418 30. .486 1. , 00 9. .04 C
ATOM 993 NE2 HIS A 136 64 , , 574 60. , 126 30. , 812 1. , 00 8. .94 N
ATOM 994 CD2 HIS A 136 65, , 796 59. , 995 31. .353 1. , 00 8. .84 C
ATOM 995 C HIS A 136 69 , , 258 62. ,399 33. .603 1. , 00 8. .98 C
ATOM 996 O HIS A 136 69 , .872 61. , 397 33. .982 1. , 00 11. .05 0
ATOM 997 N LEU A 137 69 , , 832 63. , 593 33. .557 1. , 00 9. .41 N
ATOM 998 CA LEU A 137 71 , ,313 63. , 711 33. .625 1. , 00 9. .45 C
ATOM 999 CB LEU A 137 71 , , 790 64. , 812 32. .680 1. , 00 9. .45 C
ATOM 1000 CG LEU A 137 71 , .488 64. , 536 31. .197 1. , 00 10. .52 c
ATOM 1001 CD1 LEU A 137 71 , , 891 65. , 726 30. .311 1. , 00 9. .09 C
ATOM 1002 CD2 LEU A 137 72 , , 152 63. , 258 30. .713 1. , 00 9. .69 C
ATOM 1003 C LEU A 137 71 , , 965 63. , 895 35. .022 1. , 00 9. .36 C
ATOM 1004 O LEU A 137 73 , .008 63. , 272 35. .303 1. , 00 7. .32 O
ATOM 1005 N PRO A 138 71 , ,376 64. , 744 35. .891 1. , 00 9. .50 N
ATOM 1006 CA PRO A 138 72 , , 044 65. , 010 37. .139 1. , 00 10. .29 C
ATOM 1007 CB PRO A 138 71 , , 597 66. , 438 37. .451 1. , 00 10. .28 C
ATOM 1008 CG PRO A 138 70 , .174 66. , 447 36. .998 1. , 00 10. .84 c
ATOM 1009 CD PRO A 138 70 , , 152 65. , 551 35. .774 1. , 00 10. .44 C
ATOM 1010 C PRO A 138 71 , .627 64. , 055 38. .271 1. , 00 11. .03 C
ATOM 1011 O PRO A 138 70 , .660 63. , 298 38. .143 1. , 00 10. .18 O
ATOM 1012 N SER A 139 72 , .404 64. , 072 39. .348 1. , 00 11. .65 N
ATOM 1013 CA SER A 139 72 , .102 63. ,317 40. .552 1. , 00 12. .27 C
ATOM 1014 CB SER A 139 73 , .058 62. , 126 40. .699 1. , 00 12. .48 C
ATOM 1015 OG SER A 139 73 , .018 61. , 239 39. .606 1. , 00 13. .25 O
ATOM 1016 C SER A 139 72 , .369 64. , 277 41. .717 1. , 00 13. .85 C
ATOM 1017 O SER A 139 73 , .116 65. , 243 41. .560 1. , 00 12. .37 O
ATOM 1018 N PRO A 140 71 , .846 63. , 960 42. .906 1. , 00 17. .20 N
ATOM 1019 CA PRO A 140 72 , .307 64. , 706 44. .083 1. , 00 18. .70 c
ATOM 1020 CB PRO A 140 71 , .583 64. , 009 45. .245 1. , 00 20. .40 C
ATOM 1021 CG PRO A 140 70 , .373 63. ,364 44. .600 1. , 00 17. .87 C
ATOM 1022 CD PRO A 140 70 , .911 62. , 880 43. .281 1. , 00 17. .00 C
ATOM 1023 C PRO A 140 73 , .806 64. , 557 44. .214 1. , 00 17. .28 c
ATOM 1024 O PRO A 140 74 , .338 63. , 472 44. .037 1. , 00 17. .34 O
ATOM 1025 N GLY A 141 74 , .502 65. , 643 44. .482 1. , 00 16. .43 N
ATOM 1026 CA GLY A 141 75 , .962 65. , 587 44. .489 1. , 00 18. .74 C
ATOM 1027 C GLY A 141 76 , .631 65. , 520 43. .123 1. , 00 18. .04 c
ATOM 1028 O GLY A 141 77 , .845 65. , 438 43. .067 1. , 00 22. .48 O
ATOM 1029 N MET A 142 75 , .860 65. , 535 42. .024 1. , 00 16. .24 N
ATOM 1030 CA MET A 142 76 , .437 65. , 521 40. .675 1. , 00 13. .09 C
ATOM 1031 CB MET A 142 76 , .781 64. , 093 40. .237 1. , 00 14. .71 c
ATOM 1032 CG MET A 142 77 , .649 64. , 002 38. .983 1. , 00 16. .11 C
ATOM 1033 SD MET A 142 79 , .099 65. , 060 39. .002 1. , 00 17. .36 s
ATOM 1034 CE MET A 142 80., 244 64 ,, 352 40., 174 1., 00 12., 41 C
ATOM 1035 C MET A 142 75. , 525 66 , , 196 39. ,673 1. , 00 12. , 42 C
ATOM 1036 O MET A 142 74. , 959 65, , 562 38. , 786 1. , 00 10. ,34 O
ATOM 1037 N ASP A 143 75. .377 67 , .505 39. .827 1. .00 13. .19 N
ATOM 1038 CA ASP A 143 74. , 710 68 , , 325 38. , 829 1. , 00 11. ,91 C
ATOM 1039 CB ASP A 143 73. , 987 69 , , 510 39. , 498 1. , 00 15. , 02 C
ATOM 1040 CG ASP A 143 74. , 926 70 , , 434 40. ,277 1. , 00 16. , 73 C
ATOM 1041 OD1 ASP A 143 76. , 166 70 , .322 40. .147 1. , 00 17. .79 0
ATOM 1042 OD2 ASP A 143 74. , 413 71 , , 287 41. , 030 1. , 00 16. ,51 O
ATOM 1043 C ASP A 143 75. , 750 68 , , 762 37. , 797 1. , 00 11. , 44 C
ATOM 1044 O ASP A 143 76. , 911 68 , ,347 37. , 862 1. , 00 9. ,33 O
ATOM 1045 N PHE A 144 75. ,341 69 , .565 36. .809 1. , 00 11. .16 N
ATOM 1046 CA PHE A 144 76. , 271 70 , , 003 35. , 769 1. , 00 10. ,14 C
ATOM 1047 CB PHE A 144 75. , 571 70 , , 904 34. , 742 1. , 00 11. , 49 C
ATOM 1048 CG PHE A 144 76. , 494 71 , ,361 33. ,623 1. , 00 11. , 77 C
ATOM 1049 CD1 PHE A 144 77. , 016 70 , .449 32. .730 1. , 00 11. .66 c
ATOM 1050 CE1 PHE A 144 77. , 876 70 , , 852 31. , 737 1. , 00 10. ,97 C
ATOM 1051 CZ PHE A 144 78. , 233 72 , , 182 31. ,626 1. , 00 11. ,63 C
ATOM 1052 CE2 PHE A 144 77. , 713 73 , , 111 32. , 507 1. , 00 11. , 40 c
ATOM 1053 CD2 PHE A 144 76. , 859 72 , .696 33. .499 1. , 00 10. .90 c
ATOM 1054 C PHE A 144 77. , 529 70 , , 687 36. ,323 1. , 00 10. , 74 C
ATOM 1055 O PHE A 144 78. , 641 70 , ,350 35. ,932 1. , 00 11. ,37 O
ATOM 1056 N ARG A 145 77. , 351 71 , , 644 37. , 231 1. , 00 10. , 47 N
ATOM 1057 CA ARG A 145 78. , 458 72 , .297 37. .912 1. , 00 12. .14 c
ATOM 1058 CB ARG A 145 77. , 922 73 , , 190 39. , 018 1. , 00 16. , 55 C
ATOM 1059 CG ARG A 145 77. , 857 74 , , 661 38. , 704 1. , 00 18. , 46 C
ATOM 1060 CD ARG A 145 77. .467 75 , .487 39. .948 1. .00 15. .76 c
ATOM 1061 NE ARG A 145 76. , 872 76 , , 736 39. , 487 1. , 00 21. , 12 N
ATOM 1062 CZ ARG A 145 77. , 564 77 , , 813 39. , 125 1. , 00 24. ,33 C
ATOM 1063 NH1 ARG A 145 78. , 902 77 , , 839 39. ,201 1. , 00 26. , 15 N
ATOM 1064 NH2 ARG A 145 76. ,917 78 , .889 38. .701 1. , 00 24. .96 N
ATOM 1065 C ARG A 145 79. , 431 71 , ,304 38. ,562 1. , 00 12. ,33 C
ATOM 1066 O ARG A 145 80. , 653 71 , , 465 38. , 464 1. , 00 11. , 76 O
ATOM 1067 N ASP A 146 78. , 881 70 , ,307 39. ,258 1. , 00 10. ,65 N
ATOM 1068 CA ASP A 146 79. , 705 69 , .263 39. .893 1. , 00 11. .11 C
ATOM 1069 CB ASP A 146 78. , 855 68 , , 280 40. , 714 1. , 00 12. ,33 C
ATOM 1070 CG ASP A 146 78. , 088 68 , , 968 41. , 842 1. , 00 13. ,59 C
ATOM 1071 OD1 ASP A 146 78. , 683 69 , , 833 42. , 501 1. , 00 15. ,25 O
ATOM 1072 OD2 ASP A 146 76. , 902 68 , .651 42. .041 1. , 00 14. .56 0
ATOM 1073 C ASP A 146 80. , 493 68 , , 477 38. , 848 1. , 00 10. ,17 C
ATOM 1074 O ASP A 146 81. , 654 68 , , 133 39. , 062 1. , 00 9. , 83 O
ATOM 1075 N LEU A 147 79. , 864 68 , , 184 37. , 719 1. , 00 9. , 45 N
ATOM 1076 CA LEU A 147 80. , 555 67 , .437 36. .687 1. , 00 8. .76 c
ATOM 1077 CB LEU A 147 79. , 593 67, , 022 35. , 581 1. , 00 8. , 79 C
ATOM 1078 CG LEU A 147 80. , 239 66 , ,363 34. ,341 1. , 00 7. ,88 C
ATOM 1079 CD1 LEU A 147 80., 720 64 ,.964 34..710 1., 00 7..67 C
ATOM 1080 CD2 LEU A 147 79. , 222 66 , .316 33. .185 1. , 00 7. .01 C
ATOM 1081 C LEU A 147 81. , 702 68 , .265 36. .112 1. , 00 9. .10 C
ATOM 1082 O LEU A 147 82. .817 67 , .767 35. .928 1. , 00 8. .89 O
ATOM 1083 N ALA A 148 81. , 451 69 , .528 35. .802 1. , 00 10. .28 N
ATOM 1084 CA ALA A 148 82. , 539 70 , .346 35. .279 1. , 00 10. .21 C
ATOM 1085 CB ALA A 148 82. , 043 71 , .711 34. .787 1. , 00 10. .97 C
ATOM 1086 C ALA A 148 83. , 659 70 , .478 36. .334 1. , 00 9. .79 c
ATOM 1087 O ALA A 148 84. , 838 70 , .371 35. .998 1. , 00 9. .35 O
ATOM 1088 N ALA A 149 83. ,305 70 , .636 37. .604 1. , 00 9. .63 N
ATOM 1089 CA ALA A 149 84. , 327 70 , .721 38. .679 1. , 00 9. .33 C
ATOM 1090 CB ALA A 149 83. , 714 71 , .104 40. .040 1. , 00 8. .38 c
ATOM 1091 C ALA A 149 85. , 097 69 , .389 38. .803 1. , 00 11. .27 C
ATOM 1092 O ALA A 149 86. , 296 69 , .388 39. .110 1. , 00 12. .08 O
ATOM 1093 N ALA A 150 84. , 432 68 , .255 38. .580 1. , 00 11. .83 N
ATOM 1094 CA ALA A 150 85. , 137 66 , .948 38. .658 1. , 00 14. .01 c
ATOM 1095 CB ALA A 150 84. , 173 65, .766 38. .578 1. , 00 12. .75 C
ATOM 1096 C ALA A 150 86. , 166 66 , .870 37. .546 1. , 00 13. .00 C
ATOM 1097 O ALA A 150 87. ,300 66 , .469 37. .761 1. , 00 13. .57 O
ATOM 1098 N MET A 151 85. , 791 67 , .332 36. .371 1. , 00 13. .01 N
ATOM 1099 CA MET A 151 86. , 733 67, .356 35. .256 1. , 00 13. .02 C
ATOM 1100 CB MET A 151 86. , 045 67, .768 33. .956 1. , 00 12. .26 C
ATOM 1101 CG MET A 151 85. , 024 66 , .752 33. .479 1. , 00 14. .82 c
ATOM 1102 SD MET A 151 84. , 476 67 , .113 31. .811 1. , 00 15. .18 s
ATOM 1103 CE MET A 151 83. , 191 65, .868 31. .584 1. , 00 16. .42 C
ATOM 1104 C MET A 151 87. , 886 68 , .286 35. .545 1. , 00 11. .45 C
ATOM 1105 O MET A 151 89. , 034 67 , .930 35. .310 1. , 00 13. .59 O
ATOM 1106 N HIS A 152 87. , 584 69 , .466 36. .057 1. , 00 11. .04 N
ATOM 1107 CA HIS A 152 88. , 624 70 , .454 36. .356 1. , 00 11. .67 C
ATOM 1108 CB HIS A 152 87. , 999 71 , .778 36. .792 1. , 00 11. .43 C
ATOM 1109 CG HIS A 152 87. , 153 72 , .432 35. .697 1. , 00 12. .31 c
ATOM 1110 ND1 HIS A 152 86. , 192 73 , .335 35. .960 1. , 00 12. .29 N
ATOM 1111 CE1 HIS A 152 85. , 607 73 , .730 34. .810 1. , 00 10. .76 C
ATOM 1112 NE2 HIS A 152 86. , 164 73 , .044 33. .806 1. , 00 12. .01 N
ATOM 1113 CD2 HIS A 152 87. , 133 72 , .233 34. .321 1. , 00 11. .22 C
ATOM 1114 C HIS A 152 89. , 627 69 , .974 37. .364 1. , 00 13. .31 C
ATOM 1115 O HIS A 152 90. , 830 70 , .152 37. .159 1. , 00 11. .45 O
ATOM 1116 N ARG A 153 89. , 169 69 , .345 38. .447 1. , 00 13. .29 N
ATOM 1117 CA ARG A 153 90. , 106 68 , .770 39. .427 1. , 00 17. .06 c
ATOM 1118 CB ARG A 153 89. ,358 68 , .107 40. .606 1. , 00 21. .01 C
ATOM 1119 CG ARG A 153 88. , 677 69 , .115 41. .516 1. , 00 22. .99 C
ATOM 1120 CD ARG A 153 88. , 226 68 , .456 42. .808 1. , 00 31. .60 C
ATOM 1121 NE ARG A 153 87. , 257 67 , .388 42. .550 1. , 00 29. .94 N
ATOM 1122 CZ ARG A 153 85. , 938 67, .557 42. .442 1. , 00 30. .44 C
ATOM 1123 NH1 ARG A 153 85. ,382 68 , .762 42. .568 1. , 00 31. .35 N
ATOM 1124 NH2 ARG A 153 85 ,.166 66 ,.507 42..198 1., 00 30..71 N
ATOM 1125 C ARG A 153 91 , .068 67, .758 38. .822 1. , 00 16. .63 C
ATOM 1126 O ARG A 153 92 , .159 67, .543 39. .366 1. , 00 16. .28 O
ATOM 1127 N LYS A 154 90. .670 67 , .126 37. .718 1. .00 13. .26 N
ATOM 1128 CA LYS A 154 91 , .524 66 , .177 37. .030 1. , 00 12. .93 C
ATOM 1129 CB LYS A 154 90 , .715 64, .973 36. .581 1. , 00 15. .97 C
ATOM 1130 CG LYS A 154 90 , .275 64 , .098 37. .756 1. , 00 19. .37 C
ATOM 1131 CD LYS A 154 89 , .440 62 , .932 37. .259 1. , 00 23. .42 c
ATOM 1132 CE LYS A 154 88 , .702 62 , .211 38. .383 1. , 00 30. .99 C
ATOM 1133 NZ LYS A 154 89 , .622 61 , .640 39. .392 1. , 00 27. .38 N
ATOM 1134 C LYS A 154 92 , .263 66 , .794 35. .855 1. , 00 15. .47 C
ATOM 1135 O LYS A 154 92 , .841 66 , .069 35. .038 1. , 00 14. .55 0
ATOM 1136 N GLY A 155 92 , .276 68 , .123 35. .781 1. , 00 13. .30 N
ATOM 1137 CA GLY A 155 92 , .968 68 , .802 34. .696 1. , 00 13. .43 C
ATOM 1138 C GLY A 155 92 , .278 68 , .614 33. .356 1. , 00 14. .51 C
ATOM 1139 O GLY A 155 92 , .912 68 , .790 32. .342 1. , 00 15. .46 0
ATOM 1140 N MET A 156 90 , .977 68 , .293 33. .347 1. , 00 11. .95 N
ATOM 1141 CA MET A 156 90 , .241 68 , .077 32. .090 1. , 00 11. .76 C
ATOM 1142 CB MET A 156 89 , .457 66 , .771 32. .162 1. , 00 12. .29 c
ATOM 1143 CG MET A 156 90. .355 65 , .578 32. .470 1. , 00 13. .09 c
ATOM 1144 SD MET A 156 89 , .457 64 , .061 32. .800 1. , 00 15. .41 s
ATOM 1145 CE MET A 156 89 , .132 63 , .520 31. .122 1. , 00 13. .69 C
ATOM 1146 C MET A 156 89 , .309 69 , .243 31. .716 1. , 00 11. .04 c
ATOM 1147 O MET A 156 88. .832 69 , .997 32. .595 1. , 00 13. .61 0
ATOM 1148 N LYS A 157 89 , .054 69 , .408 30. .421 1. , 00 9. .98 N
ATOM 1149 CA LYS A 157 88 , .124 70 , .444 29. .967 1. , 00 10. .72 C
ATOM 1150 CB LYS A 157 88. .680 71 , .228 28. .777 1. .00 13. .10 c
ATOM 1151 CG LYS A 157 90 , .049 71 , .891 28. .971 1. , 00 17. .30 C
ATOM 1152 CD LYS A 157 89 , .950 73 , .166 29. .769 1. , 00 17. .60 C
ATOM 1153 CE LYS A 157 91 , .171 74 , .056 29. .568 1. , 00 19. .32 C
ATOM 1154 NZ LYS A 157 92 , .405 73 , .243 29. .573 1. , 00 22. .94 N
ATOM 1155 c LYS A 157 86 , .803 69 , .801 29. .546 1. , 00 10. .51 C
ATOM 1156 0 LYS A 157 86 , .773 68 , .680 28. .992 1. , 00 9. .65 O
ATOM 1157 N LEU A 158 85, .723 70 , .537 29. .773 1. , 00 10. .14 N
ATOM 1158 CA LEU A 158 84 , .393 70 , .116 29. .372 1. , 00 9. .72 c
ATOM 1159 CB LEU A 158 83 , .385 70 , .362 30. .504 1. , 00 11. .39 C
ATOM 1160 CG LEU A 158 82 , .001 69 , .714 30. .336 1. , 00 11. .70 C
ATOM 1161 CD1 LEU A 158 81 , .338 69 , .510 31. .699 1. , 00 9. .61 C
ATOM 1162 CD2 LEU A 158 81 , .110 70 , .545 29. .399 1. , 00 9. .56 c
ATOM 1163 C LEU A 158 83 , .987 70 , .852 28. .118 1. , 00 9. .43 C
ATOM 1164 O LEU A 158 83 , .848 72 , .073 28. .132 1. , 00 9. .84 O
ATOM 1165 N VAL A 159 83 , .811 70 , .095 27. .038 1. , 00 8. .74 N
ATOM 1166 CA VAL A 159 83 , .271 70 , .592 25. .786 1. , 00 8. .60 c
ATOM 1167 CB VAL A 159 83 , .963 69 , .928 24. .552 1. , 00 8. .80 C
ATOM 1168 CGI VAL A 159 83 , .401 70 , .504 23. .248 1. , 00 9. .13 C
ATOM 1169 CG2 VAL A 159 85 ,.473 70., 131 24..591 1., 00 7..66 C
ATOM 1170 C VAL A 159 81 , .758 70. ,345 25. .754 1. , 00 9. .14 C
ATOM 1171 O VAL A 159 81 , .314 69. , 202 25. .730 1. , 00 9. .70 O
ATOM 1172 N LEU A 160 80. .970 71. , 419 25. .764 1. , 00 9. .39 N
ATOM 1173 CA LEU A 160 79 , .514 71. ,306 25. .711 1. , 00 9. .75 C
ATOM 1174 CB LEU A 160 78 , .849 72. ,395 26. .546 1. , 00 8. .89 C
ATOM 1175 CG LEU A 160 77 , .322 72. ,349 26. .614 1. , 00 10. .13 C
ATOM 1176 CD1 LEU A 160 76 , .814 71. , 088 27. .327 1. , 00 8. .66 c
ATOM 1177 CD2 LEU A 160 76 , .850 73. , 588 27. .385 1. , 00 9. .44 C
ATOM 1178 C LEU A 160 79 , .024 71. ,398 24. .264 1. , 00 9. .38 C
ATOM 1179 O LEU A 160 79 , .311 72. ,353 23. .546 1. , 00 9. .29 O
ATOM 1180 N ASP A 161 78 , .311 70. ,375 23. .839 1. , 00 9. .16 N
ATOM 1181 CA ASP A 161 77 , .601 70. ,376 22. .565 1. , 00 8. .48 C
ATOM 1182 CB ASP A 161 77 , .138 68. , 943 22. .333 1. , 00 9. .13 C
ATOM 1183 CG ASP A 161 76 , .791 68. , 627 20. .905 1. , 00 11. .27 C
ATOM 1184 OD1 ASP A 161 76 , .360 69. , 536 20. .151 1. , 00 11. .78 0
ATOM 1185 OD2 ASP A 161 76 , .950 67. , 415 20. .544 1. , 00 11. .60 O
ATOM 1186 C ASP A 161 76 , .399 71. ,311 22. .716 1. , 00 8. .96 C
ATOM 1187 O ASP A 161 75 , .605 71. , 137 23. .664 1. , 00 7. .22 O
ATOM 1188 N ILE A 162 76. .256 72. , 266 21. .796 1. , 00 7. .98 N
ATOM 1189 CA ILE A 162 75 , .198 73. , 244 21. .836 1. , 00 8. .99 C
ATOM 1190 CB ILE A 162 75 , .706 74. , 656 22. .235 1. , 00 10. .59 C
ATOM 1191 CGI ILE A 162 76 , .518 75. ,336 21. .119 1. , 00 10. .61 c
ATOM 1192 CD1 ILE A 162 76. .726 76. , 821 21. .359 1. , 00 12. .11 c
ATOM 1193 CG2 ILE A 162 76 , .572 74. , 586 23. .494 1. , 00 9. .93 C
ATOM 1194 C ILE A 162 74 , .424 73. ,353 20. .530 1. , 00 8. .62 C
ATOM 1195 O ILE A 162 74. .966 73. , 150 19. .438 1. , 00 10. .83 O
ATOM 1196 N VAL A 163 73 , .144 73. , 695 20. .653 1. , 00 7. .41 N
ATOM 1197 CA VAL A 163 72 , .280 73. , 932 19. .499 1. , 00 7. .61 C
ATOM 1198 CB VAL A 163 71 , .066 72. , 997 19. .488 1. , 00 6. .78 C
ATOM 1199 CGI VAL A 163 70 , .202 73. , 251 18. .245 1. , 00 6. .43 c
ATOM 1200 CG2 VAL A 163 71 , .546 71. , 556 19. .529 1. , 00 6. .38 C
ATOM 1201 C VAL A 163 71 , .776 75. ,376 19. .480 1. , 00 8. .62 C
ATOM 1202 O VAL A 163 71 , .347 75. , 908 20. .508 1. , 00 8. .53 O
ATOM 1203 N GLY A 164 71 , .820 75. , 994 18. .301 1. , 00 7. .93 N
ATOM 1204 CA GLY A 164 71 , .130 77. , 274 18. .081 1. , 00 7. .48 C
ATOM 1205 C GLY A 164 70 , .020 77. , 198 17. .051 1. , 00 7. .00 C
ATOM 1206 O GLY A 164 69 , .088 77. , 984 17. .083 1. , 00 7. .88 O
ATOM 1207 N ASN A 165 70 , .110 76. , 252 16. .130 1. , 00 8. .26 N
ATOM 1208 CA ASN A 165 69 , .221 76. , 215 14. .979 1. , 00 8. .67 C
ATOM 1209 CB ASN A 165 69 , .703 75. , 160 13. .969 1. , 00 8. .87 C
ATOM 1210 CG ASN A 165 68 , .745 75. , 002 12. .813 1. , 00 9. .62 c
ATOM 1211 OD1 ASN A 165 68 , .580 75. , 937 12. .041 1. , 00 9. .53 0
ATOM 1212 ND2 ASN A 165 68 , .105 73. , 814 12. .680 1. , 00 7. .11 N
ATOM 1213 C ASN A 165 67, .774 75. , 920 15. .314 1. , 00 8. .35 C
ATOM 1214 O ASN A 165 66., 854 76 ,.413 14..647 1., 00 8..60 O
ATOM 1215 N HIS A 166 67. , 559 75 , .074 16. .313 1. , 00 7. .83 N
ATOM 1216 CA HIS A 166 66. , 229 74 , .506 16. .508 1. , 00 7. .65 C
ATOM 1217 CB HIS A 166 66. .091 73 , .263 15. .628 1. , 00 8. .29 c
ATOM 1218 CG HIS A 166 67. , 172 72 , .236 15. .862 1. , 00 8. .07 C
ATOM 1219 ND1 HIS A 166 68. , 272 72 , .169 15. .098 1. , 00 8. .29 N
ATOM 1220 CE1 HIS A 166 69. , 057 71 , .155 15. .518 1. , 00 7. .93 C
ATOM 1221 NE2 HIS A 166 68. , 455 70 , .563 16. .554 1. , 00 6. .83 N
ATOM 1222 CD2 HIS A 166 67. , 272 71 , .207 16. .785 1. , 00 7. .64 C
ATOM 1223 C HIS A 166 65. , 915 74 , .135 17. .900 1. , 00 8. .08 C
ATOM 1224 O HIS A 166 66. , 823 74 , .019 18. .754 1. , 00 7. .94 O
ATOM 1225 N GLY A 167 64. , 606 74 , .009 18. .154 1. , 00 8. .96 N
ATOM 1226 CA GLY A 167 64. , 071 73 , .474 19. .403 1. , 00 8. .66 C
ATOM 1227 C GLY A 167 63. , 925 71 , .971 19. .236 1. , 00 8. .82 C
ATOM 1228 O GLY A 167 64. , 912 71 , .242 19. .214 1. , 00 9. .81 O
ATOM 1229 N SER A 168 62. , 696 71 , .518 19. .072 1. , 00 8. .21 N
ATOM 1230 CA SER A 168 62. ,386 70 , .101 19. .014 1. , 00 8. .23 C
ATOM 1231 CB SER A 168 61. , 710 69 , .691 20. .325 1. , 00 8. .48 C
ATOM 1232 OG SER A 168 60. , 578 70 , .497 20. .627 1. , 00 8. .24 O
ATOM 1233 C SER A 168 61. , 476 69 , .829 17. .829 1. , 00 8. .48 c
ATOM 1234 O SER A 168 60. , 941 70 , .772 17. .218 1. , 00 9. .27 O
ATOM 1235 N PRO A 169 61. , 282 68 , .546 17. .478 1. , 00 8. .51 N
ATOM 1236 CA PRO A 169 60. ,333 68 , .214 16. .433 1. , 00 8. .01 c
ATOM 1237 CB PRO A 169 60. , 291 66 , .691 16. .485 1. , 00 8. .56 c
ATOM 1238 CG PRO A 169 61. , 668 66 , .311 16. .939 1. , 00 8. .29 C
ATOM 1239 CD PRO A 169 62. , 009 67, .360 17. .955 1. , 00 8. .76 C
ATOM 1240 C PRO A 169 58. , 973 68 , .856 16. .740 1. , 00 8. .71 c
ATOM 1241 O PRO A 169 58. , 507 68 , .782 17. .875 1. , 00 9. .30 O
ATOM 1242 N ALA A 170 58. ,350 69 , .477 15. .742 1. , 00 8. .29 N
ATOM 1243 CA ALA A 170 57. ,331 70 , .516 16. .016 1. , 00 8. .65 C
ATOM 1244 CB ALA A 170 57. , 906 71 , .887 15. .643 1. , 00 7. .77 c
ATOM 1245 C ALA A 170 55. , 975 70 , .320 15. .342 1. , 00 8. .72 C
ATOM 1246 O ALA A 170 55. , 021 71 , .017 15. .691 1. , 00 11. .53 O
ATOM 1247 N TRP A 171 55. , 877 69 , .411 14. .377 1. , 00 9. .44 N
ATOM 1248 CA TRP A 171 54. , 588 69 , .135 13. .712 1. , 00 9. .22 c
ATOM 1249 CB TRP A 171 54. , 250 70 , .198 12. .631 1. , 00 9. .17 C
ATOM 1250 CG TRP A 171 55. , 097 70 , .062 11. .407 1. , 00 10. .08 C
ATOM 1251 CD1 TRP A 171 54. , 828 69 , .330 10. .270 1. , 00 12. .31 C
ATOM 1252 NE1 TRP A 171 55. , 861 69 , .430 9. .371 1. , 00 13. .50 N
ATOM 1253 CE2 TRP A 171 56. , 849 70 , .206 9. .869 1. , 00 11. .29 C
ATOM 1254 CD2 TRP A 171 56. , 434 70 , .628 11. .194 1. , 00 9. .86 C
ATOM 1255 CE3 TRP A 171 57. , 256 71 , .492 11. .914 1. , 00 11. .00 C
ATOM 1256 CZ3 TRP A 171 58. , 503 71 , .842 11. .364 1. , 00 10. .59 c
ATOM 1257 CH2 TRP A 171 58. , 887 71 , .395 10. .106 1. , 00 10. .19 C
ATOM 1258 CZ2 TRP A 171 58. , 063 70 , .580 9. .330 1. , 00 11. .61 C
ATOM 1259 C TRP A 171 54 ,.525 67., 751 13..157 1., 00 8..65 C
ATOM 1260 O TRP A 171 55 , .510 67. , 003 13. .202 1. , 00 10. .22 O
ATOM 1261 N GLY A 172 53 , .344 67. ,384 12. .663 1. , 00 10. .12 N
ATOM 1262 CA GLY A 172 53. .087 66. .056 12. .107 1. , 00 10. .47 C
ATOM 1263 C GLY A 172 52 , .718 65. , 028 13. .153 1. , 00 12. .29 C
ATOM 1264 O GLY A 172 52 , .660 63. , 826 12. .858 1. , 00 12. .87 O
ATOM 1265 N MET A 173 52 , .471 65. , 491 14. .374 1. , 00 12. .39 N
ATOM 1266 CA MET A 173 52 , .213 64. , 600 15. .497 1. , 00 11. .92 c
ATOM 1267 CB MET A 173 52 , .595 65. , 286 16. .817 1. , 00 11. .23 C
ATOM 1268 CG MET A 173 54 , .105 65. ,396 16. .983 1. , 00 11. .15 C
ATOM 1269 SD MET A 173 54 , .626 66. , 456 18. .324 1. , 00 11. .59 s
ATOM 1270 CE MET A 173 54 , .753 68. , 069 17. .495 1. , 00 10. .82 c
ATOM 1271 C MET A 173 50 , .739 64. , 172 15. .521 1. , 00 12. .16 C
ATOM 1272 O MET A 173 49 , .894 64. , 888 14. .996 1. , 00 10. .97 O
ATOM 1273 N ALA A 174 50 , .457 63. , 042 16. .178 1. , 00 12. .44 N
ATOM 1274 CA ALA A 174 49 , .095 62. , 508 16. .344 1. , 00 11. .08 c
ATOM 1275 CB ALA A 174 49 , .149 61. , 126 16. .988 1. , 00 11. .37 C
ATOM 1276 C ALA A 174 48 , .192 63. , 417 17. .163 1. , 00 11. .00 C
ATOM 1277 O ALA A 174 46 , .966 63. ,341 17. .051 1. , 00 12. .83 O
ATOM 1278 N PHE A 175 48. .792 64. , 249 18. .001 1. , 00 10. .60 N
ATOM 1279 CA PHE A 175 48 , .102 65. , 258 18. .798 1. , 00 11. .40 C
ATOM 1280 CB PHE A 175 47 , .381 64. , 652 20. .033 1. , 00 12. .77 C
ATOM 1281 CG PHE A 175 48 , .310 64. , 086 21. .087 1. , 00 12. .88 c
ATOM 1282 CD1 PHE A 175 48. .895 62. , 858 20. .924 1. , 00 12. .18 c
ATOM 1283 CE1 PHE A 175 49 , .759 62. ,344 21. .884 1. , 00 14. .01 C
ATOM 1284 CZ PHE A 175 50 , .025 63. , 063 23. .029 1. , 00 11. .54 C
ATOM 1285 CE2 PHE A 175 49. .427 64. , 280 23. .216 1. , 00 12. .37 c
ATOM 1286 CD2 PHE A 175 48 , .574 64. , 797 22. .257 1. , 00 12. .29 C
ATOM 1287 C PHE A 175 49 , .148 66. , 282 19. .200 1. , 00 11. .29 C
ATOM 1288 O PHE A 175 50 , .334 66. , 092 18. .933 1. , 00 11. .82 O
ATOM 1289 N ASP A 176 48 , .739 67. ,371 19. .840 1. , 00 10. .78 N
ATOM 1290 CA ASP A 176 49 , .675 68. , 429 20. .157 1. , 00 11. .84 C
ATOM 1291 CB ASP A 176 48 , .935 69. , 767 20. .323 1. , 00 13. .37 C
ATOM 1292 CG ASP A 176 49 , .875 70. , 965 20. .549 1. , 00 13. .44 C
ATOM 1293 OD1 ASP A 176 51 , .093 70. , 805 20. .750 1. , 00 12. .54 0
ATOM 1294 OD2 ASP A 176 49 , .363 72. , 102 20. .571 1. , 00 15. .40 O
ATOM 1295 C ASP A 176 50 , .460 68. , 026 21. .387 1. , 00 12. .02 C
ATOM 1296 O ASP A 176 50 , .057 68. ,314 22. .500 1. , 00 12. .00 O
ATOM 1297 N GLN A 177 51 , .591 67. ,349 21. .188 1. , 00 10. .47 N
ATOM 1298 CA GLN A 177 52 , .286 66. , 701 22. .328 1. , 00 10. .73 C
ATOM 1299 CB GLN A 177 53 , .374 65. , 735 21. .845 1. , 00 10. .09 C
ATOM 1300 CG GLN A 177 52 , .861 64. , 538 21. .071 1. , 00 8. .92 c
ATOM 1301 CD GLN A 177 53 , .962 63. , 574 20. .714 1. , 00 8. .65 c
ATOM 1302 OE1 GLN A 177 54 , .292 63. ,396 19. .525 1. , 00 8. .24 O
ATOM 1303 NE2 GLN A 177 54 , .555 62. , 936 21. .744 1. , 00 7. .40 N
ATOM 1304 C GLN A 177 52., 918 67 ,, 748 23., 254 1., 00 10..04 C
ATOM 1305 O GLN A 177 53. , 416 68 , , 776 22. , 780 1. , 00 8. .99 O
ATOM 1306 N PRO A 17 8 52. , 925 67, , 480 24. , 564 1. , 00 10. .85 N
ATOM 1307 CA PRO A 17 8 53. .580 68 , .402 25. .484 1. .00 11. .80 C
ATOM 1308 CB PRO A 17 8 53. , 253 67 , , 810 26. .860 1. , 00 13. .58 C
ATOM 1309 CG PRO A 17 8 53. , 050 66 , ,344 26. .589 1. , 00 14. .37 C
ATOM 1310 CD PRO A 17 8 52. , 284 66 , ,367 25. .294 1. , 00 13. .22 C
ATOM 1311 C PRO A 17 8 55. , 100 68 , .471 25. .223 1. , 00 11. .27 c
ATOM 1312 O PRO A 17 8 55. , 711 67, , 511 24. .724 1. , 00 8. .31 O
ATOM 1313 N LYS A 179 55. , 669 69 , , 631 25. .524 1. , 00 11. .67 N
ATOM 1314 CA LYS A 179 57. , 099 69 , , 949 25. .295 1. , 00 11. .52 C
ATOM 1315 CB LYS A 179 58. ,041 69 , .051 26. .142 1. , 00 12. .11 c
ATOM 1316 CG LYS A 179 57. , 918 69 , ,337 27. .632 1. , 00 14. .00 C
ATOM 1317 CD LYS A 179 59. , 064 68 , , 746 28. .457 1. , 00 15. .10 C
ATOM 1318 CE LYS A 179 59. , 028 69 , , 283 29. .879 1. , 00 13. .97 C
ATOM 1319 NZ LYS A 179 60. , 185 68 , .786 30. .671 1. , 00 11. .58 N
ATOM 1320 C LYS A 179 57. , 569 70 , , 059 23. .831 1. , 00 10. .28 C
ATOM 1321 O LYS A 179 58. , 251 71 , , 039 23. .490 1. , 00 10. .46 O
ATOM 1322 N PHE A 18 0 57. , 226 69 , , 099 22. .977 1. , 00 8. .82 N
ATOM 1323 CA PHE A 18 0 57. , 513 69 , .221 21. .557 1. , 00 8. .16 C
ATOM 1324 CB PHE A 18 0 56. , 907 68 , , 055 20. .771 1. , 00 8. .23 C
ATOM 1325 CG PHE A 18 0 57. , 679 66 , , 775 20. .905 1. , 00 7. .35 C
ATOM 1326 CD1 PHE A 18 0 58. , 850 66 , , 573 20. .189 1. , 00 7. .39 c
ATOM 1327 CE1 PHE A 18 0 59. , 571 65 , .398 20. .303 1. , 00 7. .17 C
ATOM 1328 CZ PHE A 18 0 59. , 124 64 , , 405 21. .162 1. , 00 8. .06 C
ATOM 1329 CE2 PHE A 18 0 57. , 961 64 , , 597 21. .890 1. , 00 7. .86 C
ATOM 1330 CD2 PHE A 18 0 57. .252 65 , .780 21. .769 1. .00 7. .55 c
ATOM 1331 C PHE A 18 0 56. , 983 70 , , 550 21. .006 1. , 00 8. .91 C
ATOM 1332 O PHE A 18 0 55. , 851 70 , , 962 21. .298 1. , 00 8. .93 O
ATOM 1333 N GLY A 18 1 57. , 811 71 , , 209 20. .209 1. , 00 7. .53 N
ATOM 1334 CA GLY A 18 1 57. , 482 72 , .482 19. .604 1. , 00 8. .62 c
ATOM 1335 C GLY A 18 1 57. , 420 73 , , 673 20. .541 1. , 00 9. .53 C
ATOM 1336 O GLY A 18 1 56. , 997 74 , , 763 20. .130 1. , 00 11. .40 O
ATOM 1337 N LYS A 18 2 57. , 833 73 , , 489 21. .794 1. , 00 9. .58 N
ATOM 1338 CA LYS A 18 2 57. , 624 74 , .521 22. .815 1. , 00 9. .35 c
ATOM 1339 CB LYS A 18 2 56. , 674 74 , , 002 23. .912 1. , 00 10. .05 C
ATOM 1340 CG LYS A 182 55. , 212 73 , , 865 23. .448 1. , 00 10. .48 C
ATOM 1341 CD LYS A 18 2 54. , 618 72 , , 529 23. .903 1. , 00 12. .92 C
ATOM 1342 CE LYS A 18 2 53. , 209 72 , .283 23. .374 1. , 00 12. .16 c
ATOM 1343 NZ LYS A 18 2 53. , 207 72 , , 193 21. .891 1. , 00 11. .08 N
ATOM 1344 C LYS A 182 58. , 952 74 , , 952 23. .408 1. , 00 8. .81 C
ATOM 1345 O LYS A 18 2 59. , 846 74 , , 140 23. .565 1. , 00 9. .49 O
ATOM 1346 N ILE A 18 3 59. , 077 76 , .241 23. .709 1. , 00 8. .75 N
ATOM 1347 CA ILE A 18 3 60. , 255 76 , , 777 24. .346 1. , 00 9. .45 C
ATOM 1348 CB ILE A 18 3 61. , 083 77 , , 623 23. .357 1. , 00 12. .55 C
ATOM 1349 CGI ILE A 183 61 ,, 710 76 ,, 659 22., 331 1., 00 15.,65 C
ATOM 1350 CD1 ILE A 183 62 , , 529 77 , , 286 21. ,266 1. , 00 22. , 71 C
ATOM 1351 CG2 ILE A 183 62 , , 171 78 , , 408 24. , 090 1. , 00 10. , 48 C
ATOM 1352 C ILE A 183 59. .853 77 , .545 25. .605 1. .00 9. .68 c
ATOM 1353 O ILE A 183 58 , , 910 78 , , 353 25. , 586 1. , 00 9. , 02 0
ATOM 1354 N TYR A 184 60 , , 577 77 , , 257 26. ,692 1. , 00 8. ,94 N
ATOM 1355 CA TYR A 184 60 , ,332 77 , , 841 27. ,983 1. , 00 10. , 44 C
ATOM 1356 CB TYR A 184 59 , .983 76 , .733 29. .003 1. , 00 10. .93 c
ATOM 1357 CG TYR A 184 58 , , 704 75 , , 963 28. ,685 1. , 00 10. , 76 C
ATOM 1358 CD1 TYR A 184 58 , , 718 74 , , 862 27. , 841 1. , 00 11. ,50 C
ATOM 1359 CE1 TYR A 184 57 , , 567 74 , , 145 27. ,562 1. , 00 10. ,91 C
ATOM 1360 CZ TYR A 184 56 , .363 74 , .538 28. .126 1. , 00 12. .16 c
ATOM 1361 OH TYR A 184 55 , , 193 73 , , 849 27. , 840 1. , 00 15. ,56 O
ATOM 1362 CE2 TYR A 184 56 , ,317 75 , , 621 28. ,978 1. , 00 12. , 75 C
ATOM 1363 CD2 TYR A 184 57 , , 486 76 , ,325 29. ,256 1. , 00 12. ,17 C
ATOM 1364 C TYR A 184 61 , .548 78 , .611 28. .467 1. , 00 10. .26 c
ATOM 1365 O TYR A 184 62 , , 674 78 , , 217 28. ,213 1. , 00 9. ,38 O
ATOM 1366 N ASP A 185 61 , ,318 79 , , 683 29. , 229 1. , 00 14. ,31 N
ATOM 1367 CA ASP A 185 62 , , 446 80 , , 446 29. , 803 1. , 00 18. , 13 c
ATOM 1368 CB ASP A 185 62. .165 81 , .960 29. .817 1. , 00 22. .31 c
ATOM 1369 CG ASP A 185 61 , , 126 82 , ,381 30. , 826 1. , 00 27. ,88 C
ATOM 1370 OD1 ASP A 185 60 , , 775 81 , , 621 31. , 779 1. , 00 27. ,23 O
ATOM 1371 OD2 ASP A 185 60 , , 649 83 , , 518 30. ,638 1. , 00 39. ,07 0
ATOM 1372 C ASP A 185 62. .897 79 , .907 31. .149 1. , 00 19. .78 c
ATOM 1373 O ASP A 185 62 , ,370 78 , , 883 31. ,635 1. , 00 15. ,62 O
ATOM 1374 N LYS A 186 63 , , 923 80 , , 538 31. , 720 1. , 00 26. , 05 N
ATOM 1375 CA LYS A 186 64. .474 80 , .086 33. .009 1. .00 28. .37 c
ATOM 1376 CB LYS A 186 65 , , 671 80 , , 947 33. , 458 1. , 00 37. ,38 C
ATOM 1377 CG LYS A 186 65, , 436 82 , , 462 33. , 452 1. , 00 50. , 77 C
ATOM 1378 CD LYS A 186 66 , , 739 83 , , 253 33. ,329 1. , 00 52. ,62 C
ATOM 1379 CE LYS A 186 66 , .503 84 , .664 32. .793 1. , 00 54. .21 c
ATOM 1380 NZ LYS A 186 67, , 681 85, , 166 32. , 024 1. , 00 56. , 86 N
ATOM 1381 C LYS A 186 63 , ,376 80 , , 052 34. , 091 1. , 00 31. ,29 C
ATOM 1382 O LYS A 186 63 , ,306 79 , , 088 34. , 838 1. , 00 29. , 02 O
ATOM 1383 N ASP A 187 62 , .486 81 , .053 34. .111 1. , 00 29. .44 N
ATOM 1384 CA ASP A 187 61 , ,306 81 , , 057 35. , 044 1. , 00 32. ,56 C
ATOM 1385 CB ASP A 187 60 , , 494 82 , , 354 34. ,913 1. , 00 33. ,92 C
ATOM 1386 CG ASP A 187 61 , , 298 83 , , 577 35. , 190 1. , 00 40. , 40 C
ATOM 1387 OD1 ASP A 187 62 , .181 83 , .493 36. .066 1. , 00 39. .07 0
ATOM 1388 OD2 ASP A 187 61 , , 048 84, , 617 34. , 527 1. , 00 50. ,00 O
ATOM 1389 C ASP A 187 60 , , 290 79 , , 935 34. , 839 1. , 00 29. , 49 C
ATOM 1390 O ASP A 187 59 , ,374 79 , , 807 35. ,625 1. , 00 37. ,65 0
ATOM 1391 N GLY A 188 60 , .404 79 , .164 33. .763 1. , 00 25. .14 N
ATOM 1392 CA GLY A 188 59 , ,374 78 , , 211 33. ,389 1. , 00 17. , 75 C
ATOM 1393 C GLY A 188 58 , , 222 78 , , 783 32. , 561 1. , 00 16. , 77 C
ATOM 1394 O GLY A 188 57., 274 78 ,, 068 32.,279 1., 00 17., 77 O
ATOM 1395 N THR A 189 58. ,312 80 , , 039 32. , 130 1. , 00 16. , 40 N
ATOM 1396 CA THR A 189 57. , 295 80 , , 646 31. ,277 1. , 00 17. ,50 C
ATOM 1397 CB THR A 189 57. .446 82. , 185 31. .246 1. , 00 19. .57 c
ATOM 1398 OG1 THR A 189 57. , 549 82 , , 670 32. , 586 1. , 00 21. , 09 O
ATOM 1399 CG2 THR A 189 56. , 249 82 , , 862 30. , 543 1. , 00 18. ,14 C
ATOM 1400 C THR A 189 57. ,374 80 , , 134 29. , 841 1. , 00 16. ,32 C
ATOM 1401 O THR A 189 58. , 469 80. , 021 29. .272 1. , 00 14. .35 0
ATOM 1402 N LEU A 190 56. , 211 79 , , 833 29. , 262 1. , 00 13. ,94 N
ATOM 1403 CA LEU A 190 56. , 107 79 , , 440 27. , 863 1. , 00 13. ,04 C
ATOM 1404 CB LEU A 190 54. , 701 78 , , 935 27. , 550 1. , 00 12. ,37 C
ATOM 1405 CG LEU A 190 54. , 466 78. , 433 26. .121 1. , 00 12. .62 c
ATOM 1406 CD1 LEU A 190 55. , 259 77 , , 176 25. , 810 1. , 00 10. , 45 C
ATOM 1407 CD2 LEU A 190 52. , 954 78 , , 196 25. ,942 1. , 00 14. ,52 C
ATOM 1408 C LEU A 190 56. ,359 80 , , 656 27. , 018 1. , 00 13. ,62 C
ATOM 1409 O LEU A 190 55. , 620 81. ,614 27. .103 1. , 00 11. .26 0
ATOM 1410 N ILE A 191 57. ,377 80 , , 619 26. , 171 1. , 00 12. ,52 N
ATOM 1411 CA ILE A 191 57. , 625 81 , , 744 25. ,309 1. , 00 12. , 76 C
ATOM 1412 CB ILE A 191 58. , 966 82 , , 411 25. ,643 1. , 00 14. ,33 C
ATOM 1413 CGI ILE A 191 60. , 124 81. , 455 25. .382 1. , 00 13. .90 c
ATOM 1414 CD1 ILE A 191 61. , 495 82 , , 098 25. ,563 1. , 00 13. , 42 C
ATOM 1415 CG2 ILE A 191 58. , 952 82 , , 952 27. , 090 1. , 00 14. , 41 C
ATOM 1416 C ILE A 191 57. , 519 81 , , 415 23. , 804 1. , 00 11. ,52 c
ATOM 1417 O ILE A 191 57. , 552 82. , 331 22. .974 1. , 00 13. .36 O
ATOM 1418 N ALA A 192 57. , 401 80 , , 138 23. , 433 1. , 00 11. , 55 N
ATOM 1419 CA ALA A 192 57. , 132 79 , , 779 22. , 010 1. , 00 9. ,87 C
ATOM 1420 CB ALA A 192 58. , 435 79. , 657 21. .198 1. , 00 8. .37 c
ATOM 1421 C ALA A 192 56. , 356 78 , , 483 21. ,973 1. , 00 10. , 48 C
ATOM 1422 O ALA A 192 56. , 614 77 , , 605 22. , 785 1. , 00 11. ,24 O
ATOM 1423 N ASP A 193 55. ,386 78 , ,370 21. , 058 1. , 00 9. ,96 N
ATOM 1424 CA ASP A 193 54. , 649 77. , 122 20. .892 1. , 00 9. .77 c
ATOM 1425 CB ASP A 193 53. , 413 77 , , 120 21. , 804 1. , 00 10. , 43 C
ATOM 1426 CG ASP A 193 52. , 706 75 , , 778 21. , 851 1. , 00 11. , 12 C
ATOM 1427 OD1 ASP A 193 53. , 020 74 , , 840 21. , 061 1. , 00 10. ,11 O
ATOM 1428 OD2 ASP A 193 51. , 810 75. , 656 22. .710 1. , 00 10. .10 0
ATOM 1429 C ASP A 193 54. , 235 76 , , 941 19. , 436 1. , 00 9. , 82 C
ATOM 1430 O ASP A 193 53. ,372 77 , , 659 18. ,946 1. , 00 8. ,98 O
ATOM 1431 N HIS A 194 54. , 844 75 , , 954 18. , 776 1. , 00 9. ,00 N
ATOM 1432 CA HIS A 194 54. , 589 75. , 627 17. .367 1. , 00 9. .00 c
ATOM 1433 CB HIS A 194 55. , 705 74 , , 662 16. , 876 1. , 00 7. ,91 C
ATOM 1434 CG HIS A 194 56. , 261 74 , , 949 15. , 488 1. , 00 8. , 06 C
ATOM 1435 ND1 HIS A 194 55. , 640 74 , , 557 14. ,337 1. , 00 8. ,17 N
ATOM 1436 CE1 HIS A 194 56. , 421 74. , 870 13. .288 1. , 00 7. .83 c
ATOM 1437 NE2 HIS A 194 57. , 551 75 , , 418 13. , 754 1. , 00 7. ,69 N
ATOM 1438 CD2 HIS A 194 57. , 483 75 , , 492 15. , 099 1. , 00 7. , 85 C
ATOM 1439 C HIS A 194 53 ,.219 74., 975 17..173 1., 00 9..75 C
ATOM 1440 O HIS A 194 52 , .753 74. , 819 16. .023 1. , 00 8. .97 O
ATOM 1441 N GLN A 195 52 , .578 74. , 573 18. .279 1. , 00 9. .77 N
ATOM 1442 CA GLN A 195 51. .189 74. .077 18. .293 1. , 00 10. .69 C
ATOM 1443 CB GLN A 195 50 , .215 75. , 248 18. .106 1. , 00 11. .53 C
ATOM 1444 CG GLN A 195 50 , .263 76. , 249 19. .232 1. , 00 12. .30 C
ATOM 1445 CD GLN A 195 49 , .330 77. , 417 18. .999 1. , 00 17. .08 C
ATOM 1446 OE1 GLN A 195 49 , .424 78. , 121 17. .999 1. , 00 14. .58 O
ATOM 1447 NE2 GLN A 195 48 , .425 77. , 632 19. .943 1. , 00 19. .44 N
ATOM 1448 C GLN A 195 50 , .893 72. , 973 17. .259 1. , 00 10. .35 C
ATOM 1449 O GLN A 195 49 , .801 72. , 906 16. .699 1. , 00 10. .47 O
ATOM 1450 N ASN A 196 51 , .887 72. , 142 16. .968 1. , 00 9. .63 N
ATOM 1451 CA ASN A 196 51 , .719 71. , 044 16. .024 1. , 00 10. .05 C
ATOM 1452 CB ASN A 196 50 , .759 69. , 975 16. .593 1. , 00 9. .67 C
ATOM 1453 CG ASN A 196 50 , .842 68. , 643 15. .861 1. , 00 9. .67 C
ATOM 1454 OD1 ASN A 196 51 , .900 68. , 263 15. .393 1. , 00 9. .77 0
ATOM 1455 ND2 ASN A 196 49 , .725 67. , 892 15. .827 1. , 00 7. .83 N
ATOM 1456 C ASN A 196 51 , .242 71. , 539 14. .668 1. , 00 9. .69 C
ATOM 1457 O ASN A 196 50 , .549 70. , 824 13. .975 1. , 00 10. .02 O
ATOM 1458 N LEU A 197 51. .639 72. , 760 14. .299 1. , 00 10. .69 N
ATOM 1459 CA LEU A 197 51 , .375 73. ,325 12. .978 1. , 00 10. .53 C
ATOM 1460 CB LEU A 197 50 , .912 74. , 779 13. .079 1. , 00 10. .60 C
ATOM 1461 CG LEU A 197 49 , .576 75. , 044 13. .780 1. , 00 12. .87 c
ATOM 1462 CD1 LEU A 197 49. .349 76. , 556 14. .059 1. , 00 11. .44 c
ATOM 1463 CD2 LEU A 197 48 , .412 74. , 409 13. .000 1. , 00 11. .47 C
ATOM 1464 C LEU A 197 52 , .642 73. , 290 12. .134 1. , 00 10. .25 C
ATOM 1465 O LEU A 197 53. .721 73. .555 12. .634 1. , 00 10. .13 O
ATOM 1466 N PRO A 198 52 , .519 72. , 944 10. .845 1. , 00 11. .95 N
ATOM 1467 CA PRO A 198 53 , .680 73. , 161 9. .967 1. , 00 12. .95 C
ATOM 1468 CB PRO A 198 53 , .197 72. , 721 8. .572 1. , 00 15. .22 C
ATOM 1469 CG PRO A 198 51 , .746 72. , 443 8. .688 1. , 00 17. .25 c
ATOM 1470 CD PRO A 198 51 , .293 72. , 566 10. .124 1. , 00 14. .07 C
ATOM 1471 C PRO A 198 54 , .091 74. , 651 9. .970 1. , 00 11. .84 C
ATOM 1472 O PRO A 198 53 , .246 75. , 521 10. .237 1. , 00 8. .86 O
ATOM 1473 N PRO A 199 55 , .379 74. ,944 9. .694 1. , 00 11. .71 N
ATOM 1474 CA PRO A 199 55 , .835 76. ,339 9. .814 1. , 00 11. .73 C
ATOM 1475 CB PRO A 199 57 , .299 76. , 270 9. .342 1. , 00 11. .77 C
ATOM 1476 CG PRO A 199 57 , .716 74. , 844 9. .631 1. , 00 11. .53 C
ATOM 1477 CD PRO A 199 56 , .482 74. , 040 9. .304 1. , 00 11. .79 c
ATOM 1478 C PRO A 199 55 , .025 77. ,334 8. .969 1. , 00 10. .05 C
ATOM 1479 O PRO A 199 54 , .747 78. , 446 9. .418 1. , 00 11. .19 O
ATOM 1480 N GLN A 200 54 , .603 76. , 904 7. .791 1. , 00 9. .70 N
ATOM 1481 CA GLN A 200 53 , .879 77. , 769 6. .872 1. , 00 11. .05 c
ATOM 1482 CB GLN A 200 53 , .707 77. , 093 5. .498 1. , 00 12. .75 C
ATOM 1483 CG GLN A 200 55 , .007 76. , 653 4. .831 1. , 00 14. .55 C
ATOM 1484 CD GLN A 200 55 ,.345 75., 174 5..079 1., 00 18..94 C
ATOM 1485 OE1 GLN A 200 55 , .220 74. , 657 6. .199 1. , 00 15. .92 O
ATOM 1486 NE2 GLN A 200 55 , .787 74. , 495 4. .025 1. , 00 17. .31 N
ATOM 1487 C GLN A 200 52. .513 78. .141 7. .419 1. , 00 10. .65 C
ATOM 1488 O GLN A 200 51 , .916 79. , 067 6. .934 1. , 00 11. .06 O
ATOM 1489 N GLN A 201 52 , .005 77. , 405 8. .399 1. , 00 9. .86 N
ATOM 1490 CA GLN A 201 50 , .724 77. , 740 9. .034 1. , 00 11. .01 C
ATOM 1491 CB GLN A 201 49 , .902 76. , 471 9. .311 1. , 00 11. .55 c
ATOM 1492 CG GLN A 201 49 , .381 75. , 828 8. .015 1. , 00 14. .28 C
ATOM 1493 CD GLN A 201 48 , .673 74. , 499 8. .267 1. , 00 16. .74 C
ATOM 1494 OE1 GLN A 201 47 , .979 74. ,334 9. .252 1. , 00 18. .13 O
ATOM 1495 NE2 GLN A 201 48 , .831 73. , 573 7. .358 1. , 00 18. .73 N
ATOM 1496 C GLN A 201 50 , .851 78. , 568 10. .306 1. , 00 11. .88 C
ATOM 1497 O GLN A 201 49 , .840 78. , 931 10. .899 1. , 00 11. .87 O
ATOM 1498 N LEU A 202 52 , .069 78. , 886 10. .738 1. , 00 11. .95 N
ATOM 1499 CA LEU A 202 52 , .211 79. , 762 11. .907 1. , 00 11. .03 c
ATOM 1500 CB LEU A 202 53 , .628 79. , 742 12. .446 1. , 00 10. .09 C
ATOM 1501 CG LEU A 202 54 , .211 78. ,382 12. .856 1. , 00 9. .78 C
ATOM 1502 CD1 LEU A 202 55 , .682 78. , 615 13. .183 1. , 00 8. .27 c
ATOM 1503 CD2 LEU A 202 53. .411 77. , 789 14. .029 1. , 00 8. .33 c
ATOM 1504 C LEU A 202 51 , .829 81. , 200 11. .527 1. , 00 12. .76 C
ATOM 1505 O LEU A 202 51 , .837 81. , 545 10. .374 1. , 00 11. .37 O
ATOM 1506 N ASP A 203 51 , .531 82. , 032 12. .519 1. , 00 13. .21 N
ATOM 1507 CA ASP A 203 51. .093 83. , 420 12. .303 1. , 00 14. .72 c
ATOM 1508 CB ASP A 203 49 , .574 83. , 457 12. .540 1. , 00 17. .99 C
ATOM 1509 CG ASP A 203 48 , .937 84. , 831 12. .271 1. , 00 22. .41 C
ATOM 1510 OD1 ASP A 203 49. .585 85. , 761 11. .748 1. , 00 21. .46 O
ATOM 1511 OD2 ASP A 203 47 , .760 84. , 946 12. .610 1. , 00 24. .92 O
ATOM 1512 C ASP A 203 51 , .844 84. ,366 13. .276 1. , 00 14. .34 C
ATOM 1513 O ASP A 203 51 , .250 84. , 909 14. .183 1. , 00 14. .43 O
ATOM 1514 N PRO A 204 53 , .168 84. , 519 13. .118 1. , 00 13. .08 N
ATOM 1515 CA PRO A 204 53 , .934 85. , 355 14. .056 1. , 00 15. .31 C
ATOM 1516 CB PRO A 204 55 , .368 85. ,309 13. .517 1. , 00 16. .29 C
ATOM 1517 CG PRO A 204 55 , .287 84. , 726 12. .134 1. , 00 14. .61 C
ATOM 1518 CD PRO A 204 53 , .999 83. , 940 12. .062 1. , 00 14. .12 c
ATOM 1519 C PRO A 204 53 , .457 86. , 801 14. .135 1. , 00 19. .16 C
ATOM 1520 O PRO A 204 53 , .633 87. , 432 15. .161 1. , 00 20. .94 O
ATOM 1521 N GLU A 205 52 , .850 87. ,317 13. .077 1. , 00 19. .31 N
ATOM 1522 CA GLU A 205 52 , .336 88. , 683 13. .112 1. , 00 23. .12 c
ATOM 1523 CB GLU A 205 51 , .748 89. , 103 11. .755 1. , 00 26. .07 C
ATOM 1524 CG GLU A 205 51 , .411 90. , 595 11. .702 1. , 00 37. .73 C
ATOM 1525 CD GLU A 205 50 , .472 90. , 976 10. .562 1. , 00 48. .65 c
ATOM 1526 OE1 GLU A 205 50 , .573 90. , 397 9. .460 1. , 00 50. .75 0
ATOM 1527 OE2 GLU A 205 49 , .629 91. , 872 10. .768 1. , 00 61. .08 O
ATOM 1528 C GLU A 205 51 , .286 88. , 836 14. .210 1. , 00 21. .84 C
ATOM 1529 O GLU A 205 51., 289 89 ,, 818 14., 920 1., 00 22.,08 O
ATOM 1530 N HIS A 206 50. , 412 87, , 857 14. ,390 1. , 00 21. , 55 N
ATOM 1531 CA HIS A 206 49. ,335 87, , 998 15. ,374 1. , 00 22. ,21 C
ATOM 1532 CB HIS A 206 47. .983 87 , .660 14. .747 1. , 00 23. .84 c
ATOM 1533 CG HIS A 206 47. , 680 88 , , 471 13. , 501 1. , 00 32. , 48 C
ATOM 1534 ND1 HIS A 206 47. , 743 87, , 949 12. ,258 1. , 00 33. ,64 N
ATOM 1535 CE1 HIS A 206 47. , 441 88 , , 906 11. ,353 1. , 00 35. ,01 C
ATOM 1536 NE2 HIS A 206 47. , 192 90 , .050 12. .022 1. , 00 34. .14 N
ATOM 1537 CD2 HIS A 206 47. ,330 89 , , 822 13. ,344 1. , 00 33. ,21 C
ATOM 1538 C HIS A 206 49. , 536 87, , 202 16. ,633 1. , 00 27. ,52 C
ATOM 1539 O HIS A 206 48. , 958 87, , 554 17. ,653 1. , 00 24. ,65 O
ATOM 1540 N ASN A 207 50. ,342 86 , .136 16. .580 1. , 00 20. .01 N
ATOM 1541 CA ASN A 207 50. , 517 85, , 228 17. , 710 1. , 00 17. ,23 C
ATOM 1542 CB ASN A 207 50. , 225 83 , , 799 17. ,280 1. , 00 17. , 75 C
ATOM 1543 CG ASN A 207 50. , 035 82 , , 879 18. , 449 1. , 00 21. ,33 C
ATOM 1544 OD1 ASN A 207 50. , 565 83 , .111 19. .566 1. , 00 21. .62 0
ATOM 1545 ND2 ASN A 207 49. , 243 81 , , 842 18. ,230 1. , 00 20. ,26 N
ATOM 1546 C ASN A 207 51. , 934 85, , 321 18. ,256 1. , 00 16. ,81 C
ATOM 1547 O ASN A 207 52. , 878 84, , 831 17. ,625 1. , 00 14. , 76 O
ATOM 1548 N PRO A 208 52. , 092 85 , .955 19. .426 1. , 00 17. .46 N
ATOM 1549 CA PRO A 208 53. ,398 86 , , 096 20. , 062 1. , 00 16. ,37 C
ATOM 1550 CB PRO A 208 53. , 059 86 , , 721 21. , 430 1. , 00 17. ,56 C
ATOM 1551 CG PRO A 208 51. , 778 87, , 451 21. ,201 1. , 00 19. , 70 c
ATOM 1552 CD PRO A 208 51. , 018 86 , .569 20. .249 1. , 00 19. .67 c
ATOM 1553 C PRO A 208 54. , 148 84, , 783 20. ,260 1. , 00 14. , 70 C
ATOM 1554 O PRO A 208 55. ,377 84, , 790 20. ,212 1. , 00 13. , 89 O
ATOM 1555 N LEU A 209 53. , 420 83 , .682 20. .488 1. , 00 12. .12 N
ATOM 1556 CA LEU A 209 54. , 036 82 , , 365 20. ,685 1. , 00 13. , 53 C
ATOM 1557 CB LEU A 209 53. , 064 81 , ,391 21. ,376 1. , 00 14. , 74 C
ATOM 1558 CG LEU A 209 52. , 626 81 , , 776 22. , 798 1. , 00 16. , 44 C
ATOM 1559 CD1 LEU A 209 51. , 472 80 , .887 23. .283 1. , 00 15. .18 c
ATOM 1560 CD2 LEU A 209 53. , 821 81 , , 684 23. , 738 1. , 00 15. ,92 C
ATOM 1561 C LEU A 209 54. , 607 81 , , 731 19. ,399 1. , 00 10. ,98 C
ATOM 1562 O LEU A 209 55. , 238 80 , , 671 19. , 468 1. , 00 9. ,53 O
ATOM 1563 N HIS A 210 54. , 429 82 , .387 18. .254 1. , 00 9. .36 N
ATOM 1564 CA HIS A 210 55. , 040 81 , .921 16. ,998 1. , 00 10. ,30 C
ATOM 1565 CB HIS A 210 54. , 016 81 , .943 15. , 859 1. , 00 10. ,35 C
ATOM 1566 CG HIS A 210 52. , 830 81 , .038 16. , 068 1. , 00 9. , 77 C
ATOM 1567 ND1 HIS A 210 51. , 738 81 , .062 15. .241 1. , 00 8. .73 N
ATOM 1568 CE1 HIS A 210 50. , 851 80 , .135 15. ,660 1. , 00 9. ,01 C
ATOM 1569 NE2 HIS A 210 51. ,343 79 , .547 16. , 755 1. , 00 10. , 12 N
ATOM 1570 CD2 HIS A 210 52. , 563 80 , .072 17. , 037 1. , 00 9. , 06 C
ATOM 1571 C HIS A 210 56. , 260 82 , .706 16. .581 1. , 00 9. .75 c
ATOM 1572 O HIS A 210 57. , 000 82 , .278 15. , 713 1. , 00 8. ,60 O
ATOM 1573 N ARG A 211 56. , 525 83 , .834 17. , 223 1. , 00 11. , 42 N
ATOM 1574 CA ARG A 211 57 ,.590 84 ,, 752 16., 792 1., 00 10., 78 C
ATOM 1575 CB ARG A 211 57 , .474 86 , , 090 17. , 544 1. , 00 13. ,67 C
ATOM 1576 CG ARG A 211 56 , .265 86 , , 842 17. , 093 1. , 00 19. ,18 C
ATOM 1577 CD ARG A 211 55. .967 88. .144 17. .845 1. .00 23. .33 c
ATOM 1578 NE ARG A 211 54 , .614 88 , , 564 17. , 454 1. , 00 26. ,69 N
ATOM 1579 CZ ARG A 211 53 , .811 89 , ,382 18. , 138 1. , 00 30. ,08 C
ATOM 1580 NH1 ARG A 211 54 , .185 89 , , 935 19. ,293 1. , 00 25. , 05 N
ATOM 1581 NH2 ARG A 211 52 , .607 89 , .639 17. .655 1. , 00 29. .36 N
ATOM 1582 C ARG A 211 59 , .013 84, , 238 16. ,936 1. , 00 9. ,58 C
ATOM 1583 O ARG A 211 59 , .919 84, , 750 16. , 294 1. , 00 9. , 05 O
ATOM 1584 N PHE A 212 59 , .216 83 , , 243 17. , 785 1. , 00 8. , 15 N
ATOM 1585 CA PHE A 212 60 , .494 82 , .589 17. .875 1. , 00 8. .77 C
ATOM 1586 CB PHE A 212 60 , .556 81 , , 688 19. , 124 1. , 00 9. ,63 C
ATOM 1587 CG PHE A 212 61 , .284 82 , ,307 20. ,272 1. , 00 11. ,98 C
ATOM 1588 CD1 PHE A 212 60 , .969 83 , , 593 20. , 694 1. , 00 14. , 73 C
ATOM 1589 CE1 PHE A 212 61 , .628 84 , .176 21. .772 1. , 00 18. .02 c
ATOM 1590 CZ PHE A 212 62 , .630 83 , , 476 22. , 414 1. , 00 15. ,52 C
ATOM 1591 CE2 PHE A 212 62 , .960 82 , , 206 21. ,997 1. , 00 14. , 74 C
ATOM 1592 CD2 PHE A 212 62 , .293 81 , , 619 20. ,926 1. , 00 15. , 43 C
ATOM 1593 C PHE A 212 60. .831 81 , .767 16. .638 1. , 00 7. .80 C
ATOM 1594 O PHE A 212 61 , .956 81 , ,358 16. , 470 1. , 00 7. ,92 O
ATOM 1595 N TYR A 213 59 , .873 81 , , 544 15. , 761 1. , 00 7. ,63 N
ATOM 1596 CA TYR A 213 60 , .067 80 , , 552 14. , 714 1. , 00 7. , 71 c
ATOM 1597 CB TYR A 213 58. .986 79 , .492 14. .842 1. , 00 7. .51 C
ATOM 1598 CG TYR A 213 58 , .994 78 , , 744 16. , 148 1. , 00 7. ,14 C
ATOM 1599 CD1 TYR A 213 60 , .172 78 , , 236 16. ,683 1. , 00 7. , 13 C
ATOM 1600 CE1 TYR A 213 60. .172 77 , .513 17. .868 1. .00 8. .13 c
ATOM 1601 CZ TYR A 213 58 , .972 77 , , 293 18. , 533 1. , 00 7. , 47 C
ATOM 1602 OH TYR A 213 58 , .958 76 , , 587 19. , 709 1. , 00 6. ,99 O
ATOM 1603 CE2 TYR A 213 57 , .789 77 , , 796 18. , 018 1. , 00 6. , 83 C
ATOM 1604 CD2 TYR A 213 57 , .803 78 , .511 16. .844 1. , 00 7. .07 c
ATOM 1605 C TYR A 213 60 , .031 81 , , 127 13. ,307 1. , 00 8. ,50 C
ATOM 1606 O TYR A 213 59 , .257 82 , , 038 13. , 008 1. , 00 7. ,98 O
ATOM 1607 N ASN A 214 60 , .852 80 , , 552 12. , 428 1. , 00 7. ,88 N
ATOM 1608 CA ASN A 214 60 , .750 80 , .821 10. .997 1. , 00 7. .95 c
ATOM 1609 CB ASN A 214 62 , .053 80 , , 428 10. ,277 1. , 00 7. , 75 C
ATOM 1610 CG ASN A 214 63 , .168 81 , ,377 10. , 587 1. , 00 7. ,21 C
ATOM 1611 OD1 ASN A 214 62 , .932 82 , , 588 10. , 795 1. , 00 6. ,20 O
ATOM 1612 ND2 ASN A 214 64 , .390 80 , .856 10. .613 1. , 00 6. .54 N
ATOM 1613 C ASN A 214 59 , .553 80 , , 050 10. , 429 1. , 00 8. ,37 C
ATOM 1614 O ASN A 214 59 , .093 79 , , 068 11. , 005 1. , 00 6. ,32 O
ATOM 1615 N THR A 215 59 , .020 80 , , 549 9. ,320 1. , 00 8. , 85 N
ATOM 1616 CA THR A 215 57 , .782 80 , .021 8. .767 1. , 00 9. .29 c
ATOM 1617 CB THR A 215 56 , .681 81 , , 100 8. , 798 1. , 00 10. ,96 C
ATOM 1618 OG1 THR A 215 57 , .057 82 , , 131 7. ,918 1. , 00 10. ,00 O
ATOM 1619 CG2 THR A 215 56 ,, 493 81 ,, 694 10., 197 1., 00 10..48 C
ATOM 1620 C THR A 215 57 , , 960 79 , , 522 7. ,335 1. , 00 10. .13 C
ATOM 1621 O THR A 215 56 , , 984 79 , , 432 6. , 578 1. , 00 11. .44 O
ATOM 1622 N VAL A 216 59. .176 79 , .146 6. .963 1. , 00 10. .50 N
ATOM 1623 CA VAL A 216 59 , , 468 78 , , 725 5. , 569 1. , 00 12. .11 C
ATOM 1624 CB VAL A 216 60 , , 973 78 , , 912 5. ,202 1. , 00 13. .76 C
ATOM 1625 CGI VAL A 216 61 , , 295 78 , ,346 3. , 811 1. , 00 12. .99 C
ATOM 1626 CG2 VAL A 216 61 , .364 80 , .405 5. .287 1. , 00 14. .35 c
ATOM 1627 C VAL A 216 59 , , 032 77 , , 279 5. ,310 1. , 00 11. .49 C
ATOM 1628 O VAL A 216 58 , , 495 76 , , 983 4. ,253 1. , 00 13. .28 O
ATOM 1629 N GLY A 217 59 , , 235 76 , ,396 6. ,286 1. , 00 11. .73 N
ATOM 1630 CA GLY A 217 59 , .054 74 , .956 6. .112 1. , 00 11. .01 c
ATOM 1631 C GLY A 217 60 , , 278 74 , ,341 5. , 441 1. , 00 10. .70 C
ATOM 1632 O GLY A 217 61 , , 156 75 , , 050 5. , 041 1. , 00 10. .37 O
ATOM 1633 N PRO A 218 60 , ,323 73 , , 017 5. , 297 1. , 00 13. .52 N
ATOM 1634 CA PRO A 218 61 , .419 72 , .322 4. .616 1. , 00 12. .51 c
ATOM 1635 CB PRO A 218 60 , , 961 70 , , 862 4. ,609 1. , 00 16. .88 C
ATOM 1636 CG PRO A 218 60 , , 072 70 , , 755 5. , 814 1. , 00 17. .49 C
ATOM 1637 CD PRO A 218 59 , ,319 72 , , 067 5. , 822 1. , 00 16. .29 C
ATOM 1638 C PRO A 218 61. .554 72 , .828 3. .207 1. , 00 11. .94 C
ATOM 1639 O PRO A 218 60 , , 555 73 , , 139 2. .573 1. , 00 10. .11 O
ATOM 1640 N VAL A 219 62 , , 781 72 , , 985 2. .741 1. , 00 11. .40 N
ATOM 1641 CA VAL A 219 63 , , 032 73 , , 494 1. .395 1. , 00 11. .36 C
ATOM 1642 CB VAL A 219 63. .920 74 , .756 1. .431 1. , 00 11. .22 C
ATOM 1643 CGI VAL A 219 64 , , 409 75 , , 122 0. .040 1. , 00 9. .95 C
ATOM 1644 CG2 VAL A 219 63 , , 154 75 , , 931 2. .060 1. , 00 8. .78 C
ATOM 1645 C VAL A 219 63. .683 72 , .367 0. .611 1. , 00 11. .48 c
ATOM 1646 O VAL A 219 64 , , 702 71 , , 824 1. .020 1. , 00 12. .35 O
ATOM 1647 N ASP A 220 63 , , 065 72 , , 003 -0. .499 1. , 00 12. .70 N
ATOM 1648 CA ASP A 220 63 , , 620 71 , , 004 -1. .444 1. , 00 14. .09 C
ATOM 1649 CB ASP A 220 62 , .564 70 , .651 -2. .512 1. , 00 19. .09 c
ATOM 1650 CG ASP A 220 63 , , 013 69 , , 504 -3. .422 1. , 00 23. .00 C
ATOM 1651 OD1 ASP A 220 62 , , 514 68 , ,395 -3. .228 1. , 00 33. .98 O
ATOM 1652 OD2 ASP A 220 63 , , 882 69 , , 697 -4. .301 1. , 00 19. .58 O
ATOM 1653 C ASP A 220 64 , .853 71 , .583 -2. .137 1. , 00 12. .59 c
ATOM 1654 O ASP A 220 64 , , 809 72 , , 725 -2. .623 1. , 00 12. .92 O
ATOM 1655 N GLY A 221 65, , 940 70 , , 810 -2. .179 1. , 00 12. .37 N
ATOM 1656 CA GLY A 221 67, , 205 71 , , 221 -2. .785 1. , 00 11. .61 C
ATOM 1657 C GLY A 221 67 , .180 71 , .643 -4. .249 1. , 00 14. .90 c
ATOM 1658 O GLY A 221 68 , , 077 72 , ,339 -4. .724 1. , 00 12. .92 O
ATOM 1659 N ALA A 222 66 , , 144 71 , , 221 -4. .970 1. , 00 15. .04 N
ATOM 1660 CA ALA A 222 65, , 919 71 , , 649 -6. .343 1. , 00 16. .06 C
ATOM 1661 CB ALA A 222 65 , .042 70 , .606 -7. .053 1. , 00 14. .37 c
ATOM 1662 C ALA A 222 65, , 259 73 , , 033 -6. .437 1. , 00 15. .61 C
ATOM 1663 O ALA A 222 65, , 190 73 , , 613 -7. .528 1. , 00 18. .27 O
ATOM 1664 N LYS A 223 64 ,, 720 73., 535 -5..326 1., 00 14..15 N
ATOM 1665 CA LYS A 223 63 , , 855 74. , 738 -5. .347 1. , 00 14. .84 C
ATOM 1666 CB LYS A 223 62 , ,384 74. ,339 -5. .086 1. , 00 17. .77 C
ATOM 1667 CG LYS A 223 61. .779 73. ,313 -6. .073 1. , 00 19. .77 c
ATOM 1668 CD LYS A 223 61 , , 871 73. , 726 -7. .547 1. , 00 20. .13 C
ATOM 1669 CE LYS A 223 61 , , 187 72. , 711 -8. .493 1. , 00 23. .78 C
ATOM 1670 NZ LYS A 223 61 , ,352 73. , 102 -9. .924 1. , 00 19. .55 N
ATOM 1671 C LYS A 223 64 , .285 75. , 799 -4. .299 1. , 00 12. .23 c
ATOM 1672 O LYS A 223 63 , , 473 76. , 600 -3. .831 1. , 00 14. .57 O
ATOM 1673 N GLY A 224 65, , 547 75. , 793 -3. .916 1. , 00 9. .38 N
ATOM 1674 CA GLY A 224 66 , , 077 76. , 781 -2. .953 1. , 00 8. .00 C
ATOM 1675 C GLY A 224 67 , .222 76. , 129 -2. .212 1. , 00 9. .36 c
ATOM 1676 O GLY A 224 67, , 511 74. , 930 -2. .420 1. , 00 8. .77 O
ATOM 1677 N SER A 225 67, , 886 76. , 883 -1. .348 1. , 00 8. .68 N
ATOM 1678 CA SER A 225 68 , , 874 76. , 286 -0. .461 1. , 00 7. .59 C
ATOM 1679 CB SER A 225 69 , .738 77. ,379 0. .190 1. , 00 7. .43 c
ATOM 1680 OG SER A 225 70 , , 619 76. , 789 1. .142 1. , 00 6. .53 O
ATOM 1681 C SER A 225 68 , , 146 75. , 534 0. .641 1. , 00 8. .56 C
ATOM 1682 O SER A 225 67, , 248 76. , 101 1. .286 1. , 00 8. .06 O
ATOM 1683 N ILE A 226 68 , .550 74. ,300 0. .928 1. , 00 7. .50 N
ATOM 1684 CA ILE A 226 67, , 912 73. , 585 2. .042 1. , 00 7. .85 C
ATOM 1685 CB ILE A 226 68 , ,368 72. , 105 2. .161 1. , 00 9. .15 C
ATOM 1686 CGI ILE A 226 69 , , 816 71. , 994 2. .658 1. , 00 10. .46 c
ATOM 1687 CD1 ILE A 226 70 , .309 70. , 556 2. .877 1. , 00 9. .97 c
ATOM 1688 CG2 ILE A 226 68 , , 147 71. ,364 0. .826 1. , 00 8. .05 C
ATOM 1689 C ILE A 226 68 , , 100 74. ,330 3. .366 1. , 00 8. .37 C
ATOM 1690 O ILE A 226 67 , .236 74. , 287 4. .236 1. , 00 7. .38 0
ATOM 1691 N PHE A 227 69 , , 187 75. , 099 3. .472 1. , 00 8. .65 N
ATOM 1692 CA PHE A 227 69 , , 505 75. , 797 4. .708 1. , 00 9. .48 C
ATOM 1693 CB PHE A 227 70 , , 965 76. , 273 4. .707 1. , 00 9. .08 C
ATOM 1694 CG PHE A 227 71 , .922 75. , 176 4. .443 1. , 00 8. .38 c
ATOM 1695 CD1 PHE A 227 71 , , 943 74. , 077 5. .252 1. , 00 8. .71 C
ATOM 1696 CE1 PHE A 227 72 , , 813 73. , 038 5. .018 1. , 00 9. .21 C
ATOM 1697 CZ PHE A 227 73 , , 664 73. , 090 3. .938 1. , 00 9. .94 C
ATOM 1698 CE2 PHE A 227 73 , .644 74. , 190 3. .120 1. , 00 10. .56 c
ATOM 1699 CD2 PHE A 227 72 , .771 75. , 225 3. .370 1. , 00 8. .98 C
ATOM 1700 C PHE A 227 68 , .595 76. , 940 5. .044 1. , 00 9. .68 C
ATOM 1701 O PHE A 227 68 , .628 77. , 417 6. .180 1. , 00 9. .62 O
ATOM 1702 N ASP A 228 67 , .765 77. , 360 4. .092 1. , 00 9. .83 N
ATOM 1703 CA ASP A 228 66 , .748 78. ,367 4. .328 1. , 00 10. .19 C
ATOM 1704 CB ASP A 228 66 , .376 79. , 108 3. .015 1. , 00 11. .95 C
ATOM 1705 CG ASP A 228 67, .518 79. , 985 2. .481 1. , 00 15. .69 C
ATOM 1706 OD1 ASP A 228 68 , .495 80. , 196 3. .224 1. , 00 15. .19 0
ATOM 1707 OD2 ASP A 228 67, .453 80. , 449 1. .318 1. , 00 17. .15 O
ATOM 1708 C ASP A 228 65, .489 77. , 796 4. .948 1. , 00 9. .82 C
ATOM 1709 O ASP A 228 64., 611 78 ,.559 5..382 1., 00 9..11 O
ATOM 1710 N GLY A 229 65. ,395 76 , .468 5. .033 1. , 00 8. .99 N
ATOM 1711 CA GLY A 229 64. , 217 75 , .831 5. .589 1. , 00 8. .16 C
ATOM 1712 C GLY A 229 64. .504 75 , .012 6. .812 1. , 00 8. .52 c
ATOM 1713 O GLY A 229 65. , 634 74 , .940 7. .273 1. , 00 8. .41 O
ATOM 1714 N ASN A 230 63. , 456 74 , .399 7. .351 1. , 00 8. .65 N
ATOM 1715 CA ASN A 230 63. , 588 73 , .541 8. .519 1. , 00 8. .57 C
ATOM 1716 CB ASN A 230 62. , 218 73 , .058 9. .001 1. , 00 9. .59 c
ATOM 1717 CG ASN A 230 61. , 289 74 , .185 9. .351 1. , 00 7. .92 C
ATOM 1718 OD1 ASN A 230 60. , 844 74 , .916 8. .444 1. , 00 7. .22 O
ATOM 1719 ND2 ASN A 230 60. , 981 74 , .360 10. .674 1. , 00 7. .00 N
ATOM 1720 C ASN A 230 64. , 449 72 , .320 8. .230 1. , 00 9. .27 c
ATOM 1721 O ASN A 230 64. , 451 71 , .807 7. .112 1. , 00 8. .03 O
ATOM 1722 N LEU A 231 65. , 184 71 , .872 9. .241 1. , 00 9. .54 N
ATOM 1723 CA LEU A 231 65. , 806 70 , .562 9. .229 1. , 00 9. .90 C
ATOM 1724 CB LEU A 231 67. , 043 70 , .567 10. .122 1. , 00 9. .26 c
ATOM 1725 CG LEU A 231 67. , 814 69 , .251 10. .327 1. , 00 10. .74 C
ATOM 1726 CD1 LEU A 231 68. , 277 68 , .661 8. .991 1. , 00 9. .80 C
ATOM 1727 CD2 LEU A 231 68. , 981 69 , .494 11. .291 1. , 00 10. .19 C
ATOM 1728 C LEU A 231 64. , 804 69 , .515 9. .734 1. , 00 10. .16 c
ATOM 1729 O LEU A 231 64. , 239 69 , .699 10. .801 1. , 00 9. .71 O
ATOM 1730 N ALA A 232 64. , 596 68 , .435 8. .971 1. , 00 10. .65 N
ATOM 1731 CA ALA A 232 63. , 612 67, .394 9. .295 1. , 00 11. .25 c
ATOM 1732 CB ALA A 232 64. ,144 66 , .460 10. .397 1. , 00 11. .26 c
ATOM 1733 C ALA A 232 62. ,311 68 , .067 9. .720 1. , 00 10. .60 C
ATOM 1734 O ALA A 232 61. , 822 68 , .893 8. .990 1. , 00 11. .68 O
ATOM 1735 N GLN A 233 61. , 748 67 , .733 10. .886 1. , 00 10. .40 N
ATOM 1736 CA GLN A 233 60. , 548 68 , .426 11. .361 1. , 00 11. .70 C
ATOM 1737 CB GLN A 233 59. ,371 67, .471 11. .600 1. , 00 13. .09 C
ATOM 1738 CG GLN A 233 59. , 068 66 , .550 10. .399 1. , 00 13. .83 C
ATOM 1739 CD GLN A 233 60. , 063 65 , .419 10. .209 1. , 00 17. .23 c
ATOM 1740 OE1 GLN A 233 60. , 507 64 , .750 11. .188 1. , 00 18. .61 O
ATOM 1741 NE2 GLN A 233 60. , 446 65, .188 8. .943 1. , 00 14. .32 N
ATOM 1742 C GLN A 233 60. , 800 69 , .278 12. .595 1. , 00 10. .17 C
ATOM 1743 O GLN A 233 59. , 869 69 , .575 13. .362 1. , 00 10. .82 O
ATOM 1744 N LEU A 234 62. , 031 69 , .735 12. .728 1. , 00 8. .49 N
ATOM 1745 CA LEU A 234 62. , 459 70 , .529 13. .869 1. , 00 8. .01 C
ATOM 1746 CB LEU A 234 63. , 982 70 , .517 13. .986 1. , 00 7. .26 C
ATOM 1747 CG LEU A 234 64. , 645 69 , .146 14. .226 1. , 00 7. .55 c
ATOM 1748 CD1 LEU A 234 66. , 147 69 , .240 13. .920 1. , 00 6. .15 C
ATOM 1749 CD2 LEU A 234 64. ,347 68 , .643 15. .646 1. , 00 6. .17 C
ATOM 1750 C LEU A 234 62. , 008 71 , .949 13. .707 1. , 00 8. .63 C
ATOM 1751 O LEU A 234 62. , 103 72 , .508 12. .615 1. , 00 10. .74 0
ATOM 1752 N SER A 235 61. , 529 72 , .547 14. .794 1. , 00 7. .87 N
ATOM 1753 CA SER A 235 61. , 231 73 , .982 14. .819 1. , 00 9. .14 C
ATOM 1754 CB SER A 235 60., 806 74., 421 16..222 1., 00 9..04 C
ATOM 1755 OG SER A 235 61. , 847 74. , 248 17. .190 1. , 00 7. .50 O
ATOM 1756 C SER A 235 62. , 479 74. , 751 14. .361 1. , 00 10. .95 C
ATOM 1757 O SER A 235 63. .584 74. , 394 14. .739 1. , 00 13. .76 0
ATOM 1758 N ASP A 236 62. ,309 75. , 765 13. .518 1. , 00 9. .21 N
ATOM 1759 CA ASP A 236 63. , 437 76. , 550 13. .094 1. , 00 8. .91 C
ATOM 1760 CB ASP A 236 63. , 436 76. , 728 11. .578 1. , 00 10. .02 C
ATOM 1761 CG ASP A 236 64. , 838 76. , 978 11. .019 1. , 00 10. .03 c
ATOM 1762 OD1 ASP A 236 65. , 716 76. , 086 11. .130 1. , 00 9. .94 O
ATOM 1763 OD2 ASP A 236 65. , 060 78. , 058 10. .476 1. , 00 10. .02 O
ATOM 1764 C ASP A 236 63. , 463 77. , 916 13. .794 1. , 00 9. .38 C
ATOM 1765 O ASP A 236 62. , 599 78. , 763 13. .561 1. , 00 9. .80 0
ATOM 1766 N LEU A 237 64. , 460 78. , 134 14. .644 1. , 00 7. .95 N
ATOM 1767 CA LEU A 237 64. , 514 79. ,368 15. .403 1. , 00 8. .32 C
ATOM 1768 CB LEU A 237 65. , 517 79. , 257 16. .557 1. , 00 8. .30 C
ATOM 1769 CG LEU A 237 65. , 179 78. , 200 17. .623 1. , 00 9. .05 c
ATOM 1770 CD1 LEU A 237 66. , 160 78. , 296 18. .787 1. , 00 7. .94 C
ATOM 1771 CD2 LEU A 237 63. , 744 78. ,343 18. .139 1. , 00 9. .53 C
ATOM 1772 C LEU A 237 64. , 831 80. , 558 14. .480 1. , 00 8. .89 C
ATOM 1773 O LEU A 237 65. , 658 80. , 467 13. .556 1. , 00 8. .00 O
ATOM 1774 N ASN A 238 64. , 130 81. , 663 14. .724 1. , 00 9. .05 N
ATOM 1775 CA ASN A 238 64. , 217 82. , 851 13. .856 1. , 00 8. .98 C
ATOM 1776 CB ASN A 238 62. , 946 83. , 700 13. .976 1. , 00 9. .29 c
ATOM 1777 CG ASN A 238 63. , 008 84. , 967 13. .146 1. , 00 9. .11 c
ATOM 1778 OD1 ASN A 238 63. , 990 85. , 223 12. .433 1. , 00 9. .95 O
ATOM 1779 ND2 ASN A 238 61. , 937 85. , 731 13. .178 1. , 00 8. .67 N
ATOM 1780 C ASN A 238 65. .439 83. , 672 14. .224 1. , 00 8. .67 c
ATOM 1781 O ASN A 238 65. , 413 84. , 447 15. .183 1. , 00 8. .71 O
ATOM 1782 N GLU A 239 66. , 515 83. , 455 13. .470 1. , 00 9. .46 N
ATOM 1783 CA GLU A 239 67. , 788 84. , 146 13. .678 1. , 00 9. .95 C
ATOM 1784 CB GLU A 239 68. , 805 83. , 725 12. .609 1. , 00 9. .75 c
ATOM 1785 CG GLU A 239 68. , 472 84. , 174 11. .178 1. , 00 10. .21 C
ATOM 1786 CD GLU A 239 67. , 484 83. , 271 10. .427 1. , 00 12. .25 C
ATOM 1787 OE1 GLU A 239 66. , 965 82. , 242 10. .962 1. , 00 11. .92 O
ATOM 1788 OE2 GLU A 239 67. ,177 83. , 629 9. .270 1. , 00 12. .19 0
ATOM 1789 C GLU A 239 67. , 685 85. , 670 13. .643 1. , 00 9. .26 C
ATOM 1790 O GLU A 239 68. , 587 86. ,346 14. .119 1. , 00 9. .01 O
ATOM 1791 N ARG A 240 66. , 628 86. , 202 13. .041 1. , 00 9. .48 N
ATOM 1792 CA ARG A 240 66. , 453 87. , 658 12. .934 1. , 00 10. .29 c
ATOM 1793 CB ARG A 240 65. , 680 88. , 024 11. .642 1. , 00 9. .90 C
ATOM 1794 CG ARG A 240 66. ,375 87. , 568 10. .355 1. , 00 10. .51 C
ATOM 1795 CD ARG A 240 65. , 548 87. , 841 9. .095 1. , 00 10. .04 C
ATOM 1796 NE ARG A 240 64. , 229 87. , 222 9. .204 1. , 00 11. .65 N
ATOM 1797 CZ ARG A 240 63. , 049 87. , 839 9. .156 1. , 00 14. .40 C
ATOM 1798 NH1 ARG A 240 62. , 939 89. , 143 8. .960 1. , 00 17. .97 N
ATOM 1799 NH2 ARG A 240 61. , 949 87. , 131 9..287 1. , 00 13..77 N
ATOM IE 100 C ARG A 240 65. , 730 88. , 262 14. .120 1. , 00 10. .21 C
ATOM IE 101 0 ARG A 240 65. , 625 89. , 477 14. .220 1. , 00 10. .26 O
ATOM IE 102 N ASN A 241 65. .225 87. , 430 15. .018 1. , 00 9. .85 N
ATOM IE 103 CA ASN A 241 64. , 440 87. , 930 16. .138 1. , 00 9. .04 C
ATOM IE 101 CB ASN A 241 63. ,365 86. , 886 16. .527 1. , 00 8. .60 C
ATOM IE 105 CG ASN A 241 62. , 496 87. ,320 17. .703 1. , 00 11. .12 C
ATOM IE 106 OD1 ASN A 241 62. , 865 88. , 212 18. .479 1. , 00 8. .46 O
ATOM IE 101 ND2 ASN A 241 61. ,340 86. , 657 17. .857 1. , 00 8. .55 N
ATOM IE 108 C ASN A 241 65. , 416 88. , 167 17. .262 1. , 00 7. .66 C
ATOM IE 109 0 ASN A 241 66. , 020 87. , 223 17. .760 1. , 00 7. .50 O
ATOM IE 110 N PRO A 242 65. , 550 89. , 412 17. .716 1. , 00 7. .70 N
ATOM IE 111 CA PRO A 242 66. , 588 89. , 627 18. .728 1. , 00 8. .44 C
ATOM 1812 CB PRO A 242 66. , 637 91. , 155 18. .869 1. , 00 8. .57 C
ATOM IE 113 CG PRO A 242 65. , 261 91. , 603 18. .501 1. , 00 9. .52 C
ATOM IE 11 CD PRO A 242 64. , 789 90. , 646 17. .421 1. , 00 8. .80 c
ATOM IE 115 C PRO A 242 66. , 298 88. , 945 20. .062 1. , 00 8. .57 C
ATOM 1816 0 PRO A 242 67. , 230 88. , 685 20. .834 1. , 00 9. .95 O
ATOM IE 111 N ASP A 243 65. , 041 88. , 622 20. .330 1. , 00 9. .65 N
ATOM IE 118 CA ASP A 243 64. , 699 87. , 818 21. .513 1. , 00 10. .05 c
ATOM IE 119 CB ASP A 243 63. , 207 87. , 645 21. .654 1. , 00 10. .85 C
ATOM IE 120 CG ASP A 243 62. , 491 88. , 910 22. .053 1. , 00 12. .95 C
ATOM IE 121 OD1 ASP A 243 63. , 098 89. , 879 22. .552 1. , 00 14. .17 O
ATOM IE 122 OD2 ASP A 243 61. , 265 88. , 884 21. .906 1. , 00 14. .33 O
ATOM IE 123 C ASP A 243 65. ,317 86. , 408 21. .443 1. , 00 10. .25 C
ATOM IE 121 0 ASP A 243 65. , 688 85. , 843 22. .485 1. , 00 8. .57 O
ATOM IE 125 N VAL A 244 65. , 394 85. , 841 20. .232 1. , 00 9. .35 N
ATOM IE 126 CA VAL A 244 66. , 012 84. , 510 20. .027 1. , 00 8. .85 C
ATOM IE 121 CB VAL A 244 65. , 807 83. , 992 18. .584 1. , 00 8. .65 C
ATOM IE 128 CGI VAL A 244 66. , 668 82. , 777 18. .279 1. , 00 6. .72 C
ATOM IE 129 CG2 VAL A 244 64. , 332 83. , 715 18. .338 1. , 00 7. .73 c
ATOM IE 130 C VAL A 244 67. , 491 84. , 556 20. .372 1. , 00 9. .96 C
ATOM IE 131 0 VAL A 244 68. , 000 83. , 725 21. .140 1. , 00 8. .97 O
ATOM IE 132 N LEU A 245 68. , 194 85. , 554 19. .852 1. , 00 8. .99 N
ATOM IE 133 CA LEU A 245 69. , 607 85. , 664 20. .140 1. , 00 8. .92 c
ATOM IE 131 CB LEU A 245 70. , 253 86. , 795 19. .296 1. , 00 9. .87 C
ATOM 1835 CG LEU A 245 71. , 749 87. , 033 19. .518 1. , 00 11. .93 C
ATOM IE 136 CD1 LEU A 245 72. , 556 85. , 781 19. .201 1. , 00 10. .23 C
ATOM IE 131 CD2 LEU A 245 72. , 230 88. , 246 18. .685 1. , 00 12. .15 c
ATOM IE 138 C LEU A 245 69. , 806 85. , 887 21. .642 1. , 00 8. .71 C
ATOM 1839 0 LEU A 245 70. , 660 85. , 240 22. .252 1. , 00 7. .81 O
ATOM IE 110 N ASP A 246 68. , 991 86. , 767 22. .236 1. , 00 8. .57 N
ATOM IE 111 CA ASP A 246 69. , 076 87. , 047 23. .676 1. , 00 8. .50 c
ATOM IE 112 CB ASP A 246 68. , 036 88. , 071 24. .127 1. , 00 8. .41 C
ATOM IE 343 CG ASP A 246 68. ,344 89. , 478 23. .683 1. , 00 9. .39 C
ATOM li 344 OD1 ASP A 246 69. , 427 89 , .766 23..094 1. , 00 11..58 O
ATOM li 345 OD2 ASP A 246 67. , 467 90 , .314 23. .934 1. , 00 9. .12 O
ATOM IE 346 C ASP A 246 68. , 848 85, .768 24. .467 1. , 00 8. .86 C
ATOM li 347 O ASP A 246 69. .564 85 , .504 25. .434 1. , 00 8. .72 O
ATOM li 348 N TYR A 247 67. , 840 84 , .988 24. .071 1. , 00 7. .85 N
ATOM li 349 CA TYR A 247 67. , 528 83 , .743 24. .778 1. , 00 8. .07 C
ATOM li 350 CB TYR A 247 66. , 283 83 , .109 24. .176 1. , 00 8. .64 C
ATOM li 351 CG TYR A 247 65. , 903 81 , .792 24. .786 1. , 00 8. .20 c
ATOM li 352 CD1 TYR A 247 66. ,344 80 , .609 24. .241 1. , 00 8. .56 C
ATOM li 353 CE1 TYR A 247 65. , 985 79 , .393 24. .796 1. , 00 8. .28 C
ATOM li 354 CZ TYR A 247 65. , 167 79 , .365 25. .895 1. , 00 7. .93 C
ATOM li 355 OH TYR A 247 64. , 830 78 , .149 26. .438 1. , 00 8. .17 0
ATOM li 356 CE2 TYR A 247 64. , 706 80 , .523 26. .448 1. , 00 8. .24 C
ATOM 1857 CD2 TYR A 247 65. , 051 81 , .735 25. .888 1. , 00 8. .63 C
ATOM li 358 C TYR A 247 68. , 729 82 , .752 24. .748 1. , 00 8. .02 C
ATOM li 359 O TYR A 247 69. , 188 82 , .258 25. .802 1. , 00 8. .17 0
ATOM li 360 N LEU A 248 69. , 267 82 , .524 23. .567 1. , 00 7. .77 N
ATOM 1861 CA LEU A 248 70. , 357 81 , .561 23. .396 1. , 00 7. .94 C
ATOM li 362 CB LEU A 248 70. , 538 81 , .235 21. .924 1. , 00 8. .51 C
ATOM li 363 CG LEU A 248 69. ,348 80 , .538 21. .227 1. , 00 8. .35 c
ATOM li 364 CD1 LEU A 248 69. , 495 80 , .612 19. .705 1. , 00 7. .39 C
ATOM li 365 CD2 LEU A 248 69. , 262 79 , . Ill 21. .735 1. , 00 7. .29 C
ATOM li 366 C LEU A 248 71. , 681 82 , .054 24. .016 1. , 00 7. .97 c
ATOM li 367 O LEU A 248 72. , 395 81 , .284 24. .641 1. , 00 8. .18 O
ATOM li 368 N VAL A 249 72. , 005 83 , .326 23. .848 1. , 00 7. .89 N
ATOM li 369 CA VAL A 249 73. , 208 83 , .868 24. .460 1. , 00 8. .40 C
ATOM li 370 CB VAL A 249 73. , 460 85 , .339 24. .076 1. , 00 8. .93 c
ATOM li 371 CGI VAL A 249 74. , 480 85 , .991 25. .022 1. , 00 8. .60 C
ATOM li 372 CG2 VAL A 249 73. , 907 85, .462 22. .627 1. , 00 8. .39 C
ATOM li 373 C VAL A 249 73. , 125 83 , .699 25. .978 1. , 00 8. .41 C
ATOM li 374 O VAL A 249 74. , 070 83 , .252 26. .595 1. , 00 9. .01 0
ATOM li 375 N GLY A 250 71. , 983 84, .008 26. .573 1. , 00 8. .12 N
ATOM li 376 CA GLY A 250 71. , 854 83 , .924 28. .026 1. , 00 8. .21 C
ATOM li 377 C GLY A 250 71. , 981 82 , .487 28. .514 1. , 00 7. .99 C
ATOM li 378 O GLY A 250 72. , 579 82 , .221 29. .545 1. , 00 7. .72 0
ATOM li 379 N ALA A 251 71. , 432 81 , .550 27. .759 1. , 00 8. .09 N
ATOM 1880 CA ALA A 251 71. , 494 80 , .131 28. .121 1. , 00 8. .04 C
ATOM li 381 CB ALA A 251 70. , 635 79 , .295 27. .169 1. , 00 7. .87 C
ATOM li 382 C ALA A 251 72. , 939 79 , .662 28. .100 1. , 00 8. .94 c
ATOM li 383 O ALA A 251 73. , 433 79 , .079 29. .069 1. , 00 8. .37 O
ATOM 1884 N TYR A 252 73. , 644 79 , .941 27. .002 1. , 00 9. .53 N
ATOM li 385 CA TYR A 252 75. , 003 79 , .460 26. .874 1. , 00 9. .09 C
ATOM li 386 CB TYR A 252 75. , 396 79 , .343 25. .395 1. , 00 9. .00 c
ATOM li 387 CG TYR A 252 74. , 508 78 , .284 24. .726 1. , 00 9. .23 C
ATOM li 388 CD1 TYR A 252 74. , 482 76 , .976 25. .209 1. , 00 9. .03 C
ATOM 1889 CE1 TYR A 252 73 ,.653 76., 026 24..660 1., 00 8..33 C
ATOM 1890 CZ TYR A 252 72 , .830 76. ,371 23. .604 1. , 00 8. .72 C
ATOM 1891 OH TYR A 252 71 , .999 75. , 425 23. .038 1. , 00 7. .85 O
ATOM 1892 CE2 TYR A 252 72. .853 77. .657 23. .103 1. , 00 8. .90 c
ATOM 1893 CD2 TYR A 252 73 , .674 78. , 601 23. .672 1. , 00 8. .75 C
ATOM 1894 C TYR A 252 76 , .019 80. , 177 27. .777 1. , 00 9. .99 C
ATOM 1895 O TYR A 252 76 , .988 79. , 541 28. .247 1. , 00 10. .36 O
ATOM 1896 N LEU A 253 75 , .769 81. , 449 28. .084 1. , 00 9. .12 N
ATOM 1897 CA LEU A 253 76 , .558 82. , 146 29. .107 1. , 00 9. .45 C
ATOM 1898 CB LEU A 253 76 , .254 83. , 656 29. .161 1. , 00 8. .84 C
ATOM 1899 CG LEU A 253 76 , .794 84. , 547 28. .022 1. , 00 9. .73 C
ATOM 1900 CD1 LEU A 253 76 , .288 85. , 995 28. .233 1. , 00 8. .83 c
ATOM 1901 CD2 LEU A 253 78 , .328 84. , 535 27. .930 1. , 00 8. .02 C
ATOM 1902 C LEU A 253 76 , .332 81. , 501 30. .482 1. , 00 8. .81 C
ATOM 1903 O LEU A 253 77 , .262 81. ,386 31. .274 1. , 00 9. .64 O
ATOM 1904 N GLN A 254 75 , .116 81. , 077 30. .772 1. , 00 8. .17 N
ATOM 1905 CA GLN A 254 74 , .864 80. ,349 32. .026 1. , 00 8. .63 C
ATOM 1906 CB GLN A 254 73 , .393 79. , 954 32. .166 1. , 00 9. .69 C
ATOM 1907 CG GLN A 254 73 , .048 79. , 230 33. .460 1. , 00 9. .28 C
ATOM 1908 CD GLN A 254 71. .595 78. , 842 33. .503 1. , 00 9. .80 c
ATOM 1909 OE1 GLN A 254 70 , .769 79. , 682 33. .718 1. , 00 10. .05 O
ATOM 1910 NE2 GLN A 254 71 , .279 77. , 561 33. .263 1. , 00 9. .63 N
ATOM 1911 C GLN A 254 75 , .723 79. , 086 32. .108 1. , 00 7. .72 c
ATOM 1912 O GLN A 254 76. .364 78. ,817 33. .127 1. , 00 7. .05 O
ATOM 1913 N TRP A 255 75 , .742 78. ,314 31. .039 1. , 00 7. .22 N
ATOM 1914 CA TRP A 255 76 , .489 77. , 064 31. .059 1. , 00 7. .92 C
ATOM 1915 CB TRP A 255 76. .096 76. , 135 29. .909 1. , 00 7. .81 c
ATOM 1916 CG TRP A 255 74 , .630 75. , 821 29. .890 1. , 00 8. .61 C
ATOM 1917 CD1 TRP A 255 73 , .755 76. , 010 28. .839 1. , 00 8. .34 C
ATOM 1918 NE1 TRP A 255 72 , .494 75. , 637 29. .180 1. , 00 9. .61 N
ATOM 1919 CE2 TRP A 255 72 , .442 75. , 205 30. .449 1. , 00 9. .34 c
ATOM 1920 CD2 TRP A 255 73 , .789 75. ,314 30. .993 1. , 00 9. .16 C
ATOM 1921 CE3 TRP A 255 74 , .013 74. , 938 32. .308 1. , 00 9. .42 C
ATOM 1922 CZ3 TRP A 255 72 , .922 74. , 497 33. .071 1. , 00 9. .28 C
ATOM 1923 CH2 TRP A 255 71 , .641 74. , 410 32. .522 1. , 00 10. .00 c
ATOM 1924 CZ2 TRP A 255 71 , .378 74. , 781 31. .205 1. , 00 9. .11 C
ATOM 1925 C TRP A 255 77 , .974 77. , 271 31. .119 1. , 00 7. .83 C
ATOM 1926 O TRP A 255 78 , .687 76. , 457 31. .736 1. , 00 7. .86 O
ATOM 1927 N ILE A 256 78 , .474 78. ,343 30. .506 1. , 00 8. .00 N
ATOM 1928 CA ILE A 256 79 , .860 78. , 725 30. .758 1. , 00 8. .64 C
ATOM 1929 CB ILE A 256 80 , .337 79. , 860 29. .848 1. , 00 9. .07 C
ATOM 1930 CGI ILE A 256 80 , .362 79. ,386 28. .396 1. , 00 9. .62 C
ATOM 1931 CD1 ILE A 256 80 , .391 80. , 512 27. .374 1. , 00 9. .62 c
ATOM 1932 CG2 ILE A 256 81 , .729 80. , 327 30. .274 1. , 00 9. .03 C
ATOM 1933 C ILE A 256 80 , .092 79. , 115 32. .224 1. , 00 9. .08 C
ATOM 1934 O ILE A 256 81 ,, 085 78 ,, 699 32., 820 1., 00 9., 19 O
ATOM 1935 N ASP A 257 79 , , 209 79 , , 914 32. , 813 1. , 00 11. , 89 N
ATOM 1936 CA ASP A 257 79 , ,341 80 , , 243 34. ,230 1. , 00 12. , 12 C
ATOM 1937 CB ASP A 257 78. .161 81. , 043 34. .791 1. , 00 16. .27 C
ATOM 1938 CG ASP A 257 78 , , 065 82 , , 429 34. , 226 1. , 00 25. ,30 C
ATOM 1939 OD1 ASP A 257 79 , , 053 82 , , 920 33. ,631 1. , 00 31. ,80 O
ATOM 1940 OD2 ASP A 257 76 , , 973 83 , , 024 34. ,362 1. , 00 34. ,61 O
ATOM 1941 C ASP A 257 79 , .434 78. , 975 35. .054 1. , 00 11. .56 c
ATOM 1942 O ASP A 257 80 , , 121 78 , , 973 36. , 041 1. , 00 11. ,65 O
ATOM 1943 N GLN A 258 78 , , 715 77 , , 928 34. ,666 1. , 00 10. , 15 N
ATOM 1944 CA GLN A 258 78 , , 734 76 , , 662 35. ,397 1. , 00 9. ,64 C
ATOM 1945 CB GLN A 258 77 , .416 75. , 915 35. .191 1. , 00 9. .24 c
ATOM 1946 CG GLN A 258 76 , , 260 76 , , 704 35. , 767 1. , 00 9. ,14 C
ATOM 1947 CD GLN A 258 74 , , 975 75 , , 917 35. , 872 1. , 00 11. ,25 C
ATOM 1948 OE1 GLN A 258 74 , , 984 74 , , 690 35. , 785 1. , 00 11. , 49 O
ATOM 1949 NE2 GLN A 258 73 , .852 76. , 631 36. .071 1. , 00 9. .01 N
ATOM 1950 C GLN A 258 79 , , 911 75 , , 757 35. , 124 1. , 00 10. , 42 C
ATOM 1951 O GLN A 258 80 , , 042 74 , , 736 35. , 802 1. , 00 10. ,37 O
ATOM 1952 N GLY A 259 80 , , 780 76 , , 111 34. , 173 1. , 00 9. , 71 N
ATOM 1953 CA GLY A 259 82. .029 75. ,373 33. .972 1. , 00 9. .16 C
ATOM 1954 C GLY A 259 82 , ,350 74 , , 824 32. ,593 1. , 00 9. , 89 C
ATOM 1955 O GLY A 259 83 , ,379 74 , , 180 32. , 436 1. , 00 9. ,39 O
ATOM 1956 N ALA A 260 81 , , 526 75 , , 110 31. ,580 1. , 00 9. ,37 N
ATOM 1957 CA ALA A 260 81. .819 74. , 691 30. .215 1. , 00 9. .37 C
ATOM 1958 CB ALA A 260 80 , , 634 74 , , 971 29. ,287 1. , 00 8. ,87 C
ATOM 1959 C ALA A 260 83 , , 061 75 , , 431 29. , 711 1. , 00 10. , 05 C
ATOM 1960 O ALA A 260 83. .136 76. , 656 29. .839 1. , 00 10. .90 0
ATOM 1961 N ASP A 261 84 , , 012 74 , , 685 29. , 146 1. , 00 8. ,84 N
ATOM 1962 CA ASP A 261 85, , 298 75 , , 218 28. ,699 1. , 00 10. ,61 C
ATOM 1963 CB ASP A 261 86 , ,396 74 , , 238 29. , 069 1. , 00 11. ,23 C
ATOM 1964 CG ASP A 261 86 , .511 74. , 070 30. .560 1. , 00 13. .42 c
ATOM 1965 OD1 ASP A 261 86 , , 750 75 , , 108 31. ,212 1. , 00 12. ,25 O
ATOM 1966 OD2 ASP A 261 86 , , 296 72 , , 942 31. , 091 1. , 00 12. , 74 O
ATOM 1967 C ASP A 261 85, ,393 75 , , 562 27. ,209 1. , 00 11. , 02 C
ATOM 1968 O ASP A 261 86 , .235 76. , 329 26. .811 1. , 00 12. .22 0
ATOM 1969 N ALA A 262 84, , 512 74 , , 987 26. , 404 1. , 00 9. ,84 N
ATOM 1970 CA ALA A 262 84, , 473 75 , , 175 24. ,968 1. , 00 8. ,24 C
ATOM 1971 CB ALA A 262 85, , 608 74 , ,384 24. ,317 1. , 00 6. ,61 C
ATOM 1972 C ALA A 262 83 , .116 74. , 648 24. .493 1. , 00 8. .23 c
ATOM 1973 O ALA A 262 82 , , 416 73 , , 922 25. ,240 1. , 00 8. ,59 O
ATOM 1974 N PHE A 263 82 , , 717 75 , , 035 23. ,284 1. , 00 7. ,94 N
ATOM 1975 CA PHE A 263 81 , , 500 74 , , 519 22. , 654 1. , 00 7. , 83 C
ATOM 1976 CB PHE A 263 80 , .557 75. , 651 22. .247 1. , 00 8. .04 c
ATOM 1977 CG PHE A 263 80 , , 028 76 , , 476 23. ,375 1. , 00 7. ,66 C
ATOM 1978 CD1 PHE A 263 79 , , 780 75 , , 934 24. ,625 1. , 00 7. , 82 C
ATOM 1979 CE1 PHE A 263 79 ,.236 76 ,.709 25..641 1., 00 7..86 C
ATOM 1980 CZ PHE A 263 78 , .902 78 , .033 25. .404 1. , 00 8. .25 C
ATOM 1981 CE2 PHE A 263 79 , .119 78 , .585 24. .158 1. , 00 7. .88 C
ATOM 1982 CD2 PHE A 263 79. .694 77. .802 23. .147 1. .00 8. .19 c
ATOM 1983 C PHE A 263 81 , .793 73 , .767 21. .364 1. , 00 8. .11 C
ATOM 1984 O PHE A 263 82 , .657 74 , .157 20. .539 1. , 00 8. .29 O
ATOM 1985 N ARG A 264 81 , .044 72 , .704 21. .186 1. , 00 8. .17 N
ATOM 1986 CA ARG A 264 80 , .885 72 , .100 19. .879 1. , 00 8. .28 c
ATOM 1987 CB ARG A 264 80 , .966 70 , .574 19. .960 1. , 00 8. .04 C
ATOM 1988 CG ARG A 264 80 , .730 69 , .848 18. .629 1. , 00 9. .74 C
ATOM 1989 CD ARG A 264 79 , .253 69 , .554 18. .350 1. , 00 10. .19 C
ATOM 1990 NE ARG A 264 79 , .056 68 , .614 17. .241 1. , 00 10. .60 N
ATOM 1991 CZ ARG A 264 77 , .870 68 , .172 16. .818 1. , 00 9. .18 C
ATOM 1992 NH1 ARG A 264 76 , .739 68 , .575 17. .374 1. , 00 8. .78 N
ATOM 1993 NH2 ARG A 264 77 , .815 67, .334 15. .810 1. , 00 11. .27 N
ATOM 1994 C ARG A 264 79 , .532 72 , .589 19. .401 1. , 00 8. .33 c
ATOM 1995 O ARG A 264 78 , .503 72 , .354 20. .088 1. , 00 7. .37 O
ATOM 1996 N ILE A 265 79 , .517 73 , .294 18. .254 1. , 00 7. .99 N
ATOM 1997 CA ILE A 265 78 , .281 73 , .894 17. .724 1. , 00 8. .22 C
ATOM 1998 CB ILE A 265 78. .541 75. .265 17. .060 1. , 00 9. .13 C
ATOM 1999 CGI ILE A 265 79 , .226 76 , .245 18. .029 1. , 00 10. .88 C
ATOM 2000 CD1 ILE A 265 79 , ,666 77 , .548 17. .385 1. , 00 11. .69 C
ATOM 2001 CG2 ILE A 265 77 , .239 75 , .892 16. .556 1. , 00 8. .65 C
ATOM 2002 C ILE A 265 77. .580 72. .987 16. .711 1. , 00 9. .45 C
ATOM 2003 O ILE A 265 78 , .026 72 , .828 15. .570 1. , 00 9. .58 O
ATOM 2004 N ASP A 266 76 , .476 72 , .405 17. .132 1. , 00 8. .42 N
ATOM 2005 CA ASP A 266 75. .616 71. .606 16. .267 1. .00 8. .74 c
ATOM 2006 CB ASP A 266 74 , .428 71 , .066 17. .084 1. , 00 9. .76 C
ATOM 2007 CG ASP A 266 73 , .624 70 , .019 16. .338 1. , 00 10. .48 C
ATOM 2008 OD1 ASP A 266 74 , .174 68 , .910 16. .102 1. , 00 10. .90 O
ATOM 2009 OD2 ASP A 266 72 , .445 70 , .298 15. .975 1. , 00 11. .82 0
ATOM 2010 C ASP A 266 75 , .063 72 , .447 15. .116 1. , 00 8. .79 C
ATOM 2011 O ASP A 266 74 , .866 73 , .657 15. .262 1. , 00 7. .60 O
ATOM 2012 N THR A 267 74 , .835 71 , .767 13. .991 1. , 00 8. .41 N
ATOM 2013 CA THR A 267 74 , .082 72 , .254 12. .846 1. , 00 8. .27 c
ATOM 2014 CB THR A 267 72 , .568 72 , .065 13. .060 1. , 00 7. .93 C
ATOM 2015 OG1 THR A 267 72 , .233 72 , .475 14. .401 1. , 00 7. .29 O
ATOM 2016 CG2 THR A 267 72 , .156 70 , .559 12. .815 1. , 00 8. .83 C
ATOM 2017 C THR A 267 74 , .423 73 , .683 12. .453 1. , 00 8. .67 c
ATOM 2018 O THR A 267 73 , .542 74 , .476 12. .203 1. , 00 9. .33 O
ATOM 2019 N ILE A 268 75 , .715 73 , .975 12. .314 1. , 00 9. .47 N
ATOM 2020 CA ILE A 268 76 , .171 75 , .349 12. .125 1. , 00 8. .75 C
ATOM 2021 CB ILE A 268 77 , .710 75 , .496 12. .374 1. , 00 8. .74 c
ATOM 2022 CGI ILE A 268 78 , .075 76 , .957 12. .691 1. , 00 7. .85 C
ATOM 2023 CD1 ILE A 268 79 , .502 77 , .162 13. .177 1. , 00 8. .43 C
ATOM 2024 CG2 ILE A 268 78 ,.516 74., 876 11..222 1., 00 7..16 C
ATOM 2025 C ILE A 268 75 , .731 75. , 905 10. .780 1. , 00 8. .85 C
ATOM 2026 O ILE A 268 75 , .455 77. , 084 10. .677 1. , 00 8. .54 O
ATOM 2027 N ALA A 269 75. .582 75. .049 9. .771 1. , 00 8. .14 N
ATOM 2028 CA ALA A 269 75 , .172 75. , 505 8. .431 1. , 00 8. .02 C
ATOM 2029 CB ALA A 269 75 , .413 74. ,392 7. .413 1. , 00 8. .12 C
ATOM 2030 C ALA A 269 73 , .715 75. , 917 8. .380 1. , 00 8. .11 C
ATOM 2031 O ALA A 269 73 , .276 76. , 556 7. .406 1. , 00 8. .13 0
ATOM 2032 N TRP A 270 72 , .952 75. , 540 9. .405 1. , 00 6. .32 N
ATOM 2033 CA TRP A 270 71 , .513 75. , 759 9. .377 1. , 00 7. .80 C
ATOM 2034 CB TRP A 270 70 , .799 74. , 603 10. .099 1. , 00 7. .06 C
ATOM 2035 CG TRP A 270 70 , .941 73. ,347 9. .295 1. , 00 7. .18 c
ATOM 2036 CD1 TRP A 270 72 , .021 72. , 457 9. .247 1. , 00 7. .64 C
ATOM 2037 NE1 TRP A 270 71 , .786 71. , 467 8. .298 1. , 00 7. .96 N
ATOM 2038 CE2 TRP A 270 70 , .579 71. , 658 7. .709 1. , 00 7. .74 C
ATOM 2039 CD2 TRP A 270 69 , .995 72. , 862 8. .290 1. , 00 7. .30 c
ATOM 2040 CE3 TRP A 270 68 , .757 73. , 284 7. .853 1. , 00 8. .03 C
ATOM 2041 CZ3 TRP A 270 68 , .102 72. , 541 6. .875 1. , 00 8. .01 C
ATOM 2042 CH2 TRP A 270 68 , .673 71. , 402 6. .327 1. , 00 8. .56 c
ATOM 2043 CZ2 TRP A 270 69. .924 70. , 932 6. .739 1. , 00 8. .33 c
ATOM 2044 C TRP A 270 71 , .055 77. , 132 9. .874 1. , 00 8. .35 C
ATOM 2045 O TRP A 270 69 , .861 77. , 425 9. .819 1. , 00 7. .81 O
ATOM 2046 N MET A 271 71 , .993 77. , 966 10. .346 1. , 00 8. .38 N
ATOM 2047 CA MET A 271 71. .730 79. ,389 10. .564 1. , 00 10. .00 c
ATOM 2048 CB MET A 271 71 , .844 79. , 731 12. .052 1. , 00 10. .84 C
ATOM 2049 CG MET A 271 70 , .960 78. , 861 12. .947 1. , 00 12. .10 C
ATOM 2050 SD MET A 271 71. .071 79. , 290 14. .704 1. , 00 13. .63 s
ATOM 2051 CE MET A 271 69 , .680 80. , 438 14. .835 1. , 00 11. .73 C
ATOM 2052 C MET A 271 72 , .749 80. , 197 9. .767 1. , 00 10. .23 C
ATOM 2053 O MET A 271 73 , .867 79. , 736 9. .544 1. , 00 8. .46 O
ATOM 2054 N PRO A 272 72 , .406 81. , 435 9. .386 1. , 00 12. .06 N
ATOM 2055 CA PRO A 272 73 , .403 82. , 176 8. .610 1. , 00 10. .33 C
ATOM 2056 CB PRO A 272 72 , .674 83. , 481 8. .232 1. , 00 12. .08 C
ATOM 2057 CG PRO A 272 71 , .208 83. , 140 8. .369 1. , 00 13. .71 C
ATOM 2058 CD PRO A 272 71 , .146 82. ,177 9. .527 1. , 00 12. .40 c
ATOM 2059 C PRO A 272 74 , .668 82. , 496 9. .405 1. , 00 10. .38 C
ATOM 2060 O PRO A 272 74 , .607 82. , 645 10. .637 1. , 00 9. .16 O
ATOM 2061 N ASP A 273 75 , .783 82. , 639 8. .690 1. , 00 9. .25 N
ATOM 2062 CA ASP A 273 77 , .058 83. , 069 9. .275 1. , 00 11. .03 c
ATOM 2063 CB ASP A 273 78 , .048 83. , 421 8. .181 1. , 00 12. .34 C
ATOM 2064 CG ASP A 273 78 , .514 82. , 236 7. .394 1. , 00 14. .80 C
ATOM 2065 OD1 ASP A 273 78 , .096 81. , 089 7. .645 1. , 00 17. .36 O
ATOM 2066 OD2 ASP A 273 79 , .332 82. , 476 6. .485 1. , 00 17. .41 0
ATOM 2067 C ASP A 273 76 , .906 84. ,340 10. .141 1. , 00 9. .98 C
ATOM 2068 O ASP A 273 77 , .574 84. , 486 11. .158 1. , 00 9. .36 O
ATOM 2069 N SER A 274 76., 055 85 ,, 261 9., 720 1., 00 9.,01 N
ATOM 2070 CA SER A 274 75. , 896 86 , , 527 10. , 445 1. , 00 10. ,29 C
ATOM 2071 CB SER A 274 75. , 022 87, , 489 9. ,650 1. , 00 12. ,00 C
ATOM 2072 OG SER A 274 73. .760 86. , 886 9. .349 1. , 00 12. .83 0
ATOM 2073 C SER A 274 75. , 279 86 , , 285 11. , 823 1. , 00 11. ,00 C
ATOM 2074 O SER A 274 75. , 561 87, , 019 12. , 766 1. , 00 8. ,84 O
ATOM 2075 N PHE A 275 74. , 426 85, , 263 11. ,944 1. , 00 9. ,61 N
ATOM 2076 CA PHE A 275 73. , 928 84. , 896 13. .266 1. , 00 8. .95 c
ATOM 2077 CB PHE A 275 72. , 783 83 , , 868 13. ,211 1. , 00 9. ,22 C
ATOM 2078 CG PHE A 275 72. , 324 83 , , 441 14. , 578 1. , 00 9. , 15 C
ATOM 2079 CD1 PHE A 275 72. , 929 82 , ,362 15. ,217 1. , 00 11. ,17 C
ATOM 2080 CE1 PHE A 275 72. , 545 81. , 984 16. .486 1. , 00 11. .39 c
ATOM 2081 CZ PHE A 275 71. , 523 82 , , 657 17. , 116 1. , 00 11. ,35 C
ATOM 2082 CE2 PHE A 275 70. , 904 83 , , 723 16. , 483 1. , 00 11. , 89 C
ATOM 2083 CD2 PHE A 275 71. , 324 84, , 123 15. , 232 1. , 00 9. ,81 C
ATOM 2084 C PHE A 275 75. ,044 84. ,370 14. .154 1. , 00 8. .45 c
ATOM 2085 O PHE A 275 75. , 152 84, , 767 15. ,315 1. , 00 9. ,57 O
ATOM 2086 N TRP A 276 75. , 857 83 , , 447 13. ,647 1. , 00 7. , 89 N
ATOM 2087 CA TRP A 276 76. , 933 82 , , 921 14. , 457 1. , 00 8. ,10 C
ATOM 2088 CB TRP A 276 77 . , 629 81. , 750 13. .759 1. , 00 7 . . 97 C
ATOM 2089 CG TRP A 276 76. , 691 80 , , 593 13. ,606 1. , 00 8. , 55 C
ATOM 2090 CD1 TRP A 276 76. , 202 80 , , 043 12. , 440 1. , 00 8. , 47 C
ATOM 2091 NE1 TRP A 276 75. ,342 79 , , 028 12. , 715 1. , 00 9. ,25 N
ATOM 2092 CE2 TRP A 276 75. , 213 78. , 859 14. .054 1. , 00 9. .45 C
ATOM 2093 CD2 TRP A 276 76. , 039 79 , , 856 14. ,686 1. , 00 9. ,04 C
ATOM 2094 CE3 TRP A 276 76. , 107 79 , , 895 16. , 077 1. , 00 10. ,69 C
ATOM 2095 CZ3 TRP A 276 75. , 332 78. , 990 16. .809 1. , 00 8. .96 c
ATOM 2096 CH2 TRP A 276 74. , 555 78 , , 028 16. , 167 1. , 00 8. ,94 C
ATOM 2097 CZ2 TRP A 276 74. , 483 77 , , 938 14. , 789 1. , 00 8. , 47 C
ATOM 2098 C TRP A 276 77. , 892 84, , 005 14. , 854 1. , 00 8. ,26 C
ATOM 2099 O TRP A 276 78. , 393 84. , 003 15. .962 1. , 00 9. .13 0
ATOM 2100 N GLN A 277 78. , 180 84, , 941 13. ,960 1. , 00 8. ,58 N
ATOM 2101 CA GLN A 277 79. , 079 86 , , 062 14. ,308 1. , 00 9. , 13 C
ATOM 2102 CB GLN A 277 79. ,352 86 , , 934 13. , 071 1. , 00 9. ,92 C
ATOM 2103 CG GLN A 277 80. , 263 88. , 139 13. .335 1. , 00 8. .79 c
ATOM 2104 CD GLN A 277 81. , 686 87, , 732 13. , 527 1. , 00 8. ,57 C
ATOM 2105 OE1 GLN A 277 82. , 090 86 , , 651 13. , 139 1. , 00 9. ,25 O
ATOM 2106 NE2 GLN A 277 82. , 463 88 , , 590 14. , 171 1. , 00 10. ,36 N
ATOM 2107 C GLN A 277 78. , 522 86. , 943 15. .434 1. , 00 9. .78 c
ATOM 2108 O GLN A 277 79. , 256 87, ,310 16. , 355 1. , 00 10. ,07 O
ATOM 2109 N ALA A 278 77. , 239 87, , 285 15. ,370 1. , 00 10. ,30 N
ATOM 2110 CA ALA A 278 76. , 564 87, , 992 16. , 499 1. , 00 11. ,00 C
ATOM 2111 CB ALA A 278 75. , 102 88. , 337 16. .144 1. , 00 9. .15 c
ATOM 2112 C ALA A 278 76. , 631 87, , 147 17. , 804 1. , 00 10. ,57 C
ATOM 2113 O ALA A 278 76. , 927 87, , 675 18. , 892 1. , 00 11. ,24 O
ATOM 2114 N PHE A 279 76 ,.417 85 ,.846 17..692 1., 00 10..59 N
ATOM 2115 CA PHE A 279 76 , .454 84, .946 18. .867 1. , 00 9. .81 C
ATOM 2116 CB PHE A 279 75 , .912 83 , .557 18. .507 1. , 00 9. .84 C
ATOM 2117 CG PHE A 279 76. .025 82. .534 19. .623 1. .00 8. .63 c
ATOM 2118 CD1 PHE A 279 75 , .045 82 , .437 20. .606 1. , 00 9. .28 C
ATOM 2119 CE1 PHE A 279 75 , .157 81 , .481 21. .635 1. , 00 10. .65 C
ATOM 2120 CZ PHE A 279 76 , .227 80 , .602 21. .654 1. , 00 9. .20 C
ATOM 2121 CE2 PHE A 279 77 , .194 80 , .681 20. .661 1. , 00 8. .93 c
ATOM 2122 CD2 PHE A 279 77 , .083 81 , .632 19. .651 1. , 00 8. .26 C
ATOM 2123 C PHE A 279 77 , .860 84, .824 19. .483 1. , 00 10. .19 C
ATOM 2124 O PHE A 279 78 , .035 85, .053 20. .694 1. , 00 10. .52 O
ATOM 2125 N THR A 280 78 , .861 84 , .476 18. .675 1. , 00 9. .66 N
ATOM 2126 CA THR A 280 80 , .201 84, .331 19. .221 1. , 00 8. .43 C
ATOM 2127 CB THR A 280 81 , .186 83 , .689 18. .227 1. , 00 8. .87 C
ATOM 2128 OG1 THR A 280 81 , .401 84, .562 17. .105 1. , 00 6. .74 O
ATOM 2129 CG2 THR A 280 80 , .643 82 , .319 17. .770 1. , 00 8. .64 c
ATOM 2130 C THR A 280 80 , .770 85, .648 19. .742 1. , 00 9. .37 C
ATOM 2131 O THR A 280 81 , .550 85, .636 20. .686 1. , 00 8. .49 O
ATOM 2132 N THR A 281 80 , .413 86 , .774 19. .137 1. , 00 9. .37 N
ATOM 2133 CA THR A 281 80. .868 88 , .082 19. .646 1. , 00 9. .96 c
ATOM 2134 CB THR A 281 80 , .388 89 , .234 18. .732 1. , 00 10. .41 C
ATOM 2135 OG1 THR A 281 81 , .007 89 , .085 17. .461 1. , 00 10. .47 O
ATOM 2136 CG2 THR A 281 80 , .728 90 , .647 19. .324 1. , 00 10. .46 c
ATOM 2137 C THR A 281 80. .344 88 , .312 21. .068 1. , 00 10. .67 c
ATOM 2138 O THR A 281 81 , .082 88 , .710 21. .965 1. , 00 10. .13 O
ATOM 2139 N ARG A 282 79 , .059 88 , .042 21. .255 1. , 00 11. .38 N
ATOM 2140 CA ARG A 282 78. .415 88 , .222 22. .544 1. .00 12. .07 c
ATOM 2141 CB ARG A 282 76 , .888 88 , .182 22. .392 1. , 00 13. .37 C
ATOM 2142 CG ARG A 282 76 , .439 89 , .573 21. .990 1. , 00 16. .82 C
ATOM 2143 CD ARG A 282 74 , .982 89 , .693 21. .745 1. , 00 21. .18 C
ATOM 2144 NE ARG A 282 74 , .212 89 , .471 22. .962 1. , 00 22. .09 N
ATOM 2145 CZ ARG A 282 72 , .896 89 , .567 22. .991 1. , 00 18. .56 C
ATOM 2146 NH1 ARG A 282 72 , .248 89 , .899 21. .891 1. , 00 21. .11 N
ATOM 2147 NH2 ARG A 282 72 , .233 89 , .301 24. .103 1. , 00 22. .68 N
ATOM 2148 C ARG A 282 78 , .926 87 , .256 23. .597 1. , 00 9. .91 c
ATOM 2149 O ARG A 282 79 , .157 87, ,666 24. .732 1. , 00 9. .98 O
ATOM 2150 N ILE A 283 79 , .186 86 , .008 23. .211 1. , 00 8. .83 N
ATOM 2151 CA ILE A 283 79 , .809 85, .034 24. .144 1. , 00 8. .69 C
ATOM 2152 CB ILE A 283 79 , .914 83 , .612 23. .560 1. , 00 8. .33 c
ATOM 2153 CGI ILE A 283 78 , .512 83 , .012 23. .332 1. , 00 8. .21 C
ATOM 2154 CD1 ILE A 283 77 , .723 82 , .744 24. .600 1. , 00 7. .73 C
ATOM 2155 CG2 ILE A 283 80 , .757 82 , .724 24. .461 1. , 00 7. .70 c
ATOM 2156 C ILE A 283 81 , .200 85 , .522 24. .533 1. , 00 9. .40 c
ATOM 2157 O ILE A 283 81 , .547 85, .519 25. .708 1. , 00 8. .82 O
ATOM 2158 N ARG A 284 81 , .986 85, .988 23. .555 1. , 00 9. .27 N
ATOM 2159 CA ARG A 284 83 ,, 368 86 ,, 323 23., 810 1., 00 9., 40 C
ATOM 2160 CB ARG A 284 84, , 219 86 , , 135 22. , 570 1. , 00 9. ,54 C
ATOM 2161 CG ARG A 284 84, , 283 84, , 672 22. , 149 1. , 00 9. , 85 C
ATOM 2162 CD ARG A 284 84 , .867 84. , 517 20. .762 1. .00 11. .05 c
ATOM 2163 NE ARG A 284 85 , , 155 83 , , 130 20. , 424 1. , 00 9. ,94 N
ATOM 2164 CZ ARG A 284 85, , 190 82 , , 648 19. , 192 1. , 00 10. , 48 C
ATOM 2165 NH1 ARG A 284 84, , 932 83 , , 437 18. , 158 1. , 00 11. ,07 N
ATOM 2166 NH2 ARG A 284 85 , .506 81. ,370 18. .982 1. , 00 9. .33 N
ATOM 2167 C ARG A 284 83 , , 561 87, , 712 24. , 418 1. , 00 10. ,50 C
ATOM 2168 O ARG A 284 84, , 616 87, , 981 24. , 964 1. , 00 9. , 41 O
ATOM 2169 N ALA A 285 82 , , 542 88 , , 558 24. ,397 1. , 00 12. ,56 N
ATOM 2170 CA ALA A 285 82 , .618 89. , 842 25. .131 1. , 00 14. .16 C
ATOM 2171 CB ALA A 285 81 , ,335 90 , , 649 24. ,977 1. , 00 13. ,34 C
ATOM 2172 C ALA A 285 82 , , 890 89 , , 560 26. ,603 1. , 00 16. , 76 C
ATOM 2173 O ALA A 285 83 , , 639 90 , , 278 27. ,241 1. , 00 14. , 83 O
ATOM 2174 N LYS A 286 82 , .289 88. , 491 27. .123 1. , 00 17. .44 N
ATOM 2175 CA LYS A 286 82 , , 434 88 , , 068 28. , 517 1. , 00 21. , 02 C
ATOM 2176 CB LYS A 286 81 , , 126 87, ,360 28. ,937 1. , 00 27. ,53 C
ATOM 2177 CG LYS A 286 79 , , 855 88 , , 212 28. , 846 1. , 00 36. ,52 C
ATOM 2178 CD LYS A 286 79 , .645 89. , 078 30. .082 1. , 00 41. .07 c
ATOM 2179 CE LYS A 286 78 , ,341 89 , , 872 29. , 999 1. , 00 48. ,98 C
ATOM 2180 NZ LYS A 286 77 , , 157 89 , , 108 30. ,506 1. , 00 43. ,98 N
ATOM 2181 C LYS A 286 83 , , 606 87, , 089 28. , 704 1. , 00 17. ,36 C
ATOM 2182 O LYS A 286 84 , .221 87. ,044 29. .750 1. , 00 16. .20 O
ATOM 2183 N HIS A 287 83 , , 890 86 , , 297 27. ,671 1. , 00 16. ,25 N
ATOM 2184 CA HIS A 287 84, , 897 85, , 226 27. , 723 1. , 00 11. ,67 C
ATOM 2185 CB HIS A 287 84 , .176 83. , 887 27. .787 1. , 00 10. .16 c
ATOM 2186 CG HIS A 287 83 , , 236 83 , , 803 28. , 965 1. , 00 11. ,21 C
ATOM 2187 ND1 HIS A 287 83 , , 671 83 , , 554 30. , 231 1. , 00 10. ,60 N
ATOM 2188 CE1 HIS A 287 82 , , 627 83 , , 636 31. , 088 1. , 00 11. ,11 C
ATOM 2189 NE2 HIS A 287 81 , .513 83. , 926 30. .365 1. , 00 11. .59 N
ATOM 2190 CD2 HIS A 287 81 , , 867 84, , 063 29. , 063 1. , 00 12. ,14 C
ATOM 2191 C HIS A 287 85, , 807 85, ,319 26. , 542 1. , 00 11. ,38 C
ATOM 2192 O HIS A 287 85, , 665 84, , 559 25. ,601 1. , 00 10. , 48 O
ATOM 2193 N PRO A 288 86 , .754 86. , 268 26. .568 1. , 00 11. .91 N
ATOM 2194 CA PRO A 288 87, , 639 86 , , 448 25. , 430 1. , 00 13. , 13 C
ATOM 2195 CB PRO A 288 88 , , 691 87, , 450 25. , 951 1. , 00 16. ,17 C
ATOM 2196 CG PRO A 288 88 , , 016 88 , , 184 27. , 054 1. , 00 15. , 44 C
ATOM 2197 CD PRO A 288 87 , .025 87. , 239 27. .652 1. , 00 14. .38 c
ATOM 2198 C PRO A 288 88 , , 294 85, , 147 24. ,969 1. , 00 12. ,37 C
ATOM 2199 O PRO A 288 88 , , 768 84, ,379 25. , 775 1. , 00 11. , 70 O
ATOM 2200 N GLY A 289 88 , ,301 84, , 912 23. , 666 1. , 00 12. ,99 N
ATOM 2201 CA GLY A 289 88 , .974 83. , 756 23. .096 1. , 00 14. .02 c
ATOM 2202 C GLY A 289 88 , , 232 82 , , 435 23. ,283 1. , 00 15. , 06 C
ATOM 2203 O GLY A 289 88 , , 771 81 , , 411 22. ,938 1. , 00 12. ,37 O
ATOM 2204 N PHE A 290 86., 999 82 ,.444 23..809 1., 00 11..44 N
ATOM 2205 CA PHE A 290 86. ,307 81 , .192 24. .102 1. , 00 10. .22 C
ATOM 2206 CB PHE A 290 84. , 875 81 , .445 24. . 620 1. , 00 9. .87 C
ATOM 2207 CG PHE A 290 84. .238 80 , .258 25. .257 1. , 00 8. .59 c
ATOM 2208 CD1 PHE A 290 84. , 499 79 , .942 26. .580 1. , 00 8. .96 C
ATOM 2209 CE1 PHE A 290 83. , 913 78 , .832 27. .166 1. , 00 8. .20 C
ATOM 2210 CZ PHE A 290 83. , 079 78 , .023 26. .439 1. , 00 9. .40 C
ATOM 2211 CE2 PHE A 290 82. , 790 78 , .343 25. .125 1. , 00 8. .96 c
ATOM 2212 CD2 PHE A 290 83. ,392 79 , .452 24. .543 1. , 00 8. .91 C
ATOM 2213 C PHE A 290 86. ,316 80 , .317 22. .836 1. , 00 9. .94 C
ATOM 2214 O PHE A 290 85. , 907 80 , .755 21. .740 1. , 00 10. .10 O
ATOM 2215 N PHE A 291 86. , 825 79 , .103 22. .976 1. , 00 9. .05 N
ATOM 2216 CA PHE A 291 87. , 013 78 , .210 21. .814 1. , 00 9. .15 C
ATOM 2217 CB PHE A 291 88. , 145 77 , .220 22. .076 1. , 00 9. .25 C
ATOM 2218 CG PHE A 291 88. , 474 76 , .347 20. .871 1. , 00 9. .99 c
ATOM 2219 CD1 PHE A 291 89. , 121 76 , .887 19. .754 1. , 00 10. .58 c
ATOM 2220 CE1 PHE A 291 89. , 433 76 , .081 18. .662 1. , 00 12. .19 C
ATOM 2221 CZ PHE A 291 89. , 133 74 , .731 18. .695 1. , 00 11. .62 C
ATOM 2222 CE2 PHE A 291 88. , 477 74 , .205 19. .785 1. , 00 11. .64 c
ATOM 2223 CD2 PHE A 291 88. , 166 75 , .019 20. .866 1. , 00 10. .63 c
ATOM 2224 C PHE A 291 85. , 735 77 , .434 21. .441 1. , 00 9. .18 C
ATOM 2225 O PHE A 291 85. , 036 76 , .887 22. .309 1. , 00 8. .66 O
ATOM 2226 N MET A 292 85. , 433 77 , .413 20. .152 1. , 00 7. .88 N
ATOM 2227 CA MET A 292 84. , 284 76 , .716 19. .632 1. , 00 9. . 77 c
ATOM 2228 CB MET A 292 83. , 108 77 , .659 19. .387 1. , 00 10. .35 C
ATOM 2229 CG MET A 292 82. , 811 78 , .590 20. .549 1. , 00 11. .54 C
ATOM 2230 SD MET A 292 81. ,378 79 , .612 20. .212 1. , 00 14. .13 s
ATOM 2231 CE MET A 292 81. , 483 80 , .824 21. .522 1. , 00 8. .87 C
ATOM 2232 C MET A 292 84. , 632 76 , .044 18. .318 1. , 00 9. .69 C
ATOM 2233 O MET A 292 85. , 452 76 , .554 17. .556 1. , 00 9. .85 O
ATOM 2234 N PHE A 293 84. , 023 74 , .887 18. .082 1. , 00 9. .21 N
ATOM 2235 CA PHE A 293 84. , 140 74 , .214 16. .792 1. , 00 9. .16 C
ATOM 2236 CB PHE A 293 85. , 195 73 , .102 16. .794 1. , 00 10. .12 C
ATOM 2237 CG PHE A 293 84. , 996 72 , .012 17. .819 1. , 00 11. .55 C
ATOM 2238 CD1 PHE A 293 85. , 522 72 , .138 19. .099 1. , 00 11. .47 c
ATOM 2239 CE1 PHE A 293 85. ,374 71 , .115 20. .041 1. , 00 12. .63 C
ATOM 2240 CZ PHE A 293 84. , 740 69 , .935 19. .688 1. , 00 12. .04 C
ATOM 2241 CE2 PHE A 293 84. , 237 69 , .786 18. .398 1. , 00 12. .95 c
ATOM 2242 CD2 PHE A 293 84. , 400 70 , .808 17. .459 1. , 00 10. .73 c
ATOM 2243 C PHE A 293 82. , 777 73 , .754 16. .279 1. , 00 9. .55 C
ATOM 2244 O PHE A 293 81. , 991 73 , .147 16. .992 1. , 00 9. .18 O
ATOM 2245 N GLY A 294 82. , 496 74 , .053 15. .021 1. , 00 9. .20 N
ATOM 2246 CA GLY A 294 81. , 203 73 , .738 14. .477 1. , 00 9. .79 c
ATOM 2247 C GLY A 294 81. , 115 72 , .512 13. .588 1. , 00 10. .37 C
ATOM 2248 O GLY A 294 82. , 108 72 , .110 12. .940 1. , 00 8. .84 O
ATOM 2249 N GLU A 295 79 ,.908 71 ,.946 13..560 1., 00 9..56 N
ATOM 2250 CA GLU A 295 79 , .588 70 , .801 12. .739 1. , 00 10. .45 C
ATOM 2251 CB GLU A 295 78 , .672 69 , .815 13. .450 1. , 00 10. .41 C
ATOM 2252 CG GLU A 295 78. .214 68. .681 12. .515 1. , 00 9. .73 c
ATOM 2253 CD GLU A 295 76 , .714 68 , .373 12. .552 1. , 00 12. .79 C
ATOM 2254 OE1 GLU A 295 75 , .900 69 , .107 13. .171 1. , 00 12. .40 O
ATOM 2255 OE2 GLU A 295 76 , .325 67, .362 11. .909 1. , 00 12. .93 O
ATOM 2256 C GLU A 295 78 , .901 71 , .281 11. .467 1. , 00 9. .52 c
ATOM 2257 O GLU A 295 77 , .705 71 , .599 11. .487 1. , 00 9. .34 O
ATOM 2258 N ALA A 296 79 , .692 71 , .360 10. .393 1. , 00 8. .43 N
ATOM 2259 CA ALA A 296 79 , .210 71 , .570 9. .030 1. , 00 10. .11 C
ATOM 2260 CB ALA A 296 79 , .970 72 , .714 8. .353 1. , 00 9. .64 c
ATOM 2261 C ALA A 296 79 , .409 70 , .251 8. .284 1. , 00 9. .91 C
ATOM 2262 O ALA A 296 80 , .539 69 , .874 7. .984 1. , 00 11. .06 O
ATOM 2263 N PHE A 297 78 , .309 69 , .545 8. .019 1. , 00 10. .07 N
ATOM 2264 CA PHE A 297 78 , .357 68 , .157 7. .504 1. , 00 10. .08 c
ATOM 2265 CB PHE A 297 77 , .140 67, .342 7. .961 1. , 00 10. .16 C
ATOM 2266 CG PHE A 297 77 , .341 65, .851 7. .936 1. , 00 11. .39 C
ATOM 2267 CD1 PHE A 297 77 , .927 65, .211 6. .851 1. , 00 11. .69 C
ATOM 2268 CE1 PHE A 297 78. .076 63 , .835 6. .822 1. , 00 12. .87 C
ATOM 2269 CZ PHE A 297 77 , .593 63 , .081 7. .863 1. , 00 11. .34 C
ATOM 2270 CE2 PHE A 297 77 , .009 63 , .700 8. .936 1. , 00 11. .87 C
ATOM 2271 CD2 PHE A 297 76 , .879 65, .078 8. .975 1. , 00 12. .63 c
ATOM 2272 C PHE A 297 78. .479 68 , .253 6. .000 1. , 00 9. .35 C
ATOM 2273 O PHE A 297 77 , .481 68 , .241 5. .255 1. , 00 7. .99 O
ATOM 2274 N ASP A 298 79 , .734 68 , .362 5. .561 1. , 00 9. .67 N
ATOM 2275 CA ASP A 298 80. .038 68 , .660 4. .167 1. , 00 9. .68 c
ATOM 2276 CB ASP A 298 79 , .778 70 , .150 3. .934 1. , 00 10. .36 C
ATOM 2277 CG ASP A 298 79 , .952 70 , .597 2. .484 1. , 00 13. .00 C
ATOM 2278 OD1 ASP A 298 80 , .669 69 , .928 1. .694 1. , 00 14. .93 O
ATOM 2279 OD2 ASP A 298 79 , .395 71 , .666 2. .141 1. , 00 12. .78 0
ATOM 2280 C ASP A 298 81 , .511 68 , .282 3. .964 1. , 00 9. .95 C
ATOM 2281 O ASP A 298 82 , .378 68 , .683 4. .741 1. , 00 10. .40 O
ATOM 2282 N TYR A 299 81 , .779 67, .466 2. .950 1. , 00 9. .57 N
ATOM 2283 CA TYR A 299 83 , .139 66 , .995 2. .653 1. , 00 11. .03 c
ATOM 2284 CB TYR A 299 83 , .053 65, .591 2. .031 1. , 00 12. .44 C
ATOM 2285 CG TYR A 299 82 , .544 64, .590 3. .060 1. , 00 11. .99 C
ATOM 2286 CD1 TYR A 299 83 , .360 64 , .170 4. .084 1. , 00 12. .22 C
ATOM 2287 CE1 TYR A 299 82 , .914 63 , .285 5. .045 1. , 00 11. .45 c
ATOM 2288 CZ TYR A 299 81 , .626 62 , .803 5. .006 1. , 00 14. .21 C
ATOM 2289 OH TYR A 299 81 , .241 61 , .901 5. .984 1. , 00 17. .09 O
ATOM 2290 CE2 TYR A 299 80 , .768 63 , .207 3. .997 1. , 00 13. .87 C
ATOM 2291 CD2 TYR A 299 81 , .237 64 , .107 3. .026 1. , 00 13. .67 c
ATOM 2292 C TYR A 299 83 , .995 67, .938 1. .809 1. , 00 11. .50 C
ATOM 2293 O TYR A 299 85, .140 67, .619 1. .474 1. , 00 11. .92 O
ATOM 2294 N ASP A 300 83 , , 457 69 , , 121 1. , 527 1. , 00 11. , 83 N
ATOM 2295 CA ASP A 300 84, , 159 70 , , 163 0. , 798 1. , 00 9. , 72 C
ATOM 2296 CB ASP A 300 83 , , 194 70 , , 903 -0. , 112 1. , 00 10. , 78 C
ATOM 2297 CG ASP A 300 83. .924 71. , 912 -0. .992 1. , 00 15. .25 c
ATOM 2298 OD1 ASP A 300 84 , , 592 71 , , 447 -1. ,917 1. , 00 17. , 17 0
ATOM 2299 OD2 ASP A 300 83 , , 918 73 , , 135 -0. , 703 1. , 00 16. ,54 O
ATOM 2300 C ASP A 300 84, , 822 71 , , 191 1. , 719 1. , 00 9. ,38 C
ATOM 2301 O ASP A 300 84 , .124 71. , 947 2. .455 1. , 00 8. .06 0
ATOM 2302 N ALA A 301 86 , , 152 71 , , 269 1. ,652 1. , 00 8. ,38 N
ATOM 2303 CA ALA A 301 86 , , 925 72 , , 136 2. , 569 1. , 00 8. ,68 C
ATOM 2304 CB ALA A 301 88 , , 423 71 , , 973 2. ,308 1. , 00 8. ,56 C
ATOM 2305 C ALA A 301 86 , .546 73. , 617 2. .480 1. , 00 9. .46 c
ATOM 2306 O ALA A 301 86 , ,364 74 , ,303 3. , 499 1. , 00 9. , 71 O
ATOM 2307 N ALA A 302 86 , , 467 74 , , 127 1. ,270 1. , 00 11. , 18 N
ATOM 2308 CA ALA A 302 86 , , 122 75 , , 529 1. , 036 1. , 00 11. , 02 C
ATOM 2309 CB ALA A 302 86 , .191 75. , 844 -0. .462 1. , 00 10. .72 c
ATOM 2310 C ALA A 302 84, , 739 75 , , 900 1. ,608 1. , 00 12. ,21 C
ATOM 2311 O ALA A 302 84, , 576 76 , , 947 2. ,212 1. , 00 11. ,22 O
ATOM 2312 N ARG A 303 83 , , 731 75 , , 055 1. , 412 1. , 00 13. ,60 N
ATOM 2313 CA ARG A 303 82 , .399 75. , 356 1. .965 1. , 00 12. .42 c
ATOM 2314 CB ARG A 303 81 , ,305 74 , , 425 1. ,397 1. , 00 13. ,39 C
ATOM 2315 CG ARG A 303 80 , , 699 74 , , 895 0. , 083 1. , 00 16. , 07 C
ATOM 2316 CD ARG A 303 79 , , 416 74 , , 164 -0. ,301 1. , 00 14. ,66 c
ATOM 2317 NE ARG A 303 79 , .670 72. , 733 -0. .348 1. , 00 16. .46 N
ATOM 2318 CZ ARG A 303 79 , , 876 71 , , 991 -1. , 440 1. , 00 14. ,22 C
ATOM 2319 NH1 ARG A 303 79 , , 811 72 , , 505 -2. , 665 1. , 00 15. , 11 N
ATOM 2320 NH2 ARG A 303 80 , .145 70. , 702 -1. .294 1. , 00 12. .11 N
ATOM 2321 C ARG A 303 82 , ,376 75 , ,344 3. , 504 1. , 00 11. ,66 C
ATOM 2322 O ARG A 303 81 , , 738 76 , , 190 4. , 091 1. , 00 12. , 84 O
ATOM 2323 N ILE A 304 83 , , 035 74 , ,399 4. , 170 1. , 00 9. , 76 N
ATOM 2324 CA ILE A 304 83 , .008 74. , 409 5. .624 1. , 00 9. .85 C
ATOM 2325 CB ILE A 304 83 , , 284 73 , , 014 6. ,263 1. , 00 9. , 85 C
ATOM 2326 CGI ILE A 304 84, , 706 72 , , 493 6. , 013 1. , 00 9. , 03 C
ATOM 2327 CD1 ILE A 304 85, , 056 71 , , 286 6. , 873 1. , 00 7. ,52 C
ATOM 2328 CG2 ILE A 304 82 , .282 71. , 965 5. .745 1. , 00 9. .80 c
ATOM 2329 C ILE A 304 83 , , 945 75 , , 488 6. , 195 1. , 00 10. ,95 C
ATOM 2330 O ILE A 304 83 , , 711 76 , , 000 7. ,314 1. , 00 9. , 86 O
ATOM 2331 N ALA A 305 84, , 961 75 , , 883 5. , 406 1. , 00 9. , 86 N
ATOM 2332 CA ALA A 305 85 , .950 76. , 848 5. .879 1. , 00 9. .62 c
ATOM 2333 CB ALA A 305 87, , 108 76 , , 978 4. , 878 1. , 00 8. ,53 C
ATOM 2334 C ALA A 305 85, ,361 78 , , 232 6. , 187 1. , 00 8. ,63 C
ATOM 2335 O ALA A 305 85, , 946 79 , , 000 6. ,948 1. , 00 7. ,54 0
ATOM 2336 N THR A 306 84 , .252 78. , 574 5. .548 1. , 00 9. .00 N
ATOM 2337 CA THR A 306 83 , , 642 79 , , 902 5. ,669 1. , 00 10. ,22 C
ATOM 2338 CB THR A 306 82 , ,323 79 , , 974 4. , 860 1. , 00 14. ,38 C
ATOM 2339 OG1 THR A 306 81 ,, 466 78., 879 5..265 1., 00 21..19 O
ATOM 2340 CG2 THR A 306 82 , , 580 79. , 887 3. .361 1. , 00 11. .24 C
ATOM 2341 C THR A 306 83 , ,358 80. , 293 7. .132 1. , 00 10. .35 C
ATOM 2342 O THR A 306 83. .494 81. , 456 7. .516 1. , 00 9. .68 O
ATOM 2343 N HIS A 307 83 , , 002 79. ,300 7. .947 1. , 00 8. .78 N
ATOM 2344 CA HIS A 307 82 , , 683 79. , 498 9. .351 1. , 00 9. .49 C
ATOM 2345 CB HIS A 307 82 , , 091 78. , 195 9. .919 1. , 00 8. .87 C
ATOM 2346 CG HIS A 307 80 , .878 77. , 723 9. .189 1. , 00 8. .30 c
ATOM 2347 ND1 HIS A 307 79 , , 697 78. ,362 9. .266 1. , 00 8. .68 N
ATOM 2348 CE1 HIS A 307 78 , , 783 77. , 731 8. .498 1. , 00 7. .50 C
ATOM 2349 NE2 HIS A 307 79 , ,384 76. , 691 7. .898 1. , 00 8. .42 N
ATOM 2350 CD2 HIS A 307 80 , .685 76. , 646 8. .309 1. , 00 9. .11 C
ATOM 2351 C HIS A 307 83 , , 871 79. , 903 10. .166 1. , 00 9. .45 C
ATOM 2352 O HIS A 307 83 , , 705 80. , 474 11. .242 1. , 00 11. .07 O
ATOM 2353 N THR A 308 85, , 081 79. , 646 9. .652 1. , 00 9. .41 N
ATOM 2354 CA THR A 308 86 , .318 79. , 958 10. .351 1. , 00 9. .31 c
ATOM 2355 CB THR A 308 87, , 501 79. , 029 9. .955 1. , 00 9. .37 C
ATOM 2356 OG1 THR A 308 87, , 961 79. ,325 8. .630 1. , 00 7. .62 O
ATOM 2357 CG2 THR A 308 87, , 113 77. , 525 10. .036 1. , 00 6. .91 C
ATOM 2358 C THR A 308 86 , .752 81. , 411 10. .137 1. , 00 10. .18 c
ATOM 2359 O THR A 308 87, , 598 81. , 896 10. .855 1. , 00 8. .94 O
ATOM 2360 N LEU A 309 86 , , 170 82. , 093 9. .162 1. , 00 10. .55 N
ATOM 2361 CA LEU A 309 86 , , 743 83. ,343 8. .688 1. , 00 11. .94 c
ATOM 2362 CB LEU A 309 86 , .366 83. , 554 7. .222 1. , 00 12. .52 c
ATOM 2363 CG LEU A 309 86 , , 850 82. , 471 6. .250 1. , 00 15. .75 C
ATOM 2364 CD1 LEU A 309 86 , ,375 82. , 827 4. .826 1. , 00 16. .29 C
ATOM 2365 CD2 LEU A 309 88 , .371 82. , 260 6. .365 1. , 00 12. .11 c
ATOM 2366 C LEU A 309 86 , .256 84. , 547 9. .488 1. , 00 13. .55 C
ATOM 2367 O LEU A 309 85, .178 84. , 488 10. .098 1. , 00 11. .41 O
ATOM 2368 N PRO A 310 87, .032 85. , 658 9. .464 1. , 00 14. .42 N
ATOM 2369 CA PRO A 310 86 , .597 86. , 860 10. .179 1. , 00 13. .98 c
ATOM 2370 CB PRO A 310 87, .711 87. , 881 9. .887 1. , 00 16. .41 C
ATOM 2371 CG PRO A 310 88 , .922 87. , 061 9. .574 1. , 00 16. .94 C
ATOM 2372 CD PRO A 310 88 , .389 85. , 810 8. .893 1. , 00 16. .27 C
ATOM 2373 C PRO A 310 85 , .244 87. , 355 9. .688 1. , 00 12. .93 c
ATOM 2374 O PRO A 310 85, .000 87. , 407 8. .480 1. , 00 11. .94 O
ATOM 2375 N GLY A 311 84, .358 87. , 675 10. .625 1. , 00 12. .00 N
ATOM 2376 CA GLY A 311 83 , .024 88. , 172 10. .296 1. , 00 11. .69 C
ATOM 2377 C GLY A 311 82 , .001 87. , 082 10. .013 1. , 00 12. .30 c
ATOM 2378 O GLY A 311 80 , .854 87. ,384 9. .832 1. , 00 12. .37 O
ATOM 2379 N HIS A 312 82 , .412 85. , 812 10. .002 1. , 00 11. .89 N
ATOM 2380 CA HIS A 312 81 , .531 84. , 718 9. .611 1. , 00 12. .94 C
ATOM 2381 CB HIS A 312 82 , .092 84. , 023 8. .373 1. , 00 12. .91 c
ATOM 2382 CG HIS A 312 81 , .940 84. , 839 7. .129 1. , 00 17. .78 C
ATOM 2383 ND1 HIS A 312 80 , .922 84. , 649 6. .259 1. , 00 19. .08 N
ATOM 2384 CE1 HIS A 312 81., 031 85 ,.530 5..238 1., 00 21..50 C
ATOM 2385 NE2 HIS A 312 82. , 108 86 , .312 5. .478 1. , 00 24. .25 N
ATOM 2386 CD2 HIS A 312 82. , 693 85, .905 6. .636 1. , 00 18. .49 C
ATOM 2387 C HIS A 312 81. .280 83 , .729 10. .729 1. , 00 11. .50 C
ATOM 2388 O HIS A 312 80. , 817 82 , .621 10. .477 1. , 00 11. .13 O
ATOM 2389 N GLY A 313 81. , 528 84, .172 11. . 963 1. , 00 10. .71 N
ATOM 2390 CA GLY A 313 81. ,391 83 , .368 13. .158 1. , 00 11. .15 C
ATOM 2391 C GLY A 313 82. , 693 83 , .135 13. .905 1. , 00 10. .67 c
ATOM 2392 O GLY A 313 82. , 700 83 , .087 15. .125 1. , 00 11. .53 O
ATOM 2393 N GLU A 314 83. , 785 82 , .962 13. .175 1. , 00 10. .32 N
ATOM 2394 CA GLU A 314 85. , 125 82 , .880 13. .757 1. , 00 12. .38 C
ATOM 2395 CB GLU A 314 85. , 483 84 , .221 14. .445 1. , 00 14. .49 c
ATOM 2396 CG GLU A 314 85. , 600 85, .376 13. .446 1. , 00 17. .46 C
ATOM 2397 CD GLU A 314 85. , 534 86 , .784 14. .058 1. , 00 20. .63 C
ATOM 2398 OE1 GLU A 314 85. , 541 86 , .931 15. .290 1. , 00 24. .13 O
ATOM 2399 OE2 GLU A 314 85. , 442 87 , .759 13. .289 1. , 00 15. .75 0
ATOM 2400 C GLU A 314 85. , 251 81 , .690 14. .700 1. , 00 10. .85 C
ATOM 2401 O GLU A 314 85. , 640 81 , .833 15. .842 1. , 00 10. .16 O
ATOM 2402 N THR A 315 84. , 842 80 , .515 14. .233 1. , 00 10. .17 N
ATOM 2403 CA THR A 315 84. , 970 79 , .309 15. .032 1. , 00 10. .01 c
ATOM 2404 CB THR A 315 83. , 609 78 , .646 15. .383 1. , 00 10. .24 C
ATOM 2405 OG1 THR A 315 82. , 962 78 , .136 14. .202 1. , 00 11. .04 O
ATOM 2406 CG2 THR A 315 82. , 663 79 , .635 16. .098 1. , 00 9. .54 c
ATOM 2407 C THR A 315 85. , 846 78 , .365 14. .214 1. , 00 11. .39 c
ATOM 2408 O THR A 315 86. , 026 78 , .550 13. .002 1. , 00 11. .64 O
ATOM 2409 N SER A 316 86. ,380 77 , .356 14. .876 1. , 00 9. .87 N
ATOM 2410 CA SER A 316 86. , 924 76 , .215 14. .178 1. , 00 9. .57 c
ATOM 2411 CB SER A 316 87. , 825 75 , .437 15. .130 1. , 00 9. .99 C
ATOM 2412 OG SER A 316 88. , 293 74 , .240 14. .532 1. , 00 13. .22 O
ATOM 2413 C SER A 316 85. , 753 75 , .372 13. .631 1. , 00 8. .57 C
ATOM 2414 O SER A 316 84. , 574 75 , .694 13. .846 1. , 00 7. .97 0
ATOM 2415 N VAL A 317 86. , 073 74 , .347 12. .857 1. , 00 8. .01 N
ATOM 2416 CA VAL A 317 85. , 083 73 , .392 12. .378 1. , 00 8. .11 C
ATOM 2417 CB VAL A 317 84. , 653 73 , . 640 10. .913 1. , 00 8. .46 C
ATOM 2418 CGI VAL A 317 83. , 855 74 , .952 10. .742 1. , 00 6. .89 c
ATOM 2419 CG2 VAL A 317 85. , 853 73 , .616 9. .961 1. , 00 7. .09 C
ATOM 2420 C VAL A 317 85. , 635 71 , .955 12. .523 1. , 00 9. .18 C
ATOM 2421 O VAL A 317 86. , 853 71 , .742 12. .629 1. , 00 8. .08 O
ATOM 2422 N LEU A 318 84. , 721 70 , .982 12. .515 1. , 00 9. .42 N
ATOM 2423 CA LEU A 318 85. , 064 69 , .589 12. .452 1. , 00 8. .66 C
ATOM 2424 CB LEU A 318 83. , 865 68 , .745 12. .843 1. , 00 9. .25 C
ATOM 2425 CG LEU A 318 83. , 636 68 , .814 14. .347 1. , 00 9. .01 C
ATOM 2426 CD1 LEU A 318 82. ,147 68 , .709 14. .755 1. , 00 8. .12 c
ATOM 2427 CD2 LEU A 318 84. , 499 67, .734 14. .991 1. , 00 7. .86 C
ATOM 2428 C LEU A 318 85. , 561 69 , .242 11. .040 1. , 00 9. .13 C
ATOM 2429 O LEU A 318 84., 936 69., 608 10..032 1., 00 8..59 O
ATOM 2430 N ASP A 319 86. , 688 68. , 547 10. .970 1. , 00 9. .15 N
ATOM 2431 CA ASP A 319 87. ,386 68. ,329 9. .709 1. , 00 9. .72 C
ATOM 2432 CB ASP A 319 88. .877 68. , 105 9. .989 1. , 00 10. .73 c
ATOM 2433 CG ASP A 319 89. , 750 68. , 169 8. .730 1. , 00 11. .92 C
ATOM 2434 OD1 ASP A 319 89. , 197 68. , 122 7. .596 1. , 00 10. .68 O
ATOM 2435 OD2 ASP A 319 91. , 003 68. , 292 8. .894 1. , 00 11. .40 O
ATOM 2436 C ASP A 319 86. , 780 67. ,147 8. .942 1. , 00 10. .86 c
ATOM 2437 O ASP A 319 87. ,364 66. , 057 8. .884 1. , 00 11. .13 O
ATOM 2438 N PHE A 320 85. , 638 67. ,383 8. .293 1. , 00 10. .86 N
ATOM 2439 CA PHE A 320 85. , 007 66. ,332 7. .492 1. , 00 9. .39 C
ATOM 2440 CB PHE A 320 83. , 595 66. , 739 7. .045 1. , 00 10. .08 c
ATOM 2441 CG PHE A 320 82. , 541 66. , 502 8. .101 1. , 00 9. .82 C
ATOM 2442 CD1 PHE A 320 82. , 409 67. ,376 9. .169 1. , 00 10. .08 C
ATOM 2443 CE1 PHE A 320 81. , 465 67. , 155 10. .163 1. , 00 10. .37 C
ATOM 2444 CZ PHE A 320 80. , 620 66. , 064 10. .093 1. , 00 10. .49 c
ATOM 2445 CE2 PHE A 320 80. , 715 65. , 184 9. .007 1. , 00 10. .69 C
ATOM 2446 CD2 PHE A 320 81. , 689 65. , 409 8. .026 1. , 00 10. .64 C
ATOM 2447 C PHE A 320 85. , 874 65. , 825 6. .333 1. , 00 10. .54 C
ATOM 2448 O PHE A 320 85. , 950 64. , 625 6. .125 1. , 00 11. .15 0
ATOM 2449 N PRO A 321 86. , 558 66. , 728 5. .598 1. , 00 10. .61 N
ATOM 2450 CA PRO A 321 87. , 447 66. , 257 4. .533 1. , 00 9. .85 C
ATOM 2451 CB PRO A 321 88. , 059 67. , 547 3. .996 1. , 00 9. .33 c
ATOM 2452 CG PRO A 321 87. , 001 68. , 576 4. .260 1. , 00 10. .73 c
ATOM 2453 CD PRO A 321 86. , 466 68. , 204 5. .620 1. , 00 9. .17 C
ATOM 2454 C PRO A 321 88. , 527 65. , 269 4. .989 1. , 00 10. .60 C
ATOM 2455 O PRO A 321 88. , 766 64. , 270 4. .334 1. , 00 9. .09 0
ATOM 2456 N MET A 322 89. , 144 65. , 523 6. .133 1. , 00 10. .53 N
ATOM 2457 CA MET A 322 90. , 122 64. , 605 6. .663 1. , 00 9. .54 C
ATOM 2458 CB MET A 322 90. , 867 65. , 253 7. .824 1. , 00 9. .97 C
ATOM 2459 CG MET A 322 91. , 833 64. , 361 8. .565 1. , 00 14. .30 c
ATOM 2460 SD MET A 322 93. , 096 63. , 690 7. .471 1. , 00 18. .22 s
ATOM 2461 CE MET A 322 94. , 130 65. , 154 7. .388 1. , 00 19. .25 C
ATOM 2462 C MET A 322 89. , 438 63. ,332 7. .112 1. , 00 10. .68 C
ATOM 2463 O MET A 322 90. , 005 62. , 250 6. .978 1. , 00 9. .25 0
ATOM 2464 N LYS A 323 88. , 212 63. , 427 7. .627 1. , 00 9. .54 N
ATOM 2465 CA LYS A 323 87. , 533 62. , 205 8. .003 1. , 00 9. .37 C
ATOM 2466 CB LYS A 323 86. , 192 62. , 493 8. .674 1. , 00 10. .72 C
ATOM 2467 CG LYS A 323 85. , 409 61. ,244 9. .004 1. , 00 12. .09 c
ATOM 2468 CD LYS A 323 84. ,308 60. , 996 7. .996 1. , 00 13. .58 C
ATOM 2469 CE LYS A 323 84. , 082 59. , 527 7. .716 1. , 00 17. .41 C
ATOM 2470 NZ LYS A 323 82. , 924 59. ,399 6. .788 1. , 00 15. .20 N
ATOM 2471 C LYS A 323 87. ,344 61. , 281 6. .807 1. , 00 8. .95 c
ATOM 2472 O LYS A 323 87. , 502 60. , 062 6. .937 1. , 00 6. .69 O
ATOM 2473 N GLN A 324 86. , 982 61. , 839 5. .655 1. , 00 9. .41 N
ATOM 2474 CA GLN A 324 86 ,, 791 61., 018 4..437 1., 00 10..60 C
ATOM 2475 CB GLN A 324 86 , , 238 61. , 873 3. .290 1. , 00 13. .02 C
ATOM 2476 CG GLN A 324 85, , 877 61. , 138 1. .997 1. , 00 18. .08 C
ATOM 2477 CD GLN A 324 85. .349 62. , 120 0. .935 1. , 00 27. .45 c
ATOM 2478 OE1 GLN A 324 86 , , 076 63. , 007 0. .482 1. , 00 34. .72 O
ATOM 2479 NE2 GLN A 324 84, , 091 61. , 979 0. .557 1. , 00 31. .23 N
ATOM 2480 C GLN A 324 88 , , 106 60. ,390 4. .008 1. , 00 9. .18 C
ATOM 2481 O GLN A 324 88 , .135 59. , 235 3. .578 1. , 00 9. .76 O
ATOM 2482 N ALA A 325 89 , , 188 61. , 152 4. .120 1. , 00 8. .67 N
ATOM 2483 CA ALA A 325 90 , , 533 60. , 633 3. .831 1. , 00 9. .60 C
ATOM 2484 CB ALA A 325 91 , , 597 61. , 752 3. .919 1. , 00 7. .32 C
ATOM 2485 C ALA A 325 90 , .863 59. , 474 4. .775 1. , 00 8. .16 c
ATOM 2486 O ALA A 325 91 , , 298 58. , 404 4. .333 1. , 00 7. .96 O
ATOM 2487 N MET A 326 90 , , 599 59. , 663 6. .059 1. , 00 9. .11 N
ATOM 2488 CA MET A 326 90 , , 811 58. , 592 7. .059 1. , 00 9. .47 C
ATOM 2489 CB MET A 326 90 , .625 59. , 102 8. .477 1. , 00 9. .63 c
ATOM 2490 CG MET A 326 91 , , 741 60. , 034 8. .883 1. , 00 10. .31 C
ATOM 2491 SD MET A 326 91 , , 694 60. , 589 10. .598 1. , 00 11. .08 s
ATOM 2492 CE MET A 326 92 , , 299 59. , 086 11. .400 1. , 00 8. .51 c
ATOM 2493 C MET A 326 89 , .982 57. ,343 6. .817 1. , 00 9. .92 c
ATOM 2494 O MET A 326 90 , , 504 56. , 216 6. .914 1. , 00 9. .99 O
ATOM 2495 N GLU A 327 88 , , 727 57. , 521 6. .425 1. , 00 11. .43 N
ATOM 2496 CA GLU A 327 87, , 881 56. ,387 6. .075 1. , 00 12. .28 c
ATOM 2497 CB GLU A 327 86 , .449 56. ,841 5. .753 1. , 00 14. .88 c
ATOM 2498 CG GLU A 327 85, , 539 55. , 675 5. .407 1. , 00 22. .11 C
ATOM 2499 CD GLU A 327 84, , 143 56. , 113 5. .048 1. , 00 29. .70 C
ATOM 2500 OE1 GLU A 327 83 , .661 57. , 127 5. .597 1. , 00 30. .92 O
ATOM 2501 OE2 GLU A 327 83 , , 538 55. , 448 4. .193 1. , 00 46. .65 O
ATOM 2502 C GLU A 327 88 , , 461 55. , 578 4. .892 1. , 00 12. .13 C
ATOM 2503 O GLU A 327 88 , , 486 54. ,347 4. .926 1. , 00 11. .15 O
ATOM 2504 N GLU A 328 88 , .907 56. , 268 3. .854 1. , 00 10. .41 N
ATOM 2505 CA GLU A 328 89 , , 497 55. , 607 2. .705 1. , 00 11. .59 C
ATOM 2506 CB GLU A 328 89 , , 628 56. , 596 1. .548 1. , 00 13. .52 C
ATOM 2507 CG GLU A 328 88 , , 243 57. , 057 1. .092 1. , 00 17. .77 C
ATOM 2508 CD GLU A 328 88 , .255 57. , 986 -0. .107 1. , 00 22. .00 c
ATOM 2509 OE1 GLU A 328 89 , ,326 58. , 226 -0. .674 1. , 00 21. .70 O
ATOM 2510 OE2 GLU A 328 87, , 175 58. , 497 -0. .467 1. , 00 27. .21 O
ATOM 2511 C GLU A 328 90 , , 837 54. , 924 3. .024 1. , 00 10. .91 C
ATOM 2512 O GLU A 328 91 , .116 53. , 811 2. .544 1. , 00 9. .33 0
ATOM 2513 N VAL A 329 91 , .636 55. , 559 3. .872 1. , 00 10. .66 N
ATOM 2514 CA VAL A 329 92 , .971 55. , 067 4. .172 1. , 00 9. .49 C
ATOM 2515 CB VAL A 329 93 , .865 56. , 225 4. .666 1. , 00 9. .29 c
ATOM 2516 CGI VAL A 329 95 , .176 55. , 693 5. .242 1. , 00 8. .33 c
ATOM 2517 CG2 VAL A 329 94 , .116 57. , 210 3. .515 1. , 00 8. .07 C
ATOM 2518 C VAL A 329 92 , .999 53. , 892 5. .166 1. , 00 10. .93 C
ATOM 2519 O VAL A 329 93 745 52 ,.937 4..975 1.00 13.97 O
ATOM 2520 N PHE A 330 92 216 53 , .993 6. .235 1.00 10.30 N
ATOM 2521 CA PHE A 330 92 216 53 , .007 7. .301 1.00 10.50 C
ATOM 2522 CB PHE A 330 92 300 53. .736 8. .663 1.00 10.10 C
ATOM 2523 CG PHE A 330 93 515 54 , .616 8. .780 1.00 9.79 C
ATOM 2524 CD1 PHE A 330 94 783 54 , .057 8. .743 1.00 9.81 C
ATOM 2525 CE1 PHE A 330 95 923 54 , .840 8. .827 1.00 8.98 C
ATOM 2526 CZ PHE A 330 95 791 56 , .201 8. .941 1.00 9.58 C
ATOM 2527 CE2 PHE A 330 94 535 56 , .770 8. .980 1.00 9.73 C
ATOM 2528 CD2 PHE A 330 93 401 55 , .981 8. .893 1.00 9.05 C
ATOM 2529 C PHE A 330 91 018 52 , .064 7. .279 1.00 12.26 C
ATOM 2530 O PHE A 330 91 043 51 , .042 7. .954 1.00 13.04 O
ATOM 2531 N GLY A 331 89 977 52 , .427 6. .531 1.00 12.02 N
ATOM 2532 CA GLY A 331 88 811 51 , .589 6. .370 1.00 13.60 C
ATOM 2533 C GLY A 331 89 013 50 , .578 5. .263 1.00 15.34 C
ATOM 2534 O GLY A 331 90 153 50 , .314 4. .815 1.00 16.11 O
ATOM 2535 N ARG A 332 87 899 50 , .022 4. .807 1.00 16.06 N
ATOM 2536 CA ARG A 332 87 940 48 , .910 3. .877 1.00 18.06 C
ATOM 2537 CB ARG A 332 86 544 48 , .342 3. .630 1.00 24.83 C
ATOM 2538 CG ARG A 332 86 143 47. .309 4. .669 1.00 37.10 C
ATOM 2539 CD ARG A 332 84 849 46 , .605 4. .285 1.00 48.85 C
ATOM 2540 NE ARG A 332 84 707 45 , .379 5. .064 1.00 59.60 N
ATOM 2541 CZ ARG A 332 85 244 44 , .210 4. .726 1.00 69.59 C
ATOM 2542 NH1 ARG A 332 85 950 44. .078 3. .602 1.00 78.32 N
ATOM 2543 NH2 ARG A 332 85 070 43 , .158 5. .512 1.00 75.92 N
ATOM 2544 C ARG A 332 88 597 49 , .241 2. .548 1.00 15.39 C
ATOM 2545 O ARG A 332 89 214 48. .388 1. .976 1.00 13.38 O
ATOM 2546 N LYS A 333 88 475 50 , .464 2. .048 1.00 14.25 N
ATOM 2547 CA LYS A 333 89 137 50 , .798 0. .780 1.00 16.66 C
ATOM 2548 CB LYS A 333 88 773 52 , .214 0. .364 1.00 20.03 C
ATOM 2549 CG LYS A 333 89 086 52 , .561 -1. .079 1.00 30.01 C
ATOM 2550 CD LYS A 333 88 438 53 , .900 -1. .449 1.00 43.02 C
ATOM 2551 CE LYS A 333 88 714 54 , .319 -2. .889 1.00 44.84 C
ATOM 2552 NZ LYS A 333 90 177 54 , .396 -3. .107 1.00 44.14 N
ATOM 2553 C LYS A 333 90 676 50 , .616 0. .857 1.00 13.92 C
ATOM 2554 O LYS A 333 91 304 50 , .272 -0. .117 1.00 11.72 O
ATOM 2555 N GLN A 334 91 266 50 , .808 2. .029 1.00 10.94 N
ATOM 2556 CA GLN A 334 92 710 50 , .663 2. .204 1.00 12.50 C
ATOM 2557 CB GLN A 334 93 165 49 , .183 2. .116 1.00 14.64 C
ATOM 2558 CG GLN A 334 92 677 48 , .272 3. .251 1.00 14.64 C
ATOM 2559 CD GLN A 334 93 307 48 , .623 4. .579 1.00 15.76 C
ATOM 2560 OE1 GLN A 334 94 497 48 , .398 4. .780 1.00 15.21 O
ATOM 2561 NE2 GLN A 334 92 509 49 , .191 5. .494 1.00 14.54 N
ATOM 2562 C GLN A 334 93 491 51 , .529 1. .192 1.00 10.81 C
ATOM 2563 O GLN A 334 94 433 51 , .069 0. .572 1.00 9.71 O
ATOM 2564 N ALA A 335 93 ,, 103 52 ,, 776 1., 032 1., 00 10..60 N
ATOM 2565 CA ALA A 335 93 , , 894 53 , , 690 0. , 186 1. , 00 11. .76 C
ATOM 2566 CB ALA A 335 93 , , 136 55 , , 015 -0. , 060 1. , 00 12. .19 C
ATOM 2567 C ALA A 335 95. .275 53 , .937 0. .832 1. , 00 9. .79 c
ATOM 2568 O ALA A 335 95 , , 458 53 , , 776 2. , 039 1. , 00 9. .13 O
ATOM 2569 N GLY A 336 96 , , 267 54 , , 247 0. , 011 1. , 00 9. .47 N
ATOM 2570 CA GLY A 336 97, , 589 54 , , 612 0. , 521 1. , 00 9. .15 C
ATOM 2571 C GLY A 336 97 , .549 55 , .985 1. .166 1. , 00 8. .00 c
ATOM 2572 O GLY A 336 96 , , 621 56 , , 775 0. ,931 1. , 00 8. .04 O
ATOM 2573 N PHE A 337 98 , , 585 56 , , 278 1. ,948 1. , 00 7. .05 N
ATOM 2574 CA PHE A 337 98 , , 704 57 , , 540 2. ,647 1. , 00 7. .59 C
ATOM 2575 CB PHE A 337 99 , .975 57 , .594 3. .487 1. , 00 7. .16 c
ATOM 2576 CG PHE A 337 99 , , 808 57 , , 104 4. , 865 1. , 00 7. .21 C
ATOM 2577 CD1 PHE A 337 99 , , 003 57 , , 795 5. , 761 1. , 00 7. .38 C
ATOM 2578 CE1 PHE A 337 98 , , 864 57 , ,374 7. , 066 1. , 00 8. .33 C
ATOM 2579 CZ PHE A 337 99 , .567 56 , .259 7. .505 1. , 00 8. .60 c
ATOM 2580 CE2 PHE A 337 100 , ,398 55 , , 564 6. ,619 1. , 00 8. .77 C
ATOM 2581 CD2 PHE A 337 100 , , 508 55 , , 990 5. ,308 1. , 00 8. .62 C
ATOM 2582 C PHE A 337 98 , , 696 58 , , 776 1. , 757 1. , 00 7. .99 C
ATOM 2583 O PHE A 337 98. .430 59 , .861 2. .268 1. , 00 7. .70 O
ATOM 2584 N GLU A 338 98 , , 992 58 , , 618 0. , 467 1. , 00 8. .15 N
ATOM 2585 CA GLU A 338 98 , , 899 59 , , 729 -0. , 491 1. , 00 9. .74 C
ATOM 2586 CB GLU A 338 99 , , 244 59 , , 294 -1. ,947 1. , 00 9. .63 c
ATOM 2587 CG GLU A 338 98. .362 58 , .191 -2. .537 1. , 00 9. .65 c
ATOM 2588 CD GLU A 338 98 , , 856 56 , , 762 -2. , 257 1. , 00 10. .53 C
ATOM 2589 OE1 GLU A 338 99 , , 721 56 , , 570 -1. ,362 1. , 00 10. .81 O
ATOM 2590 OE2 GLU A 338 98. .390 55 , .817 -2. .944 1. , 00 10. .79 O
ATOM 2591 C GLU A 338 97 , , 513 60 , , 415 -0. .436 1. , 00 10. .37 C
ATOM 2592 O GLU A 338 97, , 409 61 , , 601 -0. .684 1. , 00 10. .60 O
ATOM 2593 N ARG A 339 96 , , 468 59 , , 666 -0. .110 1. , 00 10. .40 N
ATOM 2594 CA ARG A 339 95 , .116 60 , .199 0. .001 1. , 00 10. .06 c
ATOM 2595 CB ARG A 339 94 , , 153 59 , , 043 0. .312 1. , 00 11. .83 C
ATOM 2596 CG ARG A 339 92 , , 681 59 , ,396 0. .462 1. , 00 13. .18 C
ATOM 2597 CD ARG A 339 92 , , 134 60 , , 106 -0. .771 1. , 00 14. .10 C
ATOM 2598 NE ARG A 339 90 , .753 60 , .596 -0. .578 1. , 00 13. .98 N
ATOM 2599 CZ ARG A 339 90 , , 417 61 , , 759 -0. .005 1. , 00 13. .41 C
ATOM 2600 NH1 ARG A 339 91 , ,332 62 , , 597 0. .458 1. , 00 13. .50 N
ATOM 2601 NH2 ARG A 339 89 , , 139 62 , , 104 0. .075 1. , 00 14. .68 N
ATOM 2602 C ARG A 339 95 , .008 61 , .304 1. .078 1. , 00 11. .01 c
ATOM 2603 O ARG A 339 94 , , 088 62 , , 127 1. .045 1. , 00 10. .77 O
ATOM 2604 N MET A 340 95, , 966 61 , ,358 2. .006 1. , 00 10. .31 N
ATOM 2605 CA MET A 340 96 , , 020 62 , , 457 2. .963 1. , 00 9. .26 C
ATOM 2606 CB MET A 340 96 , .993 62 , .146 4. .099 1. , 00 9. .11 c
ATOM 2607 CG MET A 340 96 , , 646 60 , , 889 4. .879 1. , 00 10. .01 C
ATOM 2608 SD MET A 340 94, , 987 60 , , 980 5. .573 1. , 00 12. .13 s
ATOM 2609 CE MET A 340 95., 008 59 ,.506 6..613 1., 00 10..09 C
ATOM 2610 C MET A 340 96. , 427 63 , .803 2. .338 1. , 00 9. .36 C
ATOM 2611 O MET A 340 96. , 074 64 , .848 2. .842 1. , 00 8. .38 O
ATOM 2612 N ILE A 341 97. .157 63. .781 1. .235 1. , 00 11. .09 N
ATOM 2613 CA ILE A 341 97. , 748 65 , .000 0. .686 1. , 00 10. .39 C
ATOM 2614 CB ILE A 341 98. , 657 64, .687 -0. .527 1. , 00 10. .79 C
ATOM 2615 CGI ILE A 341 99. , 852 63 , .876 -0. .052 1. , 00 11. .35 C
ATOM 2616 CD1 ILE A 341 100. , 648 63 , .174 -1. .145 1. , 00 11. .56 c
ATOM 2617 CG2 ILE A 341 99. , 130 65, .981 -1. .238 1. , 00 10. .01 C
ATOM 2618 C ILE A 341 96. , 722 66 , .097 0. .380 1. , 00 10. .90 C
ATOM 2619 O ILE A 341 96. , 892 67, .243 0. .799 1. , 00 12. .31 O
ATOM 2620 N PRO A 342 95. , 629 65 , .756 -0. .302 1. , 00 12. .28 N
ATOM 2621 CA PRO A 342 94. , 676 66 , .832 -0. .590 1. , 00 12. .47 C
ATOM 2622 CB PRO A 342 93. , 843 66 , .256 -1. .741 1. , 00 14. .17 C
ATOM 2623 CG PRO A 342 93. , 910 64 , .782 -1. .539 1. , 00 13. .44 C
ATOM 2624 CD PRO A 342 95. ,301 64 , .519 -1. .033 1. , 00 12. .37 c
ATOM 2625 C PRO A 342 93. , 771 67, .220 0. .601 1. , 00 11. .11 C
ATOM 2626 O PRO A 342 93. , 086 68 , .223 0. .518 1. , 00 9. .19 O
ATOM 2627 N ALA A 343 93. , 799 66 , .462 1. .705 1. , 00 10. .31 N
ATOM 2628 CA ALA A 343 92. , 963 66 , .766 2. .872 1. , 00 9. .59 c
ATOM 2629 CB ALA A 343 92. , 467 65, .479 3. .543 1. , 00 8. .88 C
ATOM 2630 C ALA A 343 93. , 700 67, .643 3. .886 1. , 00 10. .02 C
ATOM 2631 O ALA A 343 93. , 063 68 , .311 4. .708 1. , 00 10. .02 O
ATOM 2632 N LEU A 344 95. , 034 67 , .664 3. .865 1. , 00 9. .70 N
ATOM 2633 CA LEU A 344 95. , 772 68 , .381 4. .934 1. , 00 8. .81 C
ATOM 2634 CB LEU A 344 97. , 255 67, .973 4. .998 1. , 00 8. .59 C
ATOM 2635 CG LEU A 344 97. .518 66 , .577 5. .587 1. , 00 8. .97 c
ATOM 2636 CD1 LEU A 344 98. , 847 66 , .032 5. .058 1. , 00 8. .21 C
ATOM 2637 CD2 LEU A 344 97. , 450 66 , .546 7. .108 1. , 00 7. .43 C
ATOM 2638 C LEU A 344 95. , 657 69 , .916 4. .874 1. , 00 9. .98 C
ATOM 2639 O LEU A 344 95. , 549 70 , .562 5. .911 1. , 00 8. .75 0
ATOM 2640 N HIS A 345 95. , 711 70 , .505 3. .678 1. , 00 9. .35 N
ATOM 2641 CA HIS A 345 95. , 587 71 , .942 3. .535 1. , 00 9. .92 C
ATOM 2642 CB HIS A 345 94. , 129 72 , .401 3. .757 1. , 00 10. .02 C
ATOM 2643 CG HIS A 345 93. , 179 71 , .894 2. .699 1. , 00 11. .67 c
ATOM 2644 ND1 HIS A 345 93. , 205 72 , .350 1. .424 1. , 00 12. .62 N
ATOM 2645 CE1 HIS A 345 92. , 256 71 , .709 0. .692 1. , 00 13. .33 C
ATOM 2646 NE2 HIS A 345 91. , 648 70 , .814 1. .496 1. , 00 14. .96 N
ATOM 2647 CD2 HIS A 345 92. , 202 70 , .899 2. .742 1. , 00 12. .33 C
ATOM 2648 C HIS A 345 96. , 551 72 , .703 4. .428 1. , 00 9. .82 C
ATOM 2649 O HIS A 345 96. , 178 73 , .689 5. .069 1. , 00 8. .43 O
ATOM 2650 N LEU A 346 97. , 816 72 , .289 4. .423 1. , 00 9. .30 N
ATOM 2651 CA LEU A 346 98. , 818 72 , .912 5. .300 1. , 00 10. .80 c
ATOM 2652 CB LEU A 346 100. , 117 72 , .093 5. .298 1. , 00 10. .45 C
ATOM 2653 CG LEU A 346 99. , 976 70 , .661 5. .826 1. , 00 11. .00 C
ATOM 2654 CD1 LEU A 346 101 ,, 354 70 ,, 006 5., 726 1., 00 8.,88 C
ATOM 2655 CD2 LEU A 346 99 , ,373 70 , , 612 7. , 255 1. , 00 8. ,53 C
ATOM 2656 C LEU A 346 99 , , 139 74 , ,368 4. ,902 1. , 00 11. , 13 C
ATOM 2657 O LEU A 346 99. .466 75. , 186 5. .758 1. , 00 12. .08 0
ATOM 2658 N THR A 347 99 , , 068 74 , , 668 3. ,610 1. , 00 10. ,80 N
ATOM 2659 CA THR A 347 99 , , 282 76 , , 010 3. , 112 1. , 00 11. ,97 C
ATOM 2660 CB THR A 347 100 , , 670 76 , , 166 2. , 491 1. , 00 13. ,64 C
ATOM 2661 OG1 THR A 347 100 , .818 75. , 232 1. .414 1. , 00 13. .71 0
ATOM 2662 CG2 THR A 347 101 , , 794 75 , , 970 3. ,528 1. , 00 11. ,95 c
ATOM 2663 C THR A 347 98 , , 234 76 , ,304 2. , 045 1. , 00 13. ,99 C
ATOM 2664 O THR A 347 97, , 624 75 , ,386 1. , 488 1. , 00 12. ,36 0
ATOM 2665 N GLY A 348 97 , .988 77. , 590 1. .800 1. , 00 15. .02 N
ATOM 2666 CA GLY A 348 97, , 083 78 , , 010 0. , 745 1. , 00 15. ,65 C
ATOM 2667 C GLY A 348 95, , 579 77 , , 924 1. , 069 1. , 00 19. ,22 C
ATOM 2668 O GLY A 348 94 , , 776 78 , , 001 0. , 171 1. , 00 17. , 83 0
ATOM 2669 N GLY A 349 95 , .173 77. , 792 2. .333 1. , 00 18. .00 N
ATOM 2670 CA GLY A 349 93 , , 734 77 , , 951 2. ,676 1. , 00 14. ,01 C
ATOM 2671 C GLY A 349 93 , , 073 76 , , 615 2. , 480 1. , 00 14. ,38 C
ATOM 2672 O GLY A 349 93 , , 752 75 , , 667 2. , 099 1. , 00 17. ,92 0
ATOM 2673 N PRO A 350 91 , .737 76. , 537 2. .633 1. , 00 12. .78 N
ATOM 2674 CA PRO A 350 90 , , 787 77 , , 659 2. , 748 1. , 00 11. , 55 C
ATOM 2675 CB PRO A 350 89 , , 556 77 , , 130 2. , 031 1. , 00 12. ,62 C
ATOM 2676 CG PRO A 350 89 , , 586 75 , , 643 2. ,346 1. , 00 13. , 72 c
ATOM 2677 CD PRO A 350 91 , .061 75. , 279 2. .260 1. , 00 13. . 72 c
ATOM 2678 C PRO A 350 90 , , 440 78 , , 055 4. , 177 1. , 00 10. , 05 C
ATOM 2679 O PRO A 350 89 , , 797 79 , , 087 4. ,398 1. , 00 9. ,27 0
ATOM 2680 N TYR A 351 90 , .862 77. , 244 5. .135 1. , 00 9. .71 N
ATOM 2681 CA TYR A 351 90 , , 567 77 , , 499 6. , 539 1. , 00 10. ,28 c
ATOM 2682 CB TYR A 351 90 , , 875 76 , , 267 7. ,384 1. , 00 10. , 43 C
ATOM 2683 CG TYR A 351 90 , , 321 74 , , 951 6. , 885 1. , 00 11. ,04 C
ATOM 2684 CD1 TYR A 351 89 , .002 74. , 613 7. .113 1. , 00 10. .28 c
ATOM 2685 CE1 TYR A 351 88 , , 485 73 , , 407 6. ,679 1. , 00 10. , 77 c
ATOM 2686 CZ TYR A 351 89 , , 287 72 , , 508 6. , 017 1. , 00 11. ,60 C
ATOM 2687 OH TYR A 351 88 , , 757 71 , ,315 5. ,623 1. , 00 10. ,91 0
ATOM 2688 CE2 TYR A 351 90 , .616 72. , 818 5. .757 1. , 00 11. .66 c
ATOM 2689 CD2 TYR A 351 91 , , 122 74 , , 042 6. , 190 1. , 00 10. ,88 C
ATOM 2690 C TYR A 351 91 , ,384 78 , , 653 7. , 097 1. , 00 10. , 89 C
ATOM 2691 O TYR A 351 92 , , 554 78 , , 838 6. ,698 1. , 00 12. ,30 0
ATOM 2692 N ALA A 352 90 , .819 79. , 392 8. .057 1. , 00 9. .49 N
ATOM 2693 CA ALA A 352 91 , , 595 80 , , 444 8. , 729 1. , 00 10. ,18 C
ATOM 2694 CB ALA A 352 90 , , 810 81 , , 124 9. , 847 1. , 00 8. , 72 C
ATOM 2695 C ALA A 352 92 , , 863 79 , , 803 9. ,282 1. , 00 11. , 77 c
ATOM 2696 O ALA A 352 93 , .935 80. ,370 9. .171 1. , 00 11. .42 0
ATOM 2697 N ASN A 353 92 , , 752 78 , , 613 9. , 878 1. , 00 10. ,81 N
ATOM 2698 CA ASN A 353 93 , , 946 77 , , 953 10. .423 1. , 00 9. ,81 C
ATOM 2699 CB ASN A 353 94.,346 78 ,.587 11..759 1., 00 10..38 C
ATOM 2700 CG ASN A 353 95. ,666 78 , .059 12. .290 1. , 00 10. .94 C
ATOM 2701 OD1 ASN A 353 95. , 939 76 , .857 12. .206 1. , 00 10. .93 O
ATOM 2702 ND2 ASN A 353 96. , 494 78 , .954 12. .874 1. , 00 8. .59 N
ATOM 2703 C ASN A 353 93. , 701 76 , .475 10. .528 1. , 00 8. .44 C
ATOM 2704 O ASN A 353 92. , 907 76 , .019 11. .361 1. , 00 10. .00 O
ATOM 2705 N PRO A 354 94. , 354 75 , .697 9. .661 1. , 00 8. .94 N
ATOM 2706 CA PRO A 354 94. , 054 74 , .251 9. .635 1. , 00 8. .55 C
ATOM 2707 CB PRO A 354 94. , 814 73 , .777 8. .414 1. , 00 8. .43 C
ATOM 2708 CG PRO A 354 96. , 007 74 , .682 8. .355 1. , 00 8. .58 C
ATOM 2709 CD PRO A 354 95. , 475 76 , .039 8. .758 1. , 00 8. .26 C
ATOM 2710 C PRO A 354 94. , 518 73 , .521 10. .889 1. , 00 8. .60 c
ATOM 2711 O PRO A 354 94. , 028 72 , .431 11. .192 1. , 00 8. .91 O
ATOM 2712 N TYR A 355 95. , 429 74 , .128 11. .645 1. , 00 8. .49 N
ATOM 2713 CA TYR A 355 95. , 945 73 , .520 12. .889 1. , 00 7. .59 C
ATOM 2714 CB TYR A 355 97. , 283 74 , .172 13. .277 1. , 00 7. .82 c
ATOM 2715 CG TYR A 355 98. ,365 73 , .932 12. .243 1. , 00 7. .26 C
ATOM 2716 CD1 TYR A 355 99. , 166 72 , .802 12. .299 1. , 00 8. .70 C
ATOM 2717 CE1 TYR A 355 100. , 141 72 , .558 11. .356 1. , 00 8. .41 c
ATOM 2718 CZ TYR A 355 100. , 321 73 , .451 10. .338 1. , 00 8. .87 c
ATOM 2719 OH TYR A 355 101. , 272 73 , .201 9. .390 1. , 00 8. .82 O
ATOM 2720 CE2 TYR A 355 99. , 522 74 , .584 10. .250 1. , 00 8. .37 C
ATOM 2721 CD2 TYR A 355 98. , 554 74 , .801 11. .207 1. , 00 7. .94 c
ATOM 2722 C TYR A 355 94. , 928 73 , .573 14. .058 1. , 00 8. .97 c
ATOM 2723 O TYR A 355 95. , 127 72 , .918 15. .095 1. , 00 8. .72 O
ATOM 2724 N GLU A 356 93. , 824 74 , .298 13. .866 1. , 00 9. .38 N
ATOM 2725 CA GLU A 356 92. , 786 74 , .426 14. .891 1. , 00 10. .40 c
ATOM 2726 CB GLU A 356 92. ,372 75 , .889 15. .031 1. , 00 11. .74 C
ATOM 2727 CG GLU A 356 93. , 511 76 , .789 15. .465 1. , 00 14. .48 C
ATOM 2728 CD GLU A 356 93. , 179 78 , .287 15. .344 1. , 00 17. .27 C
ATOM 2729 OE1 GLU A 356 92. , 031 78 , .661 14. .974 1. , 00 18. .33 0
ATOM 2730 OE2 GLU A 356 94. , 097 79 , .084 15. .586 1. , 00 14. .75 O
ATOM 2731 C GLU A 356 91. , 545 73 , .584 14. .592 1. , 00 9. .45 C
ATOM 2732 O GLU A 356 90. , 577 73 , .580 15. .362 1. , 00 10. .71 O
ATOM 2733 N LEU A 357 91. , 567 72 , .856 13. .488 1. , 00 9. .76 N
ATOM 2734 CA LEU A 357 90. , 419 72 , .017 13. .127 1. , 00 8. .67 C
ATOM 2735 CB LEU A 357 90. , 563 71 , .482 11. .717 1. , 00 8. .92 C
ATOM 2736 CG LEU A 357 90. , 652 72 , .544 10. .624 1. , 00 9. .86 C
ATOM 2737 CD1 LEU A 357 90. , 900 71 , .810 9. .301 1. , 00 10. .02 c
ATOM 2738 CD2 LEU A 357 89. , 411 73 , .427 10. .599 1. , 00 9. .32 C
ATOM 2739 C LEU A 357 90. ,306 70 , .837 14. .078 1. , 00 9. .42 C
ATOM 2740 O LEU A 357 91. ,325 70 , .301 14. .551 1. , 00 8. .12 O
ATOM 2741 N ALA A 358 89. , 063 70 , .443 14. .359 1. , 00 9. .05 N
ATOM 2742 CA ALA A 358 88. , 783 69 , .275 15. .207 1. , 00 8. .40 C
ATOM 2743 CB ALA A 358 87. , 441 69 , .448 15. .877 1. , 00 8. .38 C
ATOM 2744 C ALA A 358 88., 776 68., 079 14..272 1., 00 9..28 C
ATOM 2745 O ALA A 358 87. , 914 67. , 989 13. .357 1. , 00 9. .07 O
ATOM 2746 N THR A 359 89. , 745 67. , 186 14. .445 1. , 00 9. .68 N
ATOM 2747 CA THR A 359 89. .871 66. , 034 13. .554 1. , 00 8. .81 C
ATOM 2748 CB THR A 359 91. ,347 65. , 763 13. .184 1. , 00 8. .14 C
ATOM 2749 OG1 THR A 359 92. , 120 65. , 685 14. .377 1. , 00 8. .36 O
ATOM 2750 CG2 THR A 359 91. , 922 66. , 886 12. .295 1. , 00 8. .45 C
ATOM 2751 C THR A 359 89. ,215 64. , 776 14. .152 1. , 00 8. .41 c
ATOM 2752 O THR A 359 89. , 174 64. , 575 15. .377 1. , 00 9. .75 O
ATOM 2753 N PHE A 360 88. , 696 63. , 931 13. .272 1. , 00 8. .08 N
ATOM 2754 CA PHE A 360 87. , 965 62. , 713 13. .634 1. , 00 7. .27 C
ATOM 2755 CB PHE A 360 86. , 542 63. , 070 14. .107 1. , 00 7. .53 c
ATOM 2756 CG PHE A 360 85. , 627 63. , 576 13. .005 1. , 00 8. .76 C
ATOM 2757 CD1 PHE A 360 85. , 772 64. , 853 12. .472 1. , 00 9. .54 C
ATOM 2758 CE1 PHE A 360 84. , 940 65. ,302 11. .459 1. , 00 9. .36 C
ATOM 2759 CZ PHE A 360 83. , 921 64. , 491 10. .976 1. , 00 9. .69 c
ATOM 2760 CE2 PHE A 360 83. , 752 63. , 231 11. .506 1. , 00 9. .12 C
ATOM 2761 CD2 PHE A 360 84. , 598 62. , 787 12. .523 1. , 00 9. .65 C
ATOM 2762 C PHE A 360 87. , 922 61. , 769 12. .425 1. , 00 7. .67 C
ATOM 2763 O PHE A 360 88. ,170 62. , 194 11. .276 1. , 00 7. .78 O
ATOM 2764 N TYR A 361 87. , 609 60. , 498 12. .668 1. , 00 7. .49 N
ATOM 2765 CA TYR A 361 87. , 405 59. , 524 11. .599 1. , 00 7. .70 C
ATOM 2766 CB TYR A 361 88. , 494 58. , 425 11. .591 1. , 00 8. .98 C
ATOM 2767 CG TYR A 361 88. , 562 57. , 653 12. .867 1. , 00 9. .92 C
ATOM 2768 CD1 TYR A 361 87. , 638 56. , 658 13. .140 1. , 00 9. .78 C
ATOM 2769 CE1 TYR A 361 87. , 670 55. , 964 14. .326 1. , 00 9. .23 C
ATOM 2770 CZ TYR A 361 88. , 631 56. , 253 15. .252 1. , 00 11. .11 c
ATOM 2771 OH TYR A 361 88. , 674 55. , 559 16. .424 1. , 00 9. .75 O
ATOM 2772 CE2 TYR A 361 89. , 580 57. , 240 15. .015 1. , 00 10. .68 C
ATOM 2773 CD2 TYR A 361 89. , 540 57. , 930 13. .820 1. , 00 10. .41 C
ATOM 2774 C TYR A 361 86. , 008 58. , 900 11. .611 1. , 00 8. .19 c
ATOM 2775 O TYR A 361 85. , 647 58. , 252 10. ,666 1. , 00 7. .67 O
ATOM 2776 N ASP A 362 85. , 234 59. , 057 12. .686 1. , 00 8. .73 N
ATOM 2777 CA ASP A 362 83. , 824 58. , 657 12. .646 1. , 00 8. .35 C
ATOM 2778 CB ASP A 362 83. , 653 57. , 128 12. .753 1. , 00 8. .07 c
ATOM 2779 CG ASP A 362 84. , 086 56. , 555 14. .113 1. , 00 8. .16 C
ATOM 2780 OD1 ASP A 362 84. , 234 57. ,344 15. .073 1. , 00 9. .60 O
ATOM 2781 OD2 ASP A 362 84. , 268 55. , 295 14. .204 1. , 00 7. .55 O
ATOM 2782 C ASP A 362 83. , 094 59. , 398 13. .755 1. , 00 8. .22 c
ATOM 2783 O ASP A 362 83. , 690 60. , 184 14. .497 1. , 00 8. .44 O
ATOM 2784 N ASN A 363 81. , 797 59. , 191 13. .826 1. , 00 8. .18 N
ATOM 2785 CA ASN A 363 80. , 986 59. , 838 14. .845 1. , 00 9. .19 C
ATOM 2786 CB ASN A 363 80. , 909 61. ,380 14. .651 1. , 00 8. .52 c
ATOM 2787 CG ASN A 363 80. , 095 61. , 816 13. .415 1. , 00 9. .77 C
ATOM 2788 OD1 ASN A 363 79. , 273 61. , 067 12. .890 1. , 00 9. .07 O
ATOM 2789 ND2 ASN A 363 80 ,.352 63 ,.063 12..944 1., 00 9..67 N
ATOM 2790 C ASN A 363 79 , .624 59 , .146 14. .955 1. , 00 7. .68 C
ATOM 2791 O ASN A 363 79 , .439 58 , .048 14. .427 1. , 00 8. .21 O
ATOM 2792 N HIS A 364 78. .701 59. .804 15. .637 1. .00 8. .34 N
ATOM 2793 CA HIS A 364 77 , .354 59 , .308 15. .913 1. , 00 8. .10 C
ATOM 2794 CB HIS A 364 76 , .786 59 , .985 17. .176 1. , 00 7. .63 C
ATOM 2795 CG HIS A 364 76 , .772 61 , .502 17. .124 1. , 00 8. .17 C
ATOM 2796 ND1 HIS A 364 77 , .735 62 , .229 16. .507 1. , 00 8. .37 N
ATOM 2797 CE1 HIS A 364 77 , .454 63 , .538 16. .646 1. , 00 8. .63 C
ATOM 2798 NE2 HIS A 364 76 , .343 63 , .653 17. .371 1. , 00 7. .82 N
ATOM 2799 CD2 HIS A 364 75 , .886 62 , .415 17. .664 1. , 00 8. .46 C
ATOM 2800 C HIS A 364 76 , .371 59 , .448 14. .799 1. , 00 8. .51 c
ATOM 2801 O HIS A 364 75 , .255 58 , .965 14. .915 1. , 00 9. .11 O
ATOM 2802 N ASP A 365 76 , .741 60 , .115 13. .712 1. , 00 9. .46 N
ATOM 2803 CA ASP A 365 75 , .817 60 , .414 12. .593 1. , 00 9. .11 C
ATOM 2804 CB ASP A 365 75 , .830 61 , .915 12. .323 1. , 00 10. .88 c
ATOM 2805 CG ASP A 365 75 , .312 62 , .724 13. .500 1. , 00 10. .65 C
ATOM 2806 OD1 ASP A 365 74 , .346 62 , .279 14. .153 1. , 00 9. .96 O
ATOM 2807 OD2 ASP A 365 75 , .844 63 , .818 13. .718 1. , 00 11. .29 O
ATOM 2808 C ASP A 365 76. .192 59. .734 11. .274 1. , 00 9. .75 c
ATOM 2809 O ASP A 365 75 , .760 60 , .176 10. .189 1. , 00 11. .96 O
ATOM 2810 N MET A 366 77 , .034 58 , .714 11. .372 1. , 00 9. .99 N
ATOM 2811 CA MET A 366 77 , .556 57 , .964 10. .237 1. , 00 10. .18 c
ATOM 2812 CB MET A 366 78. .713 58. .741 9. .578 1. , 00 9. .75 c
ATOM 2813 CG MET A 366 79 , .959 58 , .872 10. .461 1. , 00 11. .62 C
ATOM 2814 SD MET A 366 81 , .295 59 , .914 9. .803 1. , 00 13. .99 s
ATOM 2815 CE MET A 366 80. .479 61. .530 9. .682 1. .00 8. .71 c
ATOM 2816 C MET A 366 78 , .049 56 , .598 10. .762 1. , 00 9. .78 C
ATOM 2817 O MET A 366 78 , .186 56 , .414 11. .992 1. , 00 12. .65 O
ATOM 2818 N PRO A 367 78 , .325 55 , .653 9. .862 1. , 00 9. .51 N
ATOM 2819 CA PRO A 367 78 , .852 54 , .387 10. .374 1. , 00 10. .78 c
ATOM 2820 CB PRO A 367 79 , .045 53 , .527 9. .101 1. , 00 12. .65 C
ATOM 2821 CG PRO A 367 78 , .001 54 , .079 8. .125 1. , 00 13. .10 C
ATOM 2822 CD PRO A 367 77 , .978 55 , .576 8. .420 1. , 00 10. .75 C
ATOM 2823 C PRO A 367 80 , .160 54 , .582 11. .122 1. , 00 11. .09 c
ATOM 2824 O PRO A 367 80 , .975 55 , .408 10. .731 1. , 00 10. .60 O
ATOM 2825 N ARG A 368 80 , .349 53 , .859 12. .216 1. , 00 9. .20 N
ATOM 2826 CA ARG A 368 81 , .663 53 , .831 12. .827 1. , 00 9. .49 C
ATOM 2827 CB ARG A 368 81 , .643 53 , .025 14. .110 1. , 00 9. .30 c
ATOM 2828 CG ARG A 368 81 , .025 53 , .784 15. .274 1. , 00 10. .19 C
ATOM 2829 CD ARG A 368 80 , .882 52 , .871 16. .469 1. , 00 10. .27 C
ATOM 2830 NE ARG A 368 80 , .243 53 , .454 17. .672 1. , 00 9. .66 N
ATOM 2831 CZ ARG A 368 80 , .903 53 , .947 18. .717 1. , 00 9. .05 c
ATOM 2832 NH1 ARG A 368 82 , .245 54 , .025 18. .713 1. , 00 10. .32 N
ATOM 2833 NH2 ARG A 368 80 , .231 54 , .339 19. .787 1. , 00 8. .12 N
ATOM 2 i34 C ARG A 368 82., 663 53., 249 11..845 1., 00 11..61 C
ATOM 2 i 335 O ARG A 368 82. , 279 52. , 520 10. .912 1. , 00 11. .13 O
ATOM 2 i 336 N LEU A 369 83. , 943 53. , 540 12. .074 1. , 00 13. .78 N
ATOM 2 i 337 CA LEU A 369 85. .014 53. , 049 11. .196 1. , 00 12. .69 C
ATOM 2 1 338 CB LEU A 369 86. ,382 53. , 395 11. .783 1. , 00 10. . 93 C
ATOM 2 i 339 CG LEU A 369 87. , 618 53. , 010 10. .941 1. , 00 12. .41 C
ATOM 2 i 340 CD1 LEU A 369 87. , 665 53. , 862 9. .662 1. , 00 11. .24 C
ATOM 2 i 341 CD2 LEU A 369 88. , 881 53. , 165 11. .794 1. , 00 10. .18 c
ATOM 2 i 3 42 C LEU A 369 84. , 897 51. , 539 11. .041 1. , 00 13. .30 C
ATOM 2 i 343 O LEU A 369 84. , 695 50. , 826 12. .030 1. , 00 12. .35 O
ATOM 2 i 344 N ASP A 370 84. , 991 51. , 063 9. .800 1. , 00 12. .59 N
ATOM 2 i 345 CA ASP A 370 84. , 883 49. , 637 9. .504 1. , 00 12. .43 c
ATOM 2 i 346 CB ASP A 370 83. , 975 49. , 453 8. .284 1. , 00 13. .60 C
ATOM 2847 CG ASP A 370 83. , 785 47. , 969 7. .870 1. , 00 17. .03 C
ATOM 2 i 348 OD1 ASP A 370 84. , 111 47. , 032 8. .611 1. , 00 17. .52 O
ATOM 2 i 349 OD2 ASP A 370 83. , 322 47. , 754 6. .752 1. , 00 19. .77 0
ATOM 2 i 350 C ASP A 370 86. ,314 49. , 112 9. .286 1. , 00 13. .54 C
ATOM 2851 O ASP A 370 86. , 820 49. , 088 8. .175 1. , 00 11. .76 O
ATOM 2 i 352 N ALA A 371 86. , 967 48. , 720 10. .375 1. , 00 13. .86 N
ATOM 2 i 353 CA ALA A 371 88. ,379 48. , 397 10. .352 1. , 00 13. .56 c
ATOM 2 i 354 CB ALA A 371 89. , 208 49. , 607 10. .786 1. , 00 15. .22 C
ATOM 2 i 355 C ALA A 371 88. , 647 47. , 237 11. .284 1. , 00 13. .93 C
ATOM 2 i 356 O ALA A 371 87. , 898 47. , 002 12. .234 1. , 00 12. .26 O
ATOM 2 i 357 N SER A 372 89. , 720 46. , 514 10. .984 1. , 00 12. .45 N
ATOM 2 i 358 CA SER A 372 90. ,307 45. , 555 11. .892 1. , 00 11. .87 C
ATOM 2 i 359 CB SER A 372 91. , 462 44. , 823 11. .180 1. , 00 11. .70 C
ATOM 2 i 360 OG SER A 372 92. , 501 45. , 791 10. .948 1. , 00 11. .05 0
ATOM 2 1 361 C SER A 372 90. , 908 46. ,311 13. .099 1. , 00 12. .28 C
ATOM 2 i 362 O SER A 372 90. , 999 47. , 544 13. .102 1. , 00 10. .61 O
ATOM 2 i 363 N ASP A 373 91. ,346 45. , 560 14. .108 1. , 00 12. .03 N
ATOM 2 i 364 CA ASP A 373 91. , 992 46. , 153 15. .276 1. , 00 13. .25 c
ATOM 2 i 365 CB ASP A 373 92. , 475 45. , 078 16. .247 1. , 00 13. .20 C
ATOM 2 i 366 CG ASP A 373 91. ,370 44. , 539 17. .148 1. , 00 14. .89 C
ATOM 2 i 367 OD1 ASP A 373 90. , 166 44. , 703 16. .853 1. , 00 12. .62 O
ATOM 2 i 368 OD2 ASP A 373 91. , 736 43. , 978 18. .190 1. , 00 13. .86 0
ATOM 2 i 369 C ASP A 373 93. , 198 46. , 988 14. .842 1. , 00 12. .53 C
ATOM 2870 O ASP A 373 93. , 423 48. , 093 15. .337 1. , 00 10. .18 O
ATOM 2 i 371 N GLU A 374 93. , 959 46. , 427 13. .921 1. , 00 11. .93 N
ATOM 2 i 372 CA GLU A 374 95. , 127 47. , 082 13. .354 1. , 00 11. .52 c
ATOM 2 i 373 CB GLU A 374 95. , 789 46. , 152 12. .336 1. , 00 14. .06 C
ATOM 2874 CG GLU A 374 96. , 440 44. , 888 12. .889 1. , 00 17. .40 C
ATOM 2 i 375 CD GLU A 374 95. , 456 43. , 815 13. .375 1. , 00 23. .88 c
ATOM 2 i 376 OE1 GLU A 374 94. , 275 43. , 768 12. .949 1. , 00 21. .21 0
ATOM 2 i 377 OE2 GLU A 374 95. , 874 43. , 018 14. .222 1. , 00 29. .19 O
ATOM 2 i 378 C GLU A 374 94. , 777 48. , 408 12. .683 1. , 00 11. .68 C
ATOM 2879 O GLU A 374 95., 514 49 ,.379 12..804 1., 00 13..01 O
ATOM 2880 N GLY A 375 93. , 653 48 , .458 11. .971 1. , 00 9. .99 N
ATOM 2881 CA GLY A 375 93. , 163 49 , .687 11. .351 1. , 00 10. .09 C
ATOM 2882 C GLY A 375 92. .830 50 , .784 12. .365 1. , 00 10. .49 C
ATOM 2883 O GLY A 375 93. , 176 51 , .939 12. .163 1. , 00 10. .04 O
ATOM 2884 N PHE A 376 92. , 158 50 , .427 13. .452 1. , 00 10. .02 N
ATOM 2885 CA PHE A 376 91. , 932 51 , .386 14. .536 1. , 00 10. .75 C
ATOM 2886 CB PHE A 376 91. , 070 50 , .792 15. .656 1. , 00 8. .72 c
ATOM 2887 CG PHE A 376 89. , 603 50 , .807 15. .347 1. , 00 8. .79 C
ATOM 2888 CD1 PHE A 376 88. , 999 49 , .749 14. .705 1. , 00 9. .04 C
ATOM 2889 CE1 PHE A 376 87. , 652 49 , .776 14. .390 1. , 00 9. .80 C
ATOM 2890 CZ PHE A 376 86. , 894 50 , .874 14. .744 1. , 00 10. .46 c
ATOM 2891 CE2 PHE A 376 87. , 487 51 , .922 15. .413 1. , 00 9. .77 C
ATOM 2892 CD2 PHE A 376 88. , 831 51 , .886 15. .703 1. , 00 8. .64 C
ATOM 2893 C PHE A 376 93. , 267 51 , .884 15. .092 1. , 00 8. .86 C
ATOM 2894 O PHE A 376 93. , 442 53 , .081 15. .344 1. , 00 9. .11 0
ATOM 2895 N ILE A 377 94. , 204 50 , .971 15. .298 1. , 00 8. .28 N
ATOM 2896 CA ILE A 377 95. , 513 51 , .362 15. .839 1. , 00 9. .19 C
ATOM 2897 CB ILE A 377 96. , 411 50 , .113 16. .082 1. , 00 8. .69 C
ATOM 2898 CGI ILE A 377 95. , 916 49 , .352 17. .324 1. , 00 9. .02 C
ATOM 2899 CD1 ILE A 377 96. , 440 47 , .927 17. .475 1. , 00 9. .66 C
ATOM 2900 CG2 ILE A 377 97. , 866 50 , .532 16. .255 1. , 00 8. .80 C
ATOM 2901 C ILE A 377 96. , 200 52 , .391 14. .910 1. , 00 9. .14 c
ATOM 2902 O ILE A 377 96. , 706 53 , .421 15. .366 1. , 00 9. .95 O
ATOM 2903 N ASP A 378 96. , 205 52 , .098 13. .611 1. , 00 9. .15 N
ATOM 2904 CA ASP A 378 96. , 789 52 , .992 12. .601 1. , 00 10. .39 C
ATOM 2905 CB ASP A 378 96. .760 52 , .329 11. .205 1. , 00 10. .75 c
ATOM 2906 CG ASP A 378 97. , 653 51 , .067 11. .113 1. , 00 11. .43 C
ATOM 2907 OD1 ASP A 378 98. , 628 50 , .988 11. .883 1. , 00 9. .90 O
ATOM 2908 OD2 ASP A 378 97. ,370 50 , .167 10. .263 1. , 00 12. .74 O
ATOM 2909 C ASP A 378 96. , 065 54 , .361 12. .564 1. , 00 10. .57 c
ATOM 2910 O ASP A 378 96. , 704 55 , .399 12. .441 1. , 00 9. .95 O
ATOM 2911 N ALA A 379 94. , 735 54 , .346 12. .693 1. , 00 9. .88 N
ATOM 2912 CA ALA A 379 93. , 926 55 , .555 12. .618 1. , 00 8. .95 C
ATOM 2913 CB ALA A 379 92. , 416 55 , .217 12. .669 1. , 00 8. .20 c
ATOM 2914 C ALA A 379 94. ,300 56 , .446 13. .773 1. , 00 8. .88 C
ATOM 2915 O ALA A 379 94. , 473 57 , .640 13. .601 1. , 00 9. .55 O
ATOM 2916 N HIS A 380 94. , 404 55 , .870 14. .967 1. , 00 8. .76 N
ATOM 2917 CA HIS A 380 94. , 764 56 , .664 16. .139 1. , 00 9. .21 c
ATOM 2918 CB HIS A 380 94. , 550 55 , .894 17. .423 1. , 00 8. .88 C
ATOM 2919 CG HIS A 380 93. , 110 55 , .551 17. .684 1. , 00 9. .44 C
ATOM 2920 ND1 HIS A 380 92. , 734 54 , .366 18. .184 1. , 00 8. .74 N
ATOM 2921 CE1 HIS A 380 91. ,374 54 , .332 18. .275 1. , 00 10. .92 c
ATOM 2922 NE2 HIS A 380 90. , 894 55 , .509 17. .846 1. , 00 10. .69 N
ATOM 2923 CD2 HIS A 380 91. , 941 56 , .283 17. .464 1. , 00 9. .96 C
ATOM 2924 C HIS A 380 96., 188 57 ,.175 16..044 1., 00 8..44 C
ATOM 2925 O HIS A 380 96. , 453 58 , .294 16. .454 1. , 00 9. .30 O
ATOM 2926 N ASN A 381 97. , 097 56 , .384 15. .495 1. , 00 7. .47 N
ATOM 2927 CA ASN A 381 98. .466 56 , .834 15. .340 1. , 00 8. .52 C
ATOM 2928 CB ASN A 381 99. ,370 55 , .705 14. .773 1. , 00 8. .35 C
ATOM 2929 CG ASN A 381 99. , 708 54 , .645 15. .818 1. , 00 8. .34 C
ATOM 2930 OD1 ASN A 381 99. , 577 54 , .900 17. .015 1. , 00 8. .06 O
ATOM 2931 ND2 ASN A 381 100. , 163 53 , .449 15. .374 1. , 00 7. .82 N
ATOM 2932 C ASN A 381 98. , 520 58 , .093 14. .459 1. , 00 8. .41 C
ATOM 2933 O ASN A 381 99. , 241 59 , .036 14. .745 1. , 00 8. .56 O
ATOM 2934 N TRP A 382 97. , 741 58 , .109 13. .385 1. , 00 9. .15 N
ATOM 2935 CA TRP A 382 97. , 636 59 , .315 12. .526 1. , 00 8. .80 c
ATOM 2936 CB TRP A 382 96. , 884 58 , .963 11. .248 1. , 00 8. .39 C
ATOM 2937 CG TRP A 382 96. , 581 60 , .147 10. .375 1. , 00 8. .03 C
ATOM 2938 CD1 TRP A 382 95. , 403 60 , .848 10. .284 1. , 00 9. .01 C
ATOM 2939 NE1 TRP A 382 95. , 516 61 , .892 9. .374 1. , 00 8. .56 N
ATOM 2940 CE2 TRP A 382 96. , 754 61 , .914 8. .845 1. , 00 8. .18 C
ATOM 2941 CD2 TRP A 382 97. , 500 60 , .815 9. .452 1. , 00 8. .04 C
ATOM 2942 CE3 TRP A 382 98. , 818 60 , .608 9. .074 1. , 00 8. .37 c
ATOM 2943 CZ3 TRP A 382 99. , 390 61 , .482 8. .143 1. , 00 8. .26 c
ATOM 2944 CH2 TRP A 382 98. , 647 62 , .518 7. .551 1. , 00 7. .81 C
ATOM 2945 CZ2 TRP A 382 97. ,326 62 , .757 7. .894 1. , 00 8. .30 C
ATOM 2946 C TRP A 382 96. , 945 60 , .481 13. .227 1. , 00 8. .48 c
ATOM 2947 O TRP A 382 97. , 400 61 , .622 13. .177 1. , 00 8. .06 O
ATOM 2948 N LEU A 383 95. , 807 60 , .208 13. .852 1. , 00 8. .56 N
ATOM 2949 CA LEU A 383 94. , 972 61 , .257 14. .448 1. , 00 8. .01 C
ATOM 2950 CB LEU A 383 93. , 716 60 , .621 15. .076 1. , 00 7. .16 c
ATOM 2951 CG LEU A 383 92. , 510 61 , .501 15. .385 1. , 00 8. .24 C
ATOM 2952 CD1 LEU A 383 91. , 842 62 , .030 14. .103 1. , 00 7. .28 C
ATOM 2953 CD2 LEU A 383 91. , 496 60 , .788 16. .288 1. , 00 7. .85 C
ATOM 2954 C LEU A 383 95. , 748 62 , .039 15. .513 1. , 00 8. .74 c
ATOM 2955 O LEU A 383 95. , 588 63 , .255 15. .631 1. , 00 9. .24 O
ATOM 2956 N PHE A 384 96. , 605 61 , .332 16. .258 1. , 00 7. .95 N
ATOM 2957 CA PHE A 384 97. ,345 61 , .920 17. .384 1. , 00 8. .10 C
ATOM 2958 CB PHE A 384 97. , 443 60 , .918 18. .555 1. , 00 7. .93 c
ATOM 2959 CG PHE A 384 96. , 180 60 , .831 19. .337 1. , 00 8. .31 C
ATOM 2960 CD1 PHE A 384 95. , 173 59 , .968 18. .951 1. , 00 8. .85 C
ATOM 2961 CE1 PHE A 384 93. , 963 59 , .940 19. .650 1. , 00 9. .55 C
ATOM 2962 CZ PHE A 384 93. , 766 60 , .776 20. .724 1. , 00 8. .55 c
ATOM 2963 CE2 PHE A 384 94. , 770 61 , .640 21. .113 1. , 00 8. .59 C
ATOM 2964 CD2 PHE A 384 95. , 956 61 , .674 20. .415 1. , 00 8. .78 C
ATOM 2965 C PHE A 384 98. , 705 62 , .486 17. .004 1. , 00 8. .54 c
ATOM 2966 O PHE A 384 99. , 399 63 , .021 17. .872 1. , 00 8. .69 0
ATOM 2967 N THR A 385 99. , 063 62 , .415 15. .716 1. , 00 8. .38 N
ATOM 2968 CA THR A 385 100. ,333 62 , .984 15. .243 1. , 00 7. .85 C
ATOM 2969 CB THR A 3i5 101 ,, 325 61 ,, 911 14., 740 1., 00 7.,61 C
ATOM 2970 OG1 THR A 3i 15 100 , , 730 61 , , 155 13. ,682 1. , 00 8. , 05 O
ATOM 2971 CG2 THR A 3i 15 101 , , 765 60 , , 997 15. , 869 1. , 00 6. ,91 C
ATOM 2972 C THR A 3i 15 100. .211 64. , 042 14. .146 1. .00 8. .32 c
ATOM 2973 O THR A 3i 15 101 , , 047 64 , , 951 14. , 085 1. , 00 8. , 72 O
ATOM 2974 N ALA A 3i 16 99 , , 204 63 , , 910 13. ,285 1. , 00 8. ,50 N
ATOM 2975 CA ALA A 3i 16 98 , , 920 64 , , 854 12. ,205 1. , 00 8. ,68 C
ATOM 2976 CB ALA A 3i 16 98 , .028 64. , 195 11. .144 1. , 00 7. .78 c
ATOM 2977 C ALA A 3? 16 98 , , 217 66 , , 098 12. , 741 1. , 00 8. ,58 C
ATOM 2978 O ALA A 3i 16 97, , 604 66 , , 057 13. , 797 1. , 00 9. ,11 O
ATOM 2979 N ARG A 3? 11 98 , , 256 67, , 170 11. ,958 1. , 00 7. ,88 N
ATOM 2980 CA ARG A 3? 11 97 , .813 68. , 483 12. .414 1. , 00 8. .17 c
ATOM 2981 CB ARG A 3? 11 98 , , 098 69 , , 506 11. , 322 1. , 00 9. ,24 C
ATOM 2982 CG ARG A 387 97, ,347 69 , , 207 10. , 029 1. , 00 11. ,67 C
ATOM 2983 CD ARG A 3? 11 96 , , 192 70 , , 133 9. ,906 1. , 00 11. , 85 C
ATOM 2984 NE ARG A 3i 11 95 , .311 69. , 859 8. .768 1. , 00 11. .11 N
ATOM 2985 CZ ARG A 3? 11 94 , , 113 69 , , 285 8. , 840 1. , 00 10. ,57 C
ATOM 2986 NH1 ARG A 387 93 , , 655 68 , , 759 9. ,982 1. , 00 9. ,60 N
ATOM 2987 NH2 ARG A 3? 11 93 , ,364 69 , , 211 7. , 751 1. , 00 10. ,59 N
ATOM 2988 C ARG A 3i 11 96 , .325 68. , 495 12. .797 1. , 00 7. .95 C
ATOM 2989 O ARG A 3i 11 95, , 512 67, , 835 12. , 171 1. , 00 7. , 47 O
ATOM 2990 N GLY A 3i 18 95, , 983 69 , , 278 13. , 818 1. , 00 7. ,52 N
ATOM 2991 CA GLY A 3i 18 94 , , 614 69 , ,385 14. ,283 1. , 00 7. , 75 C
ATOM 2992 C GLY A 3i 18 94 , .475 68. , 905 15. .720 1. , 00 8. .33 c
ATOM 2993 O GLY A 3i 18 95, , 469 68 , , 534 16. ,369 1. , 00 10. ,59 O
ATOM 2994 N ILE A 3i 19 93 , , 230 68 , , 903 16. , 188 1. , 00 7. , 79 N
ATOM 2995 CA ILE A 3i 19 92 , .862 68. , 622 17. .562 1. .00 7. .45 c
ATOM 2996 CB ILE A 3i 19 92 , , 061 69 , , 790 18. , 178 1. , 00 7. , 55 C
ATOM 2997 CGI ILE A 3i 19 92 , , 809 71 , , 138 17. ,985 1. , 00 8. ,63 C
ATOM 2998 CD1 ILE A 3i 19 92 , , 093 72 , , 405 18. , 531 1. , 00 6. ,90 C
ATOM 2999 CG2 ILE A 3i 19 91 , .710 69. , 462 19. .643 1. , 00 8. .04 c
ATOM 3000 C ILE A 3? 19 92 , , 014 67, ,367 17. , 524 1. , 00 7. ,61 C
ATOM 3001 O ILE A 3i 19 90 , , 896 67, ,387 16. , 999 1. , 00 8. , 70 O
ATOM 3002 N PRO A 390 92 , , 541 66 , , 249 18. , 026 1. , 00 8. ,53 N
ATOM 3003 CA PRO A 390 91 , .890 64. , 949 17. .796 1. , 00 8. .74 c
ATOM 3004 CB PRO A 390 92 , , 970 63 , , 945 18. , 193 1. , 00 9. ,38 C
ATOM 3005 CG PRO A 390 93 , , 792 64, , 682 19. , 194 1. , 00 9. , 41 C
ATOM 3006 CD PRO A 390 93 , , 839 66 , , 097 18. ,692 1. , 00 8. ,91 C
ATOM 3007 C PRO A 390 90 , .653 64. , 742 18. .668 1. , 00 8. .66 c
ATOM 3008 O PRO A 390 90 , , 660 65, , 121 19. , 813 1. , 00 7. ,91 O
ATOM 3009 N VAL A 391 89 , , 625 64, , 133 18. , 085 1. , 00 9. ,23 N
ATOM 3010 CA VAL A 391 88 , ,380 63 , , 770 18. , 742 1. , 00 8. ,84 C
ATOM 3011 CB VAL A 391 87 , .182 64. , 547 18. .206 1. , 00 9. .48 c
ATOM 3012 CGI VAL A 391 85, , 918 64 , , 148 19. , 026 1. , 00 8. , 02 C
ATOM 3013 CG2 VAL A 391 87, , 473 66 , , 063 18. ,250 1. , 00 9. ,25 C
ATOM 3014 C VAL A 391 88 ,, 129 62 ,, 293 18., 503 1., 00 9.,69 C
ATOM 3015 O VAL A 391 88 , , 115 61 , , 851 17. ,345 1. , 00 10. ,62 O
ATOM 3016 N VAL A 392 87, , 953 61 , , 523 19. ,583 1. , 00 8. ,65 N
ATOM 3017 CA VAL A 392 87. .690 60. , 084 19. .472 1. , 00 8. .29 C
ATOM 3018 CB VAL A 392 88 , , 797 59 , , 266 20. , 178 1. , 00 8. , 70 C
ATOM 3019 CGI VAL A 392 88 , , 464 57 , , 773 20. , 224 1. , 00 7. ,24 C
ATOM 3020 CG2 VAL A 392 90 , , 160 59 , , 487 19. , 464 1. , 00 7. ,11 C
ATOM 3021 C VAL A 392 86 , .318 59. , 801 20. .076 1. , 00 8. .78 c
ATOM 3022 O VAL A 392 86 , , 003 60 , , 260 21. , 176 1. , 00 6. ,91 O
ATOM 3023 N TYR A 393 85, , 505 59 , , 072 19. ,325 1. , 00 7. ,24 N
ATOM 3024 CA TYR A 393 84, , 132 58 , , 760 19. , 703 1. , 00 6. ,81 C
ATOM 3025 CB TYR A 393 83 , .357 58. , 425 18. .420 1. , 00 6. .91 c
ATOM 3026 CG TYR A 393 81 , , 884 58 , , 134 18. , 514 1. , 00 7. ,60 C
ATOM 3027 CD1 TYR A 393 81 , , 064 58 , , 829 19. ,374 1. , 00 7. , 48 C
ATOM 3028 CE1 TYR A 393 79 , , 708 58 , , 555 19. , 447 1. , 00 7. ,52 C
ATOM 3029 CZ TYR A 393 79 , .162 57. , 600 18. .604 1. , 00 8. .60 c
ATOM 3030 OH TYR A 393 77 , , 816 57 , ,317 18. ,677 1. , 00 8. , 86 O
ATOM 3031 CE2 TYR A 393 79 , , 958 56 , , 909 17. , 721 1. , 00 8. ,31 C
ATOM 3032 CD2 TYR A 393 81 , ,301 57 , , 167 17. ,676 1. , 00 7. ,64 C
ATOM 3033 C TYR A 393 84 , .153 57. , 586 20. .671 1. , 00 7. .01 C
ATOM 3034 O TYR A 393 84, , 895 56 , , 602 20. , 461 1. , 00 7. ,64 O
ATOM 3035 N TYR A 394 83 , ,352 57 , , 677 21. , 736 1. , 00 6. ,60 N
ATOM 3036 CA TYR A 394 83 , , 415 56 , , 677 22. , 835 1. , 00 7. , 13 C
ATOM 3037 CB TYR A 394 82 , .275 56. , 888 23. .858 1. , 00 7. .04 C
ATOM 3038 CG TYR A 394 80 , , 931 56 , ,373 23. , 424 1. , 00 7. ,20 C
ATOM 3039 CD1 TYR A 394 80 , , 135 57 , , 102 22. , 546 1. , 00 7. ,20 C
ATOM 3040 CE1 TYR A 394 78 , .912 56. , 619 22. .135 1. , 00 7. .49 c
ATOM 3041 CZ TYR A 394 78 , , 459 55 , ,388 22. , 589 1. , 00 7. ,57 C
ATOM 3042 OH TYR A 394 77 , , 224 54 , , 883 22. , 155 1. , 00 7. , 40 O
ATOM 3043 CE2 TYR A 394 79 , , 229 54 , , 645 23. , 479 1. , 00 7. ,52 C
ATOM 3044 CD2 TYR A 394 80 , .458 55. , 143 23. .885 1. , 00 8. .12 c
ATOM 3045 C TYR A 394 83 , ,391 55 , , 264 22. ,307 1. , 00 6. ,88 C
ATOM 3046 O TYR A 394 82 , , 624 54 , , 944 21. , 408 1. , 00 6. , 13 O
ATOM 3047 N GLY A 395 84, , 237 54 , , 419 22. , 866 1. , 00 7. , 42 N
ATOM 3048 CA GLY A 395 84 , .254 53. , 018 22. .479 1. , 00 8. .26 c
ATOM 3049 C GLY A 395 85, , 251 52 , , 654 21. ,398 1. , 00 8. , 44 C
ATOM 3050 O GLY A 395 85, , 594 51 , , 475 21. ,215 1. , 00 8. , 15 O
ATOM 3051 N SER A 396 85, , 711 53 , , 658 20. ,662 1. , 00 9. ,10 N
ATOM 3052 CA SER A 396 86 , .706 53. , 441 19. .629 1. , 00 9. .33 c
ATOM 3053 CB SER A 396 87, , 121 54 , , 760 19. , 022 1. , 00 8. ,64 C
ATOM 3054 OG SER A 396 86 , , 037 55 , , 357 18. ,348 1. , 00 9. , 74 O
ATOM 3055 C SER A 396 87, , 961 52 , , 742 20. , 155 1. , 00 9. , 75 C
ATOM 3056 O SER A 396 88 , .654 52. , 066 19. .411 1. , 00 9. .65 0
ATOM 3057 N GLU A 397 88 , , 258 52 , , 961 21. , 424 1. , 00 10. ,33 N
ATOM 3058 CA GLU A 397 89 , , 432 52 , ,388 22. , 073 1. , 00 11. , 40 C
ATOM 3059 CB GLU A 397 89., 641 53 ,.011 23..474 1., 00 11..29 C
ATOM 3060 CG GLU A 397 88. , 551 52 , .770 24. .523 1. , 00 12. .39 C
ATOM 3061 CD GLU A 397 87. ,385 53 , .748 24. .508 1. , 00 11. .96 C
ATOM 3062 OE1 GLU A 397 87. .086 54 , .341 23. .452 1. , 00 13. .51 O
ATOM 3063 OE2 GLU A 397 86. , 726 53 , .892 25. .570 1. , 00 13. .49 O
ATOM 3064 C GLU A 397 89. , 403 50 , .844 22. .125 1. , 00 11. .34 C
ATOM 3065 O GLU A 397 90. ,374 50 , .224 22. .572 1. , 00 11. .52 O
ATOM 3066 N MET A 398 88. , 288 50 , .245 21. .712 1. , 00 10. .06 N
ATOM 3067 CA MET A 398 88. , 184 48 , .796 21. .499 1. , 00 10. .02 c
ATOM 3068 CB MET A 398 87. , 437 48 , .128 22. .672 1. , 00 12. .71 C
ATOM 3069 CG MET A 398 85. , 964 48 , .528 22. .786 1. , 00 12. .53 c
ATOM 3070 SD MET A 398 85. , 029 47 , .544 23. .997 1. , 00 16. .23 s
ATOM 3071 CE MET A 398 84. , 659 46 , .047 23. .032 1. , 00 14. .44 C
ATOM 3072 C MET A 398 87. , 455 48 , .473 20. .207 1. , 00 9. .47 C
ATOM 3073 O MET A 398 87. , 041 47 , .335 19. .997 1. , 00 8. .75 O
ATOM 3074 N GLY A 399 87. , 271 49 , .465 19. .333 1. , 00 9. .52 N
ATOM 3075 CA GLY A 399 86. , 505 49 , .261 18. .110 1. , 00 9. .21 C
ATOM 3076 C GLY A 399 85. , 090 48 , .803 18. .401 1. , 00 7. .96 C
ATOM 3077 O GLY A 399 84. , 568 47 , .957 17. .694 1. , 00 8. .09 O
ATOM 3078 N PHE A 400 84. , 483 49 , .386 19. .440 1. , 00 7. .58 N
ATOM 3079 CA PHE A 400 83. , 107 49 , .084 19. .861 1. , 00 8. .56 C
ATOM 3080 CB PHE A 400 82. , 767 49 , .963 21. .061 1. , 00 9. .59 C
ATOM 3081 CG PHE A 400 81. ,358 49 , .810 21. .604 1. , 00 9. .31 c
ATOM 3082 CD1 PHE A 400 80. , 966 48 , .659 22. .265 1. , 00 11. .80 c
ATOM 3083 CE1 PHE A 400 79. , 695 48 , .552 22. .820 1. , 00 11. .61 C
ATOM 3084 CZ PHE A 400 78. , 810 49 , .615 22. .714 1. , 00 11. .48 C
ATOM 3085 CE2 PHE A 400 79. , 188 50 , .765 22. .071 1. , 00 11. .37 c
ATOM 3086 CD2 PHE A 400 80. , 464 50 , .861 21. .530 1. , 00 10. .30 c
ATOM 3087 C PHE A 400 82. , 153 49 , .387 18. .722 1. , 00 9. .36 C
ATOM 3088 O PHE A 400 82. , 151 50 , .511 18. .218 1. , 00 8. .65 O
ATOM 3089 N MET A 401 81. , 350 48 , .397 18. .319 1. , 00 9. .03 N
ATOM 3090 CA MET A 401 80. , 299 48 , .602 17. .296 1. , 00 9. .83 c
ATOM 3091 CB MET A 401 79. , 216 49 , .567 17. .806 1. , 00 11. .64 C
ATOM 3092 CG MET A 401 78. , 416 49 , .024 18. .991 1. , 00 13. .59 C
ATOM 3093 SD MET A 401 77. ,307 47 , .646 18. .569 1. , 00 17. .07 s
ATOM 3094 CE MET A 401 78. , 174 46 , .251 19. .273 1. , 00 21. .13 C
ATOM 3095 C MET A 401 80. , 905 49 , .108 15. .988 1. , 00 9. .89 C
ATOM 3096 O MET A 401 80. , 275 49 , .872 15. .216 1. , 00 9. .34 O
ATOM 3097 N ARG A 402 82. , 128 48 , .656 15. .716 1. , 00 9. .32 N
ATOM 3098 CA ARG A 402 82. , 829 49 , .061 14. .503 1. , 00 9. .24 C
ATOM 3099 CB ARG A 402 84. , 162 48 , .336 14. .382 1. , 00 9. .99 C
ATOM 3100 CG ARG A 402 84. , 055 46 , .823 14. .315 1. , 00 10. .89 c
ATOM 3101 CD ARG A 402 85. , 424 46 , .171 14. .380 1. , 00 10. .30 c
ATOM 3102 NE ARG A 402 85. , 985 46 , .226 15. .733 1. , 00 10. .74 N
ATOM 3103 CZ ARG A 402 87. , 217 45 , .848 16. .064 1. , 00 12. .31 C
ATOM 3104 NH1 ARG A 402 88., 111 45 ,.453 15..137 1., 00 14..64 N
ATOM 3105 NH2 ARG A 402 87. , 580 45 , .897 17. .330 1. , 00 12. .67 N
ATOM 3106 C ARG A 402 81. , 962 48 , .772 13. .300 1. , 00 8. .71 C
ATOM 3107 O ARG A 402 81. .260 47 , .770 13. .255 1. , 00 10. .20 O
ATOM 3108 N GLY A 403 81. , 980 49 , .662 12. .331 1. , 00 8. .92 N
ATOM 3109 CA GLY A 403 81. , 199 49 , .458 11. .162 1. , 00 9. .36 C
ATOM 3110 C GLY A 403 79. , 717 49 , .743 11. .273 1. , 00 9. .18 C
ATOM 3111 O GLY A 403 79. , 048 49 , .718 10. .265 1. , 00 10. .53 0
ATOM 3112 N ARG A 404 79. , 177 50 , .032 12. .450 1. , 00 11. .12 N
ATOM 3113 CA ARG A 404 77. , 711 50 , .091 12. .601 1. , 00 11. .58 C
ATOM 3114 CB ARG A 404 77. , 268 49 , .569 13. .951 1. , 00 10. .97 C
ATOM 3115 CG ARG A 404 77. , 811 48 , .205 14. .320 1. , 00 14. .19 c
ATOM 3116 CD ARG A 404 77. , 059 47 , .080 13. .693 1. , 00 15. .25 C
ATOM 3117 NE ARG A 404 77. , 652 45 , .769 13. .974 1. , 00 17. .42 N
ATOM 3118 CZ ARG A 404 77. , 110 44 , .616 13. .569 1. , 00 16. .10 C
ATOM 3119 NH1 ARG A 404 75. , 943 44 , .593 12. .937 1. , 00 15. .16 N
ATOM 3120 NH2 ARG A 404 77. , 735 43 , .479 13. .794 1. , 00 14. .11 N
ATOM 3121 C ARG A 404 77. , 156 51 , .490 12. .419 1. , 00 13. .21 C
ATOM 3122 O ARG A 404 77. , 733 52 , .441 12. .927 1. , 00 11. .53 O
ATOM 3123 N PRO A 405 76. , 001 51 , .611 11. .730 1. , 00 12. .40 N
ATOM 3124 CA PRO A 405 75. , 357 52 , .898 11. .519 1. , 00 12. .75 C
ATOM 3125 CB PRO A 405 74. , 593 52 , .660 10. .208 1. , 00 13. .94 C
ATOM 3126 CG PRO A 405 74. , 145 51 , .261 10. .328 1. , 00 15. .91 c
ATOM 3127 CD PRO A 405 75. , 258 50 , .526 11. .060 1. , 00 13. .75 c
ATOM 3128 C PRO A 405 74. ,373 53 , .284 12. .629 1. , 00 11. .38 C
ATOM 3129 O PRO A 405 73. , 994 52 , .445 13. .492 1. , 00 9. .44 O
ATOM 3130 N GLU A 406 73. .962 54 , .540 12. .646 1. , 00 11. .11 N
ATOM 3131 CA GLU A 406 72. , 835 54 , .921 13. .495 1. , 00 11. .92 C
ATOM 3132 CB GLU A 406 72. , 447 56 , .398 13. .339 1. , 00 11. .86 C
ATOM 3133 CG GLU A 406 71. ,359 56 , .796 14. .344 1. , 00 13. .35 C
ATOM 3134 CD GLU A 406 70. , 896 58 , .242 14. .262 1. , 00 17. .58 c
ATOM 3135 OE1 GLU A 406 71. , 185 58 , .921 13. .258 1. , 00 18. .39 O
ATOM 3136 OE2 GLU A 406 70. , 248 58 , .705 15. .230 1. , 00 14. .63 O
ATOM 3137 C GLU A 406 71. , 636 54 , .026 13. .143 1. , 00 10. .51 C
ATOM 3138 O GLU A 406 71. , 390 53 , .760 11. .972 1. , 00 9. .65 0
ATOM 3139 N HIS A 407 70. , 909 53 , .600 14. .176 1. , 00 11. .75 N
ATOM 3140 CA HIS A 407 69. , 770 52 , .662 14. .105 1. , 00 10. .58 C
ATOM 3141 CB HIS A 407 68. , 723 53 , .082 13. .058 1. , 00 10. .81 C
ATOM 3142 CG HIS A 407 68. ,307 54 , .536 13. .170 1. , 00 11. .70 c
ATOM 3143 ND1 HIS A 407 68. ,332 55 , .385 12. .116 1. , 00 13. .69 N
ATOM 3144 CE1 HIS A 407 67. , 968 56 , .616 12. .521 1. , 00 13. .15 C
ATOM 3145 NE2 HIS A 407 67. , 697 56 , .560 13. .831 1. , 00 12. .12 N
ATOM 3146 CD2 HIS A 407 67. , 907 55 , .289 14. .265 1. , 00 12. .01 C
ATOM 3147 C HIS A 407 70. , 207 51 , .235 13. .911 1. , 00 11. .19 C
ATOM 3148 O HIS A 407 69. ,374 50 , .349 13. .786 1. , 00 11. .04 O
ATOM 3149 N GLY A 408 71., 516 50 ,.990 13..910 1., 00 9..43 N
ATOM 3150 CA GLY A 408 72. , 052 49 , .635 13. .812 1. , 00 9. .69 C
ATOM 3151 C GLY A 408 73. , 146 49 , .337 14. .829 1. , 00 9. .07 C
ATOM 3152 O GLY A 408 73. .943 48. .396 14. .651 1. .00 8. .47 0
ATOM 3153 N GLY A 409 73. , 225 50 , .156 15. .879 1. , 00 9. .25 N
ATOM 3154 CA GLY A 409 74. , 146 49 , .869 16. .983 1. , 00 9. .82 C
ATOM 3155 C GLY A 409 75. , 266 50 , .867 17. .219 1. , 00 9. .83 C
ATOM 3156 O GLY A 409 75. , 963 50 , .763 18. .219 1. , 00 11. .53 0
ATOM 3157 N ASN A 410 75. , 431 51 , .836 16. .319 1. , 00 9. .21 N
ATOM 3158 CA ASN A 410 76. , 487 52 , .863 16. .431 1. , 00 9. .27 C
ATOM 3159 CB ASN A 410 76. , 248 53 , .961 15. .375 1. , 00 8. .73 C
ATOM 3160 CG ASN A 410 77. , 062 55 , .217 15. .614 1. , 00 10. .22 c
ATOM 3161 OD1 ASN A 410 76. , 945 55 , .853 16. .674 1. , 00 10. .70 O
ATOM 3162 ND2 ASN A 410 77. , 864 55 , .630 14. .595 1. , 00 9. .57 N
ATOM 3163 C ASN A 410 76. , 507 53 , .459 17. .823 1. , 00 9. .80 C
ATOM 3164 O ASN A 410 77. , 570 53 , .617 18. .437 1. , 00 9. .69 0
ATOM 3165 N ARG A 411 75. ,313 53 , .769 18. .323 1. , 00 8. .55 N
ATOM 3166 CA ARG A 411 75. , 152 54 , .507 19. .587 1. , 00 9. .67 C
ATOM 3167 CB ARG A 411 74. , 069 55 , .560 19. .401 1. , 00 10. .14 C
ATOM 3168 CG ARG A 411 74. , 368 56. .565 18. .305 1. , 00 9. .41 c
ATOM 3169 CD ARG A 411 73. , 169 57 , .449 18. .073 1. , 00 9. .64 C
ATOM 3170 NE ARG A 411 73. , 414 58 , .495 17. .074 1. , 00 8. .16 N
ATOM 3171 CZ ARG A 411 72. , 728 59 , .634 17. .005 1. , 00 8. .73 c
ATOM 3172 NH1 ARG A 411 71. , 739 59. .888 17. .850 1. , 00 8. .48 N
ATOM 3173 NH2 ARG A 411 73. , 014 60 , .532 16. .079 1. , 00 9. .18 N
ATOM 3174 C ARG A 411 74. , 797 53 , .621 20. .801 1. , 00 8. .76 C
ATOM 3175 O ARG A 411 74. .301 54. .097 21. .818 1. .00 8. .13 0
ATOM 3176 N ASN A 412 75. , 067 52 , .336 20. .712 1. , 00 8. .92 N
ATOM 3177 CA ASN A 412 74. , 708 51 , .429 21. .798 1. , 00 9. .01 C
ATOM 3178 CB ASN A 412 74. , 914 49 , .949 21. .388 1. , 00 9. .99 C
ATOM 3179 CG ASN A 412 73. , 736 49 , .390 20. .568 1. , 00 10. .79 c
ATOM 3180 OD1 ASN A 412 72. , 828 50 , .126 20. .188 1. , 00 10. .43 O
ATOM 3181 ND2 ASN A 412 73. , 742 48 , .073 20. .329 1. , 00 10. .06 N
ATOM 3182 C ASN A 412 75. , 428 51 , .771 23. .113 1. , 00 9. .19 C
ATOM 3183 O ASN A 412 76. , 485 52 , .412 23. .118 1. , 00 8. .19 O
ATOM 3184 N TYR A 413 74. , 786 51 , .377 24. .218 1. , 00 9. .36 N
ATOM 3185 CA TYR A 413 75. , 240 51 , .660 25. .564 1. , 00 9. .55 C
ATOM 3186 CB TYR A 413 74. , 164 51 , .225 26. .583 1. , 00 10. .00 C
ATOM 3187 CG TYR A 413 74. , 551 51 , .420 28. .013 1. , 00 10. .43 c
ATOM 3188 CD1 TYR A 413 74. ,396 52 , .644 28. .634 1. , 00 10. .31 C
ATOM 3189 CE1 TYR A 413 74. , 767 52 , .825 29. .955 1. , 00 10. .24 C
ATOM 3190 CZ TYR A 413 75. , 292 51 , .769 30. .667 1. , 00 10. .83 C
ATOM 3191 OH TYR A 413 75. , 686 51 , .949 31. .966 1. , 00 9. .63 0
ATOM 3192 CE2 TYR A 413 75. , 461 50 , .545 30. .069 1. , 00 11. .66 C
ATOM 3193 CD2 TYR A 413 75. , 093 50 , .370 28. .751 1. , 00 11. .76 C
ATOM 3194 C TYR A 413 76 ,, 541 50 ,, 937 25., 823 1., 00 8., 78 C
ATOM 3195 O TYR A 413 76 , ,666 49 , , 767 25. ,526 1. , 00 8. ,26 O
ATOM 3196 N PHE A 414 77 , , 525 51 , , 643 26. ,374 1. , 00 9. ,51 N
ATOM 3197 CA PHE A 414 78. .899 51 , .112 26. .523 1. .00 8. .44 C
ATOM 3198 CB PHE A 414 79 , , 878 52 , , 264 26. ,303 1. , 00 8. , 41 C
ATOM 3199 CG PHE A 414 81 , ,332 51 , , 867 26. , 226 1. , 00 7. , 71 C
ATOM 3200 CD1 PHE A 414 81 , , 790 51 , , 031 25. ,238 1. , 00 8. , 09 C
ATOM 3201 CE1 PHE A 414 83 , .137 50 , .696 25. .155 1. , 00 8. .79 c
ATOM 3202 CZ PHE A 414 84, , 043 51 , , 211 26. , 046 1. , 00 8. ,29 C
ATOM 3203 CE2 PHE A 414 83 , , 599 52 , , 059 27. , 046 1. , 00 9. , 45 C
ATOM 3204 CD2 PHE A 414 82 , , 244 52 , ,383 27. , 128 1. , 00 8. ,59 C
ATOM 3205 C PHE A 414 79 , .053 50 , .487 27. .899 1. , 00 8. .90 c
ATOM 3206 O PHE A 414 79 , ,303 49 , , 293 28. , 038 1. , 00 8. ,60 O
ATOM 3207 N GLY A 415 78 , , 864 51 , , 292 28. ,939 1. , 00 10. ,68 N
ATOM 3208 CA GLY A 415 78 , , 755 50 , , 759 30. ,307 1. , 00 11. ,20 C
ATOM 3209 C GLY A 415 80 , .077 50 , .315 30. .904 1. , 00 11. .49 c
ATOM 3210 O GLY A 415 81 , , 134 50 , ,334 30. ,244 1. , 00 9. ,66 O
ATOM 3211 N THR A 416 80 , , 011 49 , , 889 32. , 157 1. , 00 12. ,37 N
ATOM 3212 CA THR A 416 81 , , 176 49 , , 297 32. , 815 1. , 00 12. ,28 C
ATOM 3213 CB THR A 416 80. .869 48 , .975 34. .282 1. , 00 12. .76 C
ATOM 3214 OG1 THR A 416 79 , , 677 48 , , 206 34. ,307 1. , 00 11. , 49 O
ATOM 3215 CG2 THR A 416 80 , ,666 50 , , 284 35. , 113 1. , 00 12. ,14 C
ATOM 3216 C THR A 416 81 , , 610 48 , , 015 32. , 106 1. , 00 12. ,64 C
ATOM 3217 O THR A 416 82. .780 47 , .681 32. .124 1. , 00 13. .25 0
ATOM 3218 N GLU A 417 80 , , 693 47 , ,318 31. , 445 1. , 00 12. ,00 N
ATOM 3219 CA GLU A 417 81 , , 080 46 , , 178 30. , 594 1. , 00 15. , 75 C
ATOM 3220 CB GLU A 417 79. .853 45 , .442 30. .043 1. .00 21. .97 c
ATOM 3221 CG GLU A 417 79 , , 130 44 , , 614 31. , 097 1. , 00 30. ,92 C
ATOM 3222 CD GLU A 417 78 , , 104 43 , , 632 30. ,520 1. , 00 41. , 78 C
ATOM 3223 OE1 GLU A 417 77 , , 862 43 , , 612 29. ,286 1. , 00 41. , 73 O
ATOM 3224 OE2 GLU A 417 77 , .530 42 , .864 31. .324 1. , 00 54. .62 0
ATOM 3225 C GLU A 417 81 , , 959 46 , , 579 29. , 418 1. , 00 12. , 75 C
ATOM 3226 O GLU A 417 82 , , 930 45 , , 894 29. , 104 1. , 00 12. ,24 O
ATOM 3227 N GLY A 418 81 , , 603 47 , , 667 28. , 730 1. , 00 10. , 82 N
ATOM 3228 CA GLY A 418 82 , .473 48 , .196 27. .668 1. , 00 10. .01 c
ATOM 3229 C GLY A 418 83 , , 852 48 , , 571 28. , 189 1. , 00 10. ,14 C
ATOM 3230 O GLY A 418 84, , 883 48 , , 261 27. , 564 1. , 00 11. ,11 O
ATOM 3231 N ILE A 419 83 , , 873 49 , , 257 29. ,329 1. , 00 10. ,51 N
ATOM 3232 CA ILE A 419 85 , .131 49 , .682 29. .955 1. , 00 11. .23 c
ATOM 3233 CB ILE A 419 84, , 879 50 , , 478 31. ,241 1. , 00 12. ,65 C
ATOM 3234 CGI ILE A 419 84, , 409 51 , , 890 30. , 885 1. , 00 13. ,62 C
ATOM 3235 CD1 ILE A 419 83 , , 646 52 , , 570 31. ,993 1. , 00 14. , 41 C
ATOM 3236 CG2 ILE A 419 86 , .141 50 , .538 32. .127 1. , 00 11. .69 c
ATOM 3237 C ILE A 419 86 , , 035 48 , , 488 30. , 232 1. , 00 12. ,04 C
ATOM 3238 O ILE A 419 87, , 226 48 , , 547 29. , 924 1. , 00 9. ,88 O
ATOM 3239 N ALA A 420 85 ,, 454 47 ,, 406 30., 768 1., 00 12.,10 N
ATOM 3240 CA ALA A 420 86 , , 203 46 , , 175 31. , 100 1. , 00 14. ,98 C
ATOM 3241 CB ALA A 420 85, ,335 45 , , 171 31. , 892 1. , 00 13. ,23 C
ATOM 3242 C ALA A 420 86. .735 45. , 541 29. .827 1. , 00 14. .21 c
ATOM 3243 O ALA A 420 87 , , 912 45 , , 168 29. , 737 1. , 00 13. ,95 O
ATOM 3244 N ALA A 421 85, , 878 45 , , 462 28. , 820 1. , 00 14. , 06 N
ATOM 3245 CA ALA A 421 86 , , 292 44 , , 905 27. , 541 1. , 00 13. ,64 C
ATOM 3246 CB ALA A 421 85 , .087 44. , 736 26. .612 1. , 00 13. .35 c
ATOM 3247 C ALA A 421 87, , 401 45 , , 760 26. ,903 1. , 00 14. , 05 C
ATOM 3248 O ALA A 421 88 , ,346 45 , , 227 26. ,345 1. , 00 15. , 44 O
ATOM 3249 N ALA A 422 87, ,319 47 , , 082 27. , 036 1. , 00 14. ,54 N
ATOM 3250 CA ALA A 422 88 , .318 47. , 973 26. .427 1. , 00 14. .27 c
ATOM 3251 CB ALA A 422 87, , 825 49 , , 423 26. , 405 1. , 00 10. ,31 C
ATOM 3252 C ALA A 422 89 , , 682 47 , , 872 27. , 112 1. , 00 14. ,26 C
ATOM 3253 O ALA A 422 90 , , 720 47 , , 955 26. , 458 1. , 00 13. , 78 O
ATOM 3254 N LYS A 423 89 , .686 47. , 668 28. .422 1. , 00 16. .41 N
ATOM 3255 CA LYS A 423 90 , , 952 47 , , 552 29. , 158 1. , 00 17. ,57 C
ATOM 3256 CB LYS A 423 90 , , 733 47 , , 489 30. ,673 1. , 00 18. ,04 C
ATOM 3257 CG LYS A 423 90 , ,364 48 , , 829 31. , 291 1. , 00 20. ,96 C
ATOM 3258 CD LYS A 423 90 , .145 48. , 702 32. .782 1. , 00 22. .90 C
ATOM 3259 CE LYS A 423 89 , , 713 50 , , 010 33. ,396 1. , 00 29. , 43 C
ATOM 3260 NZ LYS A 423 89 , , 573 49 , , 913 34. , 876 1. , 00 35. ,53 N
ATOM 3261 C LYS A 423 91 , , 729 46 , ,325 28. , 717 1. , 00 17. , 71 C
ATOM 3262 O LYS A 423 92 , .921 46. ,244 28. .948 1. , 00 18. .10 O
ATOM 3263 N ALA A 424 91 , , 055 45 , ,370 28. , 090 1. , 00 18. ,81 N
ATOM 3264 CA ALA A 424 91 , , 709 44 , , 156 27. ,629 1. , 00 17. , 13 C
ATOM 3265 CB ALA A 424 90 , .792 42. , 946 27. .888 1. , 00 17. .01 c
ATOM 3266 C ALA A 424 92 , , 084 44 , , 241 26. , 152 1. , 00 19. ,84 C
ATOM 3267 O ALA A 424 92 , , 621 43 , , 292 25. ,613 1. , 00 19. , 77 O
ATOM 3268 N SER A 425 91 , , 775 45 , ,359 25. , 485 1. , 00 17. ,11 N
ATOM 3269 CA SER A 425 92 , .008 45. , 493 24. .047 1. , 00 15. .39 c
ATOM 3270 CB SER A 425 90 , , 935 46 , , 401 23. , 432 1. , 00 17. ,94 C
ATOM 3271 OG SER A 425 91 , , 178 46 , , 615 22. , 057 1. , 00 14. ,67 O
ATOM 3272 C SER A 425 93 , ,375 46 , , 088 23. , 701 1. , 00 12. ,91 C
ATOM 3273 O SER A 425 93 , .788 47. , 099 24. .289 1. , 00 12. .79 0
ATOM 3274 N PRO A 426 94 , , 073 45 , , 498 22. , 723 1. , 00 14. ,11 N
ATOM 3275 CA PRO A 426 95, ,335 46 , , 127 22. ,274 1. , 00 14. , 44 C
ATOM 3276 CB PRO A 426 95, , 945 45 , , 104 21. ,293 1. , 00 16. ,99 C
ATOM 3277 CG PRO A 426 94 , .858 44. , 108 20. .994 1. , 00 24. .47 c
ATOM 3278 CD PRO A 426 93 , , 852 44 , , 158 22. , 129 1. , 00 18. ,07 C
ATOM 3279 C PRO A 426 95, , 153 47 , , 485 21. ,590 1. , 00 12. ,29 C
ATOM 3280 O PRO A 426 96 , , 106 48 , , 262 21. , 517 1. , 00 10. , 83 O
ATOM 3281 N ILE A 427 93 , .947 47. , 777 21. .093 1. , 00 10. .87 N
ATOM 3282 CA ILE A 427 93 , , 651 49 , , 111 20. , 544 1. , 00 9. ,18 C
ATOM 3283 CB ILE A 427 92 , , 228 49 , , 216 19. , 927 1. , 00 8. ,07 C
ATOM 3284 CGI ILE A 427 92., 028 48., 155 18..839 1., 00 8..24 C
ATOM 3285 CD1 ILE A 427 90. , 563 47. , 951 18. .430 1. , 00 8. .25 C
ATOM 3286 CG2 ILE A 427 91. , 976 50. , 608 19. .376 1. , 00 6. .93 C
ATOM 3287 C ILE A 427 93. .813 50. .155 21. .634 1. , 00 8. .85 c
ATOM 3288 O ILE A 427 94. , 454 51. , 173 21. .434 1. , 00 9. .48 O
ATOM 3289 N ARG A 428 93. , 294 49. , 886 22. .823 1. , 00 8. .60 N
ATOM 3290 CA ARG A 428 93. , 400 50. , 866 23. .894 1. , 00 8. .95 C
ATOM 3291 CB ARG A 428 92. , 500 50. , 479 25. .056 1. , 00 10. .81 c
ATOM 3292 CG ARG A 428 92. , 422 51. , 559 26. .136 1. , 00 11. .59 C
ATOM 3293 CD ARG A 428 91. , 447 51. , 164 27. .240 1. , 00 12. .02 C
ATOM 3294 NE ARG A 428 91. ,350 52. , 213 28. .252 1. , 00 12. .91 N
ATOM 3295 CZ ARG A 428 92. , 020 52. , 236 29. .398 1. , 00 13. .84 c
ATOM 3296 NH1 ARG A 428 92. , 856 51. , 263 29. .698 1. , 00 14. .05 N
ATOM 3297 NH2 ARG A 428 91. , 835 53. , 234 30. .258 1. , 00 14. .78 N
ATOM 3298 C ARG A 428 94. , 849 51. , 122 24. .363 1. , 00 9. .27 C
ATOM 3299 O ARG A 428 95. , 228 52. , 266 24. .663 1. , 00 8. .53 0
ATOM 3300 N ALA A 429 95. , 663 50. , 069 24. .391 1. , 00 9. .95 N
ATOM 3301 CA ALA A 429 97. , 056 50. , 169 24. .807 1. , 00 11. .02 C
ATOM 3302 CB ALA A 429 97. , 705 48. , 757 24. .946 1. , 00 11. .56 c
ATOM 3303 C ALA A 429 97. , 837 51. , 028 23. .809 1. , 00 10. .12 c
ATOM 3304 O ALA A 429 98. , 608 51. , 895 24. .214 1. , 00 9. .00 O
ATOM 3305 N ALA A 430 97. , 619 50. , 792 22. .512 1. , 00 8. .74 N
ATOM 3306 CA ALA A 430 98. , 286 51. , 579 21. .459 1. , 00 9. .32 c
ATOM 3307 CB ALA A 430 98. , 092 50. , 932 20. .081 1. , 00 7. .90 c
ATOM 3308 C ALA A 430 97. , 794 53. , 037 21. .466 1. , 00 9. .63 C
ATOM 3309 O ALA A 430 98. , 571 53. , 965 21. .245 1. , 00 10. .12 O
ATOM 3310 N LEU A 431 96. .505 53. .239 21. .720 1. , 00 9. .43 N
ATOM 3311 CA LEU A 431 95. , 941 54. , 610 21. .769 1. , 00 9. .39 C
ATOM 3312 CB LEU A 431 94. , 422 54. , 577 21. .980 1. , 00 8. .79 C
ATOM 3313 CG LEU A 431 93. , 674 55. , 877 22. .294 1. , 00 10. .37 C
ATOM 3314 CD1 LEU A 431 93. , 855 56. , 860 21. .155 1. , 00 10. .80 c
ATOM 3315 CD2 LEU A 431 92. , 191 55. , 572 22. .517 1. , 00 10. .19 C
ATOM 3316 C LEU A 431 96. , 590 55. ,393 22. .902 1. , 00 9. .89 C
ATOM 3317 O LEU A 431 97. , 041 56. , 529 22. .713 1. , 00 8. .92 O
ATOM 3318 N THR A 432 96. , 634 54. , 765 24. .075 1. , 00 9. .59 N
ATOM 3319 CA THR A 432 97. , 269 55. ,343 25. .242 1. , 00 9. .93 C
ATOM 3320 CB THR A 432 97. , 247 54. ,365 26. .429 1. , 00 10. .14 C
ATOM 3321 OG1 THR A 432 95. , 881 54. , 127 26. .769 1. , 00 8. .71 O
ATOM 3322 CG2 THR A 432 98. , 019 54. , 934 27. .635 1. , 00 10. .34 c
ATOM 3323 C THR A 432 98. , 696 55. , 768 24. .965 1. , 00 10. .79 C
ATOM 3324 O THR A 432 99. , 088 56. , 864 25. .351 1. , 00 10. .03 O
ATOM 3325 N ARG A 433 99. , 468 54. , 916 24. .300 1. , 00 11. .71 N
ATOM 3326 CA ARG A 433 100. , 872 55. , 225 24. .053 1. , 00 12. .68 c
ATOM 3327 CB ARG A 433 101. , 629 54. , 062 23. .396 1. , 00 16. .53 C
ATOM 3328 CG ARG A 433 102. , 155 53. , 002 24. .374 1. , 00 23. .81 C
ATOM 3329 CD ARG A 433 102 ,, 820 51 ,, 822 23., 651 1., 00 27.,35 C
ATOM 3330 NE ARG A 433 101 , , 846 50 , , 739 23. , 405 1. , 00 40. ,17 N
ATOM 3331 CZ ARG A 433 101 , , 725 50 , , 034 22. ,279 1. , 00 38. ,69 C
ATOM 3332 NH1 ARG A 433 102. .512 50 , .262 21. .224 1. .00 54. .11 N
ATOM 3333 NH2 ARG A 433 100 , , 800 49 , , 089 22. , 199 1. , 00 29. ,01 N
ATOM 3334 C ARG A 433 101 , , 003 56 , , 474 23. , 188 1. , 00 11. ,20 C
ATOM 3335 O ARG A 433 101 , , 728 57 , ,383 23. , 547 1. , 00 10. ,17 O
ATOM 3336 N ILE A 434 100 , .320 56 , .514 22. .042 1. , 00 10. .54 N
ATOM 3337 CA ILE A 434 100 , , 474 57 , , 664 21. , 136 1. , 00 9. ,22 C
ATOM 3338 CB ILE A 434 100 , , 008 57 , ,353 19. ,680 1. , 00 8. , 85 C
ATOM 3339 CGI ILE A 434 100 , , 583 58 , ,386 18. , 710 1. , 00 9. , 47 C
ATOM 3340 CD1 ILE A 434 102 , .113 58 , .485 18. .712 1. , 00 8. .53 c
ATOM 3341 CG2 ILE A 434 98 , , 482 57 , , 213 19. , 569 1. , 00 8. ,39 C
ATOM 3342 C ILE A 434 99 , , 815 58 , , 935 21. , 711 1. , 00 10. ,10 C
ATOM 3343 O ILE A 434 100 , , 351 60 , , 047 21. , 564 1. , 00 10. ,27 O
ATOM 3344 N ALA A 435 98 , .665 58 , .784 22. .377 1. , 00 9. .56 N
ATOM 3345 CA ALA A 435 97, , 962 59 , , 938 22. ,968 1. , 00 8. ,34 C
ATOM 3346 CB ALA A 435 96 , , 551 59 , , 546 23. , 477 1. , 00 7. ,35 C
ATOM 3347 C ALA A 435 98 , , 779 60 , , 590 24. , 056 1. , 00 8. ,51 C
ATOM 3348 O ALA A 435 98. .835 61 , .827 24. .143 1. , 00 9. .31 O
ATOM 3349 N GLN A 436 99 , , 441 59 , , 790 24. , 879 1. , 00 9. ,61 N
ATOM 3350 CA GLN A 436 100 , , 296 60 , ,347 25. ,919 1. , 00 11. ,37 C
ATOM 3351 CB GLN A 436 100 , , 699 59 , , 282 26. ,971 1. , 00 14. , 19 c
ATOM 3352 CG GLN A 436 99. .515 58 , .768 27. .816 1. , 00 16. .35 c
ATOM 3353 CD GLN A 436 98 , , 955 59 , , 800 28. , 825 1. , 00 22. ,66 C
ATOM 3354 OE1 GLN A 436 99 , ,375 60 , , 978 28. , 861 1. , 00 25. , 03 O
ATOM 3355 NE2 GLN A 436 97. .980 59 , .366 29. .636 1. .00 24. .58 N
ATOM 3356 C GLN A 436 101 , , 546 61 , , 037 25. ,307 1. , 00 12. ,28 C
ATOM 3357 O GLN A 436 102 , , 025 62 , , 050 25. , 850 1. , 00 11. ,97 O
ATOM 3358 N VAL A 437 102 , , 059 60 , , 513 24. , 192 1. , 00 10. ,31 N
ATOM 3359 CA VAL A 437 103 , .124 61 , .222 23. .447 1. , 00 9. .13 c
ATOM 3360 CB VAL A 437 103 , , 704 60 , ,387 22. ,288 1. , 00 10. ,04 C
ATOM 3361 CGI VAL A 437 104 , , 592 61 , , 244 21. ,390 1. , 00 9. , 42 C
ATOM 3362 CG2 VAL A 437 104 , , 483 59 , , 182 22. , 836 1. , 00 10. , 83 C
ATOM 3363 C VAL A 437 102 , .639 62 , .605 22. .973 1. , 00 8. .86 c
ATOM 3364 O VAL A 437 103 , ,320 63 , , 602 23. ,212 1. , 00 8. ,87 O
ATOM 3365 N ARG A 438 101 , , 441 62 , , 702 22. ,379 1. , 00 8. ,39 N
ATOM 3366 CA ARG A 438 100 , , 914 64 , , 002 21. ,944 1. , 00 8. ,28 C
ATOM 3367 CB ARG A 438 99 , .553 63 , .855 21. .250 1. , 00 8. .00 c
ATOM 3368 CG ARG A 438 99 , , 075 65, , 152 20. ,619 1. , 00 9. , 42 C
ATOM 3369 CD ARG A 438 97, , 660 65, , 039 20. , 072 1. , 00 8. ,50 C
ATOM 3370 NE ARG A 438 97, , 240 66 , , 241 19. ,353 1. , 00 8. ,96 N
ATOM 3371 CZ ARG A 438 96 , .840 67 , .380 19. .917 1. , 00 8. .98 c
ATOM 3372 NH1 ARG A 438 96 , , 806 67, , 526 21. , 232 1. , 00 11. , 49 N
ATOM 3373 NH2 ARG A 438 96 , , 445 68 , ,379 19. , 162 1. , 00 9. , 76 N
ATOM 3374 C ARG A 438 100., 762 64 ,, 940 23., 154 1., 00 9., 49 C
ATOM 3375 O ARG A 438 101. , 134 66 , , 103 23. , 108 1. , 00 7. ,66 O
ATOM 3376 N ALA A 439 100. , 202 64 , , 405 24. ,243 1. , 00 9. , 79 N
ATOM 3377 CA ALA A 439 99. .893 65 , .231 25. .406 1. , 00 10. .72 C
ATOM 3378 CB ALA A 439 99. , 141 64 , , 419 26. , 476 1. , 00 10. , 49 C
ATOM 3379 C ALA A 439 101. , 166 65, , 845 25. , 991 1. , 00 11. ,63 C
ATOM 3380 O ALA A 439 101. , 091 66 , , 875 26. ,602 1. , 00 9. ,51 O
ATOM 3381 N ALA A 440 102. , 326 65 , .218 25. .794 1. , 00 10. .51 N
ATOM 3382 CA ALA A 440 103. , 597 65, , 766 26. ,279 1. , 00 11. , 02 C
ATOM 3383 CB ALA A 440 104. , 458 64, , 652 26. , 889 1. , 00 10. ,64 C
ATOM 3384 C ALA A 440 104. , 416 66 , , 526 25. ,216 1. , 00 10. ,53 C
ATOM 3385 O ALA A 440 105. , 501 66 , .993 25. .519 1. , 00 10. .60 0
ATOM 3386 N SER A 441 103. , 906 66 , , 683 23. ,998 1. , 00 10. , 76 N
ATOM 3387 CA SER A 441 104. , 695 67, , 246 22. , 879 1. , 00 10. , 42 C
ATOM 3388 CB SER A 441 104. , 815 66 , , 223 21. , 732 1. , 00 11. ,33 C
ATOM 3389 OG SER A 441 105. , 390 66 , .801 20. .538 1. , 00 9. .86 0
ATOM 3390 C SER A 441 104. , 028 68 , , 493 22. , 321 1. , 00 10. ,28 C
ATOM 3391 O SER A 441 102. , 993 68 , ,395 21. ,663 1. , 00 9. , 49 O
ATOM 3392 N PRO A 442 104. , 631 69 , , 664 22. , 547 1. , 00 11. , 42 N
ATOM 3393 CA PRO A 442 104. , 121 70 , .887 21. .926 1. , 00 11. .27 c
ATOM 3394 CB PRO A 442 105. , 070 71 , , 968 22. , 430 1. , 00 13. ,81 C
ATOM 3395 CG PRO A 442 105. , 690 71 , , 414 23. , 661 1. , 00 16. ,52 C
ATOM 3396 CD PRO A 442 105. , 736 69 , , 929 23. , 484 1. , 00 14. ,07 c
ATOM 3397 C PRO A 442 104. , 142 70 , .839 20. .426 1. , 00 9. .13 c
ATOM 3398 O PRO A 442 103. , 254 71 , ,388 19. , 787 1. , 00 9. ,98 O
ATOM 3399 N ALA A 443 105. , 163 70 , , 202 19. , 856 1. , 00 8. , 71 N
ATOM 3400 CA ALA A 443 105. .228 70 , .013 18. .402 1. , 00 8. .33 c
ATOM 3401 CB ALA A 443 106. , 525 69 , , 325 18. , 013 1. , 00 7. ,21 C
ATOM 3402 C ALA A 443 104. , 034 69 , , 251 17. , 860 1. , 00 7. , 70 C
ATOM 3403 O ALA A 443 103. , 437 69 , , 661 16. , 878 1. , 00 6. , 78 O
ATOM 3404 N LEU A 444 103. , 687 68 , .125 18. .483 1. , 00 7. .83 N
ATOM 3405 CA LEU A 444 102. , 546 67, ,318 18. , 001 1. , 00 8. ,98 C
ATOM 3406 CB LEU A 444 102. , 543 65, , 917 18. ,631 1. , 00 9. , 75 C
ATOM 3407 CG LEU A 444 103. , 683 64 , , 943 18. ,311 1. , 00 9. ,68 C
ATOM 3408 CD1 LEU A 444 103. ,319 63 , .530 18. .766 1. , 00 8. .68 c
ATOM 3409 CD2 LEU A 444 103. , 926 64 , , 924 16. , 809 1. , 00 7. , 72 C
ATOM 3410 C LEU A 444 101. , 201 68 , , 010 18. ,296 1. , 00 10. ,38 C
ATOM 3411 O LEU A 444 100. , 196 67, , 754 17. , 621 1. , 00 10. ,11 O
ATOM 3412 N GLN A 445 101. ,177 68 , .889 19. .292 1. , 00 9. .45 N
ATOM 3413 CA GLN A 445 99. , 938 69 , , 615 19. , 622 1. , 00 9. , 74 C
ATOM 3414 CB GLN A 445 100. , 027 70 , , 197 21. , 040 1. , 00 10. , 47 C
ATOM 3415 CG GLN A 445 99. , 956 69 , , 140 22. , 147 1. , 00 11. ,24 c
ATOM 3416 CD GLN A 445 100. , 441 69 , .657 23. .483 1. , 00 11. .90 c
ATOM 3417 OE1 GLN A 445 100. , 443 70 , , 857 23. , 716 1. , 00 11. ,93 O
ATOM 3418 NE2 GLN A 445 100. , 841 68 , , 750 24. ,376 1. , 00 11. , 74 N
ATOM 3419 C GLN A 445 99., 647 70 ,, 741 18., 626 1., 00 9.,80 C
ATOM 3420 O GLN A 445 98. , 516 70 , , 888 18. , 142 1. , 00 10. , 73 O
ATOM 3421 N ARG A 446 100. , 680 71 , , 502 18. , 292 1. , 00 8. ,91 N
ATOM 3422 CA ARG A 446 100. .507 72 , .783 17. .632 1. , 00 8. .84 C
ATOM 3423 CB ARG A 446 100. , 285 73 , , 874 18. ,676 1. , 00 8. ,99 C
ATOM 3424 CG ARG A 446 101. , 481 74 , , 092 19. , 587 1. , 00 12. ,37 C
ATOM 3425 CD ARG A 446 101. ,339 75 , , 297 20. , 550 1. , 00 13. , 03 C
ATOM 3426 NE ARG A 446 100. ,342 75 , .023 21. .586 1. , 00 10. .64 N
ATOM 3427 CZ ARG A 446 99. , 090 75 , , 447 21. , 581 1. , 00 12. ,57 C
ATOM 3428 NH1 ARG A 446 98. , 618 76 , , 178 20. ,593 1. , 00 13. , 47 N
ATOM 3429 NH2 ARG A 446 98. , 273 75 , , 111 22. , 579 1. , 00 16. , 12 N
ATOM 3430 C ARG A 446 101. , 648 73 , .214 16. .704 1. , 00 8. .88 c
ATOM 3431 O ARG A 446 101. , 640 74 , ,350 16. ,216 1. , 00 8. ,87 O
ATOM 3432 N GLY A 447 102. , 584 72 , ,319 16. , 415 1. , 00 8. ,14 N
ATOM 3433 CA GLY A 447 103. , 690 72 , , 651 15. , 559 1. , 00 9. , 16 C
ATOM 3434 C GLY A 447 103. , 279 72 , .667 14. .105 1. , 00 8. .98 c
ATOM 3435 O GLY A 447 102. ,307 72 , , 011 13. , 721 1. , 00 8. , 78 O
ATOM 3436 N LEU A 448 104. , 020 73 , , 423 13. ,300 1. , 00 8. , 43 N
ATOM 3437 CA LEU A 448 103. , 860 73 , ,373 11. , 856 1. , 00 9. ,00 c
ATOM 3438 CB LEU A 448 104. , 858 74 , .315 11. .165 1. , 00 8. .58 c
ATOM 3439 CG LEU A 448 104. , 576 75 , .795 11. ,379 1. , 00 9. ,35 C
ATOM 3440 CD1 LEU A 448 105. , 818 76 , .659 11. , 023 1. , 00 7. ,31 C
ATOM 3441 CD2 LEU A 448 103. ,334 76 , .189 10. , 581 1. , 00 6. ,84 c
ATOM 3442 C LEU A 448 104. , 124 71 , .928 11. .378 1. , 00 8. .70 c
ATOM 3443 O LEU A 448 104. , 971 71 , .223 11. ,942 1. , 00 9. , 89 O
ATOM 3444 N GLN A 449 103. ,399 71 , .494 10. ,362 1. , 00 8. ,17 N
ATOM 3445 CA GLN A 449 103. , 636 70 , .205 9. .729 1. , 00 7. .61 c
ATOM 3446 CB GLN A 449 102. , 329 69 , .446 9. , 527 1. , 00 7. ,18 C
ATOM 3447 CG GLN A 449 102. , 507 68 , .040 8. ,926 1. , 00 6. , 89 C
ATOM 3448 CD GLN A 449 101. , 274 67, .190 9. , 103 1. , 00 7. ,36 C
ATOM 3449 OE1 GLN A 449 100. , 531 67 , .327 10. .121 1. , 00 8. .33 0
ATOM 3450 NE2 GLN A 449 101. , 034 66 , .284 8. , 150 1. , 00 7. ,23 N
ATOM 3451 C GLN A 449 104. ,356 70 , .369 8. ,397 1. , 00 7. , 73 C
ATOM 3452 O GLN A 449 103. , 955 71 , .193 7. , 558 1. , 00 10. ,39 O
ATOM 3453 N LEU A 450 105. , 424 69 , .601 8. .213 1. , 00 7. .59 N
ATOM 3454 CA LEU A 450 106. , 077 69 , .487 6. , 897 1. , 00 8. ,52 C
ATOM 3455 CB LEU A 450 107. , 499 70 , .024 6. ,941 1. , 00 8. ,31 C
ATOM 3456 CG LEU A 450 107. , 671 71 , .496 7. ,301 1. , 00 8. , 49 C
ATOM 3457 CD1 LEU A 450 109. , 155 71 , .752 7. .571 1. , 00 7. .66 c
ATOM 3458 CD2 LEU A 450 107. , 091 72 , .398 6. ,211 1. , 00 8. , 71 C
ATOM 3459 C LEU A 450 106. , 087 68 , .028 6. , 452 1. , 00 8. , 12 C
ATOM 3460 O LEU A 450 106. , 672 67, .176 7. , 119 1. , 00 8. , 15 O
ATOM 3461 N ASN A 451 105. , 422 67 , .740 5. .343 1. , 00 8. .05 N
ATOM 3462 CA ASN A 451 105. ,323 66 , .383 4. , 835 1. , 00 8. ,39 C
ATOM 3463 CB ASN A 451 104. , 189 66 , .272 3. , 804 1. , 00 8. ,84 C
ATOM 3464 CG ASN A 451 102 ,.826 66., 555 4..397 1., 00 8..85 C
ATOM 3465 OD1 ASN A 451 102 , .612 66. ,390 5. .602 1. , 00 10. .53 O
ATOM 3466 ND2 ASN A 451 101 , .893 67. , 002 3. .555 1. , 00 8. .66 N
ATOM 3467 C ASN A 451 106. .617 65. .923 4. .168 1. , 00 9. .39 C
ATOM 3468 O ASN A 451 107 , .200 66. , 659 3. .399 1. , 00 7. .88 O
ATOM 3469 N LEU A 452 107 , .040 64. , 691 4. .457 1. , 00 8. .89 N
ATOM 3470 CA LEU A 452 108 , .214 64. , 099 3. .842 1. , 00 10. .73 C
ATOM 3471 CB LEU A 452 109 , .203 63. , 640 4. .900 1. , 00 9. .84 c
ATOM 3472 CG LEU A 452 109 , .802 64. , 737 5. .795 1. , 00 12. .35 C
ATOM 3473 CD1 LEU A 452 110 , .541 64. , 105 6. .968 1. , 00 12. .21 C
ATOM 3474 CD2 LEU A 452 110 , .732 65. , 628 4. .984 1. , 00 9. .86 C
ATOM 3475 C LEU A 452 107 , .838 62. , 928 2. .942 1. , 00 10. .79 c
ATOM 3476 O LEU A 452 108 , .383 62. , 794 1. .843 1. , 00 11. .51 O
ATOM 3477 N GLU A 453 106 , .938 62. , 070 3. .416 1. , 00 8. .44 N
ATOM 3478 CA GLU A 453 106 , .483 60. , 925 2. .630 1. , 00 8. .57 C
ATOM 3479 CB GLU A 453 107 , .396 59. , 702 2. .844 1. , 00 8. .80 c
ATOM 3480 CG GLU A 453 106 , .949 58. , 484 2. .047 1. , 00 9. .90 C
ATOM 3481 CD GLU A 453 107 , .862 57. , 254 2. .128 1. , 00 10. .82 C
ATOM 3482 OE1 GLU A 453 108 , .834 57. , 219 2. .904 1. , 00 12. .72 O
ATOM 3483 OE2 GLU A 453 107. .576 56. , 293 1. .377 1. , 00 15. .14 O
ATOM 3484 C GLU A 453 105 , .066 60. , 577 3. .051 1. , 00 7. .83 C
ATOM 3485 O GLU A 453 104 , .782 60. , 470 4. .232 1. , 00 7. .09 O
ATOM 3486 N LEU A 454 104 , .178 60. , 443 2. .081 1. , 00 8. .64 N
ATOM 3487 CA LEU A 454 102. .808 59. , 973 2. .310 1. , 00 9. .02 c
ATOM 3488 CB LEU A 454 101 , .852 61. , 162 2. .398 1. , 00 9. .84 C
ATOM 3489 CG LEU A 454 101 , .953 62. , 009 3. .654 1. , 00 9. .51 C
ATOM 3490 CD1 LEU A 454 101. .219 63. .340 3. .440 1. , 00 11. .30 c
ATOM 3491 CD2 LEU A 454 101 , .411 61. , 251 4. .860 1. , 00 8. .61 C
ATOM 3492 C LEU A 454 102 , .512 59. , 116 1. .078 1. , 00 9. .20 C
ATOM 3493 O LEU A 454 102 , .124 59. , 616 0. .031 1. , 00 8. .58 O
ATOM 3494 N GLN A 455 102 , .767 57. , 827 1. .191 1. , 00 9. .05 N
ATOM 3495 CA GLN A 455 102 , .848 56. , 968 0. .030 1. , 00 9. .12 C
ATOM 3496 CB GLN A 455 104 , .235 57. , 129 -0. .599 1. , 00 9. .92 C
ATOM 3497 CG GLN A 455 104 , .454 56. ,334 -1. .868 1. , 00 12. .29 C
ATOM 3498 CD GLN A 455 105 , .791 56. , 637 -2. .564 1. , 00 11. .72 c
ATOM 3499 OE1 GLN A 455 105 , .850 56. , 679 -3. .784 1. , 00 13. .34 O
ATOM 3500 NE2 GLN A 455 106 , .850 56. , 824 -1. .794 1. , 00 12. .93 N
ATOM 3501 C GLN A 455 102 , .630 55. , 544 0. .493 1. , 00 8. .61 C
ATOM 3502 O GLN A 455 103 , .356 55. , 051 1. .350 1. , 00 7. .56 0
ATOM 3503 N GLY A 456 101 , .607 54. , 899 -0. .041 1. , 00 9. .28 N
ATOM 3504 CA GLY A 456 101 , .355 53. , 490 0. .267 1. , 00 9. .96 C
ATOM 3505 C GLY A 456 101 , .128 53. ,318 1. .753 1. , 00 9. .93 c
ATOM 3506 O GLY A 456 100 , .357 54. , 060 2. .369 1. , 00 10. .32 0
ATOM 3507 N ASN A 457 101 , .840 52. ,377 2. .337 1. , 00 9. .51 N
ATOM 3508 CA ASN A 457 101 , .719 52. , 098 3. .753 1. , 00 11. .18 C
ATOM 3509 CB ASN A 457 102., 118 50 ,, 640 4., 018 1., 00 11..06 C
ATOM 3510 CG ASN A 457 101. , 092 49 , , 650 3. , 480 1. , 00 10. .38 C
ATOM 3511 OD1 ASN A 457 101. , 446 48 , , 646 2. , 872 1. , 00 13. .80 O
ATOM 3512 ND2 ASN A 457 99. .857 49 , .946 3. .670 1. , 00 7. .53 N
ATOM 3513 C ASN A 457 102. , 536 53 , , 015 4. , 665 1. , 00 10. .84 C
ATOM 3514 O ASN A 457 102. , 499 52 , , 839 5. , 876 1. , 00 10. .92 O
ATOM 3515 N ARG A 458 103. , 218 54 , , 001 4. , 091 1. , 00 9. .12 N
ATOM 3516 CA ARG A 458 104. , 253 54 , .747 4. .781 1. , 00 9. .92 C
ATOM 3517 CB ARG A 458 105. , 590 54 , , 605 4. , 015 1. , 00 10. .01 C
ATOM 3518 CG ARG A 458 106. , 094 53 , , 171 3. , 862 1. , 00 12. .11 C
ATOM 3519 CD ARG A 458 107. , 144 53 , , 017 2. , 739 1. , 00 11. .48 C
ATOM 3520 NE ARG A 458 108. , 249 53 , .944 2. .968 1. , 00 10. .95 N
ATOM 3521 CZ ARG A 458 109. ,373 53 , , 652 3. ,616 1. , 00 13. .00 C
ATOM 3522 NH1 ARG A 458 109. , 606 52 , , 422 4. , 081 1. , 00 11. .48 N
ATOM 3523 NH2 ARG A 458 110. ,316 54 , , 593 3. , 753 1. , 00 12. .75 N
ATOM 3524 C ARG A 458 103. , 888 56 , .231 4. .906 1. , 00 9. .10 c
ATOM 3525 O ARG A 458 103. , 423 56 , , 834 3. ,947 1. , 00 10. .94 O
ATOM 3526 N ALA A 459 104. , 098 56 , , 798 6. , 089 1. , 00 8. .77 N
ATOM 3527 CA ALA A 459 104. , 010 58 , , 251 6. , 327 1. , 00 8. .88 c
ATOM 3528 CB ALA A 459 102. , 760 58 , .619 7. .089 1. , 00 7. .78 c
ATOM 3529 C ALA A 459 105. , 241 58 , , 726 7. , 096 1. , 00 9. .20 C
ATOM 3530 O ALA A 459 105. , 692 58 , , 073 8. , 044 1. , 00 9. .88 O
ATOM 3531 N ALA A 460 105. , 779 59 , , 872 6. , 695 1. , 00 9. .71 N
ATOM 3532 CA ALA A 460 106. , 843 60 , .537 7. .442 1. , 00 8. .98 c
ATOM 3533 CB ALA A 460 108. , 218 60 , , 141 6. ,902 1. , 00 7. .69 C
ATOM 3534 C ALA A 460 106. , 614 62 , , 060 7. ,331 1. , 00 9. .17 C
ATOM 3535 O ALA A 460 106. , 328 62 , .573 6. .244 1. , 00 8. .73 0
ATOM 3536 N PHE A 461 106. , 693 62 , , 767 8. .458 1. , 00 7. .86 N
ATOM 3537 CA PHE A 461 106. , 517 64 , , 207 8. .465 1. , 00 7. .58 C
ATOM 3538 CB PHE A 461 105. , 049 64 , , 579 8. .296 1. , 00 8. .14 C
ATOM 3539 CG PHE A 461 104. ,111 63 , .965 9. .316 1. , 00 8. .08 c
ATOM 3540 CD1 PHE A 461 103. , 459 62 , , 771 9. .063 1. , 00 9. .21 C
ATOM 3541 CE1 PHE A 461 102. , 552 62 , , 228 9. .988 1. , 00 9. .07 C
ATOM 3542 CZ PHE A 461 102. , 279 62 , , 906 11. .149 1. , 00 8. .95 C
ATOM 3543 CE2 PHE A 461 102. ,917 64 , .095 11. .420 1. , 00 9. .15 c
ATOM 3544 CD2 PHE A 461 103. , 817 64 , , 631 10. .493 1. , 00 8. .86 C
ATOM 3545 C PHE A 461 107. , 110 64, , 813 9. .731 1. , 00 7. .51 C
ATOM 3546 O PHE A 461 107. , 298 64 , , 110 10. .721 1. , 00 7. .73 O
ATOM 3547 N TYR A 462 107. , 487 66 , .089 9. .660 1. , 00 7. .26 N
ATOM 3548 CA TYR A 462 107. , 939 66 , , 832 10. .828 1. , 00 7. .73 C
ATOM 3549 CB TYR A 462 109. , 029 67, , 861 10. .475 1. , 00 8. .20 C
ATOM 3550 CG TYR A 462 110. , 324 67, , 288 9. .935 1. , 00 8. .39 c
ATOM 3551 CD1 TYR A 462 Ill . , 122 66 , .451 10. .705 1. , 00 8. .74 c
ATOM 3552 CE1 TYR A 462 112. ,308 65, , 951 10. .208 1. , 00 9. .16 C
ATOM 3553 CZ TYR A 462 112. , 731 66 , ,300 8. .962 1. , 00 9. .89 C
ATOM 3554 OH TYR A 462 113 ,.894 65., 795 8..465 1., 00 9..19 O
ATOM 3555 CE2 TYR A 462 111 , .973 67. , 146 8. .181 1. , 00 9. .88 C
ATOM 3556 CD2 TYR A 462 110 , .788 67. , 640 8. .681 1. , 00 9. .71 C
ATOM 3557 C TYR A 462 106. .754 67. .580 11. .438 1. , 00 7. .62 c
ATOM 3558 O TYR A 462 105 , .818 67. , 962 10. .730 1. , 00 7. .10 0
ATOM 3559 N ARG A 463 106 , .797 67. , 767 12. .755 1. , 00 6. .76 N
ATOM 3560 CA ARG A 463 106 , .007 68. , 772 13. .438 1. , 00 6. .89 C
ATOM 3561 CB ARG A 463 105 , .019 68. , 159 14. .422 1. , 00 7. .03 c
ATOM 3562 CG ARG A 463 103 , .964 67. , 236 13. .814 1. , 00 7. .68 C
ATOM 3563 CD ARG A 463 103 , .052 67. , 950 12. .831 1. , 00 7. .20 C
ATOM 3564 NE ARG A 463 102 , .264 69. , 023 13. .464 1. , 00 7. .15 N
ATOM 3565 CZ ARG A 463 101 , .142 68. , 855 14. .151 1. , 00 7. .38 c
ATOM 3566 NH1 ARG A 463 100 , .630 67. , 640 14. .360 1. , 00 8. .84 N
ATOM 3567 NH2 ARG A 463 100 , .535 69. , 908 14. .660 1. , 00 7. .17 N
ATOM 3568 C ARG A 463 107 , .014 69. , 643 14. .172 1. , 00 6. .74 C
ATOM 3569 O ARG A 463 107 , .906 69. , 115 14. .824 1. , 00 7. .05 0
ATOM 3570 N VAL A 464 106 , .906 70. , 958 14. .025 1. , 00 6. .88 N
ATOM 3571 CA VAL A 464 107 , .875 71. , 887 14. .601 1. , 00 8. .03 C
ATOM 3572 CB VAL A 464 108 , .817 72. , 497 13. .536 1. , 00 7. .08 c
ATOM 3573 CGI VAL A 464 109. .893 73. , 329 14. .217 1. , 00 6. .33 c
ATOM 3574 CG2 VAL A 464 109 , .469 71. ,395 12. .710 1. , 00 7. .39 C
ATOM 3575 C VAL A 464 107 , .219 73. , 028 15. .366 1. , 00 8. .00 C
ATOM 3576 O VAL A 464 106 , .409 73. , 742 14. .831 1. , 00 7. .76 0
ATOM 3577 N TYR A 465 107. .584 73. , 165 16. .633 1. , 00 8. .64 N
ATOM 3578 CA TYR A 465 107 , .122 74. , 276 17. .471 1. , 00 9. .47 C
ATOM 3579 CB TYR A 465 106 , .258 73. , 752 18. .600 1. , 00 10. .60 C
ATOM 3580 CG TYR A 465 105. .753 74. .825 19. .541 1. , 00 11. .90 c
ATOM 3581 CD1 TYR A 465 104 , .732 75. , 670 19. .151 1. , 00 13. .84 C
ATOM 3582 CE1 TYR A 465 104 , .254 76. , 663 19. .993 1. , 00 14. .60 C
ATOM 3583 CZ TYR A 465 104 , .779 76. , 792 21. .257 1. , 00 13. .31 C
ATOM 3584 OH TYR A 465 104 , .254 77. , 782 22. .030 1. , 00 16. .79 0
ATOM 3585 CE2 TYR A 465 105 , .806 75. , 977 21. .684 1. , 00 11. .86 C
ATOM 3586 CD2 TYR A 465 106 , .295 74. , 991 20. .825 1. , 00 12. .53 C
ATOM 3587 C TYR A 465 108 , .320 74. , 983 18. .079 1. , 00 9. .77 C
ATOM 3588 O TYR A 465 109 , .147 74. , 354 18. .740 1. , 00 11. .56 0
ATOM 3589 N GLN A 466 108 , .391 76. , 283 17. .871 1. , 00 10. .07 N
ATOM 3590 CA GLN A 466 109 , .439 77. , 128 18. .468 1. , 00 10. .77 C
ATOM 3591 CB GLN A 466 110 , .612 77. ,330 17. .505 1. , 00 10. .07 C
ATOM 3592 CG GLN A 466 111 , .717 78. , 268 18. .002 1. , 00 10. .39 c
ATOM 3593 CD GLN A 466 112 , .976 78. , 111 17. .142 1. , 00 12. .11 C
ATOM 3594 OE1 GLN A 466 113 , .598 77. , 051 17. .154 1. , 00 12. .62 O
ATOM 3595 NE2 GLN A 466 113 , .314 79. , 136 16. .361 1. , 00 10. .16 N
ATOM 3596 C GLN A 466 108 , .820 78. , 484 18. .834 1. , 00 10. .05 c
ATOM 3597 O GLN A 466 108 , .450 79. , 264 17. .963 1. , 00 9. .79 O
ATOM 3598 N HIS A 467 108 , .721 78. , 749 20. .126 1. , 00 11. .10 N
ATOM 3599 CA HIS A 467 108 ,.041 79 ,.942 20..617 1., 00 13..74 C
ATOM 3600 CB HIS A 467 106 , .521 79 , .793 20. .415 1. , 00 14. .41 C
ATOM 3601 CG HIS A 467 105 , .732 81 , .007 20. .819 1. , 00 15. .26 C
ATOM 3602 ND1 HIS A 467 105. .904 82. .213 20. .230 1. .00 15. .80 N
ATOM 3603 CE1 HIS A 467 105 , .081 83 , .117 20. .823 1. , 00 17. .11 C
ATOM 3604 NE2 HIS A 467 104 , .388 82 , .486 21. .798 1. , 00 15. .48 N
ATOM 3605 CD2 HIS A 467 104 , .760 81 , .187 21. .825 1. , 00 16. .55 C
ATOM 3606 C HIS A 467 108 , .363 80 , .097 22. .076 1. , 00 14. .42 c
ATOM 3607 O HIS A 467 108 , .351 79 , .098 22. .827 1. , 00 14. .47 O
ATOM 3608 N ASP A 468 108 , .659 81 , .329 22. .499 1. , 00 17. .70 N
ATOM 3609 CA ASP A 468 108 , .890 81 , .657 23. .942 1. , 00 22. .07 C
ATOM 3610 CB ASP A 468 107 , .587 81 , .581 24. .745 1. , 00 26. .48 c
ATOM 3611 CG ASP A 468 106 , .657 82 , .741 24. .468 1. , 00 42. .65 C
ATOM 3612 OD1 ASP A 468 107 , .077 83 , .711 23. .780 1. , 00 43. .62 O
ATOM 3613 OD2 ASP A 468 105 , .500 82 , .676 24. .953 1. , 00 44. .61 O
ATOM 3614 C ASP A 468 109 , .903 80 , .749 24. .623 1. , 00 20. .64 c
ATOM 3615 O ASP A 468 109 , .624 80 , .195 25. .680 1. , 00 26. .80 O
ATOM 3616 N GLY A 469 111 , .064 80 , .569 24. .007 1. , 00 19. .35 N
ATOM 3617 CA GLY A 469 112 , .078 79 , .708 24. .574 1. , 00 18. .61 C
ATOM 3618 C GLY A 469 111. .811 78. .209 24. .559 1. , 00 17. .77 C
ATOM 3619 O GLY A 469 112 , .649 77 , .459 25. .032 1. , 00 24. .81 O
ATOM 3620 N VAL A 470 110 , .662 77 , .754 24. .047 1. , 00 15. .70 N
ATOM 3621 CA VAL A 470 110 , .409 76 , .323 23. .886 1. , 00 14. .21 C
ATOM 3622 CB VAL A 470 108. .949 75. .970 24. .204 1. , 00 17. .62 C
ATOM 3623 CGI VAL A 470 108 , .680 74 , .490 23. .966 1. , 00 17. .53 C
ATOM 3624 CG2 VAL A 470 108 , .629 76 , .377 25. .641 1. , 00 20. .01 C
ATOM 3625 C VAL A 470 110. .718 75. .980 22. .427 1. .00 13. .68 c
ATOM 3626 O VAL A 470 110 , .289 76 , .704 21. .520 1. , 00 13. .92 O
ATOM 3627 N HIS A 471 111 , .478 74 , .909 22. .201 1. , 00 11. .87 N
ATOM 3628 CA HIS A 471 111 , .801 74 , .482 20. .838 1. , 00 11. .99 C
ATOM 3629 CB HIS A 471 113 , .095 75 , .096 20. .336 1. , 00 12. .32 c
ATOM 3630 CG HIS A 471 114 , .213 75 , .114 21. .342 1. , 00 13. .48 C
ATOM 3631 ND1 HIS A 471 114 , .667 74 , .004 21. .956 1. , 00 13. .35 N
ATOM 3632 CE1 HIS A 471 115 , .671 74 , .348 22. .782 1. , 00 16. .30 C
ATOM 3633 NE2 HIS A 471 115 , .861 75 , .671 22. .685 1. , 00 14. .13 N
ATOM 3634 CD2 HIS A 471 114 , .989 76 , .171 21. .809 1. , 00 15. .06 C
ATOM 3635 C HIS A 471 111 , .835 72 , .995 20. .698 1. , 00 12. .91 C
ATOM 3636 O HIS A 471 112 , .680 72 , .312 21. .313 1. , 00 12. .73 O
ATOM 3637 N GLN A 472 110 , .902 72 , .465 19. .900 1. , 00 9. .92 N
ATOM 3638 CA GLN A 472 110 , .824 71 , .025 19. .710 1. , 00 10. .20 C
ATOM 3639 CB GLN A 472 109 , .733 70 , .401 20. .602 1. , 00 9. .95 C
ATOM 3640 CG GLN A 472 109 , .721 68 , .875 20. .534 1. , 00 10. .88 C
ATOM 3641 CD GLN A 472 108 , .632 68 , .239 21. .380 1. , 00 10. .47 c
ATOM 3642 OE1 GLN A 472 107 , .426 68 , .462 21. .156 1. , 00 8. .61 O
ATOM 3643 NE2 GLN A 472 109 , .046 67, .401 22. .331 1. , 00 7. .85 N
ATOM 3644 C GLN A 472 110 ,.607 70 ,.692 18..240 1., 00 11..45 C
ATOM 3645 O GLN A 472 109 , .686 71 , .230 17. .602 1. , 00 10. .08 O
ATOM 3646 N ILE A 473 111 , .473 69 , .830 17. .701 1. , 00 10. .13 N
ATOM 3647 CA ILE A 473 111. .311 69. .305 16. .352 1. .00 10. .91 C
ATOM 3648 CB ILE A 473 112 , .593 69 , .498 15. .494 1. , 00 12. .38 C
ATOM 3649 CGI ILE A 473 113 , .011 70 , .960 15. .449 1. , 00 12. .96 C
ATOM 3650 CD1 ILE A 473 114 , .480 71 , .192 15. .089 1. , 00 15. .54 C
ATOM 3651 CG2 ILE A 473 112 , .369 68 , .961 14. .083 1. , 00 13. .05 c
ATOM 3652 C ILE A 473 110 , .984 67, .814 16. .477 1. , 00 10. .90 C
ATOM 3653 O ILE A 473 111 , .742 67, .053 17. .106 1. , 00 12. .78 O
ATOM 3654 N ALA A 474 109 , .837 67, .413 15. .931 1. , 00 10. .30 N
ATOM 3655 CA ALA A 474 109 , .392 66 , .012 15. .953 1. , 00 9. .91 c
ATOM 3656 CB ALA A 474 108 , .010 65, .886 16. .576 1. , 00 7. .86 C
ATOM 3657 C ALA A 474 109 , .383 65, .424 14. .551 1. , 00 9. .44 C
ATOM 3658 O ALA A 474 108 , .960 66 , .082 13. .593 1. , 00 10. .72 O
ATOM 3659 N LEU A 475 109 , .879 64 , .205 14. .441 1. , 00 8. .79 N
ATOM 3660 CA LEU A 475 109 , .812 63 , .405 13. .222 1. , 00 7. .97 C
ATOM 3661 CB LEU A 475 111 , .190 62 , .896 12. .834 1. , 00 7. .16 C
ATOM 3662 CG LEU A 475 111 , .248 61 , .869 11. .682 1. , 00 7. .43 c
ATOM 3663 CD1 LEU A 475 110. .660 62 , .450 10. .390 1. , 00 6. .53 c
ATOM 3664 CD2 LEU A 475 112 , .690 61 , .417 11. .452 1. , 00 6. .30 C
ATOM 3665 C LEU A 475 108 , .906 62 , .230 13. .500 1. , 00 8. .86 C
ATOM 3666 O LEU A 475 109 , .220 61 , .394 14. .372 1. , 00 9. .58 O
ATOM 3667 N VAL A 476 107. .794 62 , .150 12. .766 1. , 00 9. .42 N
ATOM 3668 CA VAL A 476 106 , .803 61 , .098 12. .963 1. , 00 7. .93 C
ATOM 3669 CB VAL A 476 105 , .379 61 , .686 12. .999 1. , 00 8. .69 C
ATOM 3670 CGI VAL A 476 104. .310 60 , .581 13. .034 1. .00 6. .98 c
ATOM 3671 CG2 VAL A 476 105 , .220 62 , .653 14. .164 1. , 00 7. .14 C
ATOM 3672 C VAL A 476 106 , .921 60 , .140 11. .795 1. , 00 9. .09 C
ATOM 3673 O VAL A 476 106 , .947 60 , .580 10. .632 1. , 00 9. .05 O
ATOM 3674 N LEU A 477 107 , .013 58 , .842 12. .103 1. , 00 8. .19 N
ATOM 3675 CA LEU A 477 107 , .026 57 , .787 11. .108 1. , 00 7. .85 C
ATOM 3676 CB LEU A 477 108 , .360 57 , .035 11. .113 1. , 00 7. .10 C
ATOM 3677 CG LEU A 477 109 , .662 57 , .837 11. .136 1. , 00 7. .47 C
ATOM 3678 CD1 LEU A 477 110 , .856 56 , .885 11. .282 1. , 00 7. .28 c
ATOM 3679 CD2 LEU A 477 109 , .829 58 , .741 9. .897 1. , 00 6. .57 C
ATOM 3680 C LEU A 477 105 , .882 56 , .807 11. .375 1. , 00 9. .01 C
ATOM 3681 O LEU A 477 105 , .751 56 , .303 12. .495 1. , 00 9. .23 O
ATOM 3682 N LEU A 478 105 , .074 56 , .531 10. .339 1. , 00 8. .76 N
ATOM 3683 CA LEU A 478 103 , .941 55 , .620 10. .444 1. , 00 8. .70 C
ATOM 3684 CB LEU A 478 102 , .614 56 , .382 10. .265 1. , 00 8. .48 C
ATOM 3685 CG LEU A 478 102 , .349 57 , .590 11. .188 1. , 00 9. .45 c
ATOM 3686 CD1 LEU A 478 101 , .102 58 , .392 10. .793 1. , 00 8. .74 c
ATOM 3687 CD2 LEU A 478 102 , .188 57 , .125 12. .614 1. , 00 9. .75 C
ATOM 3688 C LEU A 478 104 , .055 54 , .495 9. .410 1. , 00 8. .94 C
ATOM 3689 O LEU A 478 104 , , 468 54 , , 719 8..255 1. , 00 8..93 O
ATOM 3690 N ASN A 479 103 , , 660 53 , , 296 9. .826 1. , 00 8. .49 N
ATOM 3691 CA ASN A 479 103 , , 580 52 , , 136 8. .974 1. , 00 8. .70 C
ATOM 3692 CB ASN A 479 104. .713 51 , .185 9. .333 1. .00 10. .04 C
ATOM 3693 CG ASN A 479 104 , , 643 49 , , 865 8. .581 1. , 00 10. .21 C
ATOM 3694 OD1 ASN A 479 103 , , 847 49 , , 701 7. .652 1. , 00 8. .10 O
ATOM 3695 ND2 ASN A 479 105 , , 494 48 , , 916 8. .980 1. , 00 9. .01 N
ATOM 3696 C ASN A 479 102 , .208 51 , .476 9. .205 1. , 00 10. .92 c
ATOM 3697 O ASN A 479 101 , , 954 50 , , 865 10. .278 1. , 00 10. .14 0
ATOM 3698 N LYS A 48 iO 101 , , 313 51 , , 632 8. .235 1. , 00 9. .12 N
ATOM 3699 CA LYS A 48 10 99 , , 965 51 , , 066 8. .337 1. , 00 9. .96 C
ATOM 3700 CB LYS A 48 10 98 , .905 52 , .061 7. .801 1. , 00 10. .03 c
ATOM 3701 CG LYS A 48 !O 98 , , 904 52 , , 249 6. .286 1. , 00 10. .64 C
ATOM 3702 CD LYS A 480 97 , , 837 53 , , 234 5. .751 1. , 00 10. .86 C
ATOM 3703 CE LYS A 48 10 97 , , 583 53 , , 015 4. .250 1. , 00 11. .36 C
ATOM 3704 NZ LYS A 48 10 96 , .412 52 , .109 3. .973 1. , 00 13. .64 N
ATOM 3705 C LYS A 48 !O 99 , , 852 49 , , 725 7. .601 1. , 00 10. .86 C
ATOM 3706 O LYS A 480 98 , , 736 49 , , 272 7. .333 1. , 00 10. .55 O
ATOM 3707 N GLY A 48 il 100 , , 987 49 , , 111 7. .254 1. , 00 9. .20 N
ATOM 3708 CA GLY A 48 !l 100 , .996 47 , .815 6. .584 1. , 00 10. .33 c
ATOM 3709 C GLY A 48 !l 101 , , 345 46 , , 666 7. .520 1. , 00 11. .31 C
ATOM 3710 O GLY A 48 il 101 , , 498 46 , , 848 8. .747 1. , 00 12. .10 O
ATOM 3711 N ASP A 48 i2 101 , , 483 45 , , 489 6. .927 1. , 00 12. .13 N
ATOM 3712 CA ASP A 48 12 101 , .528 44 , .225 7. .659 1. , 00 14. .92 c
ATOM 3713 CB ASP A 48 12 100 , , 811 43 , , 119 6. .868 1. , 00 17. .05 C
ATOM 3714 CG ASP A 48 i2 99 , , 280 43 , , 268 6. .895 1. , 00 18. .25 C
ATOM 3715 OD1 ASP A 48 i2 98 , .728 43 , .944 7. .780 1. .00 17. .35 0
ATOM 3716 OD2 ASP A 48 12 98 , , 627 42 , , 699 6. .016 1. , 00 21. .98 0
ATOM 3717 C ASP A 48 12 102 , , 953 43 , , 774 7. .941 1. , 00 15. .43 C
ATOM 3718 O ASP A 48 \2 103 , , 142 42 , , 827 8. .689 1. , 00 13. .49 O
ATOM 3719 N ALA A 48 i3 103 , .937 44 , .424 7. .326 1. , 00 15. .02 N
ATOM 3720 CA ALA A 48 13 105 , , 342 44 , , 073 7. .545 1. , 00 17. .79 C
ATOM 3721 CB ALA A 48 13 105 , , 964 43 , , 459 6. .279 1. , 00 14. .25 C
ATOM 3722 C ALA A 48 13 106 , , 091 45 , , 329 7. .971 1. , 00 15. .42 C
ATOM 3723 O ALA A 48 13 105 , .643 46 , .442 7. .687 1. , 00 14. .23 0
ATOM 3724 N PRO A 48 14 107 , , 237 45 , , 159 8. .638 1. , 00 16. .34 N
ATOM 3725 CA PRO A 484 108 , , 015 46 , , 319 9. .059 1. , 00 14. .26 C
ATOM 3726 CB PRO A 48 i4 109 , , 237 45 , , 692 9. .768 1. , 00 15. .32 C
ATOM 3727 CG PRO A 48 14 108 , .774 44 , .314 10. .192 1. , 00 18. .54 c
ATOM 3728 CD PRO A 48 14 107 , , 864 43 , , 883 9. .068 1. , 00 17. .52 C
ATOM 3729 C PRO A 484 108 , , 493 47 , , 175 7. .875 1. , 00 14. .36 C
ATOM 3730 O PRO A 48 i4 108 , , 743 46 , , 645 6. .808 1. , 00 12. .69 O
ATOM 3731 N GLU A 48 15 108 , .632 48 , .480 8. .082 1. , 00 12. .15 N
ATOM 3732 CA GLU A 48 15 109 , , 123 49 , , 399 7. .059 1. , 00 13. .43 C
ATOM 3733 CB GLU A 48 15 108 , , 047 50 , , 429 6. .747 1. , 00 13. .80 C
ATOM 3734 CG GLU A 4i 5 106. , 862 49 , , 835 6. , 021 1 , .00 14..56 C
ATOM 3735 CD GLU A 4i 35 107. , 147 49 , , 536 4. , 564 1 , .00 16. .23 C
ATOM 3736 OE1 GLU A 4E 35 108. , 189 49 , , 985 4. , 037 1 , .00 13. .54 O
ATOM 3737 OE2 GLU A 4i 35 106. .298 48 , .863 3. .949 1 , .00 20. .24 0
ATOM 3738 C GLU A 4i 35 110. , 365 50 , , 132 7. , 542 1 , .00 12. .82 C
ATOM 3739 O GLU A 4i 35 110. , 428 50 , , 513 8. .711 1 , .00 13. .04 O
ATOM 3740 N HIS A 4i 36 Ill . ,336 50 , ,300 6. .645 1 , .00 12. .11 N
ATOM 3741 CA AHIS A 4i 36 112. , 585 50 , .984 6. .945 0. .50 12. .85 c
ATOM 3742 CA BHIS A 4? 36 112. , 593 51 , , 005 6. .966 0 , .50 12. .63 C
ATOM 3743 CB AHIS A 4i 36 113. , 742 50 , , 184 6. .352 0 , .50 14. .74 C
ATOM 3744 CB BHIS A 4? 36 113. , 837 50 , , 256 6. .460 0 , .50 14. .15 C
ATOM 3745 CG AHIS A 4? 36 113. , 795 48 , .744 6. .826 0 , .50 16. .29 c
ATOM 3746 CG BHIS A 4? 36 115. , 164 50 , , 829 6. .980 0 , .50 15. .34 C
ATOM 3747 NDIAHIS A 486 113. , 154 47 , , 740 6. .184 0 , .50 17. .98 N
ATOM 3748 ND1BHIS A 4? 36 115. , 935 51 , , 678 6. .246 0 , .50 15. .67 N
ATOM 3749 CE1AHIS A 4? 36 113. ,374 46 , .583 6. .829 0 , .50 17. .75 c
ATOM 3750 CEIBHIS A 4? 36 117. , 025 52 , , 030 6. .964 0 , .50 14. .82 C
ATOM 3751 NE2AHIS A 486 114. , 158 46 , , 841 7. .885 0 , .50 19. .45 N
ATOM 3752 NE2BHIS A 4? 36 116. , 963 51 , ,392 8. .162 0 , .50 16. .80 N
ATOM 3753 CD2AHIS A 4i 36 114. , 431 48 , .164 7. .919 0 , .50 17. .74 C
ATOM 3754 CD2BHIS A 4i 36 115. , 828 50 , , 657 8. .211 0 , .50 14. .06 C
ATOM 3755 C HIS A i 36 112. , 536 52 , ,389 6. .388 1 , .00 12. .98 C
ATOM 3756 O HIS A 1i 36 112. , 129 52 , , 586 5. .236 1 , .00 12. .49 O
ATOM 3757 N PHE A 4i 37 112. ,911 53 , .376 7. .200 1 , .00 11. .23 N
ATOM 3758 CA PHE A 4i 37 112. , 912 54 , , 779 6. .793 1 , .00 10. .68 C
ATOM 3759 CB PHE A 4E 37 112. , 022 55 , , 643 7. .725 1 , .00 9. .60 C
ATOM 3760 CG PHE A 4i 37 110. , 548 55 , .402 7. .559 1 , .00 9. .63 c
ATOM 3761 CD1 PHE A 4i 37 109. , 929 54 , ,307 8. .168 1 , .00 10. .45 C
ATOM 3762 CE1 PHE A 4i 37 108. , 568 54 , , 073 7. .978 1 , .00 10. .69 C
ATOM 3763 CZ PHE A 4i 37 107. , 812 54 , , 946 7. .200 1 , .00 10. .79 C
ATOM 3764 CE2 PHE A 4i 37 108. , 415 56 , .036 6. .611 1. .00 9. .55 c
ATOM 3765 CD2 PHE A 4? 37 109. , 773 56 , , 245 6. .783 1 , .00 9. .44 C
ATOM 3766 C PHE A 4i 37 114. ,371 55 , , 281 6. .848 1 , .00 10. .89 C
ATOM 3767 O PHE A 4i 37 115. , 100 54 , , 924 7. .753 1 , .00 10. .54 O
ATOM 3768 N ALA A 4? 38 114. , 766 56 , .084 5. .864 1 , .00 10. .91 N
ATOM 3769 CA ALA A 4? 38 116. , 055 56 , , 780 5. .855 1 , .00 12. .54 C
ATOM 3770 CB ALA A 4i 38 116. , 984 56 , , 210 4. .789 1 , .00 11. .00 C
ATOM 3771 C ALA A 4? 38 115. , 721 58 , , 229 5. .569 1 , .00 12. .80 C
ATOM 3772 O ALA A 4? 38 115. , 445 58 , .587 4. .428 1 , .00 14. .24 0
ATOM 3773 N VAL A 4? 39 115. , 681 59 , , 057 6. .617 1 , .00 11. .73 N
ATOM 3774 CA VAL A 489 115. , 214 60 , , 438 6. .488 1 , .00 11. .05 C
ATOM 3775 CB VAL A 4? 39 114. , 159 60 , , 787 7. .578 1 , .00 11. .13 C
ATOM 3776 CGI VAL A 4? 39 113. , 767 62 , .266 7. .524 1 , .00 9. .30 c
ATOM 3777 CG2 VAL A 4i 39 112. , 935 59 , , 885 7. .423 1 , .00 11. .83 C
ATOM 3778 C VAL A 4£ 39 116. , 409 61 , ,366 6. .587 1 , .00 10. .27 C
ATOM 3779 O VAL A 489 117., 234 61 ,, 237 7., 474 1., 00 11.,04 O
ATOM 37 80 N GLN A 490 116. , 474 62 , ,317 5. ,683 1. , 00 10. ,58 N
ATOM 37 81 CA GLN A 490 117. , 583 63 , , 256 5. ,612 1. , 00 10. , 03 C
ATOM 37 82 CB GLN A 490 118. .698 62. , 663 4. .740 1. .00 12. .01 C
ATOM 37 83 CG GLN A 490 120. , 005 63 , , 467 4. , 833 1. , 00 11. ,32 C
ATOM 37 84 CD GLN A 490 121. , 224 62 , , 666 4. , 434 1. , 00 10. ,88 C
ATOM 37 85 OE1 GLN A 490 121. , 163 61 , , 443 4. , 255 1. , 00 11. ,57 O
ATOM 37 86 NE2 GLN A 490 122. ,341 63. ,348 4. .285 1. , 00 11. .90 N
ATOM 37 87 C GLN A 490 117. , 155 64 , , 627 5. , 066 1. , 00 10. ,17 C
ATOM 37 88 O GLN A 490 117. , 786 65, , 631 5. ,362 1. , 00 8. , 72 O
ATOM 37 89 N THR A 491 116. , 085 64 , , 699 4. ,279 1. , 00 9. ,58 N
ATOM 3790 CA THR A 491 115. , 626 66. , 000 3. .792 1. , 00 11. .73 c
ATOM 3791 CB THR A 491 114. , 481 65, , 858 2. , 753 1. , 00 13. ,80 C
ATOM 3792 OG1 THR A 491 115. , 010 65, , 226 1. , 570 1. , 00 18. , 48 O
ATOM 3793 CG2 THR A 491 113. , 911 67, , 234 2. , 351 1. , 00 12. ,80 C
ATOM 3794 C THR A 491 115. , 198 66. , 859 4. .981 1. , 00 10. .05 c
ATOM 3795 O THR A 491 114. , 442 66 , , 420 5. , 827 1. , 00 10. , 42 O
ATOM 3796 N MET A 492 115. , 729 68 , , 071 5. , 031 1. , 00 9. , 45 N
ATOM 3797 CA MET A 492 115. , 485 69 , , 015 6. , 112 1. , 00 11. , 41 C
ATOM 3798 CB MET A 492 114. , 053 69. , 579 6. .012 1. , 00 10. .69 c
ATOM 3799 CG MET A 492 113. , 800 70 , ,374 4. , 730 1. , 00 10. ,20 C
ATOM 38 00 SD MET A 492 112. , 164 71 , , 124 4. ,680 1. , 00 12. ,67 s
ATOM 38 01 CE MET A 492 Ill . , 091 69 , , 704 4. ,563 1. , 00 11. ,26 c
ATOM 38 02 C MET A 492 115. , 748 68. , 465 7. .523 1. , 00 10. .13 c
ATOM 38 03 O MET A 492 115. , 215 68 , , 985 8. , 491 1. , 00 10. , 70 O
ATOM 38 04 N LEU A 493 116. , 595 67, , 453 7. ,640 1. , 00 9. ,99 N
ATOM 38 05 CA LEU A 493 116. .859 66. , 830 8. .927 1. .00 10. .70 c
ATOM 38 06 CB LEU A 493 117. ,301 65 , ,371 8. , 749 1. , 00 11. , 02 C
ATOM 38 07 CG LEU A 493 117. , 551 64 , , 643 10. , 085 1. , 00 11. ,96 C
ATOM 38 08 CD1 LEU A 493 116. , 226 64 , , 224 10. , 715 1. , 00 10. , 78 C
ATOM 38 09 CD2 LEU A 493 118. , 482 63. , 443 9. .853 1. , 00 13. .60 c
ATOM 38 10 C LEU A 493 117. , 966 67, , 606 9. ,642 1. , 00 11. , 06 C
ATOM 38 11 O LEU A 493 119. , 067 67, , 744 9. , 122 1. , 00 10. , 71 O
ATOM 38 12 N GLN A 494 117. , 685 68 , , 095 10. , 839 1. , 00 9. , 75 N
ATOM 38 13 CA GLN A 494 118. , 670 68. , 858 11. .592 1. , 00 9. .84 c
ATOM 38 14 CB GLN A 494 117. , 982 69 , , 838 12. ,523 1. , 00 10. ,66 C
ATOM 3815 CG GLN A 494 118. , 942 70 , , 688 13. , 357 1. , 00 11. ,22 C
ATOM 38 16 CD GLN A 494 118. , 237 71 , , 903 13. ,930 1. , 00 11. , 78 C
ATOM 38 17 OE1 GLN A 494 117. , 596 72. , 661 13. .193 1. , 00 9. .17 0
ATOM 38 18 NE2 GLN A 494 118. ,373 72 , , 115 15. ,241 1. , 00 10. ,94 N
ATOM 3819 C GLN A 494 119. , 533 67, , 879 12. , 405 1. , 00 9. ,56 C
ATOM 38 20 O GLN A 494 119. , 001 67, , 073 13. , 177 1. , 00 8. ,81 0
ATOM 38 21 N PRO A 495 120. , 856 67. , 935 12. .223 1. , 00 8. .63 N
ATOM 38 22 CA PRO A 495 121. , 741 67, , 115 13. , 035 1. , 00 9. ,61 C
ATOM 38 23 CB PRO A 495 123. , 150 67, , 572 12. ,592 1. , 00 9. , 42 C
ATOM 3 i24 CG PRO A 495 122., 956 68 ,, 053 11., 194 1., 00 9., 16 C
ATOM 3 i 325 CD PRO A 495 121. , 616 68 , , 731 11. ,230 1. , 00 9. , 72 C
ATOM 3 i 326 C PRO A 495 121. , 548 67, ,360 14. , 510 1. , 00 10. ,17 C
ATOM 3 i 12 1 O PRO A 495 121. .250 68. , 481 14. .914 1. , 00 11. .25 0
ATOM 3 i 328 N GLY A 496 121. , 702 66 , ,312 15. ,308 1. , 00 10. , 49 N
ATOM 3 i 329 CA GLY A 496 121. , 533 66 , , 412 16. , 733 1. , 00 11. , 03 C
ATOM 3 i 330 C GLY A 496 121. , 206 65, , 089 17. ,392 1. , 00 12. , 19 C
ATOM 3 i 331 O GLY A 496 121. , 296 64. , 004 16. .774 1. , 00 11. .67 0
ATOM 3 ? 332 N ARG A 497 120. , 821 65, , 183 18. , 659 1. , 00 12. , 78 N
ATOM 3 i 333 CA ARG A 497 120. , 500 64, , 001 19. , 430 1. , 00 14. ,38 C
ATOM 3 ? 334 CB ARG A 497 121. , 150 64 , , 096 20. , 820 1. , 00 17. , 05 C
ATOM 3 ? 335 CG ARG A 497 122. , 660 64. , 292 20. .741 1. , 00 17. .85 c
ATOM 3 ? 336 CD ARG A 497 123. ,314 64 , , 179 22. , 110 1. , 00 20. ,04 C
ATOM 3837 NE ARG A 497 124. , 776 64, , 253 22. , 074 1. , 00 18. ,23 N
ATOM 3 ? 338 CZ ARG A 497 125. , 496 65, ,361 22. ,285 1. , 00 19. ,00 C
ATOM 3 ? 339 NH1 ARG A 497 124. , 891 66. , 515 22. .517 1. , 00 21. .56 N
ATOM 3 ? 340 NH2 ARG A 497 126. , 838 65, ,329 22. ,273 1. , 00 15. , 13 N
ATOM 3841 C ARG A 497 119. , 005 63 , , 833 19. , 496 1. , 00 13. ,31 C
ATOM 3 ? 342 O ARG A 497 118. ,332 64 , , 522 20. ,247 1. , 00 17. ,01 0
ATOM 3 i 343 N TRP A 498 118. , 493 62. , 910 18. .690 1. , 00 12. .85 N
ATOM 3 i 344 CA TRP A 498 117. , 061 62 , , 667 18. , 564 1. , 00 13. , 09 C
ATOM 3 i 345 CB TRP A 498 116. , 732 62 , ,305 17. , 139 1. , 00 10. ,29 C
ATOM 3 i 346 CG TRP A 498 116. , 891 63 , , 411 16. , 133 1. , 00 8. ,66 c
ATOM 3 i 347 CD1 TRP A 498 118. , 063 63. , 884 15. .559 1. , 00 8. .76 c
ATOM 3 i 348 NE1 TRP A 498 117. , 802 64 , , 876 14. , 656 1. , 00 8. , 86 N
ATOM 3 i 349 CE2 TRP A 498 116. , 460 65, , 087 14. , 578 1. , 00 8. ,51 C
ATOM 3 i 350 CD2 TRP A 498 115. , 824 64. , 159 15. .502 1. , 00 8. .53 c
ATOM 3 i 351 CE3 TRP A 498 114. , 443 64 , , 163 15. ,619 1. , 00 9. , 16 C
ATOM 3 i 352 CZ3 TRP A 498 113. , 720 65, , 054 14. , 845 1. , 00 9. ,25 C
ATOM 3 i 353 CH2 TRP A 498 114. , 357 65, , 938 13. , 955 1. , 00 8. ,63 C
ATOM 3 i 354 CZ2 TRP A 498 115. , 732 65. , 973 13. .819 1. , 00 8. .09 c
ATOM 3 ? 355 C TRP A 498 116. , 627 61 , , 527 19. , 454 1. , 00 16. , 40 C
ATOM 3 i 356 O TRP A 498 117. , 265 60 , , 478 19. , 445 1. , 00 24. ,22 O
ATOM 3 i 357 N HIS A 499 115. , 546 61 , , 707 20. ,207 1. , 00 15. ,34 N
ATOM 3 ? 358 CA HIS A 499 115. , 081 60. , 697 21. .196 1. , 00 17. .77 c
ATOM 3 ? 359 CB HIS A 499 114. , 972 61 , ,300 22. ,618 1. , 00 21. , 46 C
ATOM 3860 CG HIS A 499 116. , 269 61 , , 968 23. , 121 1. , 00 36. , 41 C
ATOM 3 ? 361 ND1 HIS A 499 117. , 129 61 , ,359 23. ,981 1. , 00 40. ,93 N
ATOM 3 ? 362 CE1 HIS A 499 118. , 172 62. , 188 24. .239 1. , 00 39. .20 c
ATOM 3 ? 363 NE2 HIS A 499 117. , 983 63 , ,330 23. , 545 1. , 00 36. ,63 N
ATOM 3864 CD2 HIS A 499 116. , 823 63 , , 236 22. , 852 1. , 00 34. ,57 C
ATOM 3 ? 365 C HIS A 499 113. , 751 60 , , 145 20. , 771 1. , 00 15. ,91 c
ATOM 3 ? 366 O HIS A 499 112. , 801 60. , 901 20. .495 1. , 00 13. .95 0
ATOM 3 i 367 N ASP A 500 113. , 658 58 , , 826 20. ,693 1. , 00 17. , 76 N
ATOM 3 i 368 CA ASP A 500 112. , 422 58 , , 140 20. ,353 1. , 00 21. ,93 C
ATOM 3i69 CB ASP A 500 112 ,.692 56., 641 20..136 1., 00 30..24 C
ATOM 3i 370 CG ASP A 500 111 , .423 55. , 828 19. .818 1. , 00 33. .18 C
ATOM 3iill OD1 ASP A 500 110 , .286 56. ,318 19. .959 1. , 00 43. .09 O
ATOM 3i 372 OD2 ASP A 500 111. .582 54. .675 19. .391 1. , 00 42. .81 O
ATOM 3i 373 C ASP A 500 Ill , .492 58. ,312 21. .539 1. , 00 20. .91 C
ATOM 3i 374 O ASP A 500 111 , .796 57. , 809 22. .591 1. , 00 22. .20 O
ATOM 3i 375 N ALA A 501 110 , .374 59. , 008 21. .363 1. , 00 14. .99 N
ATOM 3i 376 CA ALA A 501 109 , .471 59. ,304 22. .477 1. , 00 17. .10 c
ATOM 3 ? 377 CB ALA A 501 108 , .385 60. , 274 22. .036 1. , 00 14. .69 C
ATOM 3 i 378 C ALA A 501 108 , .830 58. , 076 23. .139 1. , 00 22. .71 C
ATOM 3 ? 379 O ALA A 501 108 , .475 58. , 154 24. .303 1. , 00 35. .66 O
ATOM 3 ? 380 N I LE A 502 108 , .632 56. , 983 22. .410 1. , 00 31. .51 N
ATOM 3 ? 381 CA ILE A 502 107 , .991 55. , 756 22. .969 1. , 00 43. .49 C
ATOM 3882 CB ILE A 502 106 , .961 55. , 114 22. .023 1. , 00 43. .64 C
ATOM 3 ? 383 CGI ILE A 502 105 , .735 56. , 028 21. .853 1. , 00 44. .63 C
ATOM 3 ? 384 CD1 ILE A 502 105 , .095 55. ,971 20. .474 1. , 00 35. .62 c
ATOM 3 ? 385 CG2 ILE A 502 106 , .552 53. , 733 22. .560 1. , 00 42. .82 C
ATOM 3886 C ILE A 502 109 , .023 54. , 694 23. .328 1. , 00 46. .88 C
ATOM 3 ? 387 O ILE A 502 109 , .152 54. , 354 24. .509 1. , 00 51. .64 O
ATOM 3i 388 N GLY A 503 109. .755 54. ,174 22. .330 1. , 00 49. .47 N
ATOM 3i 389 CA GLY A 503 Ill , .057 53. , 524 22. .605 1. , 00 42. .47 C
ATOM 3i 390 C GLY A 503 111 , .819 54. , 511 23. .491 1. , 00 41. .75 C
ATOM 3i 391 O GLY A 503 111 , .240 55. , 476 23. .989 1. , 00 52. .49 O
ATOM 3i 392 N GLY A 504 113. .095 54. , 297 23. .733 1. , 00 33. .97 N
ATOM 3i 393 CA GLY A 504 113 , .853 55. , 252 24. .542 1. , 00 36. .81 C
ATOM 3i 394 C GLY A 504 115 , .188 55. , 666 23. .959 1. , 00 35. .32 C
ATOM 3i 395 O GLY A 504 115. .831 56. , 598 24. .454 1. , 00 44. .34 O
ATOM 3i 396 N GLU A 505 115 , .603 54. , 992 22. .898 1. , 00 43. .81 N
ATOM 3i 397 CA GLU A 505 116 , .913 55. , 246 22. .295 1. , 00 48. .17 C
ATOM 3i 398 CB GLU A 505 117 , .241 54. , 250 21. .160 1. , 00 54. .81 C
ATOM 3i 399 CG GLU A 505 116 , .119 53. ,974 20. .174 1. , 00 56. .61 c
ATOM 3900 CD GLU A 505 115 , .051 53. , 045 20. .748 1. , 00 79. .51 C
ATOM 3901 OE1 GLU A 505 115 , .304 51. , 822 20. .841 1. , 00 93. .44 O
ATOM 3902 OE2 GLU A 505 113 , .963 53. , 544 21. .129 1. , 00 59. .14 O
ATOM 3903 C GLU A 505 117 , .085 56. , 684 21. .798 1. , 00 42. .11 c
ATOM 3904 O GLU A 505 116 , .139 57. , 351 21. .354 1. , 00 39. .97 O
ATOM 3905 N THR A 506 118 , .317 57. , 137 21. .939 1. , 00 36. .27 N
ATOM 3906 CA THR A 506 118 , .814 58. ,333 21. .326 1. , 00 32. .81 C
ATOM 3907 CB THR A 506 119 , .746 59. , 040 22. .299 1. , 00 35. .17 c
ATOM 3908 OG1 THR A 506 118 , .999 59. ,308 23. .486 1. , 00 47. .17 O
ATOM 3909 CG2 THR A 506 120 , .294 60. ,360 21. .719 1. , 00 34. .36 C
ATOM 3910 C THR A 506 119 , .550 57. , 949 20. .052 1. , 00 25. .53 c
ATOM 3911 O THR A 506 120 , .446 57. , 110 20. .060 1. , 00 23. .32 0
ATOM 3912 N LEU A 507 119 , .133 58. , 529 18. .944 1. , 00 20. .16 N
ATOM 3913 CA LEU A 507 119 , .884 58. , 446 17. .709 1. , 00 17. .44 C
ATOM 3914 CB LEU A 507 118., 953 58 ,, 184 16..525 1., 00 18..20 C
ATOM 3915 CG LEU A 507 118. , 249 56 , , 821 16. .361 1. , 00 18. .08 C
ATOM 3916 CD1 LEU A 507 117. , 469 56 , , 808 15. .050 1. , 00 14. .73 C
ATOM 3917 CD2 LEU A 507 119. .205 55. .620 16. .388 1. , 00 21. .65 c
ATOM 3918 C LEU A 507 120. , 613 59 , , 788 17. .561 1. , 00 16. .84 C
ATOM 3919 O LEU A 507 119. , 981 60 , , 845 17. .537 1. , 00 15. .02 O
ATOM 3920 N THR A 508 121. , 941 59 , , 738 17. .498 1. , 00 15. .75 N
ATOM 3921 CA THR A 508 122. , 741 60 , .915 17. .258 1. , 00 14. .58 c
ATOM 3922 CB THR A 508 124. , 084 60 , , 861 17. .997 1. , 00 14. .89 C
ATOM 3923 OG1 THR A 508 123. , 816 60 , , 897 19. .393 1. , 00 13. .50 O
ATOM 3924 CG2 THR A 508 124. , 981 62 , , 070 17. .606 1. , 00 12. .67 C
ATOM 3925 C THR A 508 122. , 991 61 , .000 15. .776 1. , 00 14. .73 c
ATOM 3926 O THR A 508 123. , 641 60 , , 127 15. .202 1. , 00 13. .99 O
ATOM 3927 N ILE A 509 122. , 481 62 , , 055 15. .155 1. , 00 13. .45 N
ATOM 3928 CA ILE A 509 122. , 656 62 , , 225 13. .732 1. , 00 13. .06 C
ATOM 3929 CB ILE A 509 121. , 335 62 , .623 13. .045 1. , 00 13. .81 c
ATOM 3930 CGI ILE A 509 120. , 222 61 , , 628 13. .419 1. , 00 13. .63 C
ATOM 3931 CD1 ILE A 509 120. , 483 60 , , 175 13. .027 1. , 00 12. .00 C
ATOM 3932 CG2 ILE A 509 121. , 561 62 , , 728 11. .532 1. , 00 14. .78 c
ATOM 3933 C ILE A 509 123. , 699 63. .303 13. .482 1. , 00 11. .43 c
ATOM 3934 O ILE A 509 123. , 563 64 , , 418 13. .971 1. , 00 11. .35 O
ATOM 3935 N GLN A 510 124. , 739 62 , , 930 12. .744 1. , 00 10. .14 N
ATOM 3936 CA GLN A 510 125. , 805 63 , , 825 12. .332 1. , 00 10. .29 c
ATOM 3937 CB GLN A 510 127. , 129 63. .049 12. .223 1. , 00 11. .16 c
ATOM 3938 CG GLN A 510 127. , 611 62 , , 468 13. .546 1. , 00 11. .25 C
ATOM 3939 CD GLN A 510 129. , 004 61 , , 863 13. .472 1. , 00 12. .20 C
ATOM 3940 OE1 GLN A 510 129. .725 62. .031 12. .503 1. , 00 12. .77 O
ATOM 3941 NE2 GLN A 510 129. , 393 61 , , 184 14. .530 1. , 00 11. .54 N
ATOM 3942 C GLN A 510 125. , 487 64 , , 436 10. .981 1. , 00 9. .89 C
ATOM 3943 O GLN A 510 124. , 807 63 , , 805 10. .149 1. , 00 10. .54 O
ATOM 3944 N ALA A 511 126. , 006 65 , .641 10. .752 1. , 00 9. .74 N
ATOM 3945 CA ALA A 511 125. , 795 66 , .352 9. .485 1. , 00 11. .00 C
ATOM 3946 CB ALA A 511 126. , 543 67, .704 9. .468 1. , 00 9. .61 C
ATOM 3947 C ALA A 511 126. , 209 65, .478 8. .294 1. , 00 10. .27 C
ATOM 3948 O ALA A 511 127. , 285 64 , .861 8. .297 1. , 00 9. .35 0
ATOM 3949 N GLY A 512 125. ,359 65, .439 7. .274 1. , 00 9. .80 N
ATOM 3950 CA GLY A 512 125. , 614 64, .654 6. .091 1. , 00 9. .49 C
ATOM 3951 C GLY A 512 125. , 026 63 , .260 6. .153 1. , 00 11. .41 C
ATOM 3952 O GLY A 512 125. , 016 62 , .552 5. .135 1. , 00 14. .08 0
ATOM 3953 N GLU A 513 124. , 490 62 , .864 7. .311 1. , 00 12. .38 N
ATOM 3954 CA GLU A 513 123. , 914 61 , .529 7. .484 1. , 00 14. .25 C
ATOM 3955 CB GLU A 513 124. , 493 60 , .822 8. .721 1. , 00 18. .94 c
ATOM 3956 CG GLU A 513 126. , 007 60 , .915 8. .911 1. , 00 26. .20 c
ATOM 3957 CD GLU A 513 126. , 795 60 , .076 7. .930 1. , 00 37. .25 C
ATOM 3958 OE1 GLU A 513 126. , 248 59 , .070 7. .430 1. , 00 40. .40 O
ATOM 3959 OE2 GLU A 513 127 ,.975 60., 418 7..671 1., 00 45..26 O
ATOM 3960 C GLU A 513 122 , .385 61. , 579 7. .642 1. , 00 12. .91 C
ATOM 3961 O GLU A 513 121 , .811 62. , 605 7. .946 1. , 00 13. .81 O
ATOM 3962 N ALA A 514 121. .769 60. .415 7. .504 1. , 00 11. .39 N
ATOM 3963 CA ALA A 514 120 , .333 60. , 222 7. .574 1. , 00 11. .17 C
ATOM 3964 CB ALA A 514 119 , .896 59. ,309 6. .433 1. , 00 10. .91 C
ATOM 3965 C ALA A 514 119 , .979 59. , 583 8. .907 1. , 00 10. .60 C
ATOM 3966 O ALA A 514 120 , .807 58. ,914 9. .524 1. , 00 9. .46 0
ATOM 3967 N LEU A 515 118 , .738 59. , 792 9. .329 1. , 00 10. .57 N
ATOM 3968 CA LEU A 515 118 , .141 59. , 089 10. .460 1. , 00 10. .43 C
ATOM 3969 CB LEU A 515 117 , .087 59. , 965 11. .154 1. , 00 10. .29 C
ATOM 3970 CG LEU A 515 116 , .529 59. , 410 12. .461 1. , 00 11. .73 c
ATOM 3971 CD1 LEU A 515 116 , .065 60. , 553 13. .357 1. , 00 11. .08 C
ATOM 3972 CD2 LEU A 515 115 , .395 58. ,389 12. .259 1. , 00 11. .49 C
ATOM 3973 C LEU A 515 117 , .498 57. , 831 9. .909 1. , 00 10. .90 C
ATOM 3974 O LEU A 515 116 , .594 57. , 915 9. .073 1. , 00 10. .60 0
ATOM 3975 N HIS A 516 117 , .970 56. , 678 10. .370 1. , 00 10. .55 N
ATOM 3976 CA HIS A 516 117 , .477 55. ,372 9. .940 1. , 00 12. .62 C
ATOM 3977 CB HIS A 516 118 , .649 54. ,398 9. .694 1. , 00 13. .48 c
ATOM 3978 CG HIS A 516 119. .533 54. , 803 8. .567 1. , 00 17. .52 c
ATOM 3979 ND1 HIS A 516 119 , .294 54. , 427 7. .296 1. , 00 20. .78 N
ATOM 3980 CE1 HIS A 516 120 , .228 54. , 955 6. .496 1. , 00 19. .23 C
ATOM 3981 NE2 HIS A 516 121 , .070 55. , 667 7. .251 1. , 00 18. .35 N
ATOM 3982 CD2 HIS A 516 120. .660 55. , 605 8. .535 1. , 00 18. .79 C
ATOM 3983 C HIS A 516 116 , .669 54. , 795 11. .054 1. , 00 14. .02 C
ATOM 3984 O HIS A 516 117 , .119 54. , 815 12. .203 1. , 00 13. .21 O
ATOM 3985 N ALA A 517 115. .510 54. , 241 10. .719 1. , 00 13. .02 N
ATOM 3986 CA ALA A 517 114 , .621 53. , 630 11. .713 1. , 00 14. .34 C
ATOM 3987 CB ALA A 517 113 , .734 54. , 691 12. .370 1. , 00 13. .42 C
ATOM 3988 C ALA A 517 113 , .748 52. , 589 11. .037 1. , 00 14. .00 C
ATOM 3989 O ALA A 517 113 , .249 52. , 800 9. .929 1. , 00 14. .94 0
ATOM 3990 N GLU A 518 113 , .536 51. , 485 11. .723 1. , 00 13. .79 N
ATOM 3991 CA GLU A 518 112 , .557 50. , 505 11. .336 1. , 00 15. .60 C
ATOM 3992 CB GLU A 518 113 , .122 49. , 104 11. .582 1. , 00 20. .75 C
ATOM 3993 CG GLU A 518 112 , .350 47. , 992 10. .896 1. , 00 26. .36 c
ATOM 3994 CD GLU A 518 112 , .955 46. , 628 11. .199 1. , 00 31. .68 C
ATOM 3995 OE1 GLU A 518 114 , .174 46. , 471 10. .994 1. , 00 36. .57 O
ATOM 3996 OE2 GLU A 518 112 , .223 45. , 728 11. .660 1. , 00 30. .59 O
ATOM 3997 C GLU A 518 111 , .282 50. , 708 12. .160 1. , 00 14. .30 c
ATOM 3998 O GLU A 518 111 , .328 50. , 788 13. .375 1. , 00 16. .77 O
ATOM 3999 N VAL A 519 110 , .151 50. , 784 11. .483 1. , 00 11. .31 N
ATOM 4000 CA VAL A 519 108 , .854 50. , 902 12. .126 1. , 00 10. .69 c
ATOM 4001 CB VAL A 519 108 , .026 52. ,044 11. .535 1. , 00 9. .53 c
ATOM 4002 CGI VAL A 519 106 , .686 52. , 199 12. .295 1. , 00 7. .75 C
ATOM 4003 CG2 VAL A 519 108 , .850 53. ,338 11. .581 1. , 00 8. .74 C
ATOM 4004 C VAL A 519 108 ,.082 49., 589 12..008 1., 00 10..94 C
ATOM 4005 O VAL A 519 107 , .882 49. , 075 10. .897 1. , 00 9. .94 O
ATOM 4006 N PRO A 520 107 , .647 49. , 026 13. .148 1. , 00 12. .57 N
ATOM 4007 CA PRO A 520 106. .867 47. .763 13. .066 1. , 00 14. .58 C
ATOM 4008 CB PRO A 520 106 , .494 47. , 467 14. .536 1. , 00 16. .07 C
ATOM 4009 CG PRO A 520 107 , .485 48. , 235 15. .356 1. , 00 17. .24 C
ATOM 4010 CD PRO A 520 107 , .849 49. , 461 14. .545 1. , 00 16. .07 C
ATOM 4011 C PRO A 520 105 , .582 47. , 889 12. .248 1. , 00 12. .26 c
ATOM 4012 O PRO A 520 105 , .072 49. , 013 12. .053 1. , 00 11. .41 0
ATOM 4013 N ALA A 521 105 , .078 46. , 742 11. .782 1. , 00 10. .12 N
ATOM 4014 CA ALA A 521 103 , .731 46. , 642 11. .197 1. , 00 10. .96 C
ATOM 4015 CB ALA A 521 103 , .329 45. , 162 10. .990 1. , 00 10. .36 c
ATOM 4016 C ALA A 521 102 , .716 47. ,337 12. .106 1. , 00 9. .89 C
ATOM 4017 O ALA A 521 102 , .768 47. , 207 13. .323 1. , 00 9. .19 O
ATOM 4018 N HIS A 522 101 , .816 48. , 107 11. .513 1. , 00 9. .54 N
ATOM 4019 CA HIS A 522 100 , .814 48. , 843 12. .269 1. , 00 9. .64 c
ATOM 4020 CB HIS A 522 99 , .727 47. , 860 12. ,666 1. , 00 10. .73 C
ATOM 4021 CG HIS A 522 99 , .191 47. , 110 11. .469 1. , 00 10. .88 C
ATOM 4022 ND1 HIS A 522 98 , .521 47. , 744 10. .477 1. , 00 11. .26 N
ATOM 4023 CE1 HIS A 522 98. .216 46. , 859 9. .510 1. , 00 12. .49 c
ATOM 4024 NE2 HIS A 522 98 , .705 45. , 661 9. .875 1. , 00 11. .79 N
ATOM 4025 CD2 HIS A 522 99 , .310 45. , 787 11. .087 1. , 00 9. .87 C
ATOM 4026 C HIS A 522 101 , .407 49. , 618 13. .431 1. , 00 11. .25 c
ATOM 4027 O HIS A 522 100. .931 49. , 558 14. .575 1. , 00 11. .49 0
ATOM 4028 N GLY A 523 102 , .466 50. ,367 13. .114 1. , 00 11. .52 N
ATOM 4029 CA GLY A 523 103 , .316 50. , 985 14. .107 1. , 00 10. .64 C
ATOM 4030 C GLY A 523 103. .592 52. , 433 13. .862 1. , 00 8. .84 c
ATOM 4031 O GLY A 523 103 , .252 52. , 980 12. .810 1. , 00 8. .72 0
ATOM 4032 N VAL A 524 104 , .241 53. , 034 14. .854 1. , 00 8. .94 N
ATOM 4033 CA VAL A 524 104 , . 610 54. , 432 14. .845 1. , 00 8. .56 C
ATOM 4034 CB VAL A 524 103 , .520 55. , 297 15. .527 1. , 00 9. .19 c
ATOM 4035 CGI VAL A 524 103 , .171 54. , 762 16. .928 1. , 00 8. .21 C
ATOM 4036 CG2 VAL A 524 103 , .945 56. , 757 15. .591 1. , 00 7. .98 C
ATOM 4037 C VAL A 524 105 , .917 54. , 599 15. .598 1. , 00 10. .38 C
ATOM 4038 O VAL A 524 106 , .167 53. , 886 16. .593 1. , 00 10. .42 0
ATOM 4039 N ARG A 525 106 , .760 55. , 508 15. .124 1. , 00 9. .71 N
ATOM 4040 CA ARG A 525 107 , .859 56. , 028 15. .940 1. , 00 10. .60 C
ATOM 4041 CB ARG A 525 109 , .207 55. , 485 15. .455 1. , 00 13. .03 C
ATOM 4042 CG ARG A 525 109 , .297 53. , 949 15. .562 1. , 00 18. .03 c
ATOM 4043 CD ARG A 525 110 , .317 53. , 471 16. .562 1. , 00 25. .99 C
ATOM 4044 NE ARG A 525 111 , .397 52. , 876 15. .804 1. , 00 34. .69 N
ATOM 4045 CZ ARG A 525 112 , .674 52. , 854 16. .145 1. , 00 28. .95 C
ATOM 4046 NH1 ARG A 525 113 , .526 52. , 273 15. .310 1. , 00 30. .55 N
ATOM 4047 NH2 ARG A 525 113 , .114 53. ,398 17. .284 1. , 00 32. .10 N
ATOM 4048 C ARG A 525 107 , .827 57. , 551 15. .888 1. , 00 10. .27 C
ATOM 4049 O ARG A 525 107., 502 58 ,, 135 14., 849 1., 00 10.,33 O
ATOM 4050 N VAL A 526 108. , 118 58 , , 194 17. , 008 1. , 00 9. , 16 N
ATOM 4051 CA VAL A 526 108. , 174 59 , , 656 17. , 048 1. , 00 10. ,36 C
ATOM 4052 CB VAL A 526 107. .051 60 , .261 17. .917 1. .00 10. .49 C
ATOM 4053 CGI VAL A 526 107. , 197 61 , , 783 18. , 027 1. , 00 8. ,98 C
ATOM 4054 CG2 VAL A 526 105. , 665 59 , , 895 17. ,362 1. , 00 10. ,91 C
ATOM 4055 C VAL A 526 109. , 524 59 , , 987 17. ,629 1. , 00 11. ,51 C
ATOM 4056 O VAL A 526 109. , 798 59 , .616 18. .764 1. , 00 14. .38 0
ATOM 4057 N PHE A 527 110. ,383 60 , , 617 16. , 839 1. , 00 9. ,91 N
ATOM 4058 CA PHE A 527 Ill . , 660 61 , , 103 17. ,323 1. , 00 9. ,52 C
ATOM 4059 CB PHE A 527 112. , 749 60 , , 797 16. ,305 1. , 00 9. ,92 C
ATOM 4060 CG PHE A 527 112. , 995 59 , .320 16. .114 1. , 00 11. .05 c
ATOM 4061 CD1 PHE A 527 113. , 911 58 , , 643 16. ,915 1. , 00 13. ,35 C
ATOM 4062 CE1 PHE A 527 114. , 145 57 , , 276 16. , 719 1. , 00 14. ,90 C
ATOM 4063 CZ PHE A 527 113. , 448 56 , , 582 15. , 731 1. , 00 10. ,88 C
ATOM 4064 CE2 PHE A 527 112. , 567 57 , .255 14. .922 1. , 00 10. .96 c
ATOM 4065 CD2 PHE A 527 112. ,331 58 , , 614 15. , 130 1. , 00 11. , 05 C
ATOM 4066 C PHE A 527 Ill . , 614 62 , , 600 17. ,642 1. , 00 9. ,59 C
ATOM 4067 O PHE A 527 Ill . , 072 63 , ,399 16. , 877 1. , 00 10. , 13 O
ATOM 4068 N LEU A 528 112. , 197 62 , .974 18. .775 1. , 00 9. .22 N
ATOM 4069 CA LEU A 528 112. , 126 64 , ,342 19. ,272 1. , 00 9. ,68 C
ATOM 4070 CB LEU A 528 Ill . , 408 64 , ,382 20. ,634 1. , 00 9. , 09 C
ATOM 4071 CG LEU A 528 110. , 002 63 , , 775 20. ,684 1. , 00 9. , 41 c
ATOM 4072 CD1 LEU A 528 109. , 488 63 , .694 22. .124 1. , 00 9. .12 c
ATOM 4073 CD2 LEU A 528 109. , 030 64 , , 599 19. , 823 1. , 00 8. ,64 C
ATOM 4074 C LEU A 528 113. , 515 64 , , 909 19. , 449 1. , 00 9. , 76 C
ATOM 4075 O LEU A 528 114. , 362 64 , .267 20. .049 1. .00 10. .21 0
ATOM 4076 N LEU A 529 113. , 719 66 , , 115 18. , 932 1. , 00 10. ,18 N
ATOM 4077 CA LEU A 529 114. , 940 66 , , 860 19. , 071 1. , 00 10. ,08 C
ATOM 4078 CB LEU A 529 115. , 575 67, , 129 17. , 695 1. , 00 9. ,32 C
ATOM 4079 CG LEU A 529 116. , 785 68 , .085 17. .678 1. , 00 9. .62 c
ATOM 4080 CD1 LEU A 529 118. , 000 67, , 439 18. ,343 1. , 00 11. , 82 C
ATOM 4081 CD2 LEU A 529 117. , 150 68 , , 508 16. ,265 1. , 00 10. ,57 C
ATOM 4082 C LEU A 529 114. , 600 68 , , 175 19. , 799 1. , 00 11. , 06 C
ATOM 4083 O LEU A 529 113. , 727 68 , .972 19. .371 1. , 00 10. .42 0
ATOM 4084 N ASP A 530 115. , 266 68 , ,391 20. ,911 1. , 00 9. ,92 N
ATOM 4085 CA ASP A 530 115. , 041 69 , , 596 21. , 704 1. , 00 11. , 43 C
ATOM 4086 CB ASP A 530 115. , 073 69 , , 245 23. , 173 1. , 00 14. ,30 C
ATOM 4087 CG ASP A 530 114. , 798 70 , .443 24. .083 1. , 00 16. .65 c
ATOM 4088 OD1 ASP A 530 114. , 591 71 , , 602 23. ,628 1. , 00 17. ,39 O
ATOM 4089 OD2 ASP A 530 114. , 822 70 , , 196 25. ,287 1. , 00 18. ,37 O
ATOM 4090 C ASP A 530 116. , 132 70 , , 578 21. ,331 1. , 00 10. ,62 C
ATOM 4091 O ASP A 530 117. , 187 70 , .577 21. .922 1. , 00 10. .38 0
ATOM 4092 N ALA A 531 115. , 894 71 , ,359 20. , 292 1. , 00 10. ,14 N
ATOM 4093 CA ALA A 531 116. , 888 72 , ,323 19. , 825 1. , 00 10. , 79 C
ATOM 4094 CB ALA A 531 117., 950 71 ,, 649 18.,941 1., 00 9., 75 C
ATOM 4095 C ALA A 531 116. , 201 73 , , 426 19. , 042 1. , 00 10. ,66 C
ATOM 4096 O ALA A 531 115. , 168 73 , , 230 18. ,375 1. , 00 10. ,28 O
ATOM 4097 N GLN A 532 116. .791 74. , 592 19. .134 1. , 00 10. .54 N
ATOM 4098 CA GLN A 532 116. , 467 75 , , 682 18. , 253 1. , 00 15. ,30 C
ATOM 4099 CB GLN A 532 117. , 405 76 , , 857 18. , 575 1. , 00 16. , 44 C
ATOM 4100 CG GLN A 532 117. , 022 78 , , 154 17. ,900 1. , 00 23. ,33 C
ATOM 4101 CD GLN A 532 117. , 997 79. , 280 18. .191 1. , 00 25. .77 c
ATOM 4102 OE1 GLN A 532 118. , 922 79 , , 139 18. , 999 1. , 00 31. ,32 O
ATOM 4103 NE2 GLN A 532 117. , 787 80 , , 405 17. , 544 1. , 00 26. ,18 N
ATOM 4104 C GLN A 532 116. , 657 75 , , 241 16. , 781 1. , 00 13. , 76 C
ATOM 4105 O GLN A 532 117. , 622 74. , 559 16. .467 1. , 00 11. .93 0
ATOM 4106 N VAL A 533 115. , 762 75 , ,666 15. , 897 1. , 00 12. ,61 N
ATOM 4107 CA VAL A 533 115. , 898 75 , , 410 14. , 472 1. , 00 12. ,23 C
ATOM 4108 CB VAL A 533 114. , 625 75 , , 848 13. ,697 1. , 00 12. ,90 C
ATOM 4109 CGI VAL A 533 114. , 842 75. , 775 12. .192 1. , 00 10. .51 c
ATOM 4110 CG2 VAL A 533 113. , 429 74 , , 979 14. , 107 1. , 00 10. , 72 C
ATOM 4111 C VAL A 533 117. , 122 76 , , 148 13. ,920 1. , 00 13. ,56 C
ATOM 4112 O VAL A 533 117. , 261 77 , , 351 14. , 113 1. , 00 12. , 02 O
ATOM 4113 N THR A 534 118. , 018 75. , 398 13. .284 1. , 00 12. .09 N
ATOM 4114 CA THR A 534 119. , 175 75 , , 928 12. , 595 1. , 00 12. , 78 C
ATOM 4115 CB THR A 534 120. , 469 75 , , 604 13. , 351 1. , 00 14. ,58 C
ATOM 4116 OG1 THR A 534 120. , 563 74 , , 171 13. , 497 1. , 00 14. , 83 O
ATOM 4117 CG2 THR A 534 120. , 505 76. , 337 14. .719 1. , 00 13. .89 c
ATOM 4118 C THR A 534 119. ,381 75 , ,376 11. , 189 1. , 00 13. ,87 C
ATOM 4119 O THR A 534 120. , 070 76 , , 010 10. ,399 1. , 00 14. , 05 O
ATOM 4120 N GLU A 535 118. .859 74. , 196 10. .863 1. , 00 15. .17 N
ATOM 4121 CA GLU A 535 119. , 104 73 , , 617 9. , 521 1. , 00 16. , 86 C
ATOM 4122 CB GLU A 535 118. , 583 72 , , 165 9. , 452 1. , 00 20. ,66 C
ATOM 4123 CG GLU A 535 118. ,387 71 , , 568 8. , 065 1. , 00 28. , 44 C
ATOM 4124 CD GLU A 535 119. , 646 71. , 502 7. .279 1. , 00 38. .00 c
ATOM 4125 OE1 GLU A 535 120. , 694 71 , , 256 7. ,907 1. , 00 43. ,92 O
ATOM 4126 OE2 GLU A 535 119. , 584 71 , , 714 6. , 043 1. , 00 49. , 47 O
ATOM 4127 C GLU A 535 118. , 463 74 , , 530 8. , 448 1. , 00 14. ,94 C
ATOM 4128 O GLU A 535 117. , 260 74. , 784 8. .489 1. , 00 12. .32 0
ATOM 4129 N PRO A 536 119. , 269 75 , , 045 7. ,506 1. , 00 13. ,22 N
ATOM 4130 CA PRO A 536 118. , 787 76 , , 103 6. ,607 1. , 00 14. ,11 C
ATOM 4131 CB PRO A 536 119. , 994 76 , ,395 5. ,696 1. , 00 14. ,51 C
ATOM 4132 CG PRO A 536 121. ,184 75. , 893 6. .427 1. , 00 16. .92 c
ATOM 4133 CD PRO A 536 120. , 728 74 , , 833 7. ,394 1. , 00 16. ,34 C
ATOM 4134 C PRO A 536 117. , 558 75 , , 790 5. , 747 1. , 00 11. ,81 C
ATOM 4135 O PRO A 536 116. , 701 76 , , 644 5. , 572 1. , 00 12. ,20 O
ATOM 4136 N THR A 537 117. , 485 74. , 589 5. .207 1. , 00 10. .03 N
ATOM 4137 CA THR A 537 116. ,364 74 , , 177 4. ,361 1. , 00 11. , 44 C
ATOM 4138 CB THR A 537 116. , 758 72 , , 938 3. , 522 1. , 00 13. , 05 C
ATOM 4139 OG1 THR A 537 116 ,.976 71 ,.808 4..374 1., 00 16..34 O
ATOM 4140 CG2 THR A 537 118 , .049 73 , .203 2. .750 1. , 00 11. .25 C
ATOM 4141 C THR A 537 115 , .075 73 , .956 5. .196 1. , 00 10. .50 C
ATOM 4142 O THR A 537 113. .969 74. .280 4. .774 1. .00 9. .54 0
ATOM 4143 N LEU A 538 115 , .247 73 , .473 6. .411 1. , 00 9. .49 N
ATOM 4144 CA LEU A 538 114 , .154 73 , .371 7. .357 1. , 00 8. .98 C
ATOM 4145 CB LEU A 538 114 , .604 72 , .593 8. .595 1. , 00 8. .29 C
ATOM 4146 CG LEU A 538 113 , .554 72 , .443 9. .713 1. , 00 8. .54 c
ATOM 4147 CD1 LEU A 538 112 , .371 71 , .621 9. .205 1. , 00 8. .35 C
ATOM 4148 CD2 LEU A 538 114 , .163 71 , .821 10. .953 1. , 00 7. .14 C
ATOM 4149 C LEU A 538 113 , .627 74 , .792 7. .713 1. , 00 9. .27 C
ATOM 4150 O LEU A 538 112 , .419 75 , .050 7. .599 1. , 00 9. .78 0
ATOM 4151 N ALA A 539 114 , .523 75 , .699 8. .090 1. , 00 8. .43 N
ATOM 4152 CA ALA A 539 114 , .134 77 , .080 8. .399 1. , 00 10. .03 C
ATOM 4153 CB ALA A 539 115 , .338 77 , .950 8. .810 1. , 00 8. .28 C
ATOM 4154 C ALA A 539 113 , .416 77 , .714 7. .216 1. , 00 11. .94 c
ATOM 4155 O ALA A 539 112 , .420 78 , .407 7. .401 1. , 00 12. .14 O
ATOM 4156 N ALA A 540 113 , .903 77 , .479 6. .000 1. , 00 12. .69 N
ATOM 4157 CA ALA A 540 113 , .261 78 , .090 4. .829 1. , 00 13. .57 C
ATOM 4158 CB ALA A 540 114. .112 77. .914 3. .559 1. , 00 12. .36 c
ATOM 4159 C ALA A 540 Ill , .826 77 , .588 4. .624 1. , 00 12. .40 C
ATOM 4160 O ALA A 540 110 , .924 78 , .389 4. .402 1. , 00 13. .10 O
ATOM 4161 N ALA A 541 111 , .597 76 , .281 4. .744 1. , 00 12. .30 N
ATOM 4162 CA ALA A 541 110. .260 75. .730 4. .607 1. , 00 12. .06 c
ATOM 4163 CB ALA A 541 110 , .290 74 , .194 4. .601 1. , 00 12. .11 C
ATOM 4164 C ALA A 541 109 , .330 76 , .259 5. .717 1. , 00 11. .71 C
ATOM 4165 O ALA A 541 108. .167 76. .578 5. .477 1. .00 11. .89 0
ATOM 4166 N LEU A 542 109 , .861 76 , .369 6. .922 1. , 00 10. .84 N
ATOM 4167 CA LEU A 542 109 , .081 76 , .820 8. .061 1. , 00 10. .57 C
ATOM 4168 CB LEU A 542 109 , .828 76 , .539 9. .360 1. , 00 9. .37 C
ATOM 4169 CG LEU A 542 110 , .008 75 , .053 9. .792 1. , 00 9. .27 c
ATOM 4170 CD1 LEU A 542 111 , .003 74 , .968 10. .949 1. , 00 6. .79 C
ATOM 4171 CD2 LEU A 542 108 , .679 74 , .412 10. .154 1. , 00 7. .05 C
ATOM 4172 C LEU A 542 108 , .746 78 , .325 7. .946 1. , 00 12. .02 C
ATOM 4173 O LEU A 542 107 , .635 78 , .736 8. .295 1. , 00 9. .27 0
ATOM 4174 N ASP A 543 109 , .720 79 , .121 7. .481 1. , 00 10. .97 N
ATOM 4175 CA ASP A 543 109 , .503 80 , .535 7. .208 1. , 00 11. .93 C
ATOM 4176 CB ASP A 543 110 , .778 81 , .211 6. .677 1. , 00 11. .96 C
ATOM 4177 CG ASP A 543 111 , .880 81 , .335 7. .727 1. , 00 15. .31 c
ATOM 4178 OD1 ASP A 543 111 , .587 81 , .321 8. .945 1. , 00 14. .43 O
ATOM 4179 OD2 ASP A 543 113 , .056 81 , .460 7. .315 1. , 00 15. .12 O
ATOM 4180 C ASP A 543 108 , .379 80 , .721 6. .171 1. , 00 11. .78 C
ATOM 4181 O ASP A 543 107 , .494 81 , .565 6. .351 1. , 00 12. .70 0
ATOM 4182 N ALA A 544 108 , .429 79 , .949 5. .094 1. , 00 11. .45 N
ATOM 4183 CA ALA A 544 107 , .393 80 , .014 4. .057 1. , 00 13. .07 C
ATOM 4184 CB ALA A 544 107., 771 79 ,, 164 2., 840 1., 00 14..31 C
ATOM 4185 C ALA A 544 106. , 004 79 , , 630 4. , 576 1. , 00 12. .78 C
ATOM 4186 O ALA A 544 105. , 017 80 , , 249 4. , 184 1. , 00 11. .26 O
ATOM 4187 N ALA A 545 105. .929 78 , .636 5. .470 1. , 00 12. .10 N
ATOM 4188 CA ALA A 545 104. , 653 78 , , 258 6. , 101 1. , 00 11. .76 C
ATOM 4189 CB ALA A 545 104. , 766 76 , , 923 6. , 841 1. , 00 8. .95 C
ATOM 4190 C ALA A 545 104. , 122 79 , ,373 7. , 030 1. , 00 11. .06 C
ATOM 4191 O ALA A 545 102. , 924 79 , .646 7. .010 1. , 00 12. .09 0
ATOM 4192 N MET A 546 104. , 994 80 , , 051 7. , 789 1. , 00 10. .46 N
ATOM 4193 CA MET A 546 104. , 555 81 , , 173 8. ,640 1. , 00 11. .25 C
ATOM 4194 CB MET A 546 105. , 658 81 , , 652 9. ,618 1. , 00 11. .09 C
ATOM 4195 CG MET A 546 106. , 097 80 , .590 10. .628 1. , 00 11. .56 c
ATOM 4196 SD MET A 546 104. , 767 80 , , 106 11. , 774 1. , 00 12. .53 s
ATOM 4197 CE MET A 546 104. , 589 81 , , 658 12. , 694 1. , 00 10. .43 C
ATOM 4198 C MET A 546 104. , 057 82 , ,337 7. , 785 1. , 00 11. .91 C
ATOM 4199 O MET A 546 103. , 026 82 , .922 8. .103 1. , 00 13. .59 O
ATOM 4200 N ALA A 547 104. , 776 82 , , 674 6. , 719 1. , 00 12. .15 N
ATOM 4201 CA ALA A 547 104. ,346 83 , , 763 5. , 842 1. , 00 15. .42 C
ATOM 4202 CB ALA A 547 105. ,393 84, , 097 4. , 786 1. , 00 14. .98 C
ATOM 4203 C ALA A 547 102. , 997 83 , .411 5. .188 1. , 00 16. .22 c
ATOM 4204 O ALA A 547 102. , 121 84, , 251 5. , 111 1. , 00 17. .52 O
ATOM 4205 N ASP A 548 102. , 815 82 , , 159 4. , 777 1. , 00 15. .85 N
ATOM 4206 CA ASP A 548 101. , 560 81 , , 762 4. , 150 1. , 00 15. .63 c
ATOM 4207 CB ASP A 548 101. , 582 80 , .321 3. .666 1. , 00 16. .71 c
ATOM 4208 CG ASP A 548 100. , 277 79 , , 933 2. ,990 1. , 00 17. .92 C
ATOM 4209 OD1 ASP A 548 100. , 047 80 , ,384 1. .859 1. , 00 23. .54 O
ATOM 4210 OD2 ASP A 548 99. , 464 79 , .251 3. .606 1. , 00 21. .00 O
ATOM 4211 C ASP A 548 100. ,377 81 , , 945 5. .110 1. , 00 17. .65 C
ATOM 4212 O ASP A 548 99. ,316 82 , , 431 4. .704 1. , 00 13. .49 O
ATOM 4213 N ALA A 549 100. , 582 81 , , 575 6. .369 1. , 00 15. .54 N
ATOM 4214 CA ALA A 549 99. , 535 81 , .640 7. .373 1. , 00 19. .94 c
ATOM 4215 CB ALA A 549 99. , 968 80 , , 908 8. .643 1. , 00 16. .36 C
ATOM 4216 C ALA A 549 99. , 123 83 , , 086 7. .693 1. , 00 22. .92 C
ATOM 4217 O ALA A 549 98. , 055 83 , , 297 8. .260 1. , 00 20. .41 O
ATOM 4218 N ARG A 550 99. , 965 84 , .072 7. .366 1. , 00 26. .65 N
ATOM 4219 CA ARG A 550 99. , 590 85, , 480 7. .565 1. , 00 30. .18 C
ATOM 4220 CB ARG A 550 100. , 815 86 , ,373 7. .839 1. , 00 28. .79 C
ATOM 4221 CG ARG A 550 101. , 406 86 , , 152 9. .227 1. , 00 29. .17 C
ATOM 4222 CD ARG A 550 102. , 524 87 , .143 9. .534 1. , 00 39. .26 c
ATOM 4223 NE ARG A 550 103. , 742 86 , , 795 8. .804 1. , 00 47. .80 N
ATOM 4224 CZ ARG A 550 104. , 685 85, , 943 9. .230 1. , 00 51. .10 C
ATOM 4225 NH1 ARG A 550 104. , 582 85, ,330 10. .415 1. , 00 51. .89 N
ATOM 4226 NH2 ARG A 550 105. , 744 85 , .696 8. .457 1. , 00 33. .57 N
ATOM 4227 C ARG A 550 98. , 725 86 , , 069 6. .442 1. , 00 33. .84 C
ATOM 4228 O ARG A 550 98. , 128 87, , 120 6. .637 1. , 00 31. .87 O
ATOM 4229 N ARG A 551 98.619 85.392 5.301 1.00 36.87 N
ATOM 4230 CA ARG A 551 97 .786 85 .879 4 .184 1 .00 35 .97 C
ATOM 4231 CB ARG A 551 98 .063 85 .076 2 .913 1 .00 34 .44 C
ATOM 4232 CG ARG A 551 99 .484 85 .175 2 .404 1 .00 40 .33 C
ATOM 4233 CD ARG A 551 99 .671 84 .462 1 .064 1 .00 50 .67 c
ATOM 4234 NE ARG A 551 100 .913 83 .674 1 .057 1 .00 68 .59 N
ATOM 4235 CZ ARG A 551 102 .152 84 .180 1 .113 1 .00 69 .62 C
ATOM 4236 NH1 ARG A 551 102 .359 85 .496 1 .159 1 .00 70 .63 N
ATOM 4237 NH2 ARG A 551 103 .206 83 .365 1 .130 1 .00 50 .36 N
ATOM 4238 C ARG A 551 96 .283 85 .782 4 .483 1 .00 40 .67 c
ATOM 4239 O ARG A 551 95 .820 84 .797 5 .078 1 .00 41 .34 O
ATOM 4240 N SER A 552 95 .528 86 .788 4 .038 1 .00 45 .37 N
ATOM 4241 CA SER A 552 94 .055 86 .754 4 .083 1 .00 48 .64 c
ATOM 4242 CB SER A 552 93 .466 88 .064 3 .546 1 .00 45 .77 c
ATOM 4243 OG SER A 552 93 .565 89 .086 4 .511 1 .00 56 .51 O
ATOM 4244 C SER A 552 93 .465 85 .613 3 .257 1 .00 53 .97 c
ATOM 4245 O SER A 552 93 .988 85 .285 2 .195 1 .00 44 .21 O
ATOM 4246 N ARG A 553 92 .400 84 .997 3 .776 1 .00 58 .75 N
ATOM 4247 CA ARG A 553 91 .395 84 .310 2 .946 1 .00 62 .60 c
ATOM 4248 CB ARG A 553 91 .490 82 .774 2 .990 1 .00 53 .31 c
ATOM 4249 CG ARG A 553 92 .046 82 .144 4 .257 1 .00 45 .97 c
ATOM 4250 CD ARG A 553 93 .550 81 .923 4 .128 1 .00 48 .06 c
ATOM 4251 NE ARG A 553 94 .060 81 .093 5 .208 1 .00 47 .70 N
ATOM 4252 CZ ARG A 553 95 .283 80 .574 5 .267 1 .00 48 .46 c
ATOM 4253 NH1 ARG A 553 96 .176 80 .798 4 .300 1 .00 43 .23 N
ATOM 4254 NH2 ARG A 553 95 .611 79 .817 6 .313 1 .00 48 .31 N
ATOM 4255 C ARG A 553 90 .001 84 .766 3 .376 1 .00 77 .50 c
ATOM 4256 O ARG A 553 89 .820 85 .337 4 .454 1 .00 86 .21 O
ATOM 4257 OXT ARG A 553 89 .021 84 .582 2 .656 1 .00 87 .66 O
ATOM 4258 CA CA A 600 70. 802 57. 236 35. 062 1. 00 15. 98 CA
ATOM 4259 CA CA A 601 63. 208 65. 968 29. 650 1. 00 13. 29 CA
ATOM 4260 CA CA A 602 67. 621 77. 498 10. 597 1. 00 9. 99 CA
ATOM 4680 CL CL A 603 72. 381 53. 223 16. 908 1. 00 13. 61 CL
ATOM 4674 03 GOL B 1 71 .181 62 .613 17 .255 1 .00 12 .69 O
ATOM 4675 C3 GOL B 1 70 .059 63 .246 16 .665 1 .00 15 .67 c
ATOM 4676 C2 GOL B 1 68 .954 62 .285 16 .286 1 .00 21 .27 c
ATOM 4677 02 GOL B 1 69 .488 61 .026 15 .941 1 .00 26 .24 O
ATOM 4678 CI GOL B 1 68 .041 62 .832 15 .191 1 .00 22 .45 c
ATOM 4679 01 GOL B 1 67 .264 61 .777 14 .630 1 .00 26 .00 0
ATOM 4681 03 GOL B 2 72 .946 67 .251 1 .695 1 .00 15 .32 O
ATOM 4682 C3 GOL B 2 71 .767 66 .657 15 .249 1 .00 14 .09 c
ATOM 4683 C2 GOL B 2 70 .559 67 .586 15 .090 1 .00 14 .05 c
ATOM 4684 02 GOL B 2 70 .621 68 .590 16 .096 1 .00 12 .33 0
ATOM 4685 CI GOL B 2 69 .249 66 .846 15 .221 1 .00 18 .07 c
ATOM 4686 01 GOL B 2 69 .023 66 .062 14 .039 1 .00 17 .82 O
ATOM 4687 03 GOL B 3 72., 515 64., 096 10..689 1., 00 25..67 O
ATOM 4688 C3 GOL C 4 73. , 467 64. , 963 11. .307 1. , 00 26. .15 C
ATOM 4689 C2 GOL C 4 72. , 861 66. ,338 11. .544 1. , 00 25. .64 c
ATOM 4690 02 GOL C 4 73. .851 67. , 096 12. .249 1. , 00 19. .50 O
ATOM 4691 CI GOL C 4 72. , 435 67. , 029 10. .241 1. , 00 22. .26 c
ATOM 4692 01 GOL C 4 73. , 539 67. , 080 9. .298 1. , 00 23. .90 O
ATOM 4261 0 HOH W 1 73. ,301 61. , 252 25. .027 1. , 00 9. .85 O
ATOM 4262 0 HOH w 2 94. , 676 65. , 066 13. .807 1. , 00 10. .58 0
ATOM 4263 0 HOH w 3 59. , 711 76. , 459 12. .227 1. , 00 7. .99 O
ATOM 4264 0 HOH w 4 98. , 238 68. ,392 15. .962 1. , 00 11. .94 O
ATOM 4265 0 HOH w 5 97. , 088 65. , 408 16. .506 1. , 00 11. .85 O
ATOM 4266 0 HOH w 6 62. , 085 65. , 925 13. .108 1. , 00 16. .04 0
ATOM 4267 0 HOH w 7 73. , 920 61. , 551 37. .146 1. , 00 15. .54 O
ATOM 4268 0 HOH w 8 77. , 168 50. , 157 33. .169 1. , 00 15. .21 O
ATOM 4269 0 HOH w 9 61. , 919 63. , 088 23. .865 1. , 00 11. .76 O
ATOM 4270 0 HOH w 10 72. , 608 74. , 669 15. .812 1. , 00 10. .72 0
ATOM 4271 0 HOH w 11 63. , 466 64. , 051 28. .377 1. , 00 9. .82 O
ATOM 4272 0 HOH w 12 69. , 752 66. , 825 27. .140 1. , 00 12. .03 O
ATOM 4273 0 HOH w 13 96. ,364 76. , 517 4. .720 1. , 00 15. .03 O
ATOM 4274 0 HOH w 14 114. , 715 68. , 298 11. .096 1. , 00 13. .68 O
ATOM 4275 0 HOH w 15 97. , 437 71. , 403 15. .360 1. , 00 10. .85 O
ATOM 4276 0 HOH w 16 77. , 509 58. , 818 30. .992 1. , 00 10. .02 O
ATOM 4277 0 HOH w 17 90. , 586 69. , 226 5. .708 1. , 00 12. .97 O
ATOM 4278 0 HOH w 18 68. , 221 64. , 485 27. .002 1. , 00 15. .33 O
ATOM 4279 0 HOH w 19 80. ,372 45. , 787 14. .915 1. , 00 13. .14 O
ATOM 4280 0 HOH w 20 89. , 056 64. , 621 10. .452 1. , 00 13. .00 O
ATOM 4281 0 HOH w 21 71. , 497 52. , 451 19. .773 1. , 00 9. .03 O
ATOM 4282 0 HOH w 22 87. , 363 68. , 955 0. .376 1. , 00 15. .62 O
ATOM 4283 0 HOH w 23 88. , 823 64. , 449 1. .620 1. , 00 17. .84 O
ATOM 4284 0 HOH w 24 56. , 279 65. , 027 11. .407 1. , 00 19. .72 O
ATOM 4285 0 HOH w 25 87. , 136 52. , 751 3. .189 1. , 00 17. .97 0
ATOM 4286 0 HOH w 26 69. , 509 54. , 464 17. .311 1. , 00 13. .33 O
ATOM 4287 0 HOH w 27 82. , 485 70. , 854 9. .739 1. , 00 9. .26 O
ATOM 4288 0 HOH w 28 72. , 417 49. , 514 23. .793 1. , 00 10. .68 O
ATOM 4289 0 HOH w 29 75. , 569 72. , 013 9. .882 1. , 00 7. .94 0
ATOM 4290 0 HOH w 30 61. , 004 77. , 739 8. .577 1. , 00 10. .78 O
ATOM 4291 0 HOH w 31 96. , 117 69. , 727 1. .018 1. , 00 16. .27 O
ATOM 4292 0 HOH w 32 74. ,350 72. , 855 37. .663 1. , 00 26. .67 O
ATOM 4293 0 HOH w 33 63. , 557 56. , 952 14. .842 1. , 00 22. .65 0
ATOM 4294 0 HOH w 34 87. , 254 72. , 718 -1. .207 1. , 00 18. .26 O
ATOM 4295 0 HOH w 35 63. , 640 78. , 182 7. .997 1. , 00 12. .81 O
ATOM 4296 0 HOH w 36 85. , 296 51. , 423 5. .220 1. , 00 14. .66 O
ATOM 4297 0 HOH w 37 97. , 165 53. ,677 18. .048 1. , 00 10. .53 0
ATOM 4298 0 HOH w 38 64. , 992 73. , 401 11. .693 1. , 00 8. .79 O
ATOM 4299 0 HOH w 39 101. , 191 51. , 144 16. .856 1. , 00 15. .00 O
ATOM 4300 O HOH W 40 75., 284 85 ,.427 6..931 1., 00 17..36 O
ATOM 4301 O HOH w 41 101. , 467 75 , .016 7. .535 1. , 00 10. .33 O
ATOM 4302 O HOH w 42 72. , 548 75 , .644 -0. .400 1. , 00 10. .93 O
ATOM 4303 O HOH w 43 61. .115 84 , .329 10. .013 1. , 00 12. .37 O
ATOM 4304 O HOH w 44 69. , 051 57 , .185 17. .220 1. , 00 11. .51 O
ATOM 4305 O HOH w 45 73. , 187 63 , .785 15. .880 1. , 00 11. .17 O
ATOM 4306 O HOH w 46 65. , 091 72 , .566 4. .534 1. , 00 7. .86 O
ATOM 4307 O HOH w 47 57. , 598 82 , .971 20. .246 1. , 00 17. .55 O
ATOM 4308 O HOH w 48 74. , 831 78 , .095 6. .016 1. , 00 23. .98 O
ATOM 4309 O HOH w 49 94. , 448 64, .970 10. .381 1. , 00 53. .92 O
ATOM 4310 0 HOH w 50 68. , 059 48 , .033 21. .658 1. , 00 14. .51 O
ATOM 4311 0 HOH w 51 69. , 864 50 , .679 30. .531 1. , 00 23. .84 O
ATOM 4312 0 HOH w 52 101. , 668 62 , .523 28. .476 1. , 00 16. .04 O
ATOM 4313 0 HOH w 53 60. , 457 82 , .754 8. .003 1. , 00 10. .43 O
ATOM 4314 0 HOH w 54 98. , 841 70 , .632 2. .293 1. , 00 18. .12 O
ATOM 4315 0 HOH w 55 67. , 848 79 , .785 10. .036 1. , 00 9. .14 O
ATOM 4316 0 HOH w 56 74. , 745 56 , .274 10. .581 1. , 00 18. .03 O
ATOM 4317 0 HOH w 57 107. , 555 65, .751 24. .148 1. , 00 17. .39 O
ATOM 4318 0 HOH w 58 81. , 728 45 , .851 19. .575 1. , 00 20. .99 O
ATOM 4319 0 HOH w 59 74. , 805 89 , .197 26. .239 1. , 00 32. .84 O
ATOM 4320 0 HOH w 60 78. , 732 79 , .384 5. .490 1. , 00 19. .88 O
ATOM 4321 0 HOH w 61 99. , 553 53 , .549 -2. .727 1. , 00 19. .88 O
ATOM 4322 0 HOH w 62 94. , 758 52 , .611 18. .965 1. , 00 11. .05 O
ATOM 4323 0 HOH w 63 67. , 287 76 , .740 8. .446 1. , 00 11. .29 O
ATOM 4324 0 HOH w 64 83. , 036 49 , .847 4. .981 1. , 00 34. .91 O
ATOM 4325 0 HOH w 65 87. , 761 78 , .169 25. .477 1. , 00 13. .59 O
ATOM 4326 0 HOH w 66 121. .197 67 , .660 19. .855 1. , 00 22. .88 O
ATOM 4327 0 HOH w 67 68. , 256 60 , .005 12. .705 1. , 00 15. .28 O
ATOM 4328 0 HOH w 68 67. ,303 78 , .440 12. .806 1. , 00 8. .06 O
ATOM 4329 0 HOH w 69 66. , 845 79 , .531 -1. .065 1. , 00 11. .75 O
ATOM 4330 0 HOH w 70 55. , 069 60 , .199 32. .523 1. , 00 23. .75 0
ATOM 4331 0 HOH w 71 74. , 053 46 , .715 12. .581 1. , 00 15. .95 O
ATOM 4332 0 HOH w 72 105. , 148 61 , .097 -0. .647 1. , 00 17. .00 O
ATOM 4333 0 HOH w 73 64. , 494 84, .335 9. .317 1. , 00 12. .26 O
ATOM 4334 0 HOH w 74 100. , 602 53 , .757 19. .330 1. , 00 10. .19 0
ATOM 4335 0 HOH w 75 67. , 110 83 , .095 1. .371 1. , 00 20. .23 O
ATOM 4336 0 HOH w 76 103. , 582 73 , .620 6. .383 1. , 00 13. .21 O
ATOM 4337 0 HOH w 77 63. , 044 58 , .060 30. .204 1. , 00 10. .84 O
ATOM 4338 0 HOH w 78 83. , 815 86 , .086 17. .713 1. , 00 20. .16 0
ATOM 4339 0 HOH w 79 123. , 110 58 , .293 10. .793 1. , 00 22. .58 O
ATOM 4340 0 HOH w 80 90. , 442 77 , .202 11. .004 1. , 00 12. .85 O
ATOM 4341 0 HOH w 81 57. , 217 86 , .597 21. .382 1. , 00 23. .04 O
ATOM 4342 0 HOH w 82 53. ,312 61 , .917 29. .205 1. , 00 18. .81 0
ATOM 4343 0 HOH w 83 Ill . , 915 70 , .094 23. .094 1. , 00 25. .06 O
ATOM 4344 0 HOH w 84 89. , 625 78 , .270 13. .483 1. , 00 18. .96 O
ATOM 4345 O HOH W 85 96., 516 63., 335 23..952 1., 00 14..76 O
ATOM 4346 O HOH w 86 58. ,334 56. , 896 7. .380 1. , 00 27. .79 O
ATOM 4347 O HOH w 87 99. , 460 74. , 622 25. .157 1. , 00 22. .95 O
ATOM 4348 O HOH w 88 104. .112 48. , 169 5. .268 1. , 00 18. .97 O
ATOM 4349 O HOH w 89 99. , 458 68. , 003 1. .873 1. , 00 15. .08 O
ATOM 4350 O HOH w 90 88. , 400 80. , 289 2. .682 1. , 00 17. .29 O
ATOM 4351 O HOH w 91 75. , 965 70. , 423 4. .608 1. , 00 14. .51 O
ATOM 4352 O HOH w 92 53. , 399 62. , 729 10. .490 1. , 00 24. .51 O
ATOM 4353 O HOH w 93 68. , 022 85. , 521 7. .670 1. , 00 17. .66 O
ATOM 4354 O HOH w 94 105. , 909 78. , 173 24. .075 1. , 00 26. .48 O
ATOM 4355 O HOH w 95 101. , 655 73. , 227 23. .203 1. , 00 23. .79 O
ATOM 4356 O HOH w 96 103. , 701 50. , 954 0. .929 1. , 00 23. .11 O
ATOM 4357 O HOH w 97 59. ,352 53. , 719 26. .555 1. , 00 23. .04 O
ATOM 4358 O HOH w 98 101. , 037 80. , 238 12. .096 1. , 00 14. .80 O
ATOM 4359 O HOH w 99 104. , 659 69. , 936 3. .574 1. , 00 14. .04 O
ATOM 4360 O HOH w 100 103. , 458 57. , 078 25. .658 1. , 00 18. .60 O
ATOM 4361 O HOH w 101 53. , 805 58. , 863 12. .913 1. , 00 19. .64 O
ATOM 4362 O HOH w 102 99. , 745 78. ,300 10. .665 1. , 00 14. .90 O
ATOM 4363 O HOH w 103 77. , 791 89. , 583 25. .879 1. , 00 23. .54 O
ATOM 4364 O HOH w 104 67. , 035 91. , 326 12. .980 1. , 00 13. .78 O
ATOM 4365 O HOH w 105 98. , 933 81. , 009 13. .428 1. , 00 19. .24 O
ATOM 4366 O HOH w 106 75. , 580 70. , 650 7. .511 1. , 00 11. .53 O
ATOM 4367 O HOH w 107 59. , 161 75. ,386 2. .056 1. , 00 24. .13 O
ATOM 4368 O HOH w 108 66. , 095 86. , 539 5. .769 1. , 00 49. .22 O
ATOM 4369 O HOH w 109 70. , 790 48. , 831 21. .889 1. , 00 17. .12 O
ATOM 4370 O HOH w 110 97. , 251 66. , 013 23. .525 1. , 00 13. .20 O
ATOM 4371 O HOH w 111 90. , 643 65. , 822 0. .466 1. , 00 17. .40 0
ATOM 4372 0 HOH w 112 77. , 830 47. , 523 31. .224 1. , 00 18. .87 O
ATOM 4373 0 HOH w 113 75. , 241 65. , 944 15. .456 1. , 00 9. .63 O
ATOM 4374 0 HOH w 114 60. , 785 50. , 522 27. .718 1. , 00 31. .80 O
ATOM 4375 0 HOH w 115 64. , 668 86. , 557 24. .982 1. , 00 23. .71 0
ATOM 4376 0 HOH w 116 101. , 098 77. , 558 8. .202 1. , 00 22. .22 O
ATOM 4377 0 HOH w 117 106. , 242 44. , 285 12. .903 1. , 00 21. .32 O
ATOM 4378 0 HOH w 118 123. , 211 57. , 118 17. .184 1. , 00 30. .89 O
ATOM 4379 0 HOH w 119 49. ,666 66. , 238 12. .695 1. , 00 21. .19 0
ATOM 4380 0 HOH w 120 71. , 195 75. ,320 36. .843 1. , 00 25. .49 O
ATOM 4381 0 HOH w 121 81. , 772 73. , 968 38. .012 1. , 00 18. .19 O
ATOM 4382 0 HOH w 122 106. , 975 54. , 241 19. .306 1. , 00 35. .36 O
ATOM 4383 0 HOH w 123 95. , 547 54. , 279 -2. .956 1. , 00 27. .88 0
ATOM 4384 0 HOH w 124 87. , 005 79. , 057 18. .173 1. , 00 14. .23 O
ATOM 4385 0 HOH w 125 79. ,393 54. , 321 37. .906 1. , 00 37. .60 O
ATOM 4386 0 HOH w 126 94. , 507 47. ,383 26. .763 1. , 00 23. .32 O
ATOM 4387 0 HOH w 127 84. , 879 52. , 791 7. .568 1. , 00 17. .75 0
ATOM 4388 0 HOH w 128 55. , 781 62. , 401 29. .883 1. , 00 18. .64 O
ATOM 4389 0 HOH w 129 110. , 793 64. ,343 1. .131 1. , 00 31. .33 O
ATOM 4390 O HOH W 130 74 ,.853 47., 091 10..115 1., 00 25..36 O
ATOM 4391 O HOH w 131 68 , .626 48. , 003 15. .032 1. , 00 19. .30 O
ATOM 4392 O HOH w 132 52 , .375 86. , 184 10. .402 1. , 00 20. .12 O
ATOM 4393 O HOH w 133 61. .832 62. .184 10. .783 1. , 00 14. .63 O
ATOM 4394 O HOH w 134 59 , .866 87. , 352 20. .211 1. , 00 18. .08 O
ATOM 4395 O HOH w 135 53 , .092 75. , 022 28. .267 1. , 00 29. .34 O
ATOM 4396 O HOH w 136 71 , .633 51. , 059 32. .204 1. , 00 27. .11 O
ATOM 4397 O HOH w 137 65 , .108 44. , 264 19. .073 1. , 00 15. .72 O
ATOM 4398 O HOH w 138 94 , .400 49. , 865 27. .884 1. , 00 30. .99 O
ATOM 4399 O HOH w 139 50 , .035 63. , 741 26. .686 1. , 00 25. .35 O
ATOM 4400 O HOH w 140 46 , .992 72. , 601 19. .528 1. , 00 23. .56 O
ATOM 4401 O HOH w 141 58 , .833 62. , 898 33. .585 1. , 00 24. .60 O
ATOM 4402 O HOH w 142 44 , .590 64. , 528 17. .631 1. , 00 22. .37 O
ATOM 4403 O HOH w 143 111 , .934 67. , 262 22. .863 1. , 00 21. .36 O
ATOM 4404 O HOH w 144 120 , .388 70. ,373 16. .578 1. , 00 14. .72 O
ATOM 4405 O HOH w 145 79 , .376 74. , 934 5. .640 1. , 00 26. .11 O
ATOM 4406 O HOH w 146 90 , .390 44. , 528 20. .624 1. , 00 19. .00 O
ATOM 4407 O HOH w 147 76 , .952 72. , 281 2. .898 1. , 00 17. .25 O
ATOM 4408 0 HOH w 148 62 , .874 55. , 052 36. .871 1. , 00 21. .58 O
ATOM 4409 0 HOH w 149 109. .178 61. , 001 0. .007 0. , 50 12. .71 O
ATOM 4410 0 HOH w 150 56 , .846 65. , 884 26. .764 1. , 00 16. .72 O
ATOM 4411 0 HOH w 151 71 , .021 82. ,380 33. .675 1. , 00 35. .41 O
ATOM 4412 0 HOH w 152 87, .683 55. , 474 36. .044 1. , 00 25. .03 O
ATOM 4413 0 HOH w 153 123. .483 60. , 711 3. .281 1. , 00 21. .36 O
ATOM 4414 0 HOH w 154 120 , .007 74. , 411 17. .561 1. , 00 23. .51 O
ATOM 4415 0 HOH w 155 104 , .565 50. , 945 16. .980 1. , 00 26. .60 O
ATOM 4416 0 HOH w 156 50. .788 68. , 696 12. .297 1. , 00 13. .87 O
ATOM 4417 0 HOH w 157 64 , .965 81. , 214 5. .914 1. , 00 19. .59 O
ATOM 4418 0 HOH w 158 77 , .322 74. , 978 4. .042 1. , 00 30. .33 O
ATOM 4419 0 HOH w 159 90 , .308 65. , 171 -2. .317 1. , 00 25. .76 O
ATOM 4420 0 HOH w 160 60 , .365 48. , 834 20. .546 0. , 50 16. .06 0
ATOM 4421 0 HOH w 161 106 , .038 63. , 576 23. .717 1. , 00 17. .05 O
ATOM 4422 0 HOH w 162 72 , .774 83. , 675 31. .704 1. , 00 24. .11 O
ATOM 4423 0 HOH w 163 115 , .099 80. , 456 11. .855 1. , 00 21. .57 O
ATOM 4424 0 HOH w 164 83 , .838 59. , 796 3. .811 1. , 00 32. .77 0
ATOM 4425 0 HOH w 165 96 , .519 81. , 214 9. .486 1. , 00 24. .49 O
ATOM 4426 0 HOH w 166 110 , .093 46. , 455 13. .328 1. , 00 25. .75 O
ATOM 4427 0 HOH w 167 88 , .789 80. , 875 13. .101 1. , 00 22. .74 O
ATOM 4428 0 HOH w 168 93 , .993 42. , 594 18. .292 1. , 00 35. .15 0
ATOM 4429 0 HOH w 169 114 , .470 62. , 504 3. .385 1. , 00 17. .97 O
ATOM 4430 0 HOH w 170 78 , .601 75. , 108 -3. .396 1. , 00 18. .66 O
ATOM 4431 0 HOH w 171 93 , .427 74. , 507 26. .479 1. , 00 18. .67 O
ATOM 4432 0 HOH w 172 97 , .843 81. , 288 16. .078 1. , 00 23. .16 0
ATOM 4433 0 HOH w 173 54 , .469 71. ,301 27. .298 1. , 00 15. .33 O
ATOM 4434 0 HOH w 174 93 , .660 76. ,333 5. .563 1. , 00 17. .81 O
ATOM 4435 O HOH W 175 84., 853 68 ,.817 -2..631 1., 00 22..36 O
ATOM 4436 O HOH w 176 121. , 180 70 , .239 19. .170 1. , 00 29. .32 O
ATOM 4437 O HOH w 177 108. , 298 83 , .418 12. .138 1. , 00 25. .06 O
ATOM 4438 O HOH w 178 83. .712 78 , .028 32. .000 1. , 00 20. .46 O
ATOM 4439 O HOH w 179 67. , 141 81 , .712 7. .552 1. , 00 21. .89 O
ATOM 4440 O HOH w 180 62. , 541 68 , .408 33. .461 1. , 00 29. .28 O
ATOM 4441 O HOH w 181 65. , 408 70 , .134 3. .389 1. , 00 27. .44 O
ATOM 4442 O HOH w 182 113. , 444 63 , .788 0. .119 1. , 00 33. .32 O
ATOM 4443 O HOH w 183 99. , 202 79 , .019 23. .664 1. , 00 33. .88 O
ATOM 4444 O HOH w 184 81. , 811 56 , .321 8. .309 1. , 00 18. .01 O
ATOM 4445 O HOH w 185 104. , 695 48 , .686 1. .991 1. , 00 29. .99 O
ATOM 4446 O HOH w 186 91. , 166 47 , .945 8. .668 1. , 00 22. .78 O
ATOM 4447 O HOH w 187 76. , 861 76 , .447 -1. .977 1. , 00 29. .90 O
ATOM 4448 0 HOH w 188 96. ,399 56 , .342 -4. .528 1. , 00 36. .18 O
ATOM 4449 0 HOH w 189 121. , 474 73 , .020 15. .923 1. , 00 25. .26 O
ATOM 4450 0 HOH w 190 75. , 992 79 , .362 8. .074 1. , 00 15. .32 O
ATOM 4451 0 HOH w 191 54. , 002 55 , .741 33. .109 1. , 00 27. .06 O
ATOM 4452 0 HOH w 192 57. , 016 67, .916 7. .091 1. , 00 27. .85 O
ATOM 4453 0 HOH w 193 65. , 809 67, .983 6. .395 1. , 00 26. .22 O
ATOM 4454 0 HOH w 194 66. , 750 47 , .235 25. .205 1. , 00 21. .34 O
ATOM 4455 0 HOH w 195 95. , 184 73 , .954 0. .422 1. , 00 22. .08 O
ATOM 4456 0 HOH w 196 75. ,333 75 , .333 -0. .000 0. , 50 13. .56 O
ATOM 4457 0 HOH w 197 81. , 217 56 , .678 3. .652 1. , 00 37. .14 O
ATOM 4458 0 HOH w 198 125. , 030 60 , .203 11. .952 1. , 00 17. .94 O
ATOM 4459 0 HOH w 199 76. , 598 89 , .434 12. .516 1. , 00 22. .60 O
ATOM 4460 0 HOH w 200 112. , 635 73 , .309 24. .326 1. , 00 20. .41 O
ATOM 4461 0 HOH w 201 84. , 628 74 , .145 37. .911 1. , 00 29. .11 0
ATOM 4462 0 HOH w 202 72. , 195 61 , .400 12. .843 1. , 00 21. .93 0
ATOM 4463 0 HOH w 203 102. , 947 60 , .372 -2. .389 1. , 00 19. .32 O
ATOM 4464 0 HOH w 204 82. , 537 67, .759 41. .630 1. , 00 21. .13 O
ATOM 4465 0 HOH w 205 69. , 650 90 , .264 20. .249 1. , 00 25. .92 0
ATOM 4466 0 HOH w 206 75. , 713 82 , .524 5. .830 1. , 00 19. .46 O
ATOM 4467 0 HOH w 207 47. , 260 68 , .787 16. .787 1. , 00 23. .79 O
ATOM 4468 0 HOH w 208 57. , 044 54 , .098 29. .348 1. , 00 27. .99 O
ATOM 4469 0 HOH w 209 101. , 703 64 , .874 29. .441 1. , 00 35. .49 0
ATOM 4470 0 HOH w 210 87. , 050 58 , .665 37. .767 1. , 00 28. .29 O
ATOM 4471 0 HOH w 211 84. , 684 88 , .336 19. .151 1. , 00 28. .61 O
ATOM 4472 0 HOH w 212 73. , 249 79 , .393 37. .007 1. , 00 36. .47 O
ATOM 4473 0 HOH w 213 59. , 038 83 , .563 5. .692 1. , 00 35. .37 0
ATOM 4474 0 HOH w 214 67. ,372 56 , .095 9. .127 1. , 00 31. .02 O
ATOM 4475 0 HOH w 215 83. , 573 74 , .823 -2. .801 1. , 00 19. .22 O
ATOM 4476 0 HOH w 216 68. , 447 47 , .207 26. .924 1. , 00 42. .51 0
ATOM 4477 0 HOH w 217 81. , 010 91 , .223 14. .982 1. , 00 30. .59 0
ATOM 4478 0 HOH w 218 87. ,303 68 , .371 -2. .354 1. , 00 27. .16 O
ATOM 4479 0 HOH w 219 110. , 154 50 , .325 17. .049 1. , 00 47. .88 O
ATOM 4480 O HOH W 220 60., 787 73 ,.335 -1..445 1., 00 24..21 O
ATOM 4481 O HOH w 221 123. , 179 57 , .853 6. .627 1. , 00 26. .28 O
ATOM 4482 O HOH w 222 50. , 620 61 , .766 13. .066 1. , 00 28. .43 O
ATOM 4483 O HOH w 223 94. .515 48. .871 -1. .271 1. , 00 39. .99 O
ATOM 4484 O HOH w 224 65. , 325 42 , .750 11. .974 1. , 00 26. .08 O
ATOM 4485 O HOH w 225 90. , 620 42 , .752 14. .084 1. , 00 28. .44 O
ATOM 4486 O HOH w 226 97. , 230 50 , .969 1. .379 1. , 00 26. .27 O
ATOM 4487 O HOH w 227 87. , 263 60 , .639 -1. .472 1. , 00 22. .95 O
ATOM 4488 O HOH w 228 71. , 204 48 , .313 26. .160 1. , 00 37. .66 O
ATOM 4489 O HOH w 229 47. , 164 73 , .476 16. .870 1. , 00 27. .02 O
ATOM 4490 O HOH w 230 57. , 026 82 , .251 5. .185 1. , 00 35. .48 O
ATOM 4491 O HOH w 231 121. , 634 72 , .538 11. .669 1. , 00 22. .09 O
ATOM 4492 O HOH w 232 63. , 830 58 , .417 37. .824 1. , 00 22. .78 O
ATOM 4493 O HOH w 233 109. , 779 64, .696 25. .610 1. , 00 27. .71 O
ATOM 4494 O HOH w 234 127. , 112 62 , .797 3. .298 1. , 00 21. .77 O
ATOM 4495 O HOH w 235 92. , 909 70 , .007 29. .855 1. , 00 19. .69 O
ATOM 4496 O HOH w 236 53. , 860 79 , .833 30. .827 1. , 00 25. .84 O
ATOM 4497 0 HOH w 237 76. , 482 90 , .500 18. .681 1. , 00 25. .12 O
ATOM 4498 0 HOH w 238 90. , 052 68 , .736 1. .081 1. , 00 19. .11 O
ATOM 4499 0 HOH w 239 86. , 447 86 , .342 17. .699 1. , 00 49. .63 O
ATOM 4500 0 HOH w 240 96. , 085 44 , .177 17. .449 1. , 00 42. .22 O
ATOM 4501 0 HOH w 241 66. , 922 49 , .221 7. .857 1. , 00 47. .05 O
ATOM 4502 0 HOH w 242 106. , 498 61 , .383 25. .442 1. , 00 28. .21 O
ATOM 4503 0 HOH w 243 100. , 486 45 , .422 4. .123 1. , 00 27. .75 O
ATOM 4504 0 HOH w 244 58. ,307 51 , .452 28. .048 1. , 00 35. .59 O
ATOM 4505 0 HOH w 245 85. , 984 79 , .272 1. .854 1. , 00 26. .64 O
ATOM 4506 0 HOH w 246 84. .234 69 , .367 -5. .526 1. , 00 29. .58 0
ATOM 4507 0 HOH w 247 87. , 115 52 , .747 35. .439 1. , 00 34. .73 0
ATOM 4508 0 HOH w 248 70. , 792 87, .469 26. .838 1. , 00 26. .97 O
ATOM 4509 0 HOH w 249 96. , 459 49 , .952 5. .974 1. , 00 21. .53 O
ATOM 4510 0 HOH w 250 47. , 767 70 , .550 13. .770 1. , 00 29. .95 0
ATOM 4511 0 HOH w 251 86. , 560 73 , .668 39. .293 1. , 00 47. .51 O
ATOM 4512 0 HOH w 252 45. , 832 61 , .785 15. .244 1. , 00 21. .66 O
ATOM 4513 0 HOH w 253 108. , 580 44 , .328 14. .260 1. , 00 35. .28 O
ATOM 4514 0 HOH w 254 95. , 902 46 , .926 2. .969 1. , 00 34. .16 0
ATOM 4515 0 HOH w 255 56. , 550 63 , .161 32. .245 1. , 00 37. .13 O
ATOM 4516 0 HOH w 256 60. , 187 76 , .165 -0. .446 1. , 00 35. .06 O
ATOM 4517 0 HOH w 257 72. , 684 70 , .326 36. .840 1. , 00 25. .08 O
ATOM 4518 0 HOH w 258 54. , 750 60 , .357 10. .750 1. , 00 18. .22 0
ATOM 4519 0 HOH w 259 52. , 763 72 , .610 5. .100 1. , 00 50. .83 O
ATOM 4520 0 HOH w 260 117. , 089 79 , .182 5. .585 1. , 00 22. .93 O
ATOM 4521 0 HOH w 261 103. , 172 74 , .524 24. .878 1. , 00 41. .61 0
ATOM 4522 0 HOH w 262 72. , 037 43 , .960 19. .954 1. , 00 26. .38 0
ATOM 4523 0 HOH w 263 79. , 804 50 , .175 7. .642 1. , 00 31. .24 O
ATOM 4524 0 HOH w 264 84. , 826 46 , .412 11. .054 1. , 00 32. .09 O
ATOM 4525 O HOH W 265 98., 104 77., 912 6..547 1., 00 30..32 O
ATOM 4526 O HOH w 266 62. , 993 82. , 801 7. .209 1. , 00 17. .64 O
ATOM 4527 O HOH w 267 67. , 160 45. , 671 23. .036 1. , 00 17. .48 O
ATOM 4528 O HOH w 268 88. .333 80. , 250 27. .199 1. , 00 21. .70 O
ATOM 4529 O HOH w 269 65. , 119 65. , 664 5. .972 1. , 00 42. .90 O
ATOM 4530 O HOH w 270 97. ,353 78. , 743 9. .238 1. , 00 21. .99 O
ATOM 4531 O HOH w 271 73. , 949 91. ,306 18. .938 1. , 00 36. .21 O
ATOM 4532 O HOH w 272 91. , 649 76. , 339 26. .681 1. , 00 22. .47 O
ATOM 4533 O HOH w 273 69. , 562 90. , 168 17. .110 1. , 00 32. .94 O
ATOM 4534 O HOH w 274 125. , 965 68. , 961 22. .198 1. , 00 53. .30 O
ATOM 4535 O HOH w 275 89. , 524 60. , 937 -4. .011 1. , 00 36. .66 O
ATOM 4536 O HOH w 276 73. , 712 71. , 887 25. .613 1. , 00 11. .78 O
ATOM 4537 O HOH w 277 71. , 697 72. , 645 23. .573 1. , 00 11. .89 O
ATOM 4538 O HOH w 278 69. , 689 71. , 131 22. .527 1. , 00 9. .20 O
ATOM 4539 O HOH w 279 67. , 801 71. , 824 20. .327 1. , 00 11. .05 O
ATOM 4540 O HOH w 280 83. , 110 63. , 463 16. .520 1. , 00 14. .63 O
ATOM 4541 O HOH w 281 84. , 730 61. , 296 16. .825 1. , 00 13. .02 O
ATOM 4542 O HOH w 282 82. ,330 64. , 752 14. .305 1. , 00 11. .14 O
ATOM 4543 O HOH w 283 79. , 849 65. , 958 14. .376 1. , 00 12. .95 O
ATOM 4544 O HOH w 284 77. , 982 64. , 918 12. .545 1. , 00 9. .14 O
ATOM 4545 O HOH w 285 87. , 321 60. , 005 15. .505 1. , 00 10. .28 O
ATOM 4546 O HOH w 286 86. ,315 57. , 787 16. .752 1. , 00 7. .53 O
ATOM 4547 O HOH w 287 84. , 249 54. , 156 16. .640 1. , 00 12. .46 O
ATOM 4548 O HOH w 288 84. , 416 51. , 453 16. .932 1. , 00 13. .71 O
ATOM 4549 O HOH w 289 54. ,396 71. , 736 18. .322 1. , 00 9. .78 O
ATOM 4550 O HOH w 290 53. , 429 69. , 712 19. .975 1. , 00 14. .49 O
ATOM 4551 O HOH w 291 60. , 518 74. , 256 19. .629 1. , 00 12. .46 O
ATOM 4552 0 HOH w 292 61. , 215 72. , 779 21. .734 1. , 00 10. .28 O
ATOM 4553 0 HOH w 293 73. , 011 64. , 617 26. .681 1. , 00 11. .58 O
ATOM 4554 0 HOH w 294 70. , 799 63. , 140 27. .201 1. , 00 11. .06 O
ATOM 4555 0 HOH w 295 75. , 251 65. , 494 28. .347 1. , 00 10. .71 0
ATOM 4556 0 HOH w 296 70. , 115 59. ,305 30. .735 1. , 00 16. .03 O
ATOM 4557 0 HOH w 297 69. , 647 58. , 706 33. .452 1. , 00 10. .27 O
ATOM 4558 0 HOH w 298 99. , 000 55. , 080 10. .817 1. , 00 11. .69 O
ATOM 4559 0 HOH w 299 100. , 425 53. , 056 12. .524 1. , 00 12. .45 0
ATOM 4560 0 HOH w 300 66. , 242 65. , 428 14. .776 1. , 00 16. .43 O
ATOM 4561 0 HOH w 301 81. , 405 80. , 063 12. .880 1. , 00 12. .57 O
ATOM 4562 0 HOH w 302 79. , 412 80. ,396 11. .081 1. , 00 10. .27 O
ATOM 4563 0 HOH w 303 68. , 824 60. , 209 41. .465 1. , 00 31. .92 0
ATOM 4564 0 HOH w 304 59. , 552 55. , 604 29. .512 1. , 00 17. .67 O
ATOM 4565 0 HOH w 305 60. , 083 55. , 562 36. .754 1. , 00 33. .95 O
ATOM 4566 0 HOH w 306 69. , 237 44. ,346 23. .441 1. , 00 24. .40 O
ATOM 4567 0 HOH w 307 69. , 770 40. , 875 12. .372 1. , 00 26. .72 0
ATOM 4568 0 HOH w 308 95. , 206 62. , 760 30. .095 1. , 00 25. .02 O
ATOM 4569 0 HOH w 309 95. , 950 59. , 484 34. .012 1. , 00 27. .04 O
ATOM 4570 O HOH W 310 56 ,, 711 66 ,, 265 31., 461 1., 00 30.,50 O
ATOM 4571 O HOH w 311 55 , , 298 65, , 451 28. ,963 1. , 00 25. , 06 O
ATOM 4572 O HOH w 312 90 , , 647 71 , , 911 33. , 467 1. , 00 23. , 47 O
ATOM 4573 O HOH w 313 50. .077 73 , .625 22. .368 1. , 00 19. .98 O
ATOM 4574 O HOH w 314 49 , ,389 72 , .201 24. , 596 1. , 00 35. , 15 O
ATOM 4575 O HOH w 315 58 , , 934 84, .452 11. , 723 1. , 00 22. ,56 O
ATOM 4576 O HOH w 316 69 , , 713 73 , .707 -3. , 172 1. , 00 14. ,53 O
ATOM 4577 O HOH w 317 71 , .077 73 , .328 -0. .554 1. , 00 16. .15 O
ATOM 4578 O HOH w 318 64 , , 017 79 , .240 0. , 019 1. , 00 26. ,81 O
ATOM 4579 O HOH w 319 79 , , 020 83 , .776 31. , 548 1. , 00 29. , 75 O
ATOM 4580 O HOH w 320 85, , 533 76 , .663 33. ,378 1. , 00 30. , 19 O
ATOM 4581 O HOH w 321 73 , .452 69 , .182 7. .589 1. , 00 17. .09 O
ATOM 4582 O HOH w 322 71 , , 448 86 , .128 10. , 831 1. , 00 19. ,34 O
ATOM 4583 O HOH w 323 83 , , 503 89 , .728 21. , 490 1. , 00 22. ,84 O
ATOM 4584 O HOH w 324 87, , 715 86 , .894 21. , 851 1. , 00 23. ,54 O
ATOM 4585 O HOH w 325 79 , .121 60 , .500 5. .937 1. , 00 27. .41 O
ATOM 4586 O HOH w 326 86 , ,368 65, .385 0. ,562 1. , 00 27. ,35 O
ATOM 4587 O HOH w 327 95, , 444 50 , .430 8. , 518 1. , 00 15. ,26 O
ATOM 4588 O HOH w 328 94 , ,301 57 , .944 -3. ,286 1. , 00 29. ,04 O
ATOM 4589 0 HOH w 329 116 , .537 52 , .721 3. .638 1. , 00 27. .13 O
ATOM 4590 0 HOH w 330 76 , , 275 59 , .378 7. ,337 1. , 00 40. , 19 O
ATOM 4591 0 HOH w 331 70 , , 747 63 , .822 13. , 042 1. , 00 28. ,63 O
ATOM 4592 0 HOH w 332 84, , 440 47 , .986 34. , 116 1. , 00 18. , 43 O
ATOM 4593 0 HOH w 333 83 , .154 43 , .293 29. .434 1. , 00 25. .96 O
ATOM 4594 0 HOH w 334 98 , , 697 47 , .505 21. , 570 1. , 00 27. , 02 O
ATOM 4595 0 HOH w 335 102 , , 772 52 , .282 20. , 064 1. , 00 32. , 70 O
ATOM 4596 0 HOH w 336 110 , .632 58 , .836 4. .205 1. , 00 26. .47 0
ATOM 4597 0 HOH w 337 111 , , 849 61 , .745 3. , 152 1. , 00 44. ,94 O
ATOM 4598 0 HOH w 338 101 , , 125 49 , .787 -0. ,988 1. , 00 33. ,96 O
ATOM 4599 0 HOH w 339 117 , , 159 66 , .545 21. , 836 1. , 00 21. ,84 O
ATOM 4600 0 HOH w 340 120 , .244 56 , .609 12. .191 1. , 00 23. .32 0
ATOM 4601 0 HOH w 341 115 , , 170 51 , .278 14. , 257 1. , 00 30. , 44 O
ATOM 4602 0 HOH w 342 111 , , 491 80 , .833 3. , 165 1. , 00 29. , 82 O
ATOM 4603 0 HOH w 343 101 , , 953 82 , .610 10. , 773 1. , 00 25. , 43 O
ATOM 4604 0 HOH w 344 46 , .011 67 , .990 20. .238 1. , 00 19. .80 0
ATOM 4605 0 HOH w 345 115 , , 554 79 , .243 14. , 726 1. , 00 22. ,39 O
ATOM 4606 0 HOH w 346 87, , 673 44 , .927 20. , 726 1. , 00 20. , 86 O
ATOM 4607 0 HOH w 347 57 , , 443 78 , .585 2. ,560 1. , 00 23. , 83 O
ATOM 4608 0 HOH w 348 60 , .383 74 , .826 31. .955 1. , 00 23. .36 0
ATOM 4609 0 HOH w 349 108 , , 102 83 , .941 7. , 708 1. , 00 25. , 74 O
ATOM 4610 0 HOH w 350 86 , , 983 47 , .544 34. , 355 1. , 00 26. ,63 O
ATOM 4611 0 HOH w 351 82 , , 239 52 , .554 8. ,350 1. , 00 23. , 73 O
ATOM 4612 0 HOH w 352 79 , .115 47 , .275 26. .274 1. , 00 27. .40 0
ATOM 4613 0 HOH w 353 50 , , 652 70 , .037 24. , 709 1. , 00 25. , 78 O
ATOM 4614 0 HOH w 354 114 , , 054 65, .476 22. , 808 1. , 00 25. ,31 O
ATOM 4615 O HOH W 355 111., 136 48 ,, 957 4., 228 1., 00 26., 53 O
ATOM 4616 O HOH w 356 104. , 951 81 , , 706 2. , 014 1. , 00 23. ,31 O
ATOM 4617 O HOH w 357 117. , 608 69 , , 017 3. ,208 1. , 00 26. ,80 O
ATOM 4618 O HOH w 358 60. .255 54. , 205 15. .233 1. , 00 21. .92 O
ATOM 4619 O HOH w 359 125. , 742 61 , , 809 20. , 938 1. , 00 23. ,68 O
ATOM 4620 O HOH w 360 71. ,318 45 , , 857 21. , 866 1. , 00 28. , 16 O
ATOM 4621 O HOH w 361 64. ,336 48 , , 484 29. , 783 1. , 00 25. ,84 O
ATOM 4622 O HOH w 362 64. , 410 70. , 021 33. .218 1. , 00 28. .32 O
ATOM 4623 O HOH w 363 99. , 662 51 , , 728 26. , 853 1. , 00 25. ,37 O
ATOM 4624 O HOH w 364 57. , 619 85, , 004 23. , 472 1. , 00 26. , 78 O
ATOM 4625 O HOH w 365 109. , 513 85, , 026 14. , 107 1. , 00 31. , 48 O
ATOM 4626 O HOH w 366 71. , 356 78. ,874 7. .358 1. , 00 27. .32 O
ATOM 4627 O HOH w 367 56. , 265 89 , , 128 21. ,290 1. , 00 31. , 78 O
ATOM 4628 O HOH w 368 115. , 927 67, , 059 -0. ,659 1. , 00 28. ,23 O
ATOM 4629 O HOH w 369 86. ,347 87, , 208 6. ,215 1. , 00 28. ,36 O
ATOM 4630 O HOH w 370 100. , 399 47. ,343 16. .164 1. , 00 28. .80 O
ATOM 4631 O HOH w 371 125. , 082 66 , , 017 15. , 408 1. , 00 29. , 06 O
ATOM 4632 O HOH w 372 106. , 891 75 , , 678 3. , 454 1. , 00 25. ,59 O
ATOM 4633 O HOH w 373 88. , 447 43 , , 131 24. ,920 1. , 00 29. ,14 O
ATOM 4634 O HOH w 374 48. , 419 79. , 853 13. .172 1. , 00 26. .52 O
ATOM 4635 O HOH w 375 47. , 642 75 , , 779 22. ,363 1. , 00 34. , 45 O
ATOM 4636 O HOH w 376 114. , 861 82 , , 439 9. , 087 1. , 00 35. ,35 O
ATOM 4637 O HOH w 377 74. , 958 81 , , 956 35. ,600 1. , 00 28. ,35 O
ATOM 4638 O HOH w 378 Ill . , 202 79. , 501 1. .078 1. , 00 32. .07 O
ATOM 4639 O HOH w 379 97. , 594 63 , , 105 29. ,216 1. , 00 30. , 45 O
ATOM 4640 O HOH w 380 89. , 237 84, , 051 11. ,598 1. , 00 28. ,28 O
ATOM 4641 O HOH w 381 97. , 163 60. , 858 31. .713 1. , 00 37. .20 0
ATOM 4642 0 HOH w 382 Ill . , 105 76 , , 736 1. , 164 1. , 00 37. ,60 O
ATOM 4643 0 HOH w 383 89. , 108 80 , , 551 20. , 138 1. , 00 31. ,32 O
ATOM 4644 0 HOH w 384 109. , 208 54 , , 650 0. , 012 0. , 50 20. , 78 O
ATOM 4645 0 HOH w 385 56. , 533 53. , 431 9. .966 1. , 00 29. .03 0
ATOM 4646 0 HOH w 386 70. , 988 87, , 241 12. ,976 1. , 00 30. , 02 O
ATOM 4647 0 HOH w 387 59. , 625 58 , , 605 35. , 175 1. , 00 33. , 71 O
ATOM 4648 0 HOH w 388 86. , 544 43 , , 124 22. , 806 1. , 00 33. ,84 O
ATOM 4649 0 HOH w 389 87. ,274 64. , 588 39. .751 1. , 00 33. .06 0
ATOM 4650 0 HOH w 390 72. , 877 87, , 061 6. , 624 1. , 00 32. ,33 O
ATOM 4651 0 HOH w 391 71. , 897 80 , , 278 5. ,628 1. , 00 26. , 70 O
ATOM 4652 0 HOH w 392 69. , 516 42 , , 838 9. , 000 1. , 00 32. ,63 O
ATOM 4653 0 HOH w 393 109. , 629 83. , 070 9. .904 1. , 00 30. .86 0
ATOM 4654 0 HOH w 394 78. , 125 92 , , 023 24. , 704 1. , 00 30. ,52 O
ATOM 4655 0 HOH w 395 78. , 953 91 , , 893 22. ,367 1. , 00 27. ,36 O
ATOM 4656 0 HOH w 396 77. , 655 92 , , 424 20. , 194 1. , 00 27. , 78 O
ATOM 4657 0 HOH w 397 95. , 558 64. , 394 32. .011 1. , 00 41. .48 0
ATOM 4658 0 HOH w 398 68. , 942 73 , , 097 35. ,691 1. , 00 31. ,00 O
ATOM 4659 0 HOH w 399 123. , 235 58 , , 210 19. ,901 1. , 00 37. , 19 O
ATOM 4660 0 HOH W 400 81 ,.026 75., 783 39..765 1 00 33.48 O
ATOM 4661 0 HOH w 401 89 , .218 44. , 167 31. .825 1 00 29.29 O
ATOM 4662 0 HOH w 402 75 , .739 46. , 600 22. .208 1 00 32.70 O
ATOM 4663 0 HOH w 403 93. .604 58. .674 35. .287 1 00 33.41 O
ATOM 4664 0 HOH w 404 114 , .143 53. ,316 2. .795 1 00 36.89 O
ATOM 4665 0 HOH w 405 62 , .314 77. , 901 -1. .690 1 00 31.79 O
ATOM 4666 0 HOH w 406 119 , .132 74. , 672 20. .486 1 00 38.01 O
ATOM 4667 0 HOH w 407 89 , .827 72. , 251 40. .058 1 00 37.03 O
ATOM 4668 0 HOH w 408 62 , .078 85. , 872 25. .358 1 00 38.89 O
ATOM 4669 0 HOH w 409 66 , .149 86. , 250 27. .163 1 00 39.99 O
ATOM 4670 0 HOH w 410 111 , .238 48. , 481 15. .078 1 00 38.99 O
ATOM 4671 0 HOH w 411 122 , .333 65. , 281 8. .119 1 00 26.63 O
ATOM 4672 0 HOH w 412 104 , .767 59. ,363 26. .560 1 00 33.01 O
ATOM 4673 0 HOH w 413 91 , .741 81. , 149 13. .569 1 00 36.22 O
Claims
1 . An isolated nucleic acid comprising the nucleotide sequence of SEQ I D NO: 1 or of a degenerate variant of SEQ ID NO: 1 .
2. An isolated nucleic acid comprising a sequence that encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3.
3. An isolated nucleic acid comprising a sequence that hybridizes under stringent conditions to a hybridization probe the nucleotide sequence of which consists of SEQ I D NO: 1 , or the complement of SEQ ID NO: 1 .
4. An isolated nucleic acid comprising a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ ID NO: 1 .
5. The isolated nucleic acid of claim 4 wherein the nucleic acid encodes a polypeptide that has starch hydrolysis activity.
6. An isolated nucleic acid comprising a sequence that encodes a polypeptide at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ I D NO: 3, wherein the polypeptide has starch hydrolysis activity.
7. An isolated nucleic acid comprising a sequence that encodes a polypeptide comprising the sequence of SEQ ID NO: 3, or SEQ ID NO: 3 with up to 50 conservative amino acid substitutions, wherein the polypeptide has starch hydrolysis activity.
8. A purified polypeptide, the amino acid sequence of which comprises a sequence at least 66%, 67%, 68%, 69%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% identical to SEQ I D NO: 3.
9. A purified polypeptide comprising the amino acid sequence of SEQ I D NO: 3, but with 0 to 20 conservative amino acid substitutions.
10. An expression vector comprising the nucleic acid sequence of any of claims 1 -6 operably linked to an expression control sequence.
11 . A cultured cell comprising the vector of claim 10.
12. A cultured cell comprising the nucleic acid of any of claims 1 -6 operably linked to an expression control sequence.
13. A cultured cell tranfected with the vector of claim 10, or a progeny of said cell, wherein the cell expresses the nucleic acid to form a polypeptide.
14. A method of producing a protein, the method comprising culturing the cell of claim 1 1 under conditions permitting expression of the polypeptide.
15. A method of using the polypeptide of any of claims 8 or 9, the method comprising including the polypeptide in any of: starch liquefaction, starch saccharification, textile desizing, starch modification in the paper and pulp industry, brewing, baking, production of syrups for the food industry, production of feedstocks for fermentation processes, animal feed, and, removal of starchy soils and/or stains during dishwashing and/or laundry washing.
16. A composition comprising the polypeptide of any of claims 8 or 9, and at least one accessory enzyme selected from the group consisting of phytase, protease, puiiuianase, β-amylase, isoamylase, a different amylase, alpha-glucosidase, cellulase, xylanase, hemicellulase, beta-glucosidase, transferase, pectinase, lipase, cutinase, esterase, choline oxidases, peroxidases/oxidases, pectate lyases, mannanases, cutinases, laccases, phospholipases, lysophospholipases, acyltransferases, perhydrolases, arylesterases, and redox enzymes.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP13811646.2A EP2922951B1 (en) | 2012-11-20 | 2013-11-15 | Amylase with maltogenic properties |
US14/441,617 US20180112203A1 (en) | 2012-11-20 | 2013-11-15 | Amylase with maltogenic properties |
CN201380059512.9A CN104822829A (en) | 2012-11-20 | 2013-11-15 | Amylase with maltogenic properties |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNPCT/CN2012/084883 | 2012-11-20 | ||
CN2012084883 | 2012-11-20 |
Publications (1)
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WO2014081622A1 true WO2014081622A1 (en) | 2014-05-30 |
Family
ID=49876966
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2013/070237 WO2014081622A1 (en) | 2012-11-20 | 2013-11-15 | Amylase with maltogenic properties |
Country Status (3)
Country | Link |
---|---|
US (1) | US20180112203A1 (en) |
EP (1) | EP2922951B1 (en) |
WO (1) | WO2014081622A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10017751B2 (en) * | 2014-12-05 | 2018-07-10 | Godo Shusei Co., Ltd. | Heat-resistant isoamylase |
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