KR20090038757A - Lipases and esterases isolated from deep sea sediment - Google Patents

Abstract

Provided is lipase and esterase isolated from deep sea mass-produced by genes coding the lipase and esterase. An esterase gene codes esterase having amino acid sequence of SEQ ID:NO 2, 4, 6, 10, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34. A method for preparing the esterase comprises the following steps of: cultivating a transformed cell with a recombinant vector; and isolating the esterase from the cultivated cells. The recombinant vector includes sequence number of 1, 3, 5, 9, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33. The lipase gene codes lipase having amino acid sequence of SEQ ID:NO 8 or 12. The method for preparing the lipase consists of the following steps of: cultivating a transformed cell with a recombinant vector; and isolating the lipase from the cultivated cells. The recombinant vector has sequence number of 7 or 11.

Description

Lipases and Esterases Isolated from Deep Sea Sediment

The present invention relates to novel lipases, genes encoding them and methods for mass production of lipases. The invention also relates to novel esterases, genes encoding them and methods of mass production of esterases.

Lipases (lipases, glycerol ester hydrolases) are enzymes that catalyze the hydrolysis or synthesis of a wide range of water-soluble or non-aqueous carboxylic acid esters or amides. Participates in various biocatalytic reactions such as alcoholysis, acidolysis, esterification, aminolysis, etc., maintains activity in organic solvents as well as aqueous solutions, and assists catalysis It is one of the high industrially applicable enzymes because it does not require a cofactor, exhibits substrate specificity for various substrates, and has isomer selectivity that shows activity for only one of the optical isomers. To date, many kinds of animals, plants and microorganisms have been known to produce lipases, and research into the biochemical properties of lipases and the acquisition of lipase genes has been carried out, especially food, detergents, fine chemicals and cosmetics. Researches to use lipases are being actively conducted throughout the industry, such as and environmental materials industry.

In the field of fine chemistry that synthesizes high value-added leading substances including pharmaceuticals, the synthesis of ester compounds by conventional chemical methods requires a lot of side reactions that consume energy and adversely affect the quality because they are synthesized at high temperatures and pressures. Not only does this occur, there is a difficulty in producing high-purity fine chemicals due to low conversion and purity of specific optical isomers. In recent years, in order to supplement these problems, the reaction using a lipase showing position specificity and optical activity specificity as a biocatalyst has been actively studied, but the application of the lipase has been limited due to the disadvantage of low activity at low temperatures.

In addition, lipase has been studied as an additive for detergents and bleaching agents because it has a function of facilitating the action of a surfactant by hydrolyzing fatty components with fatty acids or glycerol, which are well soluble in water, but the results of use are still poor. This is because of the disadvantage of poor activity of the lipase at low cleaning temperatures, poor or incomplete removal of fats or fats and oils.

Esterases are enzymes that break down ester bonds to produce low-molecular fats and alcohols. They are only active against glycerolesters with up to 10 carbonyl acyl chains. To date, many kinds of esterases have been found in various organisms including plants, including bacteria, yeasts and higher organisms, and research on biochemical properties and the esterase genes themselves has been actively conducted. Esterases are hydrolysis of lipids, acidolysis (replacement of esterified fatty acids with free fatty acids), transesterification, exchange of triglyceride fatty acids, It is used in the synthesis of esters and the like, and is an industrially important enzyme. In particular, it can be used in the detergent industry because it can decompose fat components in laundry stains into easily removable hydrophilic substances, and margarine with anti-aging or natural butter flavors for cheese production and baking, such as the addition and flavor of cheese. It can be used in the food and beverage industry, such as manufacturing, in the garbage disposal industry, and in the pharmaceutical industry as a major component of the extinguishing agent. In addition, esterases have unique characteristics such as substrate specificity, optical activity specificity, and site specificity, and thus, their use has been attracting attention in developing various bioactive chiral drugs.

The production of chiral compounds useful for the production of such optically active drugs is attracting attention as a task that has a great influence on securing the competitiveness of the pharmaceutical industry and promoting the health of the public.

This important reaction is sometimes carried out effectively under high temperatures and organic solvents. In particular, bee tallow and palm oil, which are mainly used for the production of free fatty acids in the food industry, remain solid at normal enzymatic reaction temperatures, so a high temperature reaction environment is required for enzymatic processing of these materials. do. Therefore, there is an urgent need for heat resistant esterases that are stable at high temperatures.

The present invention provides novel lipase and esterase enzymes.

The present invention provides a method for producing novel lipase and esterase enzymes.

The present invention provides a lipase enzyme and a method of making the same. The present invention also provides an esterase enzyme and a method of preparing the same.

The present invention provides an isolated DNA sequence encoding a lipase enzyme and a recombinant vector containing the same. The present invention also provides an isolated DNA sequence encoding an esterase enzyme and a recombinant vector containing the same.

The present invention provides a lipase enzyme and a method of making the same. The present invention also provides an esterase enzyme and a method of preparing the same. The manufacturing method is not limited thereto, but is preferably a manufacturing method by a genetic engineering method.

In a first aspect, the invention provides nucleic acid sequences encoding novel lipase enzymes having SEQ ID NO: 7 or 11 and equivalent nucleic acid sequences to the sequence. And a nucleic acid sequence encoding novel esterase enzymes having the sequence of SEQ ID NOs: 1, 3, 5, 9, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 Equivalent nucleic acid sequences are provided.

"Equivalent nucleic acid sequences" include codon degenerate sequences of the lipase enzyme and esterase sequences.

By "codon degenerate sequence" is meant a nucleic acid sequence that encodes a polypeptide of the same sequence as the naturally occurring lipase and esterase enzyme that is different from the naturally occurring sequence described above.

In a second aspect, the present invention provides a lipase enzyme having a amino acid sequence of SEQ ID NO: 8 or 12 and functional equivalents thereof. The present invention also provides esterase enzymes having functional amino acid sequences of SEQ ID NOs: 2, 4, 6, 10, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, or 34 and functional equivalents thereof. to provide.

“Functional equivalents” of the invention include lipases of SEQ ID NO: 8 or 12 and S of SEQ ID NOs: 2, 4, 6, 10, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34 Amino acid sequence variants in which some or all of the terase enzymes are substituted, or in which a portion of the amino acids are deleted or added, include both maintaining the enzyme function. Substitutions of amino acids are preferably conservative substitutions. Examples of conservative substitutions of amino acids present in nature are as follows; Aliphatic amino acids (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn ) And sulfur-containing amino acids (Cys, Met). Deletion of amino acids is preferably located in portions not directly involved in the activity of lipase and esterase enzymes.

In a third aspect, the present invention provides a recombinant vector comprising a DNA segment of SEQ ID NO: 7 or 11 encoding the lipase enzyme. In addition, a recombinant vector comprising a DNA segment of the isolated SEQ ID NO: 1, 3, 5, 9, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 encoding the esterase enzyme To provide.

The term vector, as used herein, refers to a nucleic acid molecule capable of binding and transferring another nucleic acid thereto. The term expression vector includes plasmids, cosmids or phages capable of synthesizing proteins encoded by each recombinant gene carried by the vector. Preferred vectors are vectors capable of self-replicating and expressing the nucleic acid to which they are bound.

In a fourth aspect of the present invention, there is provided a cell transformed with a recombinant vector comprising an isolated DNA segment of SEQ ID NO: 7 or 11 encoding a lipase enzyme. In addition, a recombinant vector comprising a DNA segment of the isolated SEQ ID NO: 1, 3, 5, 9, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 encoding the esterase enzyme To give the transformed cells.

The term 'transformation' used in the present invention means that foreign DNA or RNA is absorbed into a cell and the genotype of the cell is changed.

Suitable transformed cells include, but are not limited to, prokaryotes, fungi, plants, animal cells, and the like. Most preferably, E. coli is used.

 In a fifth aspect of the present invention, there is provided a method for producing a lipase using a cell transformed with a recombinant vector comprising an isolated DNA segment of SEQ ID NO: 7 or 11 encoding a lipase enzyme. In addition, a recombinant vector comprising a DNA segment of the isolated SEQ ID NO: 1, 3, 5, 9, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 encoding the esterase enzyme It provides a method for producing an esterase enzyme using the transformed cells.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following Examples are intended to illustrate the present invention, but the content of the present invention is not limited by the Examples.

< Example  1> Gene Isolation and Microorganisms

The genes for producing the novel lipase and esterase of the present invention extracted the environmental DNA from the sediment up to 1,400 meters deep in the Southwest Pacific, and purified to build a library using a Fosmid vector. The average insert DNA size of the library was about 32 kb, with about 8,000 clones constructed, corresponding to about 280 mb base pairs. From the constructed metagenome library, it was isolated from clones showing activity in Tricaprylin plate media.

< Example  2> Lipase  And Esterase  Gene Cloning  And amino acid sequencing

After obtaining a large amount of fosmid DNA of a tricaprylnin-degrading clone found from the metagenome library, it was cut to 2-4 kb using a nebulizer, followed by end-repairing. Enzymes are used to make each DNA fragment blunt-end to prepare a small size of DNA to be inserted. The pBluescript SK vector was cut with the restriction enzyme Echoalo (EcoRV), treated with calf intestinal phosphatase, and then ligated with the DNA to be inserted and transformed into E. coli DH5α. This was used as a tricaprylin plate medium with ampicillin (1% tryptone, 0.5% yeast extract, 1% sodium chloride (NaCl), 1% tricaprylin )) And colonies were formed by incubating at 37 ° C. for 24 hours to decompose tricaprylnin to form a transparent ring. This was confirmed by culturing the plasmid was isolated. Sequencing of the inserted DNA was performed by designing a sequenase kit, a T7 primer, a T3 primer, and a primer for sequencing. Termination (Sanger dideoxy chain termination) was determined by the method. The assembly was performed using a vector enti (NTI) program, and an open reading frame ( ORF ) was found using an NCF OF finder.

As a result, EM2L4 and EM2L7 appeared as new lipases, EM2L1, EM2L2, EM2L3, EM2L6, ATL1, ATL3, ATL5, ATL6, ATL7, ATL11, KTL1, KTL3, KTL4, KTL7 and KTL9 as new esterases ( Table 1 ).

New Lipases and Esterases Clone name Lipase / esterase (bp) Expression vector Enzyme site amino acid characteristic activation pH pH range Temperature Temperature range temperament EM2L1 780 (SEQ ID NO: 1) pET 24a (+) NdeI and XhoI 259 (SEQ ID NO: 2) 8 7-10 20 5-40 C6 Esterase EM2L2 1215 (SEQ ID NO: 3) pET 24a (+) NdeI and XhoI 404 (SEQ ID NO: 4) 9 7-10 30 10-40 C6 Esterase EM2L3 900 (SEQ ID NO: 5) pET 24a (+) NdeI and XhoI 299 (SEQ ID NO: 6) 8.5 6.5-9.5 15 5-60 C6 Esterase EM2L4 993 (SEQ ID NO: 7) pET 24a (+) NdeI and XhoI 330 (SEQ ID NO: 8) Lipase EM2L6 807 (SEQ ID NO: 9) pET 24a (+) NdeI and XhoI 268 (SEQ ID NO: 10) Esterase EM2L7 834 (SEQ ID NO: 11) pET 24a (+) NdeI and XhoI 277 (SEQ ID NO: 12) Lipase ATL1 912 (SEQ ID NO: 13) pET 24d (+) NcoI and Not I 303 (SEQ ID NO: 14) 8 7.0-9.0 35 20-50 C4 Esterase ATL3 918 (SEQ ID NO: 15) pET 24a (+) NdeI and XhoI 305 (SEQ ID NO: 16) 8.5 6.5-10 20 5-30 C4 Esterase ATL5 1080 (SEQ ID NO: 17) pET 24a (+) NdeI and XhoI 359 (SEQ ID NO: 18) Esterase ATL6 870 (SEQ ID NO: 19) pET 24a (+) NdeI and XhoI 289 (SEQ ID NO: 20) 8 6.5-10 40 20-50 C6 Esterase ATL7 1248 (SEQ ID NO: 21) pET 24a (+) NdeI and XhoI 416 (SEQ ID NO: 22) Esterase ATL11 939 (SEQ ID NO: 23) pET 24a (+) NdeI and XhoI 312 (SEQ ID NO: 24) 8 7.5-8.5 35 25-40 C6 Esterase KTL1 1566 (SEQ ID NO: 25) pET 24a (+) NdeI and XhoI 521 (SEQ ID NO: 26) 8 6.5-10 30 5-45 C4 Esterase KTL3 1500 (SEQ ID NO: 27) pET 24a (+) NdeI and Not I 499 (SEQ ID NO: 28) Esterase KTL4 1059 (SEQ ID NO: 29) pET 24a (+) NdeI and Not I 352 (SEQ ID NO: 30) 8.5 6.5-10 20 5-25 C6 Esterase KTL7 951 (SEQ ID NO: 31) pET 24d (+) NcoI and Not I 316 (SEQ ID NO: 32) 8 7-9 30 15-35 C6 Esterase KTL9 1119 (SEQ ID NO: 33) pET 24a (+) NdeI and XhoI 372 (SEQ ID NO: 34) 8 6.5-10 30 5-45 C6 Esterase

< Example  3> Lipase  And Esterase  Development of recombinant plasmid for mass production

Recombinant plasmids for mass production were developed to produce large amounts of new lipases and esterases found from the metagenome library. DNA containing lipase and esterase genes was used as template DNA and nucleic acid amplification reaction (PCR) was performed using the synthesized two primers. Restriction enzyme was developed after each cut the DNA ends synthesized so as to have a recognition site, Li recombinant plasmid for mass production, by ligation (ligation) with a vector for mass production process with the same restriction enzymes as shown in Table 1 (Table 1 ).

Recombinant plasmids have a strong T7 promotor and translational signal inherent, enabling the production of desired lipases when using E. coli BL21 (DE3), which contains the T7 RNA polymerase gene, as a host.

The activity of the lipase was measured by a pH stat method in which olive oil was hydrolyzed by enzymes at a constant pH to titrate the amount of fatty acids produced with sodium hydroxide (NaOH), and one unit of enzyme was 1 per minute. It was defined as the amount of enzyme that produced μmol of fatty acid.

S. atmospheres activity of the p- nitrophenyl butyrate was measured by using;; (pNPEs p -nitrophenyl esters) as the substrate (p -nitrophenyl butyrate pNPB) or other p- nitro petil ester. The reaction mixture consisted of an appropriate amount of enzyme in 0.01 ml of 10 mM substrate, 0.04 ml ethanol and 0.95 ml of 50 mM Tris-HCl buffer (pH 8.0) dissolved in acetonitrile. Enzyme reaction was performed at 35 ° C. for 3 minutes. The amount of p-nitrophenol released during the reaction was measured at 405 nm. One unit of enzyme was defined as the amount of enzyme that required 1 μmol of p-nitrophenol per minute at 35 ° C.,

< Example  4> Lipase  And Esterase In E. coli  Expression and Production

Escherichia coli (E. coli BL21 (DE3)) with a recombinant plasmid was transferred to a liquid medium (LB) containing 50 micrograms (ug) per ml of kanamycin (1% tryptone, 0.5% yeast extract). Incubate in yeast extract, 1% sodium chloride (NaCl) until the absorbance audio (OD600 nm) is 0.6, and then add Iptage (IPTG) to a final concentration of 1 mM and incubate at 25 ° C. for 12 hours. Lapase and esterase enzymes were obtained.

< Example  5> Lipase  And Esterase  Separation Purification Method

Lipases and esterases produced by E. coli BL21 (DE3) were purely separated as follows: Mass-produced cells were recovered by centrifugation. Only the supernatant obtained by 12,000 × g, 4 ° C., 10 min) was used as coenzyme solution.His Bind column was bound with binding buffer (500 mM NaCl, 20 mM Tris-HCl, 40 mM imidazole, pH 7.9). After equilibration, the crude enzyme solution obtained above is passed through the column, washed with washing buffer solution (500 mM NaCl, 20 mM Tris-HCl, 60 mM imidazole, pH 7.9), and the imidazole concentration is 100 at the same buffer conditions. The lipase and esterase were purified by raising to mM.

The present invention provides a novel lipase and esterase and a gene encoding the same, a transformed Escherichia coli producing a new lipase and esterase, a method for culturing the mass produced lipase and esterase and the produced lipase and esterase As described above, as described above, the novel lipase and esterase activity was excellent, has stability at a wide temperature, and can be usefully used in high value-added fine chemical industry, cosmetics, detergent industry, and the like.

1 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 2 (EM2L1).

Figure 2 shows the esterase base sequence and amino acid sequence of SEQ ID NO: 3 and SEQ ID NO: 4 (EM2L2).

Figure 3 shows the esterase nucleotide sequence and amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 (EM2L3).

Figure 4 shows the lipase base sequence and amino acid sequence of SEQ ID NO: 7 and SEQ ID NO: 8 (EM2L4).

Figure 5 shows the esterase base sequence and amino acid sequence of SEQ ID NO: 9 and SEQ ID NO: 10 (EM2L6).

Figure 6 shows the lipase base sequence and amino acid sequence of SEQ ID NO: 11 and SEQ ID NO: 12 (EM2L7).

7 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 13 and SEQ ID NO: 14 (ATL1).

8 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 15 and SEQ ID NO: 16 (ATL3).

9 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 17 and SEQ ID NO: 18 (ATL5).

10 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 19 and SEQ ID NO: 20 (ATL6).

11 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 21 and SEQ ID NO: 22 (ATL7).

12 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 23 and SEQ ID NO: 24 (ATL11).

13 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 25 and SEQ ID NO: 26 (KTL1).

14 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 27 and SEQ ID NO: 28 (KTL3).

15 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 29 and SEQ ID NO: 30 (KTL4).

Figure 16 shows the esterase nucleotide sequence and amino acid sequence of SEQ ID NO: 31 and SEQ ID NO: 32 (KTL7).

17 shows the esterase nucleotide sequence and the amino acid sequence of SEQ ID NO: 33 and SEQ ID NO: 34 (KTL9).

<110> Korea Ocean Research & Development Institute <120> lipases and esterases isolated from deep sea sediment <160> 34 <170> KopatentIn 1.71 <210> 1 <211> 780 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 1 atgcttatcc cttccgatgg tctggaactg gccgtagagt gttttggcga gggaccgtca 60 ctgattttcg cgcacgggct gacgggcaac cgccacttct cgcggcaaca gcttgcacca 120 ctggcggacc aatatacgat catcatctac gaccagcgcg gccactgcga ctcttccccc 180 gtcaccgacc cggctttgta cgatcccgtg cgcatggccg aagatatgac ggccgttctc 240 gatacactcg gtatcaaaaa agcgatcgtt ggcggcgaat caatgggcgc ggcgacgacg 300 ctgctttttg ccctgcgcca cccgcaacgt gtagaaaaat tactgctcac tgctcccgcc 360 tttggcgata cccgcagcag cgaagccgct ggactgcaac agatggggca gcggattgcc 420 gccattggca tcgaggcgtt tttagcggaa tcggcggtga gccagcgtga ggaactgggt 480 tggccggaac ctgtcatcac tgccgtggcc gccatgcaag gttcgcacga aacccacagt 540 atagcgaccg cctgccagag tgtcattgag tggacgctgg atttatcgcc gctgtctacc 600 atcgcttgcc ccacctgcat tattgcctgg gaagacgacc cgctgcaccc gctggcgttg 660 gcccaaagat atgctgctga aattcctaac gcccggctgg aagttctgcc gtcgctggca 720 gaatttttct ctaatccagc aattattggg gggatatacg gccgtttttt ggcaaactga 780                                                                          780 <210> 2 <211> 259 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 2 Met Leu Ile Pro Ser Asp Gly Leu Glu Leu Ala Val Glu Cys Phe Gly   1 5 10 15 Glu Gly Pro Ser Leu Ile Phe Ala His Gly Leu Thr Gly Asn Arg His              20 25 30 Phe Ser Arg Gln Gln Leu Ala Pro Leu Ala Asp Gln Tyr Thr Ile Ile          35 40 45 Ile Tyr Asp Gln Arg Gly His Cys Asp Ser Ser Pro Val Thr Asp Pro      50 55 60 Ala Leu Tyr Asp Pro Val Arg Met Ala Glu Asp Met Thr Ala Val Leu  65 70 75 80 Asp Thr Leu Gly Ile Lys Lys Ala Ile Val Gly Gly Glu Ser Met Gly                  85 90 95 Ala Ala Thr Thr Leu Leu Phe Ala Leu Arg His Pro Gln Arg Val Glu             100 105 110 Lys Leu Leu Leu Thr Ala Pro Ala Phe Gly Asp Thr Arg Ser Ser Glu         115 120 125 Ala Ala Gly Leu Gln Gln Met Gly Gln Arg Ile Ala Ala Ile Gly Ile     130 135 140 Glu Ala Phe Leu Ala Glu Ser Ala Val Ser Gln Arg Glu Glu Leu Gly 145 150 155 160 Trp Pro Glu Pro Val Ile Thr Ala Val Ala Ala Met Gln Gly Ser His                 165 170 175 Glu Thr His Ser Ile Ala Thr Ala Cys Gln Ser Val Ile Glu Trp Thr             180 185 190 Leu Asp Leu Ser Pro Leu Ser Thr Ile Ala Cys Pro Thr Cys Ile Ile         195 200 205 Ala Trp Glu Asp Asp Pro Leu His Pro Leu Ala Leu Ala Gln Arg Tyr     210 215 220 Ala Ala Glu Ile Pro Asn Ala Arg Leu Glu Val Leu Pro Ser Leu Ala 225 230 235 240 Glu Phe Phe Ser Asn Pro Ala Ile Ile Gly Gly Ile Tyr Gly Arg Phe                 245 250 255 Leu Ala Asn             <210> 3 <211> 1215 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 3 atgaacagaa tgacaatagg cttctcacca atagtactgc tgggttttgc gctgattgcc 60 gcttgtggtt ccggtcatga tgacgatgac gtaaccggcg cggtaaaccc ctttgtgggc 120 tatacaagcg agcaatacag caacagtgcg aattggctat gtcatgcggg tcttgcgaca 180 gccgataatg tatgcgccgc aaacctcgat gcgacgatcg tctttgcgga cggaacgacg 240 gagctggaac ctttcgaacc gctggccgag ccgcctgtcg attgctttta cgtctatccc 300 acggtgagtt cggacagcag cggtaattcg gacctgatcc ccgatgagga gcgatttgtc 360 gtggtcagtc aggcggctcg ttacagtgcc gtatgtcgcg tatttgcgcc tctctaccgg 420 caggtgacgg tgaccgcgtt gttcagcgat atcgagggcg attttgaact cgcctatggc 480 gatgtggtcg atgcgttcaa acattacatg gccaatgaga ataacggtcg gggctttgtg 540 cttatcggcc actcccaggg cacaggacac cttcgccgct tgattgccga ggaaattgaa 600 accgacagtt atctattgga tcggctgatc gcggcgcacc tgataggcct gactgtcgag 660 ataccgaaag gcgcggatgt cggtgccacc ttccagcagg tgcccgtgtg tcgaaaagca 720 gatgaaaccg cctgtgtcgt cagctatgcc agtttccgca ataccgaccc cctgttggct 780 gatgccgtct ttggtgtcac cgacgacgaa agtaccgagg ctgtatgcac caatcccgcc 840 gcgcttgccg gcggcagcgc gactctcgat gcctatttca ataacgaatc catccccgca 900 ctggacgccg tcctgattca acgggcaccg ggtggtccct ttgccgatga aagcttgcgg 960 gatactatca ccacgccgtt ctataaaatg ccggattttt tactgggcga atgtaaaatc 1020 tcgcccgggg gcgcaaacat tctgcaggtg accgcgcaag tcgatgccga tgacccccgt 1080 gcggatgatt tcaacggcga gttcactatc ggcccgggtt ggggcctgca tctggtggat 1140 atcaatctgg gcatgggaaa cctggtcgag ctggccgcaa cgcaaagcca atcctggttg 1200 caagacaata attag 1215 <210> 4 <211> 404 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 4 Met Asn Arg Met Thr Ile Gly Phe Ser Pro Ile Val Leu Leu Gly Phe   1 5 10 15 Ala Leu Ile Ala Ala Cys Gly Ser Gly His Asp Asp Asp Asp Val Thr              20 25 30 Gly Ala Val Asn Pro Phe Val Gly Tyr Thr Ser Glu Gln Tyr Ser Asn          35 40 45 Ser Ala Asn Trp Leu Cys His Ala Gly Leu Ala Thr Ala Asp Asn Val      50 55 60 Cys Ala Ala Asn Leu Asp Ala Thr Ile Val Phe Ala Asp Gly Thr Thr  65 70 75 80 Glu Leu Glu Pro Phe Glu Pro Leu Ala Glu Pro Pro Val Asp Cys Phe                  85 90 95 Tyr Val Tyr Pro Thr Val Ser Ser Asp Ser Ser Gly Asn Ser Asp Leu             100 105 110 Ile Pro Asp Glu Glu Arg Phe Val Val Val Ser Gln Ala Ala Arg Tyr         115 120 125 Ser Ala Val Cys Arg Val Phe Ala Pro Leu Tyr Arg Gln Val Thr Val     130 135 140 Thr Ala Leu Phe Ser Asp Ile Glu Gly Asp Phe Glu Leu Ala Tyr Gly 145 150 155 160 Asp Val Val Asp Ala Phe Lys His Tyr Met Ala Asn Glu Asn Asn Gly                 165 170 175 Arg Gly Phe Val Leu Ile Gly His Ser Gln Gly Thr Gly His Leu Arg             180 185 190 Arg Leu Ile Ala Glu Glu Ile Glu Thr Asp Ser Tyr Leu Leu Asp Arg         195 200 205 Leu Ile Ala Ala His Leu Ile Gly Leu Thr Val Glu Ile Pro Lys Gly     210 215 220 Ala Asp Val Gly Ala Thr Phe Gln Gln Val Pro Val Cys Arg Lys Ala 225 230 235 240 Asp Glu Thr Ala Cys Val Val Ser Tyr Ala Ser Phe Arg Asn Thr Asp                 245 250 255 Pro Leu Leu Ala Asp Ala Val Phe Gly Val Thr Asp Asp Glu Ser Thr             260 265 270 Glu Ala Val Cys Thr Asn Pro Ala Ala Leu Ala Gly Gly Ser Ala Thr         275 280 285 Leu Asp Ala Tyr Phe Asn Asn Glu Ser Ile Pro Ala Leu Asp Ala Val     290 295 300 Leu Ile Gln Arg Ala Pro Gly Gly Pro Phe Ala Asp Glu Ser Leu Arg 305 310 315 320 Asp Thr Ile Thr Thr Pro Phe Tyr Lys Met Pro Asp Phe Leu Leu Gly                 325 330 335 Glu Cys Lys Ile Ser Pro Gly Gly Ala Asn Ile Leu Gln Val Thr Ala             340 345 350 Gln Val Asp Ala Asp Asp Pro Arg Ala Asp Asp Phe Asn Gly Glu Phe         355 360 365 Thr Ile Gly Pro Gly Trp Gly Leu His Leu Val Asp Ile Asn Leu Gly     370 375 380 Met Gly Asn Leu Val Glu Leu Ala Ala Thr Gln Ser Gln Ser Trp Leu 385 390 395 400 Gln Asp Asn Asn                 <210> 5 <211> 900 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 5 gtgatccttt ttagcttggt cggtgtctat ttcctatttc ctgaagtaac gtctgatctc 60 ctgctaaagg tggagcgaag caccgggggt cttaagcagc gcagcatcga agtggatggg 120 ctgcggattg aatacctgga aggaggaaaa ggcgatgtcc ttgtactact ccacggcttc 180 agtgccaaca aggacaactg ggcgcggatt ggaaaatatc tcacacccta tttcagggtg 240 attgcaccgg atctgcccgg tttcggtgaa agcagccttg accctggcgg tgactatagc 300 atcggcgttc aggttgagcg ggtgaaagca tttatccggg ccctgggaat caagtcgttg 360 catctcggcg gaagttccat ggggggcggc atcgcaggcg cgtatgcggc aaggtatcca 420 gacgatctca aaagcctcct gctgatctcc ccgggcggcg ttgcctcttc tgaaccaagt 480 gagatgtttc gtctgttgaa gaaggataag ctaaatccgc tgattgcaaa gaacgccgaa 540 gactacgagt acttgctgga tttcgttttt gtgaaaaagc ctttcatacc cggccccgtc 600 aaaaagtctc taattcaggt ggctatcgag catgagccgc tctatcaaaa gatatttaaa 660 caactcctca gctctacgga ttcacccctg gagatcgttc tggaagggct gccggttccc 720 acccttatca tctggggcgc acaggacagg gtgctgcatg tatccggtgc aaaaattctt 780 gaatcggtta taccgaaggc caaggtagaa ataattgatg ccgtgggcca ccttcccatg 840 attgaaaaac cggaagaaac agcgaagttg tacttgaatt ttcttgggat taagacataa 900                                                                          900 <210> 6 <211> 299 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 6 Val Ile Leu Phe Ser Leu Val Gly Val Tyr Phe Leu Phe Pro Glu Val   1 5 10 15 Thr Ser Asp Leu Leu Leu Lys Val Glu Arg Ser Thr Gly Gly Leu Lys              20 25 30 Gln Arg Ser Ile Glu Val Asp Gly Leu Arg Ile Glu Tyr Leu Glu Gly          35 40 45 Gly Lys Gly Asp Val Leu Val Leu Leu His Gly Phe Ser Ala Asn Lys      50 55 60 Asp Asn Trp Ala Arg Ile Gly Lys Tyr Leu Thr Pro Tyr Phe Arg Val  65 70 75 80 Ile Ala Pro Asp Leu Pro Gly Phe Gly Glu Ser Ser Leu Asp Pro Gly                  85 90 95 Gly Asp Tyr Ser Ile Gly Val Gln Val Glu Arg Val Lys Ala Phe Ile             100 105 110 Arg Ala Leu Gly Ile Lys Ser Leu His Leu Gly Gly Ser Ser Met Gly         115 120 125 Gly Gly Ile Ala Gly Ala Tyr Ala Ala Arg Tyr Pro Asp Asp Leu Lys     130 135 140 Ser Leu Leu Leu Ile Ser Pro Gly Gly Val Ala Ser Ser Glu Pro Ser 145 150 155 160 Glu Met Phe Arg Leu Leu Lys Lys Asp Lys Leu Asn Pro Leu Ile Ala                 165 170 175 Lys Asn Ala Glu Asp Tyr Glu Tyr Leu Leu Asp Phe Val Phe Val Lys             180 185 190 Lys Pro Phe Ile Pro Gly Pro Val Lys Lys Ser Leu Ile Gln Val Ala         195 200 205 Ile Glu His Glu Pro Leu Tyr Gln Lys Ile Phe Lys Gln Leu Leu Ser     210 215 220 Ser Thr Asp Ser Pro Leu Glu Ile Val Leu Glu Gly Leu Pro Val Pro 225 230 235 240 Thr Leu Ile Ile Trp Gly Ala Gln Asp Arg Val Leu His Val Ser Gly                 245 250 255 Ala Lys Ile Leu Glu Ser Val Ile Pro Lys Ala Lys Val Glu Ile Ile             260 265 270 Asp Ala Val Gly His Leu Pro Met Ile Glu Lys Pro Glu Glu Thr Ala         275 280 285 Lys Leu Tyr Leu Asn Phe Leu Gly Ile Lys Thr     290 295 <210> 7 <211> 993 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 7 atgactggta gaattgggcg tatcatattg gtggttgtcg tggcgatcgt gttggtgctg 60 tgcattgccg cgatggtgac ctattggatg gtgggtccga ccccgcgctg cgcactgccg 120 gcggaggggc cgtctgggcc aggctggtcg gcccggaccc tggactcggg tggccgtgag 180 cgctgttacc atctctacgt gccgccgggg tacaatcccg cccagccagc gccagtggtc 240 gtgagtttcc atggcttcct gtccaacccc aacagccacg cgctcatcag cggctggcac 300 aagctcgccg agcaggaggg ctttctggtg gcctatcccc agggcacgtc gttcccgcta 360 cgatgggacg ccggctcgac gtggggagcg ccgggggtca acgacgtgca gttcttccgc 420 gatatggtcg acgacatggc caatgtggcg gccgtcgacc gctcgcgggt gtatgtcaat 480 ggcttttcca acggcggcgg catggccgtg cgtatcggct gcgaggctgc cggagaagtg 540 gccgcgctgg gttcggttgc gggcgccgtc gtcgagatag aggagtgcag cccatcacgc 600 cctgtgccgg tcatggccta ccacgggacg gcggaccctg ttgtgccgta cgagggcggc 660 gagatgcaag gttggctgct gcgctggggg gccggcgctg ttgacgcgcc cgtctacttt 720 gtgggcgccg aggactgggt ctccctctgg gcggcgggga atggctgcga tcccacgccg 780 gacgtggtcc cggcgcaagg ggatgcgagt ggcatccgct acaccgggtg cgaccaggac 840 gccgaggtgt ccttgtatac cattgacggc ggcgggcata cctggcccgg gggctggccg 900 atccccgccg tgggcaagac aagtaccgac atcaatgcga ccggggagat gtggcagttc 960 ttccagagat accaactgga ggagcagccg tag 993 <210> 8 <211> 330 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 8 Met Thr Gly Arg Ile Gly Arg Ile Ile Leu Val Val Val Val Ala Ile   1 5 10 15 Val Leu Val Leu Cys Ile Ala Ala Met Val Thr Tyr Trp Met Val Gly              20 25 30 Pro Thr Pro Arg Cys Ala Leu Pro Ala Glu Gly Pro Ser Gly Pro Gly          35 40 45 Trp Ser Ala Arg Thr Leu Asp Ser Gly Gly Arg Glu Arg Cys Tyr His      50 55 60 Leu Tyr Val Pro Pro Gly Tyr Asn Pro Ala Gln Pro Ala Pro Val Val  65 70 75 80 Val Ser Phe His Gly Phe Leu Ser Asn Pro Asn Ser His Ala Leu Ile                  85 90 95 Ser Gly Trp His Lys Leu Ala Glu Gln Glu Gly Phe Leu Val Ala Tyr             100 105 110 Pro Gln Gly Thr Ser Phe Pro Leu Arg Trp Asp Ala Gly Ser Thr Trp         115 120 125 Gly Ala Pro Gly Val Asn Asp Val Gln Phe Phe Arg Asp Met Val Asp     130 135 140 Asp Met Ala Asn Val Ala Ala Val Asp Arg Ser Arg Val Tyr Val Asn 145 150 155 160 Gly Phe Ser Asn Gly Gly Gly Met Ala Val Arg Ile Gly Cys Glu Ala                 165 170 175 Ala Gly Glu Val Ala Ala Leu Gly Ser Val Ala Gly Ala Val Val Glu             180 185 190 Ile Glu Glu Cys Ser Pro Ser Arg Pro Val Pro Val Met Ala Tyr His         195 200 205 Gly Thr Ala Asp Pro Val Val Pro Tyr Glu Gly Gly Glu Met Gln Gly     210 215 220 Trp Leu Leu Arg Trp Gly Ala Gly Ala Val Asp Ala Pro Val Tyr Phe 225 230 235 240 Val Gly Ala Glu Asp Trp Val Ser Leu Trp Ala Ala Gly Asn Gly Cys                 245 250 255 Asp Pro Thr Pro Asp Val Val Pro Ala Gln Gly Asp Ala Ser Gly Ile             260 265 270 Arg Tyr Thr Gly Cys Asp Gln Asp Ala Glu Val Ser Leu Tyr Thr Ile         275 280 285 Asp Gly Gly Gly His Thr Trp Pro Gly Gly Trp Pro Ile Pro Ala Val     290 295 300 Gly Lys Thr Ser Thr Asp Ile Asn Ala Thr Gly Glu Met Trp Gln Phe 305 310 315 320 Phe Gln Arg Tyr Gln Leu Glu Glu Gln Pro                 325 330 <210> 9 <211> 807 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 9 atgaaatgca tcccatcaga cggcctgcaa ctggccgtag aaacctttgg cagcggcccg 60 ccgctgatca tggcacatgg attaaccggc aaccggaaca tgagccgcca acaattggcg 120 ccgctggccg acagctaccg cgtcattgtc tttgaccagc gcgggcacaa cgaatccacc 180 cctgtcaccg atcccgccct ctacgacccg ctgcgcatgg cggccgatat cggtaacatc 240 ctcgacgctt ttaacgtgga acgggccatc atcggcggcg aatcaatggg cgcagccact 300 gccttaacct ttgccctcaa ctggccgcag cgtgtcgaaa cattattact caccgcccct 360 gcttttggcg acaagccaaa cccggagtcc gagggcattc gagaaatggg cgctgccatt 420 gccactgtcg gcattgaagc ctttctcgac gccgccgagg tgaagtggcg cgaagaatta 480 gagtggccac aagttgtgat agaccatgtg cgggccatgc aaagctgcca tgacgcggcc 540 agcctcgccg tcgcctgtca gacagtgatt gattggcagc tatttacaaa cctgtcggtt 600 gtgagtcagt tggagatgcc ggtacgcatc gtggcctggg agaatgacga tctgcatccg 660 ttaatcctcg cccagcgcat ggcggcgtta atcccggatg tgaagatgac agtgctggct 720 tcgctcacgg agatgttttc caacccggcc gtcatcacta tccattaccg gcgctttctg 780 gaggaattag gctcgaataa tgattag 807 <210> 10 <211> 268 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 10 Met Lys Cys Ile Pro Ser Asp Gly Leu Gln Leu Ala Val Glu Thr Phe   1 5 10 15 Gly Ser Gly Pro Pro Leu Ile Met Ala His Gly Leu Thr Gly Asn Arg              20 25 30 Asn Met Ser Arg Gln Gln Leu Ala Pro Leu Ala Asp Ser Tyr Arg Val          35 40 45 Ile Val Phe Asp Gln Arg Gly His Asn Glu Ser Thr Pro Val Thr Asp      50 55 60 Pro Ala Leu Tyr Asp Pro Leu Arg Met Ala Ala Asp Ile Gly Asn Ile  65 70 75 80 Leu Asp Ala Phe Asn Val Glu Arg Ala Ile Gle Gly Glu Ser Met                  85 90 95 Gly Ala Ala Thr Ala Leu Thr Phe Ala Leu Asn Trp Pro Gln Arg Val             100 105 110 Glu Thr Leu Leu Leu Thr Ala Pro Ala Phe Gly Asp Lys Pro Asn Pro         115 120 125 Glu Ser Glu Gly Ile Arg Glu Met Gly Ala Ala Ile Ala Thr Val Gly     130 135 140 Ile Glu Ala Phe Leu Asp Ala Ala Glu Val Lys Trp Arg Glu Glu Leu 145 150 155 160 Glu Trp Pro Gln Val Val Ile Asp His Val Arg Ala Met Gln Ser Cys                 165 170 175 His Asp Ala Ala Ser Leu Ala Val Ala Cys Gln Thr Val Ile Asp Trp             180 185 190 Gln Leu Phe Thr Asn Leu Ser Val Val Ser Gln Leu Glu Met Pro Val         195 200 205 Arg Ile Val Ala Trp Glu Asn Asp Asp Leu His Pro Leu Ile Leu Ala     210 215 220 Gln Arg Met Ala Ala Leu Ile Pro Asp Val Lys Met Thr Val Leu Ala 225 230 235 240 Ser Leu Thr Glu Met Phe Ser Asn Pro Ala Val Ile Thr Ile His Tyr                 245 250 255 Arg Arg Phe Leu Glu Glu Leu Gly Ser Asn Asn Asp             260 265 <210> 11 <211> 834 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 11 atgacttatc cgattgtgct cgcccatggc gtgtgtcgtt ttgacgcctt gttgaacgac 60 gccctggacc tcgacaataa cgatgatccg gagcttgaca acctgcacta tttcaaaggc 120 atacggacga tgctaaagca gaaaggatat atcgtttatc attccaacgt tgcctgggca 180 gcaggggtag agaaacgggc cgatgacctg aaagaaaacc tgttgaagat tcttaacgaa 240 acacgaacgg aaaaggtcaa cattatcgca cacagcatgg gtggactgga tgcgcgtcac 300 atgatgttta atgatcgtaa tgatgaagaa atccaccaac gtatagcttc tcttactact 360 atctctacac ctcacgaggg atctccattc gccgactggg ggcttgataa cctcacgcct 420 ctgcatttat taatccaaaa gctgggtgtg gacttgagcg ctttgaagga tctaaggacg 480 gatacatgca aaacgttcaa tgaacatcag gatgtcaaag aattcgaaat ggcatgtaag 540 ggtacgataa agttccagac ctatgctgga agacaggatt tttggggtgt ctttagtgga 600 ctcaaaattc cctttgagat aatcaggaag gcagaaggtg aaaatgacgg tatggtttca 660 gttaggtcag ccacatggag agatgagtat ttcaagggaa cactcgataa aactgaccat 720 ctcaatgaga tggggtggtg ggcccctgat caacttttag ctgccgagag tccggacgag 780 ttgcttgccc gtatacacag cttttatgcc ggaatcgcgg aacaactacc ttaa 834 <210> 12 <211> 277 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 12 Met Thr Tyr Pro Ile Val Leu Ala His Gly Val Cys Arg Phe Asp Ala   1 5 10 15 Leu Leu Asn Asp Ala Leu Asp Leu Asp Asn Asn Asp Asp Pro Glu Leu              20 25 30 Asp Asn Leu His Tyr Phe Lys Gly Ile Arg Thr Met Leu Lys Gln Lys          35 40 45 Gly Tyr Ile Val Tyr His Ser Asn Val Ala Trp Ala Ala Gly Val Glu      50 55 60 Lys Arg Ala Asp Asp Leu Lys Glu Asn Leu Leu Lys Ile Leu Asn Glu  65 70 75 80 Thr Arg Thr Glu Lys Val Asn Ile Ile Ala His Ser Met Gly Gly Leu                  85 90 95 Asp Ala Arg His Met Met Phe Asn Asp Arg Asn Asp Glu Glu Ile His             100 105 110 Gln Arg Ile Ala Ser Leu Thr Thr Ile Ser Thr Pro His Glu Gly Ser         115 120 125 Pro Phe Ala Asp Trp Gly Leu Asp Asn Leu Thr Pro Leu His Leu Leu     130 135 140 Ile Gln Lys Leu Gly Val Asp Leu Ser Ala Leu Lys Asp Leu Arg Thr 145 150 155 160 Asp Thr Cys Lys Thr Phe Asn Glu His Gln Asp Val Lys Glu Phe Glu                 165 170 175 Met Ala Cys Lys Gly Thr Ile Lys Phe Gln Thr Tyr Ala Gly Arg Gln             180 185 190 Asp Phe Trp Gly Val Phe Ser Gly Leu Lys Ile Pro Phe Glu Ile Ile         195 200 205 Arg Lys Ala Glu Gly Glu Asn Asp Gly Met Val Ser Val Arg Ser Ala     210 215 220 Thr Trp Arg Asp Glu Tyr Phe Lys Gly Thr Leu Asp Lys Thr Asp His 225 230 235 240 Leu Asn Glu Met Gly Trp Trp Ala Pro Asp Gln Leu Leu Ala Ala Glu                 245 250 255 Ser Pro Asp Glu Leu Leu Ala Arg Ile His Ser Phe Tyr Ala Gly Ile             260 265 270 Ala Glu Gln Leu Pro         275 <210> 13 <211> 912 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 13 atgacgatga gccgccgtgc aaaatttgtt cgctggttaa ccagcggata catgcaaaaa 60 ctggacgtgg cgaatgtgcc ggtcgacaca gcgagaaagc acctcgaaac ggtcgcccgg 120 acgttgctgg tccgagcgac tggcgtccat gttgaacagt cgcaacttgc cgggattgac 180 gttgactggc taagaccgaa agaggcacgc aaggacaagg tcctgtttta tttgcatggc 240 ggcgcttacg tcctgggcag tcgtcggaca caccgccagc tagtgagtca tatggcaagg 300 gaggccggca tcaccgcggt acttccagag tatcgactgg ctcccgagca cccgtttcct 360 gcggcgatcg acgatgcggt agctgtgtac cgggcgcttc tcgaatccgg cttcaaaccg 420 gaggacatcg ttatctcggg agattcggct ggtgggggct tgtcggtcgc gacgctgctc 480 gcacttcgac acgccggaga tcccctccct gcggctgccg tgttgctttc gccatttctc 540 gatgtcacag cgagcggcga atcggcgacg acgcgagcgg accaggaccc ctggtttgat 600 gtgcatgaca tggaggtcgt ggcgcgctac tactgtgcgg atgagagcaa gtggcgcgac 660 ccactgatat caccggtgtt cgccaacgtc tcgggtcttc cgcccatgct tattcaggta 720 ggtgacgacg aaatactcct cgacgactcg acgcgactgg cagaaaaact cgaggcggca 780 ggcattgacg tcgagatcga gatttggccg aacatgtggc acgttttcca gatgttcatt 840 ggcaagatgc cggaaagccg caaggccgtc gagaagatag ggaagtacat cgaccaggcg 900 gtttcgaagt ag 912 <210> 14 <211> 303 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 14 Met Thr Met Ser Arg Arg Ala Lys Phe Val Arg Trp Leu Thr Ser Gly   1 5 10 15 Tyr Met Gln Lys Leu Asp Val Ala Asn Val Pro Val Asp Thr Ala Arg              20 25 30 Lys His Leu Glu Thr Val Ala Arg Thr Leu Leu Val Arg Ala Thr Gly          35 40 45 Val His Val Glu Gln Ser Gln Leu Ala Gly Ile Asp Val Asp Trp Leu      50 55 60 Arg Pro Lys Glu Ala Arg Lys Asp Lys Val Leu Phe Tyr Leu His Gly  65 70 75 80 Gly Ala Tyr Val Leu Gly Ser Arg Arg Thr His Arg Gln Leu Val Ser                  85 90 95 His Met Ala Arg Glu Ala Gly Ile Thr Ala Val Leu Pro Glu Tyr Arg             100 105 110 Leu Ala Pro Glu His Pro Phe Pro Ala Ala Ile Asp Asp Ala Val Ala         115 120 125 Val Tyr Arg Ala Leu Leu Glu Ser Gly Phe Lys Pro Glu Asp Ile Val     130 135 140 Ile Ser Gly Asp Ser Ala Gly Gly Gly Leu Ser Val Ala Thr Leu Leu 145 150 155 160 Ala Leu Arg His Ala Gly Asp Pro Leu Pro Ala Ala Ala Val Leu Leu                 165 170 175 Ser Pro Phe Leu Asp Val Thr Ala Ser Gly Glu Ser Ala Thr Thr Arg             180 185 190 Ala Asp Gln Asp Pro Trp Phe Asp Val His Asp Met Glu Val Val Ala         195 200 205 Arg Tyr Tyr Cys Ala Asp Glu Ser Lys Trp Arg Asp Pro Leu Ile Ser     210 215 220 Pro Val Phe Ala Asn Val Ser Gly Leu Pro Pro Met Leu Ile Gln Val 225 230 235 240 Gly Asp Asp Glu Ile Leu Leu Asp Asp Ser Thr Arg Leu Ala Glu Lys                 245 250 255 Leu Glu Ala Ala Gly Ile Asp Val Glu Ile Glu Ile Trp Pro Asn Met             260 265 270 Trp His Val Phe Gln Met Phe Ile Gly Lys Met Pro Glu Ser Arg Lys         275 280 285 Ala Val Glu Lys Ile Gly Lys Tyr Ile Asp Gln Ala Val Ser Lys     290 295 300 <210> 15 <211> 918 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 15 atgaaggctg tctggtacac cgtgctggca atgcttctgc ttttgacctc ctgcgttaag 60 gttccggatg gaacgcttca ggatgatctg cgaaacaggg aaataaagtt tgcggtctgg 120 gtgcctgaga gtacagaacc tgccccattg atagttctca gtcatggcag cggcggccac 180 tacagtaatt tcaactggct gacccgtgtt ctggttgagc atggctacgt cgtggcggcc 240 gtcaaccacc ctttcaacac gacgggaaat gacacaccgg agggcgtggc aagggcctgg 300 gaccggccgc ctgatctgtc actgctgatc tctgaattgc tatccaatcc tgaatgggcc 360 gctgcaatag acagtactcg tataggtgca actggctttt cttcgggggg gtatacggtt 420 ctggcgctgg cgggcgccgt ctatgatatt gatcaaatgc atgcctattg caatagcact 480 gaagctggtc ccgagtgtgg tctggcagtc gcactgcctg cggctgatcc ggatgcttcc 540 tccttattaa aggacccgag aataaaggcc gtatttgcca tggcgccggg atggggtgca 600 gctaccaggt ctgatagtct tgaagccatc gatattccgg tcaagatagt tgcggctgag 660 gatgatgaaa tacttatacc cgcgcatcat gcggtttatt ttgatgaatt aattcctgat 720 tcagtgctcg tgatgctgcc caagggtggt cactttatat ttcttagctg cagccccatg 780 acgcgagttg cggactggtt tatcgataga ttcaaccttt gcggcattgg gattgatgtc 840 gatcgcgatg ccgtgcaaaa agaaactgca gccctggcgg tacagttttt cgaccagcat 900 ctgcgcgctg tggagtga 918 <210> 16 <211> 305 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 16 Met Lys Ala Val Trp Tyr Thr Val Leu Ala Met Leu Leu Leu Leu Thr   1 5 10 15 Ser Cys Val Lys Val Pro Asp Gly Thr Leu Gln Asp Asp Leu Arg Asn              20 25 30 Arg Glu Ile Lys Phe Ala Val Trp Val Pro Glu Ser Thr Glu Pro Ala          35 40 45 Pro Leu Ile Val Leu Ser His Gly Ser Gly Gly His Tyr Ser Asn Phe      50 55 60 Asn Trp Leu Thr Arg Val Leu Val Glu His Gly Tyr Val Val Ala Ala  65 70 75 80 Val Asn His Pro Phe Asn Thr Thr Gly Asn Asp Thr Pro Glu Gly Val                  85 90 95 Ala Arg Ala Trp Asp Arg Pro Pro Asp Leu Ser Leu Leu Ile Ser Glu             100 105 110 Leu Leu Ser Asn Pro Glu Trp Ala Ala Ala Ile Asp Ser Thr Arg Ile         115 120 125 Gly Ala Thr Gly Phe Ser Ser Gly Gly Tyr Thr Val Leu Ala Leu Ala     130 135 140 Gly Ala Val Tyr Asp Ile Asp Gln Met His Ala Tyr Cys Asn Ser Thr 145 150 155 160 Glu Ala Gly Pro Glu Cys Gly Leu Ala Val Ala Leu Pro Ala Ala Asp                 165 170 175 Pro Asp Ala Ser Ser Le Leu Leu Lys Asp Pro Arg Ile Lys Ala Val Phe             180 185 190 Ala Met Ala Pro Gly Trp Gly Ala Ala Thr Arg Ser Asp Ser Leu Glu         195 200 205 Ala Ile Asp Ile Pro Val Lys Ile Val Ala Ala Glu Asp Asp Glu Ile     210 215 220 Leu Ile Pro Ala His His Ala Val Tyr Phe Asp Glu Leu Ile Pro Asp 225 230 235 240 Ser Val Leu Val Met Leu Pro Lys Gly Gly His Phe Ile Phe Leu Ser                 245 250 255 Cys Ser Pro Met Thr Arg Val Ala Asp Trp Phe Ile Asp Arg Phe Asn             260 265 270 Leu Cys Gly Ile Gly Ile Asp Val Asp Arg Asp Ala Val Gln Lys Glu         275 280 285 Thr Ala Ala Leu Ala Val Gln Phe Phe Asp Gln His Leu Arg Ala Val     290 295 300 Glu 305 <210> 17 <211> 1080 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 17 atgaaacgta atctttttta ccttgccgct ggcctgatgt gcgtaacttc gacagttagt 60 cttcatgctg agaatacgcc ggtgcggcca gtttttgacg aagcaggcac ggtccatgtt 120 cccgctttcg agctgccgcc ttcagtattt tcaagtcagg aggctcagga ttttatgaag 180 cttcgtgcaa gaatgcccag cgaattgccg acaaaggagc ctgatattgt aaagtcgcgc 240 aaggggcttg aggcaatgtt ggcacctcaa gtggcgggga tgttgaagtt gcatcctgtc 300 gatgtggtcg atgagacaat caatggcgtt cctacacgaa ttgtcacgcc caagggcaag 360 gatgttgatc ctgatcgtgt cctcatcaac ctgcatggcg gcgcttttac gacatgctgg 420 gagtcctgtt caatgctgga atcggcaccc atcgcgtcgc ttggtggcta caaagttgtc 480 tctgttaatt accgcatggc gccagaggct actcatccgg cgggagttga agatgttgcc 540 aaggtctaca gtgagcttct gaagacttac gagtccaaac atatcggtat atatggctgt 600 tcggcaggtg gcgcgcttac cgctcagaca gcggcctggc taccaacaca caacctgccc 660 caggctggcg ctgtcggcat tttcggtgca ggtggagtgc gcttcagcga aggggattcg 720 gcttatatcg ctggttatat cgatggttcc tttccgccgc caccgaaacc gggtgaaacg 780 aaaattgaca tgactcgcgg ctatttcgct aacgctgata tgtcggacgc aattatttcc 840 cctgcacttc atcccagtgt actcaagaac ttcccgccgg ctctcattat taccggcaca 900 agggcaatgg acatgagtcc tgcaattgtc accaattcaa atctcattag ggcaggtata 960 cgctcgacgt tgatcgtagg tgaaggcatg ggccactgct atatttatat ggcgcagcta 1020 ccggaagcgc aggacgccca ctcggcgatc gtcgcctttt ttgacgaaaa cctgcgctag 1080                                                                         1080 <210> 18 <211> 359 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 18 Met Lys Arg Asn Leu Phe Tyr Leu Ala Ala Gly Leu Met Cys Val Thr   1 5 10 15 Ser Thr Val Ser Leu His Ala Glu Asn Thr Pro Val Arg Pro Val Phe              20 25 30 Asp Glu Ala Gly Thr Val His Val Pro Ala Phe Glu Leu Pro Pro Ser          35 40 45 Val Phe Ser Ser Gln Glu Ala Gln Asp Phe Met Lys Leu Arg Ala Arg      50 55 60 Met Pro Ser Glu Leu Pro Thr Lys Glu Pro Asp Ile Val Lys Ser Arg  65 70 75 80 Lys Gly Leu Glu Ala Met Leu Ala Pro Gln Val Ala Gly Met Leu Lys                  85 90 95 Leu His Pro Val Asp Val Val Asp Glu Thr Ile Asn Gly Val Pro Thr             100 105 110 Arg Ile Val Thr Pro Lys Gly Lys Asp Val Asp Pro Asp Arg Val Leu         115 120 125 Ile Asn Leu His Gly Gly Ala Phe Thr Thr Cys Trp Glu Ser Cys Ser     130 135 140 Met Leu Glu Ser Ala Pro Ile Ala Ser Leu Gly Gly Tyr Lys Val Val 145 150 155 160 Ser Val Asn Tyr Arg Met Ala Pro Glu Ala Thr His Pro Ala Gly Val                 165 170 175 Glu Asp Val Ala Lys Val Tyr Ser Glu Leu Leu Lys Thr Tyr Glu Ser             180 185 190 Lys His Ile Gly Ile Tyr Gly Cys Ser Ala Gly Gly Ala Leu Thr Ala         195 200 205 Gln Thr Ala Ala Trp Leu Pro Thr His Asn Leu Pro Gln Ala Gly Ala     210 215 220 Val Gly Ile Phe Gly Ala Gly Gly Val Arg Phe Ser Glu Gly Asp Ser 225 230 235 240 Ala Tyr Ile Ala Gly Tyr Ile Asp Gly Ser Phe Pro Pro Pro Lys                 245 250 255 Pro Gly Glu Thr Lys Ile Asp Met Thr Arg Gly Tyr Phe Ala Asn Ala             260 265 270 Asp Met Ser Asp Ala Ile Ile Ser Pro Ala Leu His Pro Ser Val Leu         275 280 285 Lys Asn Phe Pro Pro Ala Leu Ile Ile Thr Gly Thr Arg Ala Met Asp     290 295 300 Met Ser Pro Ala Ile Val Thr Asn Ser Asn Leu Ile Arg Ala Gly Ile 305 310 315 320 Arg Ser Thr Leu Ile Val Gly Glu Gly Met Gly His Cys Tyr Ile Tyr                 325 330 335 Met Ala Gln Leu Pro Glu Ala Gln Asp Ala His Ser Ala Ile Val Ala             340 345 350 Phe Phe Asp Glu Asn Leu Arg         355 <210> 19 <211> 870 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 19 atgaaacaaa aagtagcact tgttttgtcg ggagggggtg cccgtggcat tgcccatatc 60 ggagtgattg aggaactcga aagtcaggga ttcgagatca cttccattgc cggaacctcc 120 atgggctcgg tggtgggagg gatctatgca tcaggcaaac tggatgtttt aaagaagtgg 180 ctctttaccc tggacaaact gaaggttctg aatatggtgg acttttcatt tagcaaacag 240 ggactggtga aaggagatcg tgtaagtggc aggatgaaga ggcttatggc tgacacccag 300 attgaggatc ttagaattcc ctttgctgca gtggcagtgg atctggtcca ttcgaaggag 360 gtagtcttta gaagcgggag catgtatgag gcccttcggg cttctgtttc cattccgacc 420 attttcaccc ctgtgaaaac agatgatgga ttgttggtgg atggaggtgt gatgaataat 480 gtacccatcg acagggtgga aagaactccc ggtgatatat tggtggctgt agatgtaaat 540 gcaagtgtcc ctgtaaataa acctgccatt acaaaaaaag aggaaaaggc cagacaatcc 600 ctttatcgaa aaaaaatgtt agagtttcag caacatttgc gtggtagcgg tcccaagagc 660 agcggtccca acctgggata cttcgatctg attaataaaa ccctcagttt aatgacccat 720 cacacggccc gcttactcat ggagaaacat aagccggata tccttataga ggtatctcat 780 gaatcctgtg aaatgtatga cttctacaag gcagaagaga tggtggagat tgggaggcat 840 gcagcatcgc tcaagctctc ttcgagatag 870 <210> 20 <211> 289 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 20 Met Lys Gln Lys Val Ala Leu Val Leu Ser Gly Gly Gly Ala Arg Gly   1 5 10 15 Ile Ala His Ile Gly Val Ile Glu Glu Leu Glu Ser Gln Gly Phe Glu              20 25 30 Ile Thr Ser Ile Ala Gly Thr Ser Met Gly Ser Val Val Gly Gly Ile          35 40 45 Tyr Ala Ser Gly Lys Leu Asp Val Leu Lys Lys Trp Leu Phe Thr Leu      50 55 60 Asp Lys Leu Lys Val Leu Asn Met Val Asp Phe Ser Phe Ser Lys Gln  65 70 75 80 Gly Leu Val Lys Gly Asp Arg Val Ser Gly Arg Met Lys Arg Leu Met                  85 90 95 Ala Asp Thr Gln Ile Glu Asp Leu Arg Ile Pro Phe Ala Ala Val Ala             100 105 110 Val Asp Leu Val His Ser Lys Glu Val Val Phe Arg Ser Gly Ser Met         115 120 125 Tyr Glu Ala Leu Arg Ala Ser Val Ser Ile Pro Thr Ile Phe Thr Pro     130 135 140 Val Lys Thr Asp Asp Gly Leu Leu Val Asp Gly Gly Val Met Asn Asn 145 150 155 160 Val Pro Ile Asp Arg Val Glu Arg Thr Pro Gly Asp Ile Leu Val Ala                 165 170 175 Val Asp Val Asn Ala Ser Val Pro Val Asn Lys Pro Ala Ile Thr Lys             180 185 190 Lys Glu Glu Lys Ala Arg Gln Ser Leu Tyr Arg Lys Lys Met Leu Glu         195 200 205 Phe Gln Gln His Leu Arg Gly Ser Gly Pro Lys Ser Ser Gly Pro Asn     210 215 220 Leu Gly Tyr Phe Asp Leu Ile Asn Lys Thr Leu Ser Leu Met Thr His 225 230 235 240 His Thr Ala Arg Leu Leu Met Glu Lys His Lys Pro Asp Ile Leu Ile                 245 250 255 Glu Val Ser His Glu Ser Cys Glu Met Tyr Asp Phe Tyr Lys Ala Glu             260 265 270 Glu Met Val Glu Ile Gly Arg His Ala Ala Ser Leu Lys Leu Ser Ser         275 280 285 Arg     <210> 21 <211> 1248 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 21 atggccggac tatattttac gctcgcagtg ctgggcgcac tgggtacatt tacagccctg 60 gtgcaggcac gtcgcctggg ctggctagtg ccagtgtact ttctcagcgc gtggctgacg 120 ggagagctgg ccatgattca cctgctgctg cagttgacac taaccgccct gatggctttt 180 ggaggtgtgt tttccgatcc acttgcccag accgggctcg gtattttcgc attatcctgg 240 ctcggtctga tttacctgca catccaggct atggatacgc cgcagtattt acgcgcggca 300 ctgcaccggg ccctgggagc ggactatcgc aatagcatcc cggaggagcg acgggaggtg 360 ttgagtgaga atattgtcag ccggcactgg atgaagccgt ttcattttcg ccgggatggc 420 gttcgagagc acaacaatat cagttatggc gaggcgggta agcgcaactt gctggatgtc 480 tatcagcccg taaatccgcg gtcgggtggt gccccggtcc tgctgcaggt acacggtggc 540 gcctggatca ttggtgagaa agaacaacag gccaagccgc tcatgtacca catgtcccag 600 cggggctggg tgtgtgtggc tatcaattat cgactgagcc ccaaggaagc ttttcccgcc 660 catatcgtcg atgttaaaaa agccatcgcc tgggtcaggg aaaatattgc ccagtacggg 720 ggcgacccca actttattgc tattaccggg ggctccgctg ggggccacct cagttcgctg 780 gccgcactga cgcccaaccg tgcccagtgg cagccggggt ttgaagacgc ggacacgacg 840 atacaggccg cggttccgtt ctacggtgtt tatgattttc tggatcggta tgacatacgc 900 ccggacatgt ccatggaaga tatcatcgcc gacaaggtgt tgcagtgcgc caaggaggat 960 aaccaccagt tgtgggagga tggttctccg atgagccaca taggggctca cgccccaccc 1020 atgtatgtaa tccagggtac ccacgattcc ctcgtttggg tggaagaagc acggactttc 1080 gtcgccgctt tacaggaagt tgccacccat cctgttgcct atgctgagct gcctggcgca 1140 cagcatgcgt ttgagatctt tcactcggta cgtaccgacc acacggtaaa ttctgttgct 1200 gatttcctcg aatggagtca cgccggatgg ctacaaaata agggctga 1248 <210> 22 <211> 415 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 22 Met Ala Gly Leu Tyr Phe Thr Leu Ala Val Leu Gly Ala Leu Gly Thr   1 5 10 15 Phe Thr Ala Leu Val Gln Ala Arg Arg Leu Gly Trp Leu Val Pro Val              20 25 30 Tyr Phe Leu Ser Ala Trp Leu Thr Gly Glu Leu Ala Met Ile His Leu          35 40 45 Leu Leu Gln Leu Thr Leu Thr Ala Leu Met Ala Phe Gly Gly Val Phe      50 55 60 Ser Asp Pro Leu Ala Gln Thr Gly Leu Gly Ile Phe Ala Leu Ser Trp  65 70 75 80 Leu Gly Leu Ile Tyr Leu His Ile Gln Ala Met Asp Thr Pro Gln Tyr                  85 90 95 Leu Arg Ala Ala Leu His Arg Ala Leu Gly Ala Asp Tyr Arg Asn Ser             100 105 110 Ile Pro Glu Glu Arg Arg Glu Val Leu Ser Glu Asn Ile Val Ser Arg         115 120 125 His Trp Met Lys Pro Phe His Phe Arg Arg Asp Gly Val Arg Glu His     130 135 140 Asn Asn Ile Ser Tyr Gly Glu Ala Gly Lys Arg Asn Leu Leu Asp Val 145 150 155 160 Tyr Gln Pro Val Asn Pro Arg Ser Gly Gly Ala Pro Val Leu Leu Gln                 165 170 175 Val His Gly Gly Ala Trp Ile Ile Gly Glu Lys Glu Gln Gln Ala Lys             180 185 190 Pro Leu Met Tyr His Met Ser Gln Arg Gly Trp Val Cys Val Ala Ile         195 200 205 Asn Tyr Arg Leu Ser Pro Lys Glu Ala Phe Pro Ala His Ile Val Asp     210 215 220 Val Lys Lys Ala Ile Ala Trp Val Arg Glu Asn Ile Ala Gln Tyr Gly 225 230 235 240 Gly Asp Pro Asn Phe Ile Ala Ile Thr Gly Gly Ser Ala Gly Gly His                 245 250 255 Leu Ser Ser Leu Ala Ala Leu Thr Pro Asn Arg Ala Gln Trp Gln Pro             260 265 270 Gly Phe Glu Asp Ala Asp Thr Thr Ile Gln Ala Ala Val Pro Phe Tyr         275 280 285 Gly Val Tyr Asp Phe Leu Asp Arg Tyr Asp Ile Arg Pro Asp Met Ser     290 295 300 Met Glu Asp Ile Ile Ala Asp Lys Val Leu Gln Cys Ala Lys Glu Asp 305 310 315 320 Asn His Gln Leu Trp Glu Asp Gly Ser Pro Met Ser His Ile Gly Ala                 325 330 335 His Ala Pro Pro Met Tyr Val Ile Gln Gly Thr His Asp Ser Leu Val             340 345 350 Trp Val Glu Glu Ala Arg Thr Phe Val Ala Ala Leu Gln Glu Val Ala         355 360 365 Thr His Pro Val Ala Tyr Ala Glu Leu Pro Gly Ala Gln His Ala Phe     370 375 380 Glu Ile Phe His Ser Val Arg Thr Asp His Thr Val Asn Ser Val Ala 385 390 395 400 Asp Phe Leu Glu Trp Ser His Ala Gly Trp Leu Gln Asn Lys Gly                 405 410 415 <210> 23 <211> 939 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 23 atgattagtc taagaggccg catgattcgc ttcatgtcga agcagttctt taaacgcatc 60 agacccgata gcgatattca taagctgcga actgaattcg aagcaattgg tacgaaaatg 120 cgcccggcag aaggcgtgca ggtcaggcat acaaaaattg ccggcatcga atgtgactgg 180 ctggtgccgg aaggatgtga tggcgcaccg atactttatt acctgcatgg cggtgcctac 240 atgatgggca gcccgaaaac gcatcggcgc atggtaagcc atattatcaa gagggccgga 300 atgcgggcgt tgctgccgga ctatcgcctc gccccggaaa acccgttccc ggcgtcgctc 360 gaagactcga ccagggtcta tcgtgcgctg attgaggccg gcacagattc gtcgaccatg 420 gcattcggtg gcgattcagc cggaggaaac ctggccatgg caacactgct ggcgctacgc 480 gatgcaggtg atcccttgcc ggcaacctgt ttcctgctgt caccatggct cgatctcgcg 540 gcccagggcg agtcacacga atcgcgcgcc gagcacgatc cgtggttcag agcagcagac 600 atgccggaga tcgtcatgaa gttttgttcc gaattcgacg taaaaaatcc gttggtgtcg 660 cctgtctacg ccgatgcatc tgatctgccg ccgatgctga ttcaggtagg ggatcatgag 720 attttgctca gcgattcaac ccgacttgca gataatattg caaaggcggg cggtgaggtt 780 acgttacagg tctggcccga tatgtggcat gtatttcagt tttttatagg ccagatgccg 840 gagagcaaaa aggcgatcaa aggaatagca gagtatttgc agaagcggtt tggtgcgcag 900 gaggtagtgc agcagccgca agaggatcag gctgcgtga 939 <210> 24 <211> 312 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 24 Met Ile Ser Leu Arg Gly Arg Met Ile Arg Phe Met Ser Lys Gln Phe   1 5 10 15 Phe Lys Arg Ile Arg Pro Asp Ser Asp Ile His Lys Leu Arg Thr Glu              20 25 30 Phe Glu Ala Ile Gly Thr Lys Met Arg Pro Ala Glu Gly Val Gln Val          35 40 45 Arg His Thr Lys Ile Ala Gly Ile Glu Cys Asp Trp Leu Val Pro Glu      50 55 60 Gly Cys Asp Gly Ala Pro Ile Leu Tyr Tyr Leu His Gly Gly Ala Tyr  65 70 75 80 Met Met Gly Ser Pro Lys Thr His Arg Arg Met Val Ser His Ile Ile                  85 90 95 Lys Arg Ala Gly Met Arg Ala Leu Leu Pro Asp Tyr Arg Leu Ala Pro             100 105 110 Glu Asn Pro Phe Pro Ala Ser Leu Glu Asp Ser Thr Arg Val Tyr Arg         115 120 125 Ala Leu Ile Glu Ala Gly Thr Asp Ser Ser Thr Met Ala Phe Gly Gly     130 135 140 Asp Ser Ala Gly Gly Asn Leu Ala Met Ala Thr Leu Leu Ala Leu Arg 145 150 155 160 Asp Ala Gly Asp Pro Leu Pro Ala Thr Cys Phe Leu Leu Ser Pro Trp                 165 170 175 Leu Asp Leu Ala Ala Gln Gly Glu Ser His Glu Ser Arg Ala Glu His             180 185 190 Asp Pro Trp Phe Arg Ala Ala Asp Met Pro Glu Ile Val Met Lys Phe         195 200 205 Cys Ser Glu Phe Asp Val Lys Asn Pro Leu Val Ser Pro Val Tyr Ala     210 215 220 Asp Ala Ser Asp Leu Pro Pro Met Leu Ile Gln Val Gly Asp His Glu 225 230 235 240 Ile Leu Leu Ser Asp Ser Thr Arg Leu Ala Asp Asn Ile Ala Lys Ala                 245 250 255 Gly Gly Glu Val Thr Leu Gln Val Trp Pro Asp Met Trp His Val Phe             260 265 270 Gln Phe Phe Ile Gly Gln Met Pro Glu Ser Lys Lys Ala Ile Lys Gly         275 280 285 Ile Ala Glu Tyr Leu Gln Lys Arg Phe Gly Ala Gln Glu Val Val Gln     290 295 300 Gln Pro Gln Glu Asp Gln Ala Ala 305 310 <210> 25 <211> 1566 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 25 atgaaaaaag ccactgcggt atttattttg atcgtaagcg tttatttccc atcatatgtt 60 tttgctggta gttttattca gagtgatcaa tgtaacatcg aaggacttcc ttccttacct 120 gacgtcaaca tcatatctgc cacccaagag atatcacctg ctccacactg caaggtcgca 180 ggcgtgattg gccccgaaat ccattttgaa ttactcttgc cagaaaaatg gaacggaaaa 240 ttcgtcatgg gtggcggtgg tggttatgtc ggttcagttg tgaatgtcgc agaagactta 300 tatggcgcag tgagctctgg ctacgccaca gccggcacag atactggcca ccaggcacac 360 agtctagatg ccagctgggc acagaacaat atggagcgga ttgtcaattt cggccatcaa 420 gccgtgcacc gtaccaaggt cactgcgaag gcattgatcg agtcctatta ccagcaagat 480 atctcacgta attactttat cggttgttca cgcggtggcg gccaaggtct gatggcagcg 540 caacgttatc ccgatgattt tgatgccatt gctgccggag ctccggccta caactggacc 600 ggccagcttg gcgccgccat ggttcaggtc cagcaagcca tgtatccgga cccagacaat 660 cttcaggaag cgattatcgg gccgaaagaa caggcgctga tagaatcttc ctatctggcc 720 gcttgtgatg cgctggacgg gattaaagat ggcattctga acgatcccag agattgtaag 780 tttgatgtgg gagccctgct gtgtaaggag actgatgggc aatgcctcag tcagcagcaa 840 ctagatgcag tccgcgttta ttatgaagga cccaaagata ataagggaaa ctcactctat 900 tatggctatc cccttggcgg cgaaaccagt ccgaccggtt ggtccttgtg gttaacaggc 960 ggtttgaaat atacggataa ttccggcgac ttccaggcag gaacccgctc tgattttgag 1020 gcgccggtga cgcccaatgc aatgttcccg tttagtacaa acttcctgaa atatatggtt 1080 tatcacaacc cggactgggc ttatacggga tacaactggg acaactatgt tgaagacagc 1140 acgttgatcg gtgcaaccct aaatgcgact aatcctgacc tgtctaagtt tcgcaagaag 1200 ggcggcaagc tgctgatgtt taccggttgg accgatgccg gaatttcccc cctagcaact 1260 ctcgggtact atgaaagcgt aatcgagcac gataatacag cggctgaaga tgttcgtcta 1320 ttcatgatgc ccggtgtgga ccactgcttg ggtggggttg gaccttcatt tgccaatttt 1380 ctaaccgatc tcgataagtg ggtagagtca ggcaaggcgc ccgaacaaac taccgcattt 1440 tggctcgatg aaaagtttca acccagcggc tccaggcttt tgtgtgctta tcccgaagtt 1500 gcccaatacg acggcaaagg ggatacgcgc gatgcagcca gctttagctg tgtgagccca 1560 gactag 1566 <210> 26 <211> 521 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 26 Met Lys Lys Ala Thr Ala Val Phe Ile Leu Ile Val Ser Val Tyr Phe   1 5 10 15 Pro Ser Tyr Val Phe Ala Gly Ser Phe Ile Gln Ser Asp Gln Cys Asn              20 25 30 Ile Glu Gly Leu Pro Ser Leu Pro Asp Val Asn Ile Ile Ser Ala Thr          35 40 45 Gln Glu Ile Ser Pro Ala Pro His Cys Lys Val Ala Gly Val Ile Gly      50 55 60 Pro Glu Ile His Phe Glu Leu Leu Leu Pro Glu Lys Trp Asn Gly Lys  65 70 75 80 Phe Val Met Gly Gly Gly Gly Gly Tyr Val Gly Ser Val Val Asn Val                  85 90 95 Ala Glu Asp Leu Tyr Gly Ala Val Ser Ser Gly Tyr Ala Thr Ala Gly             100 105 110 Thr Asp Thr Gly His Gln Ala His Ser Leu Asp Ala Ser Trp Ala Gln         115 120 125 Asn Asn Met Glu Arg Ile Val Asn Phe Gly His Gln Ala Val His Arg     130 135 140 Thr Lys Val Thr Ala Lys Ala Leu Ile Glu Ser Tyr Tyr Gln Gln Asp 145 150 155 160 Ile Ser Arg Asn Tyr Phe Ile Gly Cys Ser Arg Gly Gly Gly Gln Gly                 165 170 175 Leu Met Ala Ala Gln Arg Tyr Pro Asp Asp Phe Asp Ala Ile Ala Ala             180 185 190 Gly Ala Pro Ala Tyr Asn Trp Thr Gly Gln Leu Gly Ala Ala Met Val         195 200 205 Gln Val Gln Gln Ala Met Tyr Pro Asp Pro Asp Asn Leu Gln Glu Ala     210 215 220 Ile Ile Gly Pro Lys Glu Gln Ala Leu Ile Glu Ser Ser Tyr Leu Ala 225 230 235 240 Ala Cys Asp Ala Leu Asp Gly Ile Lys Asp Gly Ile Leu Asn Asp Pro                 245 250 255 Arg Asp Cys Lys Phe Asp Val Gly Ala Leu Leu Cys Lys Glu Thr Asp             260 265 270 Gly Gln Cys Leu Ser Gln Gln Gln Leu Asp Ala Val Arg Val Tyr Tyr         275 280 285 Glu Gly Pro Lys Asp Asn Lys Gly Asn Ser Leu Tyr Tyr Gly Tyr Pro     290 295 300 Leu Gly Gly Glu Thr Ser Pro Thr Gly Trp Ser Leu Trp Leu Thr Gly 305 310 315 320 Gly Leu Lys Tyr Thr Asp Asn Ser Gly Asp Phe Gln Ala Gly Thr Arg                 325 330 335 Ser Asp Phe Glu Ala Pro Val Thr Pro Asn Ala Met Phe Pro Phe Ser             340 345 350 Thr Asn Phe Leu Lys Tyr Met Val Tyr His Asn Pro Asp Trp Ala Tyr         355 360 365 Thr Gly Tyr Asn Trp Asp Asn Tyr Val Glu Asp Ser Thr Leu Ile Gly     370 375 380 Ala Thr Leu Asn Ala Thr Asn Pro Asp Leu Ser Lys Phe Arg Lys Lys 385 390 395 400 Gly Gly Lys Leu Leu Met Phe Thr Gly Trp Thr Asp Ala Gly Ile Ser                 405 410 415 Pro Leu Ala Thr Leu Gly Tyr Tyr Glu Ser Val Ile Glu His Asp Asn             420 425 430 Thr Ala Ala Glu Asp Val Arg Leu Phe Met Met Pro Gly Val Asp His         435 440 445 Cys Leu Gly Gly Val Gly Pro Ser Phe Ala Asn Phe Leu Thr Asp Leu     450 455 460 Asp Lys Trp Val Glu Ser Gly Lys Ala Pro Glu Gln Thr Thr Ala Phe 465 470 475 480 Trp Leu Asp Glu Lys Phe Gln Pro Ser Gly Ser Arg Leu Leu Cys Ala                 485 490 495 Tyr Pro Glu Val Ala Gln Tyr Asp Gly Lys Gly Asp Thr Arg Asp Ala             500 505 510 Ala Ser Phe Ser Cys Val Ser Pro Asp         515 520 <210> 27 <211> 1500 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 27 atggcgatgt taaattgctc gagcagactg ccgatggtaa gttcaagcta caacaacgaa 60 aagcctacat tcatacacgg tcgaggattt tcgaaatttc tgatcctcct aaaagtgaat 120 tatcaggttt cgacgaacct tagaagccgc tattactcgc gcttcaaaat aggggaaata 180 acaatgaaaa agctaatgca gtcattcttt atcgtcattc tgtttgcatg gggcagcgct 240 gctgcggaca aaccgccggg ctatttcgtc gatgagagta aactcccgtt ccccgcactg 300 ccaagtgcca cggccatgtg gggagtgcat aaaggtgccg gctatcgtat tgaagtgccg 360 gataactgga acgggacgct ggtcgtctgg gcgcacggtt tccgcggcac cggacttgag 420 ctgaccgtcg acaaccaccc cctgcgcgaa tatctgattc ccaacggtta cgcttgggcg 480 gcatcgagtt acagtcgcaa cgattacgat attacgactg gcgttcagga tacgcacgcg 540 cttacccagc gcttcaacgg catcgtcggc aagcccgaca aagtttatct gaccggtgcg 600 tcaatgggcg gtcatgtcac ggccgtagcg atcgagcagt atccaaacac ctacgacgcg 660 gccttgccca tttgcggcgt gctcggagac tacgaattgt tcgattactt cctggatttc 720 aatgtggctg cgcagcagct gggtgtaggc ggttcgcagt acccggtcga tccgattccg 780 tatcttttcg gaacagtgcc ggcaataaaa ggtaacttgg agcaattccc cgggggatgg 840 cctgtggatt tgaatgcaca gggaacccag ctgaagtcgc tgacagagct tcgcagtggc 900 ggcaagcggc caaatttcga tgaggcctgg tttttctgga atacttttgc aaactttgaa 960 tccggccccg gcaatttctt gttcgatcta gcgatcggcg acggcactct gccacgcaca 1020 cccggtgttg gagtcgataa cagcgatacg gtctatcagt tcgacaccga cccgtccctg 1080 agcaccgaag aagaaaactt caacaatgat gttgtccgcg tcgcagccga tccacagggc 1140 cgaaatccga acggcttgtc tcaggtacca gtgatctcag gagatatcca gattccggta 1200 ctcacgttgc acaatctagg cgatttgttc gtgccggttc ttaacgaagt cgaatatgca 1260 tcccgcgtag cggcgaatgg aaaaagtgac cttctggtac agcgcgcaat tcgcggcgtg 1320 cttcactgcg acttcactcc agttgaattt gtaacggcat tcgcggaatt agcagcctgg 1380 gagtccttcg gagtgaaacc tgctggcgat gactggctcg atccagcggc agtagccgat 1440 ccattgttcg gctgcgcctt cacggacggg ccccacctct tcggcacacc ctgtccgtaa 1500                                                                         1500 <210> 28 <211> 499 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 28 Met Ala Met Leu Asn Cys Ser Ser Arg Leu Pro Met Val Ser Ser Ser   1 5 10 15 Tyr Asn Asn Glu Lys Pro Thr Phe Ile His Gly Arg Gly Phe Ser Lys              20 25 30 Phe Leu Ile Leu Leu Lys Val Asn Tyr Gln Val Ser Thr Asn Leu Arg          35 40 45 Ser Arg Tyr Tyr Ser Arg Phe Lys Ile Gly Glu Ile Thr Met Lys Lys      50 55 60 Leu Met Gln Ser Phe Phe Ile Val Ile Leu Phe Ala Trp Gly Ser Ala  65 70 75 80 Ala Ala Asp Lys Pro Pro Gly Tyr Phe Val Asp Glu Ser Lys Leu Pro                  85 90 95 Phe Pro Ala Leu Pro Ser Ala Thr Ala Met Trp Gly Val His Lys Gly             100 105 110 Ala Gly Tyr Arg Ile Glu Val Pro Asp Asn Trp Asn Gly Thr Leu Val         115 120 125 Val Trp Ala His Gly Phe Arg Gly Thr Gly Leu Glu Leu Thr Val Asp     130 135 140 Asn His Pro Leu Arg Glu Tyr Leu Ile Pro Asn Gly Tyr Ala Trp Ala 145 150 155 160 Ala Ser Ser Tyr Ser Arg Asn Asp Tyr Asp Ile Thr Thr Gly Val Gln                 165 170 175 Asp Thr His Ala Leu Thr Gln Arg Phe Asn Gly Ile Val Gly Lys Pro             180 185 190 Asp Lys Val Tyr Leu Thr Gly Ala Ser Met Gly Gly His Val Thr Ala         195 200 205 Val Ala Ile Glu Gln Tyr Pro Asn Thr Tyr Asp Ala Ala Leu Pro Ile     210 215 220 Cys Gly Val Leu Gly Asp Tyr Glu Leu Phe Asp Tyr Phe Leu Asp Phe 225 230 235 240 Asn Val Ala Ala Gln Gln Leu Gly Val Gly Gly Ser Gln Tyr Pro Val                 245 250 255 Asp Pro Ile Pro Tyr Leu Phe Gly Thr Val Pro Ala Ile Lys Gly Asn             260 265 270 Leu Glu Gln Phe Pro Gly Gly Trp Pro Val Asp Leu Asn Ala Gln Gly         275 280 285 Thr Gln Leu Lys Ser Leu Thr Glu Leu Arg Ser Gly Gly Lys Arg Pro     290 295 300 Asn Phe Asp Glu Ala Trp Phe Phe Trp Asn Thr Phe Ala Asn Phe Glu 305 310 315 320 Ser Gly Pro Gly Asn Phe Leu Phe Asp Leu Ala Ile Gly Asp Gly Thr                 325 330 335 Leu Pro Arg Thr Pro Gly Val Gly Val Asp Asn Ser Asp Thr Val Tyr             340 345 350 Gln Phe Asp Thr Asp Pro Ser Leu Ser Thr Glu Glu Glu Asn Phe Asn         355 360 365 Asn Asp Val Val Arg Val Ala Ala Asp Pro Gln Gly Arg Asn Pro Asn     370 375 380 Gly Leu Ser Gln Val Pro Val Ile Ser Gly Asp Ile Gln Ile Pro Val 385 390 395 400 Leu Thr Leu His Asn Leu Gly Asp Leu Phe Val Pro Val Leu Asn Glu                 405 410 415 Val Glu Tyr Ala Ser Arg Val Ala Ala Asn Gly Lys Ser Asp Leu Leu             420 425 430 Val Gln Arg Ala Ile Arg Gly Val Leu His Cys Asp Phe Thr Pro Val         435 440 445 Glu Phe Val Thr Ala Phe Ala Glu Leu Ala Ala Trp Glu Ser Phe Gly     450 455 460 Val Lys Pro Ala Gly Asp Asp Trp Leu Asp Pro Ala Ala Val Ala Asp 465 470 475 480 Pro Leu Phe Gly Cys Ala Phe Thr Asp Gly Pro His Leu Phe Gly Thr                 485 490 495 Pro Cys Pro             <210> 29 <211> 1059 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 29 atgaattcac gactcgcgat cattcttgtt gccgttttgg cagctggttc cctaacacca 60 ctcgcttcgt cggctcaaga ttcctccgac tcaaagacca tggaacatcg ctggatcccg 120 acaccggcag gggtctccca agagttacag gccttcattg ccggcaaggt gatcccgccg 180 ccaatgcccg ttcccaaaac ccgagaggag tggttgacgc ttcagcgtac gaaggacgca 240 ggtgacgcca agcaggccga ggaattcgcc aagaaactcg gagcgagttg gaaggcgatg 300 aaggtggggg gagtacagtg ttttcttgta acacccaaaa agatcgggga acggttcaaa 360 gatcgctggc tggtccacat ccatggaggc gcgtttgtgt tcaatggtgg tgaaggatgt 420 gtgaccgagg ctatttggat agcagatgca tgcgagtctc aggtgctttc aatcgattac 480 cgacggccgc ccgttcatcc tttcccagcc gcgattgatg acgctgtggc agtgtggaaa 540 gccgtaatcg agaagcagga tgcgtccgcc acggcgctat tcggcacttc tgcgggtggt 600 aaccttacgc tggcgacgac gctcaaactc aagcaactcg gacttgagct tcccggtgcc 660 ttgttcgcgg gtacgccagc aactgacctc gagaagattg gagacacgtg gtacacgctg 720 gaaggtcttg actcgcttgg tcgatacgat ggtttcctcg ccgcctgctt cgaggcctac 780 ttcccttcaa aggattcctc taatccgctg gtctctcctg tccgtggtga cgtcaagggc 840 ttcccaccta cgattctgat ttcaggcacg cgggacttac ttctcagcga caccgcacga 900 atgcaccgcg cgctgcgcga ggccgaagtt gaagccgaac tccacgtcta cgaggggcaa 960 tctcacggtg attatctcat gggcactctg gttgatgcgc cagaatcaaa agacgcgcag 1020 aaggagatct tccgattctt tgacaagcat ctaaagtga 1059 <210> 30 <211> 352 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 30 Met Asn Ser Arg Leu Ala Ile Ile Leu Val Ala Val Leu Ala Ala Gly   1 5 10 15 Ser Leu Thr Pro Leu Ala Ser Ser Ala Gln Asp Ser Ser Asp Ser Lys              20 25 30 Thr Met Glu His Arg Trp Ile Pro Thr Pro Ala Gly Val Ser Gln Glu          35 40 45 Leu Gln Ala Phe Ile Ala Gly Lys Val Ile Pro Pro Pro Met Pro Val      50 55 60 Pro Lys Thr Arg Glu Glu Trp Leu Thr Leu Gln Arg Thr Lys Asp Ala  65 70 75 80 Gly Asp Ala Lys Gln Ala Glu Glu Phe Ala Lys Lys Leu Gly Ala Ser                  85 90 95 Trp Lys Ala Met Lys Val Gly Gly Val Gln Cys Phe Leu Val Thr Pro             100 105 110 Lys Lys Ile Gly Glu Arg Phe Lys Asp Arg Trp Leu Val His Ile His         115 120 125 Gly Gly Ala Phe Val Phe Asn Gly Gly Glu Gly Cys Val Thr Glu Ala     130 135 140 Ile Trp Ile Ala Asp Ala Cys Glu Ser Gln Val Leu Ser Ile Asp Tyr 145 150 155 160 Arg Arg Pro Pro Val His Pro Phe Pro Ala Ala Ile Asp Asp Ala Val                 165 170 175 Ala Val Trp Lys Ala Val Ile Glu Lys Gln Asp Ala Ser Ala Thr Ala             180 185 190 Leu Phe Gly Thr Ser Ala Gly Gly Asn Leu Thr Leu Ala Thr Thr Leu         195 200 205 Lys Leu Lys Gln Leu Gly Leu Glu Leu Pro Gly Ala Leu Phe Ala Gly     210 215 220 Thr Pro Ala Thr Asp Leu Glu Lys Ile Gly Asp Thr Trp Tyr Thr Leu 225 230 235 240 Glu Gly Leu Asp Ser Leu Gly Arg Tyr Asp Gly Phe Leu Ala Ala Cys                 245 250 255 Phe Glu Ala Tyr Phe Pro Ser Lys Asp Ser Ser Asn Pro Leu Val Ser             260 265 270 Pro Val Arg Gly Asp Val Lys Gly Phe Pro Pro Thr Ile Leu Ile Ser         275 280 285 Gly Thr Arg Asp Leu Leu Leu Ser Asp Thr Ala Arg Met His Arg Ala     290 295 300 Leu Arg Glu Ala Glu Val Glu Ala Glu Leu His Val Tyr Glu Gly Gln 305 310 315 320 Ser His Gly Asp Tyr Leu Met Gly Thr Leu Val Asp Ala Pro Glu Ser                 325 330 335 Lys Asp Ala Gln Lys Glu Ile Phe Arg Phe Phe Asp Lys His Leu Lys             340 345 350 <210> 31 <211> 951 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 31 atggattatg ttcctgccac agaagaggag tggcttgtca atatcaaaga aaaagatgct 60 ggggctgcgg tagtagcgca gtcaattgcc gacgcgctta atatagctat caagaaagat 120 aggctcgcag gcgtgaatgt gcattgggtg acccctcctg atatcgatgc ggagcataaa 180 ggtcacctgt tcatttatat ccacggtggt gcatatgttt acaacggagg gatggctggg 240 ctagtggagc catcacttat tgctcaccac ctgaaaatcc aggttgtttc aatcgattac 300 agaatgccgc ccaaacatcc tgcgccttcc gccactgacg atgtgattac cgtttggaaa 360 gagctgctga agactcgagc gcacacctct atggcaatgg gggggacatc agcgggagga 420 aatatcgcgc ttgcgtccag ccaacgcttt gtaggccttg gaatggctgt tccagcagcg 480 ctctacatcg gcacgcccac cgtcgatttg aatgcagaag ctggggacag ccgttacctg 540 aatgaagggg ctgaccgagt tctgatcaag atgggcgact ttggcgcagc tgcgatgaac 600 ctatatgctg ggaaactggg tctgaaccat ccttatgttt ctccaatctt tggtgatttc 660 acagggctac ctccaactta cctcatcacc ggcacaagag atcttcttct gagtgacact 720 gtgcgagcgc accgtgcgct gcgacgagcc ggcatagcgg cagagctaca tgtgtatgaa 780 ggccaaggtc acgcggatta tattgttgca atgaaaacac cggaatctgc tgaacattac 840 gctgaactga atcggtttgt acttgagcat ctgtcggctc ctttggtgac tgaatctatg 900 atttccgggg aacagattcc aaaggagatt ccaaagtccg ttggatacta g 951 <210> 32 <211> 316 <212> PRT <213> Unknown <220> <223> DNA isolated from metagenome library <400> 32 Met Asp Tyr Val Pro Ala Thr Glu Glu Glu Trp Leu Val Asn Ile Lys   1 5 10 15 Glu Lys Asp Ala Gly Ala Ala Val Val Ala Gln Ser Ile Ala Asp Ala              20 25 30 Leu Asn Ile Ala Ile Lys Lys Asp Arg Leu Ala Gly Val Asn Val His          35 40 45 Trp Val Thr Pro Pro Asp Ile Asp Ala Glu His Lys Gly His Leu Phe      50 55 60 Ile Tyr Ile His Gly Gly Ala Tyr Val Tyr Asn Gly Met Ala Gly  65 70 75 80 Leu Val Glu Pro Ser Leu Ile Ala His His Leu Lys Ile Gln Val Val                  85 90 95 Ser Ile Asp Tyr Arg Met Pro Pro Lys His Pro Ala Pro Ser Ala Thr             100 105 110 Asp Asp Val Ile Thr Val Trp Lys Glu Leu Leu Lys Thr Arg Ala His         115 120 125 Thr Ser Met Ala Met Gly Gly Thr Ser Ala Gly Gly Asn Ile Ala Leu     130 135 140 Ala Ser Ser Gln Arg Phe Val Gly Leu Gly Met Ala Val Pro Ala Ala 145 150 155 160 Leu Tyr Ile Gly Thr Pro Thr Val Asp Leu Asn Ala Glu Ala Gly Asp                 165 170 175 Ser Arg Tyr Leu Asn Glu Gly Ala Asp Arg Val Leu Ile Lys Met Gly             180 185 190 Asp Phe Gly Ala Ala Ala Met Asn Leu Tyr Ala Gly Lys Leu Gly Leu         195 200 205 Asn His Pro Tyr Val Ser Pro Ile Phe Gly Asp Phe Thr Gly Leu Pro     210 215 220 Pro Thr Tyr Leu Ile Thr Gly Thr Arg Asp Leu Leu Leu Ser Asp Thr 225 230 235 240 Val Arg Ala His Arg Ala Leu Arg Arg Ala Gly Ile Ala Ala Glu Leu                 245 250 255 His Val Tyr Glu Gly Gln Gly His Ala Asp Tyr Ile Val Ala Met Lys             260 265 270 Thr Pro Glu Ser Ala Glu His Tyr Ala Glu Leu Asn Arg Phe Val Leu         275 280 285 Glu His Leu Ser Ala Pro Leu Val Thr Glu Ser Met Ile Ser Gly Glu     290 295 300 Gln Ile Pro Lys Glu Ile Pro Lys Ser Val Gly Tyr 305 310 315 <210> 33 <211> 1119 <212> DNA <213> Unknown <220> <223> DNA isolated from metagenome library <400> 33 atgaaatcaa atctcatcag aatacaggct tttgtctttg catttatgtc acttgcagtg 60 atagcaaatg cactggcgaa agagaatatt tggacgattg gaccgagaat acttccgcca 120 tccgtggatg tgagcgatga attccgtgag gtaatgttga atactccggc accggatgta 180 aaagcggtga aaaagaatgt tcctgaaacc accgaacaat gggtaaccat gattcgggcg 240 ttggatgata aaacggcagc ggtggcacgg aaactggcac agacgctttc ggtaaatgtc 300 cagcatgaca caattgccgg ggtgaatgtc taccgggtca tgccgccgga aattgctccg 360 gaacatcaaa aacatctttt ctttcacgtg catgggggcg gatacatctt aaacggcggt 420 gaggccggca cctacgaagc ggttttgatt gccagccacc tgaagatagg tgttgtttca 480 attgattatc gaatgccgcc aaagcacccg gcgcctgctg ctatggagga tgtcgttttg 540 gtttggaaag agttactgaa agagcgttct ccagctctaa tggccatggg gggtacttcc 600 ggcggagcag gccttacctt gtcgtcagtc cagcatttta aagaacttgg gttagcgctg 660 cctcaggcgc ttctcctggg cacaccctgc gtagatgccg gcaaagtggg tgatgctcgc 720 tacattaatg acggcatcga cacgagcctg gccagctggg acggactttt agagcaagcg 780 ctgaaaatgt atgcagtcag ttacgatctt acacaccccc atgtttcacc catttacgga 840 gattttgagg gttttccgcc aagttatctg atcagcggaa caagggatct tatgttgagt 900 gataccgttc gcgtacaccg gaaattgcga agggccggtg tggatgcgga cctgcacgta 960 tatgaaggcc aatcacatgc tgactatgtc aaggttatta atgcgcccga gtcctttgag 1020 cacttcgccg aactgaatgc attcttgcta aaacatctcc agtatccgtt gctgtctaag 1080 ccagtcgaga cgttgaagga tattgaggtc ccgaaataa 1119 <210> 34 <211> 372 <212> PRT <213> Unknown <220> <223> protein isolated from metagenome library <400> 34 Met Lys Ser Asn Leu Ile Arg Ile Gln Ala Phe Val Phe Ala Phe Met   1 5 10 15 Ser Leu Ala Val Ile Ala Asn Ala Leu Ala Lys Glu Asn Ile Trp Thr              20 25 30 Ile Gly Pro Arg Ile Leu Pro Pro Ser Val Asp Val Ser Asp Glu Phe          35 40 45 Arg Glu Val Met Leu Asn Thr Pro Ala Pro Asp Val Lys Ala Val Lys      50 55 60 Lys Asn Val Pro Glu Thr Thr Glu Gln Trp Val Thr Met Ile Arg Ala  65 70 75 80 Leu Asp Asp Lys Thr Ala Ala Val Ala Arg Lys Leu Ala Gln Thr Leu                  85 90 95 Ser Val Asn Val Gln His Asp Thr Ile Ala Gly Val Asn Val Tyr Arg             100 105 110 Val Met Pro Pro Glu Ile Ala Pro Glu His Gln Lys His Leu Phe Phe         115 120 125 His Val His Gly Gly Gly Tyr Ile Leu Asn Gly Gly Glu Ala Gly Thr     130 135 140 Tyr Glu Ala Val Leu Ile Ala Ser His Leu Lys Ile Gly Val Val Ser 145 150 155 160 Ile Asp Tyr Arg Met Pro Pro Lys His Pro Ala Pro Ala Ala Met Glu                 165 170 175 Asp Val Val Leu Val Trp Lys Glu Leu Leu Lys Glu Arg Ser Pro Ala             180 185 190 Leu Met Ala Met Gly Gly Thr Ser Gly Gly Ala Gly Leu Thr Leu Ser         195 200 205 Ser Val Gln His Phe Lys Glu Leu Gly Leu Ala Leu Pro Gln Ala Leu     210 215 220 Leu Leu Gly Thr Pro Cys Val Asp Ala Gly Lys Val Gly Asp Ala Arg 225 230 235 240 Tyr Ile Asn Asp Gly Ile Asp Thr Ser Leu Ala Ser Trp Asp Gly Leu                 245 250 255 Leu Glu Gln Ala Leu Lys Met Tyr Ala Val Ser Tyr Asp Leu Thr His             260 265 270 Pro His Val Ser Pro Ile Tyr Gly Asp Phe Glu Gly Phe Pro Pro Ser         275 280 285 Tyr Leu Ile Ser Gly Thr Arg Asp Leu Met Leu Ser Asp Thr Val Arg     290 295 300 Val His Arg Lys Leu Arg Arg Ala Gly Val Asp Ala Asp Leu His Val 305 310 315 320 Tyr Glu Gly Gln Ser His Ala Asp Tyr Val Lys Val Ile Asn Ala Pro                 325 330 335 Glu Ser Phe Glu His Phe Ala Glu Leu Asn Ala Phe Leu Leu Lys His             340 345 350 Leu Gln Tyr Pro Leu Leu Ser Lys Pro Val Glu Thr Leu Lys Asp Ile         355 360 365 Glu val pro lys     370  

Claims (119)

A protein having the amino acid sequence of SEQ ID NO. Gene encoding the protein of claim 1. Esterase having the amino acid sequence of SEQ ID NO: 2. Esterase gene, characterized in that encoding the esterase of claim 3. Recombinant vector comprising SEQ ID NO: 1. A cell transformed with the recombinant vector according to claim 5. Esterase enzyme production method characterized by culturing the transformed cells according to claim 6, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. Gene encoding the protein of claim 8. Esterase having the amino acid sequence of SEQ ID NO: 4. Esterase gene, characterized in that encoding the esterase of claim 10. Recombinant vector comprising SEQ ID NO: 3. A cell transformed with the recombinant vector according to claim 11. Esterase enzyme production method comprising culturing the transformed cells according to claim 13, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 15. Esterase having the amino acid sequence of SEQ ID NO: 6. Esterase gene, characterized in that encoding the esterase of claim 17. Recombinant vector comprising SEQ ID NO: 5. A cell transformed with the recombinant vector according to claim 19. Esterase enzyme production method comprising culturing the transformed cells according to claim 20, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 22. Lipase having the amino acid sequence of SEQ ID NO: 8. A lipase gene, characterized by encoding the lipase of claim 24. Recombinant vector comprising SEQ ID NO: 7. A cell transformed with the recombinant vector according to claim 26. A method for producing a lipase enzyme, comprising culturing the transformed cell according to claim 27 and separating the lipase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 29. Esterase having the amino acid sequence of SEQ ID NO: 10. An esterase gene characterized by encoding the esterase of claim 31. Recombinant vector comprising SEQ ID NO: 9. A cell transformed with the recombinant vector according to claim 33. Esterase enzyme production method comprising culturing the transformed cells according to claim 34, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 36. Lipase having the amino acid sequence of SEQ ID NO: 12. The lipase gene, which encodes the lipase of claim 38. Recombinant vector comprising SEQ ID NO: 11. A cell transformed with the recombinant vector according to claim 40. A method for producing a lipase enzyme, comprising culturing the transformed cell according to claim 41 and separating the lipase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 43. Esterase having the amino acid sequence of SEQ ID NO: 14. Esterase gene, characterized by encoding the esterase of claim 45. Recombinant vector comprising SEQ ID NO: 13. A cell transformed with the recombinant vector according to claim 47. Esterase enzyme production method comprising culturing the transformed cells according to claim 48, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 50. Esterase having the amino acid sequence of SEQ ID NO: 16. The esterase gene, which encodes the esterase of claim 52. Recombinant vector comprising SEQ ID NO: 15. A cell transformed with the recombinant vector according to claim 54. Esterase enzyme production method characterized by culturing the transformed cells according to claim 55, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 57. Esterase having the amino acid sequence of SEQ ID NO: 18. An esterase gene characterized by encoding the esterase of claim 59. Recombinant vector comprising SEQ ID NO: 17. A cell transformed with the recombinant vector according to claim 61. The method of producing an esterase enzyme, comprising culturing the transformed cell according to claim 62 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 64. Esterase having the amino acid sequence of SEQ ID NO: 20. An esterase gene characterized by encoding the esterase of claim 66. Recombinant vector comprising SEQ ID NO: 19. A cell transformed with the recombinant vector according to claim 68. The method of producing an esterase enzyme, comprising culturing the transformed cell according to claim 69 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 71. Esterase having the amino acid sequence of SEQ ID NO: 22. Esterase gene, characterized in that encoding the esterase of claim 73. A recombinant vector comprising SEQ ID NO: 21. A cell transformed with the recombinant vector according to claim 75. The method of producing an esterase enzyme, comprising culturing the transformed cell according to claim 76 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 78. Esterase having amino acid sequence of SEQ ID NO: 24. Esterase gene, characterized in that for encoding the esterase of claim 80. Recombinant vector comprising SEQ ID NO: 23. The cell transformed with the recombinant vector according to claim 82. Esterase enzyme production method characterized by culturing the transformed cells according to claim 83, and separating the esterase enzyme from the cultured cells. A protein having the amino acid sequence of SEQ ID NO. The gene encoding the protein of claim 85. An esterase having the amino acid sequence of SEQ ID NO: 26. Esterase gene, characterized in that for encoding the esterase of claim 87. A recombinant vector comprising SEQ ID NO: 25. A cell transformed with the recombinant vector according to claim 89. The method of producing an esterase enzyme, comprising culturing the transformed cell according to claim 90 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. The gene encoding the protein of claim 92. Esterase having the amino acid sequence of SEQ ID NO: 28. An esterase gene characterized by encoding the esterase of claim 94. A recombinant vector comprising SEQ ID NO: 27. 97. A cell transformed with the recombinant vector according to claim 96. 97. A method for producing an esterase enzyme, comprising culturing the transformed cell according to claim 97 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID 30. The gene encoding the protein of claim 99. Esterase having the amino acid sequence of SEQ ID NO: 30. Esterase gene, characterized by encoding the esterase of claim 101. Recombinant vector comprising SEQ ID NO: 29. 103. A cell transformed with the recombinant vector according to claim 103. 104. A method for producing an esterase enzyme, comprising culturing the transformed cell according to claim 104 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID 32. 108. A gene encoding the protein of claim 106. Esterase having amino acid sequence of SEQ ID NO: 32. Esterase gene, characterized in that for encoding the esterase of claim 108. Recombinant vector comprising SEQ ID NO: 31. A cell transformed with the recombinant vector according to claim 110. A method of producing an esterase enzyme, comprising culturing the transformed cell according to claim 111 and separating an esterase enzyme from the cultured cell. A protein having the amino acid sequence of SEQ ID NO. A gene encoding the protein of claim 113. Esterase having amino acid sequence of SEQ ID NO: 34. Esterase gene, characterized by encoding the esterase of claim 115. A recombinant vector comprising SEQ ID NO: 33. 117. A cell transformed with the recombinant vector according to claim 117. 118. A method for producing an esterase enzyme, comprising culturing the transformed cell according to claim 118 and separating an esterase enzyme from the cultured cell.

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