WO2006037655A2 - Bioactive compositions, natural methods of producing them and computational methods for designing natural production processes - Google Patents
Bioactive compositions, natural methods of producing them and computational methods for designing natural production processes Download PDFInfo
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- WO2006037655A2 WO2006037655A2 PCT/EP2005/010837 EP2005010837W WO2006037655A2 WO 2006037655 A2 WO2006037655 A2 WO 2006037655A2 EP 2005010837 W EP2005010837 W EP 2005010837W WO 2006037655 A2 WO2006037655 A2 WO 2006037655A2
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/74—Rubiaceae (Madder family)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/168—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- bioactive molecules occur naturally in food stuffs as part of larger precursor molecules. Because they are components of foods that are commonly eaten, such agents have the potential to be less toxic to humans and animals. If methods could be found to release these compounds from their precursor food sources these molecules could be utilized to enrich foods or could be isolated and used as nutritional or therapeutic agents.
- PYY is a high affinity positive agonist of Y2 G-protein- cpupled. receptors (GPCR ⁇ _ and__representS-_a. xelati ⁇ ely_ne.w_class__ of. iherapeutic treatment for obesity, among other diseases. It is a natural hormone produced by specialized endocrine L-cells in the gut in proportion to the calorie content of a meal. PYY operates by reducing appetite and food intake by modulating appetite circuits in the hypothalamus. The agent has been shown to reduce caloric intake by 30% two hours after subjects, either obese or nonobese, received a 90-minute intravenous infusion. These subjects also experienced a significant decrease in their cumulative 24-hour caloric intake, hi other studies, obese individuals have been observed to have lower levels of circulating PYY.
- the present invention provides methods for the isolation of naturally occurring bioactive agents such as PYY and is directed to the resulting inventive compositions themselves. It also covers processes for developing these methods.
- the present invention provides methods for producing functional foods enriched in functional biomo'lecules, procedures for developing those methods and the compositions prepared by those methods.
- the procedure first involves identifying the functional biomolecules to be generated. Attributes within these biomolecules that are necessary for function are then identified by analyzing the relationship between the biomolecule's structure and function. Precursor molecules containing this consensus motif, with all of the chemical and structural attributes required for activity, are then identified by electronically searching genomic databases of a variety of precursor food sources. Once a potential precursor molecule and food source is identified, another database search is carried out to identify organisms or enzymes that can be used to release from the precursor molecule a smaller compound containing all the attributes identified as necessary for function.
- the functional molecule can then be generated by treating the precursor with the processing agent(s) and releasing the functional biomolecule from its precursor.
- Functional molecules include bioactive molecules or biomaterials wherein the functions are bioactivity and self-organization and assembly, respectively.
- Precursor food sources include food stuffs or food grade proteins and their mixtures.
- the invention provides a method for generating a bioactive molecule from a precursor food or protein source.
- the method includes the steps of identifying a bioactive molecule, identifying the attributes within the bioactive molecule that are necessary for bioactivity, identifying a precursor food source that contains a precursor molecule with the attributes necessary for bioactivity, identifying an agent that can be used to release from the precursor molecule a compound comprising the attributes necessary for bioactivity, and treating the precursor food source with the processing agent to thereby release from the precursor molecule a molecule that contains the attributes necessary for bioactivity.
- the bioactive molecule can be a peptide, protein or nucleic acid polymer.
- the method further comprises identifying chemical attributes within the bioactive molecule that are necessary for bioactivity.
- the method further comprises identifying chemical attributes within a biomaterial that are necessary for self-organization/assembly into higher order structures.
- the method further comprises identifying topological (two dimensional) and/or structural (three dimensional) attributes within the bioactive molecule that are necessary for bioactivity or self-organization/assembly.
- the precursor food source can be derived from a plant such as rice, soy, maize, potato, coffee, milk, meat, and the like.
- the method involves preparing a biomolecule using an agent that is a cell and the cell can be a lactobacteria, including Lactobacillus johnsonii (LaI) or bifidobacteria, such as Bifidobacterium longum (B129) for example.
- LaI Lactobacillus johnsonii
- B129 Bifidobacterium longum
- the method involves preparing a biomolecule using an agent that is an enzyme.
- the method involves preparing a biomolecule using an agent that is an enzyme such as a protease, glycosidase, nuclease, oxidase or lipase or- their- combinations- — — - —
- an enzyme such as a protease, glycosidase, nuclease, oxidase or lipase or- their- combinations- — — - —
- biomolecule binds to a receptor.
- Certain embodiments of the invention are directed to a bioactive agent comprising a peptide generated from a food stuff or food grade protein comprising the amino acid sequence LNLV[TS][RK]X[RK][YFW], where X can be any naturally occurring amino acid and brackets denote the logical OR operation, and F, H, K, L, Q, R, W, Y are the standard amino acid abbreviations.
- the functional agent is isolated from a food stuff and the food stuff is rice.
- the food stuff is Oryza sativa.
- the bioactive agent is a peptide or protein that includes the sequence YSCRYFGYLVSKKKY (SEQ ID No. 1) derived from Arabidopsis thaliana amylogenin protein RGP or its closest analogue peptide sequences from the edible plant Oryza sativa, HSCRYFGYLVSRKKY (SEQ ID No. 2) found in the protein RGP2, or SACRCFGYMVSKKKY (SEQ ID No. 3) in the protein RGPl, or FDGVDFSEPLTRARF (SEQ ID No. 4) in the BiP protein.
- SEQ ID No. 1 derived from Arabidopsis thaliana amylogenin protein RGP or its closest analogue peptide sequences from the edible plant Oryza sativa
- HSCRYFGYLVSRKKY SEQ ID No. 2
- SACRCFGYMVSKKKY SEQ ID No. 3
- FDGVDFSEPLTRARF SEQ ID No. 4
- the bioactive agent is a peptide or protein that includes the sequence VWEKPWMDFK (SEQ ID No. 5), PWMDFK (SEQ ID No. 6), PWMDFKELQEFK (SEQ ID NO. 7), PWMDF (SEQ ID No. 8), or VWEKPWMDF (SEQ ID No. 9) all derived from Oryza sativa oryzacystatin.
- the functional agent is generated by releasing it from a precursor molecule.
- the functional agent is generated by releasing it from a precursor molecule wherein the precursor molecule is in rice.
- bioactive molecule and biomaterial are both considered to be functional biomolecules.
- the terms can be used interchangeably-so-long-as-their-unique-funetions-are kept4n-mind ⁇
- bioactive molecules are equally applicable to biomaterials.
- Biomaterials are generally considered to be molecules that are capable of self-organization and assembly.
- biomaterials can be peptides that can form filaments and fibrils, hydrogels, surfactants and peptide hybrids when released from their precursor molecules.
- the present invention provides methods for producing functional foods that are enriched in biomaterials or bioactive molecules such as bioactive peptides, proteins or nucleic acid polymers using natural food sources and food grade proteins, hi addition, the present invention relates to the foods so designed.
- the present invention is directed to methods for developing natural bioprocessing procedures for producing enriched foods upon treatment with naturally occurring biological molecules, processing activities present in select organisms or extracts of such organisms or other natural processing agents.
- the invention also covers the various food compositions that can be prepared by the procedures, in addition to the bioactive molecules and methods for their production.
- the method generally involves identifying a bioactive molecule for production, identifying the attributes within the bioactive molecule that are necessary for bioactivity, identifying a precursor food source that contains a precursor molecule with those attributes, identifying an agent or agents that can be used to release from the precursor molecule a compound comprising the attributes necessary for bioactivity. Once a suitable processing method is identified, the method can be carried out by treating the precursor food source with the processing agent(s) to cause the release of a molecule that contains the attributes necessary for bioactivity.
- the method also desirably includes steps for assessing the abundance of the potential precursor molecules.
- the method also desirably includes an assessment of the stability of the potential precursor proteins towards peptidases and other enzymes or treatments that can be used to release it.
- the functional agent can be any suitable biological molecule having a desired activity ⁇ especially-peptides,-proteins r nuclei& acid-polymers-in addition-to-their- - derivatives which can be with lipids, saccharides, peptides and nucleic acids or their combinations.
- Functional agents can be excised from food stuffs, including food grade proteins, or other agricultural sources by the present methods.
- the desired bioactive molecules will specifically bind to a target receptor and activate or deactivate the receptor at concentrations that are obtainable from food processing.
- concentrations of the functional agent are lower than desired, chosen functional agents can be concentrated in the food source.
- extracts of the processed food stuff having elevated concentrations of functional agents can be prepared.
- Purification methods can also be used to purify agents partially or to substantially pure form. Methods for preparing concentrates, extracts, and for purification of functional agents necessarily vary depending on the nature of the target functional agent but these methods are well known in the art and can be easily carried out by one of skill in the art.
- Precursor molecules can also be concentrated or purified and treated to release the bioactive molecules which can then be added back to food stuffs, as desired.
- a variety of methods can be used to identify the attributes within the bioactive molecule that are necessary for bioactivity. For example, a literature search can be used and information can be gathered to determine how variations in the peptide, protein or nucleic acid polymer structure affect the desired activity and to identify all conserved amino acids, or other structures such as carbohydrate structures, nucleotides, or lipids that exist in the peptide or protein active agent. In addition, variations can often be generated by mutagenesis and the activity of the resulting compounds determined. Ultimately, a consensus motif that is responsible for activity can be obtained. The consensus motif will desirably contain all of the attributes within the bioactive molecule that are necessary for bioactivity and defines the simplest molecular framework that is known to be active. The molecular framework can include the chemical attributes within the molecule that are required for activity and structural attributes such as primary, secondary and tertiary structural requirements, in addition to modifications, such as glycosylation, acylation and the like.
- [0034J-A- precursor— food source- that— contains-a— molecule— having- all— the attributes necessary for bioactivity can then be identified.
- Such food sources can be identified by searching for the occurrence of the consensus motif at both sequence and structural levels in large collections of genomic information including for example, ENSEMBL®, Nestle genomic databases, and public genomic sequencing projects on food raw materials such as rice, soy, maize, potato, coffee and bovine milk.
- Precursor molecules found within these food sources can be proteins, peptides or nucleic acid polymers and their derivatives, depending on the nature of the desired biomolecule.
- the agent is a peptide
- a search is performed using BLAST by searching for short nearly exact matches and/or by EMBOSS pattern matching module "patmatdb" by searching for short nearly exact matches.
- patterndb EMBOSS pattern matching module
- these structures can be found using structure prediction software, as is known in the art.
- a number of potential precursor molecules may be identified by this method. In such cases, it can be desirable to determine the abundance of the precursor molecules in order to identify precursors that will yield suitable amounts of the desired agent.
- a processing agent or agents can then be identified that can be used to release a compound from the precursor molecule such that the released compound contains all of the attributes necessary for bioactivity.
- the selected agents will, of course, depend on the nature of the functional agent to be released and the characteristics of the matrix in which it is found.
- Potential agents include microorganisms, extracts of microorganisms, proteolytic, glycolytic, nucleolytic, and lipolytic enzymes, oxidases, glycosidases, and chemical agents. In some instances these activities can be found in the genome repertoire of cells, such as probiotic bacteria.
- PeptideCutter the automated service offered by ExPASy website termed PeptideCutter
- the module DIGEST from the bioinformatics software package EMBOSS and/or the MEROPS protease database can be used to identify suitable enzyme activities.
- PeptideCutter knowledge-based -algorithm-ean-be-used-to-identify cleavage sites-produced- by- a panei-of-mor&-tiian ⁇ 20- - different proteases on protein sequences.
- Certain methods utilize microorganisms to release the functional agent from its matrix.
- the sequences of the enzymes thought to be useful can be compared to known bacterial genomes to identify similar protease sequences in those genomes.
- the Bifidobacterium longum (B129) genome contains 74 protein sequences annotated as proteases or peptidases. One of them is highly similar to an Arg-C proteinase for which cleavage outcomes are computationally predictable by the PeptideCutter model.
- B129 Bifidobacterium longum
- the Bifidobacterium longum (B129) genome also has a sequence highly similar to sedolisin. It can be used in situations where both activities would be useful in releasing the bioactive agent from its precursor food source. This method is equally applicable with other enzyme activities.
- probiotic bacterial strains can be identified and utilized without specific knowledge of the nature of the enzymes involved. At least with respect to proteases both intracellular and extracellular activity profiles have been reported. In a similar manner glycolytic, nucleolytic, and lipolytic activities can also be screened and the entire NCC bacterial collection evaluated to select the most promising strains.
- the method can be carried out by treating the food source with the processing agent(s) to release from the precursor molecules, molecules that contain the consensus motif.
- the food source is treated with agents that are sufficient to release the functional agent in a single fermentation or treatment.
- multiple processing agents are required and they are incompatible or require distinct environments for use, multiple processing steps can be undertaken.
- the functional agent can be derived from a protein or peptide sequence.
- Variable or noncritical amino acids and critical amino acids in the sequence can be identified by analysis of published structure-activity relationships and by aligning the sequences of all known similar sequences. With this consensus motif a database containing all known plant proteins can be searched to identify the consensus motif within larger precursor proteins.
- a computational assessment of the tertiary structure can also be used to identify internal sequences in the identified precursors that will adopt a structure that is required for activity of the bioactive agent.
- peptide sequences can be analyzed by a variety of known modeling methods to determine whether their predicted structures are likely to bind to the receptor. The list of precursor proteins is then evaluated. An analysis can then be done by known methods, such as microarrays for genes differentially expressed in seeds and crops, to determine if the precursor molecule is sufficiently abundant to enable the preparation of biologically relevant amounts of the functional agent from the precursor. In addition, the sensitivity of the precursor molecule to proteases can be determined in order to evaluate the potential for releasing the target bioactive agent. Peptide synthesis of identified peptide sequences can be carried out and the peptides can then be tested for activity.
- the generation of analogues to the bioactive peptide PYY3-36 can be selected.
- This compound is a ligand of the GPCR peptide hormone receptor Y, existing as subtypes Yl, Y2, Y4, and Y5.
- the receptor is involved in the regulation of satiety, the feeling of hunger.
- the PYY3-36 peptide activates its target receptors Yl and Y2 at concentrations of about 0.5 nM.
- the method would also be useful in the identification and generation of other regulatory peptides against diseases such as diabetes and obesity, such as cholecystokinin (CCK), human growth hormone (HGH), and melanocortin, for example.
- CCK cholecystokinin
- HGH human growth hormone
- melanocortin melanocortin
- examples of the present irtventiort can-no w-be-set-forths —
- PYY3-36 is a ligand of the GPCR peptide hormone receptor Y, existing as subtypes Yl, Y2, Y4, and Y5.
- the receptor is involved in the regulation of satiety, the feeling of hunger, and blood pressure. It activates its target receptors Yl and Y2 at concentrations of about 0.5 nM.
- NMR structure determination techniques have shown that the PYY3-36 peptide adopts a very particular three-dimensional shape known as the PP-turn type or fold.
- the N-terminal region is unstructured, while the C-terminal region forms a characteristic ⁇ -helical structure.
- peptides having similar sequences and tertiary structures were identified to find potential homologues to the natural PYY3-36 ligand peptide.
- Table 1 also identifies three potential bioactive peptides resulting from a search of known ESTs using the EMBOSS program. These sequences include two peptides from the protein Amylogenin, one from Arabidopsis thaliana (AtRGPT), the other from Oryza sativa (OsRGPl). The other potential peptide sequence was from a binding protein found in Oryza sativa (Oi 1 BiP).
- Amylogenin is thought to be responsible for starch biosynthesis in plants and is also known as reversibly glycosylated protein (RGP).
- RGP reversibly glycosylated protein
- the peptides derived from the proteins ⁇ tRGP2, OyRGPl and OsBiP were synthesized in 5 mg quantities at and purified to a purity of above about 90%. The peptides were tested in a competitive binding assay against p ⁇ 22 - 36 f or binding to the GPCR receptor.
- proteases that could release the p ⁇ 3"36 analogue.
- Arg- C proteinase- and sedolisirt were-identified as-having activities-that- could- release-the- bioactive agent from its native matrix.
- the sequences of the proteases used by PeptideCutter, MEROPS and Digest were compared to several bacterial genomes to check for the occurrence of highly similar sequences. For instance, in the Bifidobacterium longum genome 74 protein sequences are annotated as proteases or peptidases.
- the one derived from AtRGPl protein was able to inhibit binding of PYY 22'36 by 20-40% when 10 ⁇ M concentration of both the p ⁇ 22 - 36 an( j QyBip peptide were used in the binding assay. It is expected that the close analogue of the peptide derived from the Arabidopsis Thaliana protein ⁇ 4/RGPl found in the rice protein OsRGPl, differing by only one chemically equivalent amino acid, should have similar activity. Using this technique OsBIP protein was also shown to bind the Y2 receptor the Y2 receptor. In vivo tissue-based screening on cultures of rat colon cells can be carried out according to known protocols, such as described in Dumont et al., Eur. J. Pharmacol. 238, 37 (1993).
- CCK receptor subtype A is thought to be expressed in the gut and to be involved in the regulation of satiety, and the feeling of hunger.
- CCK receptor subtype B also known as gastrin receptor, is thought to be involved in the secretion of gastric acid, and in the development of pathological conditions, such as of gastric ulcer and cancer. Antagonists of the CCK-B receptor can be used to treat these conditions.
- a common strategy in the design of antagonists is to mimic a positive agonist and to introduce molecular structure changes that enhance binding to the cognate receptor such that the antagonist occupies the binding site thereby preventing agonist binding.
- An extensive literature search and sequence comparison was used to determine that only a small sequence in the C-terminal part of human CCK peptides is essential for receptor binding. A summary of this information is provided in Table 2.
- the amino acids in bold in Table 2 are highly conserved amino acids that constitute the simplest bioactive framework for CCK peptides.
- Table 2 also identifies two potential bioactive peptides from a search of known ESTs using the EMBOSS program. These two sequences include two fragment peptides from the protein oryzacystatin, from the organism Oryza sativa.
- Oryzacystatin is a cystein proteinase inhibitor protein. Analysis of literature demonstrated that this protein is produced in high quantities in rice crops, that could reach lmg of protein per kilogram of crops.
- the peptides VWEKPWMDFK, PWMDFK and PWMDFKELQEFK were synthesized in 5 mg quantities and purified to a purity of above about 90%. The peptides were tested in a competitive binding assay against CCK-8 for binding to the CCK-B GPCR receptor. Identifying target bioactive molecular frameworks. Search for adequate proteolytic enzymes: [0059] Commercial proteolytic enzymes such as trypsin or carboxypeptidase have been used to generate the active fragments from Oryza sativa rice. In vitro tests for bioactivity:
- Hydrolysates were generated from the recombinantly expressed and purified oryzacystatin protein which was isolated from Oryza sativa. Trypsin was used to hydrolyze 1 Omg of each sample using standard conditions. Two peptides were identified and purified from these hydrolysates, namely the VWEKPWMDFK and PWMDFK peptides, as these exhibited the highest activities in the first screen.
- hydrolysate and the two purified peptides were submitted to receptor-based screening,- The- hydrolysate- itself 4nhibited-GCK-& binding- to-the-GGK-B-reeeptor- by- 21% at a 100 ⁇ M hydrolysate concentration, while the purified peptides, VWEKPWMDFK and PWMDFK, inhibited CCK-8 binding by 16%, and 14% at a 10 ⁇ M concentrations.
- Trypsin digests were carried out by adding 96 mg urea to 200 ⁇ l oryzacystatin protein (2mg) and incubating at room temperature until the urea dissolves (final concentration: ca. 4 M urea).
- 900 ⁇ l of 10OmM ammonium bicarbonate in ImM CaCl 2 is added followed by 120 ⁇ l of acetonitrile.
- 40 ⁇ g of trypsin 40 ⁇ g Promega sequencing grade trypsin in Promega resuspension buffer
- a second aliquot of lO ⁇ g trypsin was added followed by incubation for 3 h at 37 0 C.
- 50% of the digestion solution of oryzacystatin is stored at -20°C for desalting by solid phase extraction using Cl 8 reverse phase material and 50% of the digestion solution of oryzacystatin is subjected to carboxypeptiodase digestion.
- Carboxypeptidase Y digestion was carried out on 720 ⁇ l (ca. lmg) of the tryptic digestion solution of oryzacystatin to which 1080 ⁇ l of 200 mM acetate buffer, pH 5 in 10% acetonitrile was added. lOO ⁇ g carboxypeptidase Y (Sigma Aldrich, carboxypeptidase Y solution was prepared three days before. It is good for one week according to Sigma Aldrich) is added and the solution incubated for 30 min at room temperaturefoUowed by the addition of 910 ⁇ l of 1% formic acid
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AU2005291359A AU2005291359B2 (en) | 2004-10-07 | 2005-10-07 | Bioactive compositions, natural methods of producing them and computational methods for designing natural production processes |
CN2005800386881A CN101056987B (en) | 2004-10-07 | 2005-10-07 | Bioactive compositions, natural methods of producing them and computational methods for designing natural production processes |
US11/576,902 US20070286935A1 (en) | 2004-10-07 | 2005-10-07 | Bioactive Compositions, Natural Methods of Producing Them and Computational Methods for Designing Natural Production Processes |
CA002583220A CA2583220A1 (en) | 2004-10-07 | 2005-10-07 | Bioactive compositions, natural methods of producing them and computational methods for designing natural production processes |
AT05796122T ATE491804T1 (en) | 2004-10-07 | 2005-10-07 | METHOD FOR PRODUCING BIOACTIVE COMPOSITIONS |
DE602005025381T DE602005025381D1 (en) | 2004-10-07 | 2005-10-07 | PROCESS FOR PREPARING BIOACTIVE COMPOSITIONS |
JP2007535106A JP2008515841A (en) | 2004-10-07 | 2005-10-07 | Bioactive compositions, natural methods for producing them, and computer methods for designing natural production processes |
EP05796122A EP1799838B1 (en) | 2004-10-07 | 2005-10-07 | Methods of producing bioactive compositions |
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EP0567661A1 (en) * | 1992-04-25 | 1993-11-03 | Societe Des Produits Nestle S.A. | Modified lipase, method of modification and applications |
EP0749694A1 (en) * | 1995-06-20 | 1996-12-27 | Societe Des Produits Nestle S.A. | Treatment enzymatic of cocoa |
WO1998043498A1 (en) * | 1997-04-01 | 1998-10-08 | Nichimo Co., Ltd. | Product comprising health-promotive ingredient and process for producing the same |
NZ532427A (en) * | 2001-09-24 | 2008-07-31 | Imp Innovations Ltd | Modification of feeding behavior using PYY (peptide YY) and its agonists such as NPY (neuropeptide Y) |
EP1358807A3 (en) * | 2002-05-03 | 2004-03-17 | Agro-korn a/s | Use of stillage from alcohol production as functional ingredients in food |
-
2005
- 2005-10-07 JP JP2007535106A patent/JP2008515841A/en not_active Ceased
- 2005-10-07 CA CA002583220A patent/CA2583220A1/en not_active Abandoned
- 2005-10-07 AU AU2005291359A patent/AU2005291359B2/en not_active Ceased
- 2005-10-07 DE DE602005025381T patent/DE602005025381D1/en active Active
- 2005-10-07 EP EP05796122A patent/EP1799838B1/en not_active Not-in-force
- 2005-10-07 CN CN2005800386881A patent/CN101056987B/en not_active Expired - Fee Related
- 2005-10-07 WO PCT/EP2005/010837 patent/WO2006037655A2/en active Application Filing
- 2005-10-07 US US11/245,562 patent/US20060210655A1/en not_active Abandoned
- 2005-10-07 AT AT05796122T patent/ATE491804T1/en not_active IP Right Cessation
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Also Published As
Publication number | Publication date |
---|---|
AU2005291359B2 (en) | 2011-10-13 |
CN101056987A (en) | 2007-10-17 |
ATE491804T1 (en) | 2011-01-15 |
EP1799838B1 (en) | 2010-12-15 |
CN101056987B (en) | 2012-08-29 |
US20060210655A1 (en) | 2006-09-21 |
DE602005025381D1 (en) | 2011-01-27 |
WO2006037655A3 (en) | 2007-01-04 |
JP2008515841A (en) | 2008-05-15 |
EP1799838A2 (en) | 2007-06-27 |
CA2583220A1 (en) | 2006-04-13 |
AU2005291359A1 (en) | 2006-04-13 |
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