D-psicose 3-epimerase for bioconversion of D-fructose to D-psicose

Abstract

Provided herein are methods for identifying and isolating polynucleotides encoding polypeptides having D-psicose 3-epimerase activity from a wide variety of microorganisms. Also provided are nucleic acid constructs, vectors, and recombinant host cells comprising polynucleotides encoding D-psicose 3-epimerase activity, and methods for producing psicose from fructose using the recombinant host cells having D-psicose 3-epimerase activity or the D-psicose 3-epimerase enzyme of the recombinant host cells having D-psicose 3-epimerase activity.

Description

[0001] Related Applications

[0002] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 63 / 022,617, filed on May 11, 2020, entitled “D-ALLULOSE 3-EPIMERASES FOR BIOCONVERSION OF D-FRUCTOSE TO D-ALLULOSE,” the contents of which are hereby incorporated by reference in their entirety. Field of the Invention

[0003] The present invention relates to a method for producing D-psicose from fructose using a D-psicose 3-epimerase enzyme. The present invention provides a polypeptide having D-psicose 3-epimerase activity, and a nucleic acid molecule encoding the polypeptide. The present invention also relates to a recombinant nucleic acid construct comprising a vector and a recombinant host cell comprising the recombinant nucleic acid construct. Background Art

[0004] D-allulose, also known as D-psicose, is a class of sugars that are structurally similar to fructose, a naturally occurring sugar in fruit. D-psicose is available in granular form and looks like sucrose crystals. Psicose is a low-calorie sweetener with 70% of the sweetness of sucrose. According to the U.S. Food and Drug Administration (FDA), Psicose provides about 0.4 calories per gram, significantly less than the 4 calories per gram provided by sucrose. Although the human body has the ability to absorb Psicose, it lacks the ability to metabolize it. Therefore, Psicose has little effect on blood sugar or insulin levels. For this reason, Psicose is considered a low-calorie sweetener.

[0005] Psicose occurs naturally in some foods, such as dried fruits, brown sugar, and maple syrup, in very small amounts. Psicose is difficult to chemically synthesize, and there is a need in the art to produce Psicose from readily available sugars, such as fructose, using recombinant microorganisms. Summary of the invention

[0006] Enzymes from the ketose 3-epimerase family, including D-allulose 3-epimerase (DAE; also known as D-psicose 3-epimerase (DPE)), can catalyze the reversible conversion of D-fructose into D-psicose. D-psicose 3-epimerase can have high activity for the bioconversion of D-fructose into D-psicose. The present invention particularly provides a method for producing D-psicose from D-fructose using a recombinant polypeptide having D-psicose 3-epimerase activity.

[0007] In one embodiment, the method of the present invention involves contacting a fructose substrate with a reaction mixture comprising an enzyme system under conditions such that the D-fructose substrate is converted to D-psicose, the enzyme system comprising a recombinant polypeptide having D-psicose 3-epimerase enzyme activity. In one aspect of the present invention, the enzyme system provides a recombinant polypeptide having D-psicose 3-epimerase enzyme activity and D-fructose as a substrate to produce D-psicose as a product. In another aspect of the present invention, the method of the present invention may include contacting a fructose substrate with a reaction mixture, the reaction mixture comprising a host cell capable of expressing an exogenous D-psicose 3-epimerase or a lysate of the host cell as a source of the D-psicose 3-epimerase enzyme. In certain aspects of this embodiment, the D-psicose 3-epimerase enzyme present in the enzyme system is a purified enzyme, and it can be derived from a lysate of a host cell expressing the D-psicose 3-epimerase enzyme using one or more biochemical techniques known in the art. In another aspect of the present invention, a purified form of D-psicose 3-epimerase is immobilized on a solid support. In various embodiments, the inventive method may further include adding at least one type of metal ion to the reaction system. For example, the metal ion may be selected from the group consisting of magnesium ion, manganese ion, copper ion, zinc ion, nickel ion, cobalt ion, iron ion, aluminum ion and calcium ion. In some preferred embodiments, manganese ion and / or magnesium ion are added to the reaction system. The metal ion may be added at a concentration of about 0.01 mM to about 5 mM (e.g., 0.01 mM, 0.05 mM, 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM or 5 mM). In various aspects, the inventive method may include removing a product stream containing psicose without residual fructose in the reaction system.

[0008] In another embodiment, the present invention provides a polynucleotide sequence encoding a polypeptide having D-psicose 3-epimerase activity. In one aspect of this embodiment, the polynucleotide encoding the polypeptide having D-psicose 3-epimerase activity is cloned into an appropriate plasmid expression vector having required regulatory elements such as a promoter and a terminator. In one aspect, the plasmid expression vector has an inducible promoter so that the expression of D-psicose 3-epimerase enzyme activity can be induced by using certain chemicals.

[0009] In another embodiment, an expression plasmid vector comprising a polynucleotide sequence encoding a polypeptide having D-psicose 3-epimerase activity is used to transform a host cell, including a prokaryotic microbial cell, a eukaryotic microbial cell, a eukaryotic animal cell, or a eukaryotic plant cell. In one aspect, the expression plasmid vector in the transformed host cell exists as a self-replicating nucleic acid entity. In another aspect of this embodiment, the expression plasmid vector is integrated into the host chromosomal DNA.

[0010] In another embodiment, the present invention provides a method for recovering D-psicose without D-fructose as a substrate in a reaction involving D-psicose 3-epimerase.

[0011] In some embodiments, the method for producing psicose described herein includes contacting a fructose substrate with a reaction mixture under conditions such that the fructose substrate is converted into psicose, the reaction mixture comprising:

[0012] (a) an enzyme system comprising a D-psicose 3-epimerase enzyme;

[0013] (b) a host cell transformed with a recombinant vector comprising a nucleic acid molecule encoding a D-psicose 3-epimerase enzyme; and / or

[0014] (c) a lysate of the host cells of (b), wherein the D-psicose 3-epimerase enzyme comprises an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, and SEQ ID NO: 43. In some embodiments, the reaction mixture comprises a D-psicose 3-epimerase enzyme comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 25).

[0015] In some embodiments, the D-psicose 3-epimerase enzyme comprises an amino acid sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 00%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 25.

[0016] In some embodiments, the D-psicose 3-epimerase enzyme comprises an amino acid sequence having at least 95% (e.g., 95%, 96%, 97%, 98%, 00%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 25.

[0017] In some embodiments, the D-psicose 3-epimerase enzyme comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, or SEQ ID NO: 25. In some embodiments, the D-psicose 3-epimerase enzyme comprises the amino acid sequence of SEQ ID NO: 5.

[0018] In some embodiments, the reaction mixture comprises a D-psicose 3-epimerase enzyme comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO:3, SEQ ID NO:11, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, and SEQ ID NO:43.

[0019] In some embodiments, the D-psicose 3-epimerase enzyme comprises an amino acid sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 00%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO:3, SEQ ID NO:11, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, and SEQ ID NO:43.

[0020] In some embodiments, the D-psicose 3-epimerase enzyme comprises an amino acid sequence having at least 95% (e.g., 95%, 96%, 97%, 98%, 00%, or 100%) sequence identity to a polypeptide selected from the group consisting of SEQ ID NO:3, SEQ ID NO:11, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, and SEQ ID NO:43.

[0021] In some embodiments, the D-psicose 3-epimerase enzyme comprises the amino acid sequence of SEQ ID NO:3, SEQ ID NO:11, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, and SEQ ID NO:43.

[0022] In some embodiments, the conditions include maintaining the enzyme system and fructose substrate at a temperature between 25°C and 75°C (e.g., 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, or 75°C).

[0023] In some embodiments, the conditions include maintaining the enzyme system and fructose substrate at a pH between 4 and 10 (e.g., 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10).

[0024] In some embodiments, the D-psicose 3-epimerase enzyme is in isolated form. In some embodiments, the D-psicose 3-epimerase enzyme is immobilized on a solid matrix.

[0025] In some embodiments, the reaction mixture comprises a host cell transformed with a recombinant vector comprising a nucleic acid molecule encoding a D-psicose 3-epimerase enzyme, or a lysate of the host cell, wherein the nucleic acid molecule encoding the D-psicose 3-epimerase enzyme comprises a residue selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, and SEQ ID NO:43. In some embodiments, the nucleic acid molecule encoding the D-psicose 3-epimerase enzyme comprises the polynucleotide sequence of SEQ ID NO:44.

[0026] In some embodiments, the host cell is selected from the group consisting of: yeast cells, filamentous fungal cells, bacterial cells, mammalian cells, plant cells and Labryinthulomycetes cells. In some embodiments, the host cell is Escherichia coli (E.coli) or Pichia pastoris (P.pastoris). In some embodiments, the recombinant vector exists as a self-replicating nucleic acid molecule in the host cell. In some embodiments, the recombinant vector is integrated into the host cell chromosome.

[0027] In some embodiments, the method further comprises adding at least one type of metal ion to the reaction system, wherein the at least one type of metal ion is selected from the group consisting of magnesium ion, manganese ion, copper ion, zinc ion, nickel ion, cobalt ion, iron ion, aluminum ion and calcium ion. In some embodiments, the metal ion is added at a concentration of about 0.01 mM to about 5 mM (e.g., 0.01 mM, 0.05 mM, 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM or 5 mM). In some embodiments, the method further comprises removing a product stream comprising psicose from the reaction system when the fructose substrate is converted to psicose. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The following drawings constitute a part of this specification and are included to further demonstrate certain aspects of the present disclosure, which aspects may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0029] Figure 1 .D-fructose is bioconverted into D-psicose by D-psicose 3-epimerase (DAE).

[0030] Figure 2A-2B . Enzymatic screening of D-psicose 3-epimerase candidates. Purified candidate recombinant polypeptides with D-psicose 3-epimerase activity were dialyzed against 50 mM phosphate buffer (pH 7.2), and D-psicose synthesis was assayed using D-fructose as a substrate. Typically, recombinant polypeptides (5-10 μg) were tested in a 200 μl in vitro reaction system. The reaction system contained 20 mM Tris-HCl buffer pH 8.0, 3 mM MgCl 2 or MnSO4, 20 g / L D-fructose. The reaction was carried out at 60°C and for 2 hours ( Figure 2A ) and 16 hours ( Figure 2B The reaction was terminated by heating for 10 min. The samples were analyzed by HPLC to determine the amounts of D-fructose and D-psicose.

[0031] Figure 3 .Map of pHKA-AL39 plasmid construct.

[0032] Figure 4A-4B .Identification and characterization of the AL enzyme produced by an engineered Pichia pastoris strain. Figure 4ASDS-PAGE (16%) analysis of secreted AL39 enzyme from the culture medium of an engineered Pichia pastoris strain is shown. The AL39 enzyme is indicated by an arrow. M: Protein ladder; S: culture medium sample (12 μL) of an engineered Pichia pastoris strain. Figure 4B DAE activity is shown. The presence of fructose and psicose in the medium was analyzed after 1 hour of reaction.

[0033] Figure 5 . Comparison of DAE activity of immobilized AL39 enzyme in repeated use cycles (1-5). DETAILED DESCRIPTION

[0034] The method for producing D-psicose according to the present invention comprises contacting a fructose substrate with a reaction mixture comprising a polypeptide having D-psicose 3-epimerase activity. D-fructose is a preferred substrate and D-psicose 3-epimerase is a preferred enzyme. The reaction mixture according to the present invention may further comprise one or more cofactors. In the reaction mixture according to the present invention, divalent cations are preferred cofactors. Manganese and magnesium are preferred divalent cations and they may be present in the form of MgCl 2 and MnSO 4 in the form of. Other suitable divalent cations may include copper ions, zinc ions, nickel ions, cobalt ions, iron ions, aluminum ions and calcium ions. In one embodiment of the present invention, the D-psicose 3-epimerase used according to the present invention is provided as a highly purified homologous recombinant protein in a suitable buffer. In another embodiment of the present invention, the D-psicose 3-epimerase used according to the present invention is provided as a highly purified recombinant protein immobilized on a solid support. In another embodiment of the present invention, the reaction mixture comprises a recombinant prokaryotic cell or a recombinant eukaryotic cell expressing the recombinant D-psicose 3-epimerase and the recombinant prokaryotic cell or the recombinant eukaryotic cell is used as a source of the D-psicose 3-epimerase.

[0035] The host cell according to the present invention is any cell suitable for expressing a heterologous protein having D-psicose 3-epimerase activity. Such a host cell expressing a heterologous protein having 3-epimerase activity is obtained by transforming the host cell with a recombinant plasmid comprising a polynucleotide sequence encoding a polypeptide having D-psicose 3-epimerase activity, and such a host cell is referred to as a recombinant host cell in the present invention. The list of host cells suitable for the present invention includes, but is not limited to, bacterial cells, yeast cells, plant cells, animal cells, and Labyrinthus cells. In some embodiments, the cell system comprises bacterial cells, yeast cells, or a combination thereof. Bacterial cells suitable for use in the present invention include, but are not limited to, Escherichia spp., Streptomyces spp., Zymomonas spp., Acetobacter spp., Citrobacter spp., Synechocystis spp., Rhizobium spp., Clostridium spp., Corynebacterium spp., Streptococcus spp., Xanthomonas spp., Lactobacillus spp., Lactococcus spp., Bacillus spp., Alcaligenes spp. spp.), Pseudomonas spp., Aeromonas spp., Azotobacter spp., Comamonas spp., Mycobacterium spp., Rhodococcus spp., Gluconobacter spp., Ralstonia spp., Acidithiobacillus spp., Microlunatus spp., Geobacter spp., Geobacillus spp., Arthrobacter spp., Flavobacterium spp.), Serratias spp., Saccharopolyspora spp., Thermus spp., Stenotrophomonas spp., Chromobacterium spp., Sinorhizobium spp., Saccharopolyspora spp., Agrobacterium spp., Pantoea spp., and Vibrionatriegens. Yeast cells suitable for use in the present invention include, but are not limited to, engineered Saccharomyces spp., Schizosaccharomyces, Hansenula, Candida, Kluyveromyces, Yarrowia, Candida boidinii, and Pichia.

[0036] The term cell culture refers to any one or more cells in culture, including recombinant host cells. Cultivation is the process of growing cells under controlled conditions, usually outside their natural environment. For example, cells, such as yeast cells, can be grown as a cell suspension in a liquid nutrient broth. Cell cultures include, but are not limited to, bacterial cell cultures, yeast cell cultures, plant cell cultures, and animal cell cultures.

[0037] In some embodiments, the cells are cultured at a temperature of 16° C. to 40° C. For example, the cells can be cultured at a temperature of 16° C., 17° C., 18° C., 19° C., 20° C., 21° C., 22° C., 23° C., 24° C., 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., 35° C., 36° C., 37° C., 38° C., 39° C., or 40° C.

[0038] In some embodiments, the cells are cultured for a period of 12 hours to 72 hours or longer. For example, the cells may be cultured for a period of 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, or 72 hours. Typically, cells, such as bacterial cells, are cultured for a period of 12 to 24 hours. In some embodiments, the cells are cultured at a temperature of 37° C. for 12 to 24 hours. In some embodiments, the cells are cultured at a temperature of 16° C. for 12 to 24 hours.

[0039] In some embodiments, cells are cultured to a concentration of 1 x 10 8 (OD 600 <1) to 2x10 11 (ODˉ200) viable cells. In some embodiments, cells are cultured to a density of 1x10 8 , 2x10 8 , 3x10 8 , 4x10 8 , 5x10 8 , 6x10 8 , 7x10 8 , 8x10 8 , 9x10 8 , 1x10 9 , 2x10 9 , 3x10 9 , 4x10 9 , 5x10 9 , 6x10 9 , 7x10 9 , 8x10 9 , 9x10 9 , 1x10 10 , 2x10 10 , 3x10 10 , 4x10 10 , 5x10 10 , 6x10 10 , 7x10 10 , 8x10 10 , 9x10 10 , 1x10 11 or 2x10 11 The density of viable cells / ml. (Conversion factor: OD 1 = 8x10 8 cells / mL).

[0040] To induce the host cells to express proteins, 0.5 mM isopropyl β-D-1-thiogalactoside (IPTG) was added and the culture was further grown at 16°C for 22 hr. The cells were harvested by centrifugation (3,000 x g; 10 min; 4°C). The cell pellets were collected and used immediately or stored at -80°C.

[0041] In some embodiments, a cell pellet is harvested from a host cell expressing a polypeptide having D-psicose 3-epimerase activity. In some embodiments, the host cell pellet can be resuspended at various concentrations. In some embodiments, the host cell pellet is resuspended at a concentration of 1 g / L to 250 g / L. In some embodiments, the host cell pellet harvested from the cell system is resuspended at a concentration of 1 g / L, 10 g / L, 25 g / L, 50 g / L, 75 g / L, 100 g / L, 125 g / L, 150 g / L, 175 g / L, 200 g / L, 225 g / L or 250 g / L.

[0042] As used herein, the terms "incubating" and "incubation" refer to a process of mixing two or more chemical or biological entities or at least one chemical entity and at least one biological entity (such as a compound and an enzyme) and allowing them to interact under conditions that are favorable for the production of a product as described herein, such as D-psicose.

[0043] The term "downstream separation process" refers to the recovery of the final product D-psicose from the original substrate D-fructose. When fructose is used as a substrate in an enzymatic reaction to produce D-psicose, the fructose is not completely consumed in the reaction, and a large amount of fructose is still present in the reaction medium at the end of the enzymatic reaction. D-fructose and D-psicose have similar physical and chemical properties, and it is difficult to separate them at the end of an enzymatic reaction involving D-psicose 3-epimerase and fructose as a substrate. One possible way to recover D-psicose without D-fructose is to convert the remaining fructose into mannitol and separate D-psicose from mannitol. NADPH or NADH-dependent mannitol dehydrogenase can be used to convert D-fructose into mannitol. Mannitol dehydrogenase can be used in a dual enzyme system together with formate dehydrogenase to regenerate the reducing cofactor NADH required for the action of mannitol dehydrogenase. Formate dehydrogenase converts formate into carbon dioxide and reduces NAD to NADH. Mannitol can be crystallized from aqueous solution by cold crystallization, and D-psicose can be recovered in a D-fructose-free form. (Saha, BC and Racine, FM (2011) Biotechnological production of mannitol and its applications. Appl Microbiol Biotechnol. 89: 879–891; U.S. Patent No. 10266862B2).

[0044] The term immobilization refers to binding a host cell expressing D-psicose 3-epimerase or a purified polypeptide having D-psicose 3-epimerase activity to a solid support using one or other methods well known in the art. Sodium alginate derived from seaweed can be used as a solid support to immobilize a host cell expressing D-psicose 3-epimerase or a purified polypeptide having D-psicose 3-epimerase activity, as described in detail in patent document US2019169591A1.

[0045] The terms "nucleic acid" and "nucleotide" are used according to their respective ordinary and customary meanings understood by those of ordinary skill in the art, and are used without limitation to refer to deoxyribonucleotides or ribonucleotides and polymers thereof in single-stranded or double-stranded form. Unless otherwise specified, the term encompasses nucleic acids containing known analogs of natural nucleotides, which have similar binding properties to the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise specified, a specific nucleic acid sequence also implicitly encompasses conservative modifications or degenerate variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as explicitly specified sequences.

[0046] The terms "polypeptide", "protein" and "peptide" are used according to their respective ordinary and customary meanings as understood by those of ordinary skill in the art; these three terms are sometimes used interchangeably and are used without restriction to refer to amino acid polymers or amino acid analogs, regardless of their size or function. Although "protein" is often used with respect to relatively large polypeptides, and "peptide" is often used with respect to small polypeptides, the usage of these terms in the art is overlapping and varied. As used herein, the term "polypeptide" refers to peptides, polypeptides and proteins, unless otherwise indicated. When referring to polypeptide products, the terms "protein", "polypeptide" and "peptide" are used interchangeably herein. Therefore, exemplary polypeptides include polypeptide products, naturally occurring proteins, homologs, orthologs, paralogs, fragments, and other equivalents, variants and analogs of the foregoing.

[0047] The term "functional fragment" of a polypeptide or protein refers to a peptide fragment that is a portion of the full-length polypeptide or protein and has substantially the same biological activity as the full-length polypeptide or protein, or performs substantially the same function (e.g., performs the same enzymatic reaction).

[0048] The term "functional variant" also includes conservative substitution variants. The term "conservative substitution variant" refers to a peptide having an amino acid sequence that differs from a reference peptide by one or more conservative amino acid substitutions and retains some or all of the activity of the reference peptide. "Conservative amino acid substitution" is the replacement of an amino acid residue with a functionally similar residue. Examples of conservative substitutions include replacing another residue with a non-polar (hydrophobic) residue (such as isoleucine, valine, leucine or methionine); replacing another residue with a charged or polar (hydrophilic) residue, such as between arginine and lysine, between glutamine and asparagine, between threonine and serine; replacing another residue with a basic residue (such as lysine or arginine); or replacing another residue with an acidic residue (such as aspartic acid or glutamic acid); or replacing another residue with an aromatic residue (such as phenylalanine, tyrosine or tryptophan). Such substitutions are expected to have little effect on the apparent molecular weight or isoelectric point of a protein or polypeptide. The phrase "conservatively substituted variants" also includes peptides in which residues are replaced with chemically derivatized residues, provided that the resulting peptide retains some or all of the activity of the reference peptide as described herein.

[0049] The term "variant" in relation to polypeptides of the subject technology also includes functionally active polypeptides having an amino acid sequence that is at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, and even 100% identical to the amino acid sequence of the reference polypeptide.

[0050] The term "homologous" in all its grammatical forms and spelling variations refers to the relationship between polynucleotides or polypeptides having a "common evolutionary origin", including polynucleotides or polypeptides from superfamilies and homologous polynucleotides or proteins from different species (Reeck et al., Cell 50:667, 1987). Such polynucleotides or polypeptides have sequence homology, as reflected by their sequence similarity, whether in terms of percent identity or the presence of specific amino acids or motifs at conserved positions. For example, two homologous polypeptides can have amino acid sequences that are at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, and even 100% identical.

[0051] "Percent (%) amino acid sequence identity" with respect to variant polypeptide sequences of the subject technology refers to the percentage of amino acid residues in the candidate sequence that are identical with the amino acid residues of a reference polypeptide, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity.

[0052] Comparison for determining amino acid sequence identity percentage can be achieved in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for measuring the comparison, including any algorithm required for maximum comparison over the full length of the compared sequences. For example, the sequence comparison program NCBI-BLAST2 can be used to determine % amino acid sequence identity. The NCBI-BLAST2 sequence comparison program can be downloaded from ncbi.nlm.nih.gov. NCBI BLAST2 uses several search parameters, all of which are set to default values, including, for example, unmask yes, strand=all, expected occurrences 10, minimum low complexity length=15 / 5, multi-pass e-value=0.01, constant for multi-pass=25, drop off for final gapped alignment=25, and scoring matrix=BLOSUM62. In the case of amino acid sequence comparison using NCBI-BLAST2, the % amino acid sequence identity of a given amino acid sequence A with, with, or to a given amino acid sequence B (which can be alternatively expressed as a given amino acid sequence A having or including a certain % amino acid sequence identity with, with, or to a given amino acid sequence B) is calculated as follows: 100 multiplied by a fraction X / Y, where X is the number of amino acid residues that are scored as identical pairs by the NCBI-BLAST2 program when A and B are aligned by the sequence alignment program; and where Y is the total number of amino acid residues in B. It should be understood that when the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A.

[0053] The technology for determining the "similarity" of amino acid sequences is well known in the art. In general, "similarity" refers to the precise amino acid to amino acid comparison of two or more polypeptides in the appropriate position, wherein the amino acids are identical or have similar chemical and / or physical properties, such as charge or hydrophobicity. Then the "similarity percentage" between the compared polypeptide sequences can be determined. The technology for determining the identity of nucleic acid and amino acid sequences is also well known in the art, and includes determining the nucleotide sequence of the mRNA of the gene (usually through a cDNA intermediate) and determining the amino acid sequence encoded therein, and comparing it with a second amino acid sequence. In general, "identity" refers to the precise nucleotide to nucleotide or amino acid to amino acid correspondence of two polynucleotides or polypeptide sequences. Two or more polynucleotide sequences can be compared by determining their "identity percentage", and two or more amino acid sequences can also be like this. The available programs in Wisconsin Sequence Analysis Software Package Version 8 (available from Genetics Computer Group, Madison, Wis.), such as the GAP program, can calculate the identity between two polynucleotides and the identity and similarity between two polypeptide sequences, respectively. Other programs for calculating the identity or similarity between sequences are known to those skilled in the art.

[0054] An amino acid position that "corresponds to" a reference position refers to a position that is aligned with a reference sequence, as identified by aligning the amino acid sequences. Such alignment can be done manually, or can be done using well-known sequence alignment programs (such as ClustalW2, Blast 2, etc.).

[0055] Unless otherwise indicated, the percent identity of two polypeptide or polynucleotide sequences refers to the percentage of identical amino acid residues or nucleotides over the entire length of the shorter of the two sequences.

[0056] As used herein, the term "expression" is used according to its ordinary and customary meaning as understood by those of ordinary skill in the art, and is used without limitation to refer to the transcription and stable accumulation of sense (mRNA) or antisense RNA of nucleic acid fragments derived from the subject technology. "Overexpression" refers to the production of a gene product in a transgenic or recombinant organism that exceeds the level of production in a normal or non-transformed organism.

[0057] "Transformation" is used according to its common and customary meaning as understood by those of ordinary skill in the art, and is used without limitation to refer to the transfer of a polynucleotide into a target cell. The transferred polynucleotide may be incorporated into the genome or chromosomal DNA of the target cell, thereby resulting in genetically stable inheritance, or it may replicate independently of the host chromosomal DNA. Host organisms containing the transformed nucleic acid fragments are referred to as "transgenic" or "recombinant" or "transformed" organisms.

[0058] The terms "transformed," "transgenic," and "recombinant," when used herein in conjunction with a host cell, are used according to their ordinary and customary meaning as understood by those of ordinary skill in the art, and are used without limitation to refer to a cell of a host organism, such as a plant or microbial cell, into which a heterologous nucleic acid molecule has been introduced. The nucleic acid molecule may be stably integrated into the genome of the host cell, or the nucleic acid molecule may exist as an extrachromosomal molecule. Such extrachromosomal molecules may self-replicate. A transformed cell, tissue, or subject is understood to include not only the final product of the transformation process, but also its transgenic progeny.

[0059] The terms "recombinant," "heterologous," and "exogenous," when used herein in conjunction with polynucleotides, are used according to their ordinary and customary meanings as understood by those of ordinary skill in the art, and are used without limitation to refer to a polynucleotide (e.g., a DNA sequence or gene) that is derived from a source foreign to a particular host cell or, if derived from the same source, modified from its original form. Thus, a heterologous gene in a host cell includes a gene that is endogenous to a particular host cell but has been modified, for example, using site-directed mutagenesis or other recombinant techniques. The term also includes non-naturally occurring multiple copies of a naturally occurring DNA sequence. Thus, the term refers to a DNA segment that is foreign or heterologous to a cell, or homologous to a cell but is in a position or form within the host cell where the element is not normally found.

[0060] Similarly, the terms "recombinant," "heterologous," and "exogenous," when used herein in conjunction with a polypeptide or amino acid sequence, refer to a polypeptide or amino acid sequence that is derived from a source foreign to the particular host cell, or, if derived from the same source, has been modified from its original form. Thus, a recombinant DNA segment can be expressed in a host cell to produce a recombinant polypeptide.

[0061] The terms "plasmid", "vector" and "box" are used according to their common and customary meanings as understood by those of ordinary skill in the art, and are used without limitation to refer to extrachromosomal elements that often carry genes that are not part of the central metabolism of the cell and are usually in the form of circular double-stranded DNA molecules. Such elements can be nucleotide sequences (linear or circular) of autonomously replicating sequences, genome integration sequences, bacteriophages, or single-stranded or double-stranded DNA or RNA derived from any source, wherein multiple nucleotide sequences have been connected or recombined into a unique structure that is capable of introducing a promoter fragment and DNA sequence of a selected gene product into a cell together with appropriate 3' non-translated sequences. "Transformation cassette" refers to a specific vector that contains foreign genes and, in addition to foreign genes, has elements that are beneficial to transforming a specific host cell. "Expression cassette" refers to a specific vector that contains foreign genes and, in addition to foreign genes, has elements that enhance the expression of the gene in a foreign host.

[0062] Standard recombinant DNA and molecular cloning techniques used herein are well known in the art and are described in, for example, Sambrook, J., Fritsch, EF, and Maniatis, T. Molecular Cloning: A Laboratory Manual, 2nd ed.; Cold Spring Harbor Laboratory: Cold Spring Harbor, NY, 1989 (hereinafter "Maniatis"); and Silhavy, TJ, Bennan, ML, and Enquist, LW Experiments with Gene Fusions; Cold Spring Harbor Laboratory: Cold Spring Harbor, NY, 1984; and Ausubel, FM et al., in Current Protocols in Molecular Biology, published by Greene Publishing and Wiley-Interscience, 1987; the entire contents of each of which are hereby incorporated herein by reference to the extent consistent with this invention.

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, preferred methods and materials are described below.

[0064] Example

[0065] Embodiment 1: Construction of a plasmid vector for expressing D-psicose 3-epimerase

[0066] D-psicose 3-epimerase (DAE) has the ability to convert D-fructose into D-psicose. Figure 1 In order to identify a specific D-psicose 3-epimerase enzyme that produces D-psicose using D-fructose as a substrate, polynucleotide sequences encoding polypeptides having D-psicose 3-epimerase activity were identified using polygenetic and BLAST analysis. Full-length DNA fragments of all candidate D-psicose 3-epimerase genes were commercially synthesized. Almost all cDNA codons were changed to those preferred by Escherichia coli (E. coli) (Twist Bioscience, CA). The synthetic DNA was cloned into a bacterial expression vector to generate an expression construct.

[0067] Embodiment 2: Measurement of D-psicose 3-epimerase activity in recombinant Escherichia coli cells

[0068] Each expression vector construct having a polynucleotide encoding a polypeptide having D-psicose 3-epimerase activity prepared as in Example 1 was transformed into E. coli T7 expression cells (Biolabs, MA), which were then grown in Terrific Broth medium containing 50 μg / mL ampicillin at 37° C. until an OD600 of 0.8-1.0 was reached. Protein expression was induced by adding 0.5 mM isopropyl β-D-1-thiogalactoside (IPTG), and the culture was further grown at 16° C. for 22 hr. Cells were harvested by centrifugation (3,000×g; 10 min; 4° C.). Cell pellets were collected and used immediately or stored at −80° C.

[0069] The cell pellet was extracted with extraction buffer (BugBuster Master Mix, EMD Millipore, US). After centrifugation, the supernatant was added to the reaction mixture for activity determination. Typically, the supernatant (5 μl) was tested in a 200 μl in vitro reaction system. The reaction system contained 20 mM Tris-HCl buffer pH 8.0, 1 mM MgCl 2 or MnSO4, 10 g / L D-fructose. The reaction was carried out at 60°C and terminated by heating for 10 min. The samples were analyzed by HPLC.

[0070] The concentrations of D-fructose and D-psicose were determined by an HPLC system (Vanquish, Thermo Scientific, USA) equipped with a RefractorMax521 detector (IDEX Health & Science KK, Japan). 2+ The column (100x7.8mm, Phenomenex, CA) was used for chromatographic separation. The column was eluted with water at a flow rate of 0.5ml / min at 80°C. Through the analysis of D-psicose 3-epimerase activity, a total of 21 candidate D-psicose 3-epimerase enzymes with appropriate enzyme activity for bioconverting D-fructose into D-psicose were identified (Table 1).

[0071] Embodiment 3: Purification of recombinant D-psicose 3-epimerase

[0072] To purify the recombinant protein, the cell pellet is usually resuspended in a lysis buffer (50 mM potassium phosphate buffer pH 7.2, 25 mg / ml lysozyme, 5 mg / ml DNase I, 20 mM imidazole, 500 mM NaCl, 10% glycerol, and 0.4% Triton X-100). The cells are disrupted by ultrasound at 4°C, and the cell debris is clarified by centrifugation (18,000 x g; 30 min). The supernatant is loaded onto an affinity column Ni-NTA (Qiagen) equilibrated with an equilibration buffer containing 50 mM potassium phosphate buffer pH 7.2, 20 mM imidazole, 500 mM NaCl, and 10% glycerol. After loading the protein sample, the column is washed with an equilibration buffer to remove unbound contaminating proteins. The His-tagged recombinant polypeptide having D-psicose 3-epimerase activity is eluted with an equilibration buffer containing 250 mM imidazole.

[0073] The purified candidate recombinant polypeptide having D-psicose 3-epimerase activity was dialyzed against 50 mM phosphate buffer (pH 7.2), and D-psicose synthesis was measured using D-fructose as a substrate. Typically, the recombinant polypeptide (5-10 μg) was tested in a 200 μl in vitro reaction system. The reaction system contained 20 mM Tris-HCl buffer pH 8.0, 3 mM MgCl 2 or MnSO4, 20 g / L D-fructose. The reaction was carried out at 60°C and terminated at 2 hours and 16 hours by heating for 10 min. The samples were analyzed by HPLC.

[0074] These candidate recombinant polypeptides having D-psicose 3-epimerase activity have different enzymatic activities for producing psicose and have different preferences for divalent metal factors. Figure 2A-2BAs shown, all selected candidates had D-psicose 3-epimerase activity at 2 hr ( Figure 2A ) and more D-psicose can be produced when the reaction is extended to 16 hr ( Figure 2B ). With the addition of Mg 2+ Compared with the addition of Mn 2+ It can increase the activities of most enzymes (AL28, 35, 39, 42, 50, and 51), indicating that these enzymes prefer Mn 2+ Cofactor.

[0075] Embodiment 4: Generation of an engineered Pichia pastoris strain to produce DAE enzyme

[0076] A full-length DNA fragment of the AL39 gene (SEQ ID NO:43) was synthesized for transformation of Pichia pastoris cells. Specifically, the cDNA was codon optimized for Pichia pastoris expression to produce the AL39 enzyme (SEQ ID NO:5). The AL39 gene was inserted into a frame with the alpha mating factor signal peptide. The synthesized fusion gene was cloned into the pHKA vector (modified Pichia pastoris expression vector) using EcoRI and NotI restriction digests. In the expression plasmid (pHKA-AL39, Figure 3 ), the expression cassette contains AOX1 promoter, α-mating factor signal peptide, AL39 gene and AOX1 transcription terminator.

[0077] The linearized pHKA-AL39 plasmid was transformed into Pichia pastoris (GS115) cells using methods known in the art (Lin-Cereghino et al., Biotechniques, 38(1):44-48, 2005), and the expression cassette was integrated into the HIS4 locus in the Pichia pastoris genome.

[0078] To demonstrate the production of secretory AL39 enzyme, a single colony of Pichia pastoris strain pHKA-AL39 was inoculated in BMGY medium in a baffled flask and grown in a shaking incubator (250-300 rpm) at 28-30°C until the culture reached an OD600 of 2-6 (logarithmic phase growth). pHKA-AL39 cells were harvested by centrifugation and resuspended in BMMY medium to an OD600 of 1.0 to induce expression. 100% methanol was added to the BMMY medium every 24 hours to a final concentration of 1% methanol to maintain induction of expression. The culture medium from pHKA-AL39 culture was harvested by centrifugation and subjected to DAE activity analysis and SDS-PAGE analysis as described below.

[0079] After centrifugation, the supernatant is added to the reaction mixture for activity determination. Usually, the supernatant (65 μl) is tested in a 200 μl in vitro reaction system. The reaction system contains 20mM Tris-HCl buffer pH 8.0, 1mM MnSO4, 30g / LD-fructose. The reaction is carried out at 55-60°C and the reaction is terminated by heating for 10min. Samples are analyzed by HPLC.

[0080] The concentrations of D-fructose and D-psicose were determined by an HPLC system (Vanquish, Thermo Scientific, USA) equipped with a RefractorMax521 detector (IDEX Health & Science KK, Japan). 2+ Chromatographic separation was performed on a 100 x 7.8 mm column (Phenomenex, CA). The column was eluted with water at 80 °C at a flow rate of 0.5 ml / min.

[0081] The presence of AL39 protein was determined by SDS-PAGE analysis ( Figure 4A ). The culture medium samples were separated by SDS-PAGE using methods known in the art (Laemmli, Nature, 227(5259):680-685, 1970). The produced AL39 protein was visible after staining with a staining solution. Based on SDS-PAGE analysis, the produced AL39 was the major protein in the culture medium samples ( Figure 4A ).

[0082] After SDS-PAGE and DAE activity screening, the best strain was identified as having high secretory AL39 enzyme production. The produced AL39 enzyme has the activity to bioconvert D-fructose to D-psicose. Figure 4A As shown, the produced AL39 protein can be detected in the fermentation medium by SDS-PAGE analysis, indicating that the produced AL39 enzyme can be secreted into the extracellular space of the engineered Pichia pastoris strain. After removing the cell pellet by centrifugation, the produced enzyme can be easily collected and concentrated from the fermentation medium. DAE activity can be detected in the culture medium samples. Figure 4B As shown, about 25% of fructose could be converted into psicose after 1 hr of reaction.

[0083] Example 5 : Bioconversion using immobilized DAE enzyme

[0084] Enzyme immobilization provides an excellent basis for increasing the availability of enzymes to their substrates and having greater turnover over a considerable period of time. A method for efficiently producing immobilized D-psicose epimerase having high activity and excellent persistence for the bioconversion of fructose to psicose is described below.

[0085] AL39 protein was concentrated by filtration. The extracted AL39 was dissolved in 20 mM phosphate buffer (pH 8.0) for immobilization. One unit of DAE activity was defined as the amount of enzyme that catalyzes the formation of 1 μmol of D-psicose per minute at pH 8.0 and 55°C. The specific activity of the immobilized DAE enzyme was defined as units per milligram of enzyme.

[0086] The prepared AL39 protein was mixed with water-pretreated ion exchange resin (LXP-505, SUNRESIN) in 20 mM phosphate buffer (pH 8.0) and slowly stirred in a reactor at 25° C. for 24 hours. Subsequently, the supernatant was removed, and the resulting mixture was washed with 20 mM phosphate buffer (pH 8.0) to obtain immobilized DAE enzyme.

[0087] The immobilized DAE enzyme was added to the reaction mixture for activity determination. The immobilized enzyme was tested in a 1 ml in vitro reaction system. The reaction system contained 20 mM phosphate buffer pH 8.0, 1 mM MnSO4, 500 g / L D-fructose. The reaction was carried out at 55°C and terminated by heating for 10 min. The samples were analyzed by HPLC. After 3 hours, the immobilized enzyme converted more than 23% of fructose into psicose.

[0088] In order to determine whether AL39 can maintain activity after use, the repeated use cycle of immobilized AL39 was tested. 180 mg of immobilized AL39 was added to 1 L of reaction solution containing 20 mM phosphate buffer (pH 8.0), 1 mM MnSO4, and 500 g / L D-fructose. The reaction was carried out at 55°C. After 5 hours of reaction, the immobilized enzyme was collected by filtration. The supernatant was analyzed by HPLC to measure the production of psicose, and the immobilized enzyme was washed 5 times with 20 mM phosphate buffer (pH 8.0) for the next cycle reaction. After 5 cycles, the activity of each cycle was compared. Figure 5 As shown, the activity of immobilized AL39 lost only 10.76% in the fifth cycle compared with the first cycle, indicating that immobilized AL39 was stable for the bioconversion of psicose.

[0089] References:

[0090] 1. Lin-Cereghino J, Wong WW, Xiong S, et al. Condensed protocol for competitive cell preparation and transformation of the methylotrophic yeast Pichia pastoris. Biotechniques. 2005; 38(1): 44-48.

[0091] 2. Laemmli, UK (1970). "Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4". Nature. 227 (5259): 680-685.

[0092]

[0093]

[0094] Sequence information

[0095] SEQ ID NO: 1; Protein; Thermus silvanus

[0096] MPRFGAHAFIWAAEWNPEAAEKVIQGATRVGLDFVEIPLLHPESFDVELTRRLLNGYGVGCTCSLGLPREASLPEHPEAATRFLIQALNVAHQIGSEVLTGVTYATLGTLSGRAPREADYQAVVKALKPAARHAAALGMRL GIEPVNRYETYLINLAAQGLELIRRLDEPNVFLHLDTYHMNIEEKGFRGPIVEAGAHLGYIHLSESDRGTPGTGNVHWDAVFAGLREIGFSGDLVMESFVALNPDIARATCMMWRDVVGDPQALVQEGLAFLRGKASEYGLLG

[0097] SEQ ID NO:2; DNA; Thermus silvanus

[0098] ATGCCACGCTTCGGAGCTCATGCGTTCATTTGGGCCGCTGAGTGGAACCCGGAAGCGGCAGAGAAAGTGATTCAGGGTGCAACCCGTGTTGGCCTCGACTTCGTTGAAATTCCGCTCTTGCACCCGGAATCCTTCGATGTCGAGCTCACACGTCGTTTACTAAACGGGTACGGCGTCGGTTGTACGTGTTCTCTGGGCTTGCCGCGTGAAGCGTCCCTGCCGGAGCACCCCGAAGCCGCGACTCGTTTTCTGATTCAGGCCCTGAATGTTGCACATCAGATTGGCTCAGAGGTGCTGACCGGGGTAACCTACGCGACTTTAGGCACTTTATCTGGCCGGGCGCCGCGCGAAGCTGATTATCAAGCAGTCGTAAAAGCACTGAAACCGGCGGCTAGACATGCAGCAGCACTGGGTATGCGCCTCGGCATTGAACCCGTTAATCGTTATGAAACTTATTTAATTAACCTGGCCGCTCAGGGCCTGGAGTTAATCCGTCGTCTGGACGAACCGAATGTGTTCCTGCACCTGGATACGTACCACATGAATATTGAAGAGAAAGGTTTCCGGGGCCCGATCGTGGAAGCGGGCGCCCACCTGGGGTATATTCATCTGAGCGAGAGTGATCGCGGCACCCCTGGAACGGGCAACGTCCATTGGGACGCGGTGTTCGCGGGGCTGCGTGAAATTGGCTTCTCAGGCGATTTGGTCATGGAAAGCTTCGTCGCACTGAACCCTGATATTGCGCGCGCTACATGCATGTGGCGGGACGTCGTTGGCGATCCCCAGGCGCTGGTCCAGGAAGGCTTAGCATTTCTCCGTGGCAAGGCTAGCGAGTATGGCCTACTCGGTtaa

[0099] SEQ ID NO:3; Protein; Deep-sea hydrothermal vent metagenome

[0100] MEGFGVHTSMWTMSWDKKGAEYAVSQAVRYQMDFLEIALLSPLDVDAQHSRRLLEKNHMRAICSLGLPEGAWLSNNPQAGVEYLKIALEKTAEMGCEALSGVIYGGIGERTGFPPTEKELDNVVRALGEAASYAKSLGLLLGIEPVNRYESHLINTGRQSVEMIEKVGASNMFVHLDTYHMNIEEKGVAQGILDAREHIKYIHLSESDRGTPGAGTCDWDEIFAVLAAINFKGGLAMESFVNMPPEIAYGLSIWRPVAKSADEVMENGLPFLRNKARQYQLL

[0101] SEQ ID NO:4; DNA; Deep-sea hydrothermal vent metagenome

[0102] ATGGAAGGTTTTGGAGTGCACACGAGCATGTGGACTATGAGTTGGGATAAGAAGGGTGCGGAGTATGCTGTAAGCCAGGCGGTGCGCTATCAGATGGATTTTCTTGAAATCGCTCTGCTGAGTCCACTGGATGTCGATGCCCAGCATAGTCGCCGTCTGCTAGAGAAGAACCACATGCGCGCGATTTGCAGCCTTGGGTTACCGGAAGGTGCGTGGCTCAGTAATAACCCTCAGGCCGGAGTGGAATACCTGAAGATCGCCCTGGAAAAGACTGCGGAGATGGGTTGCGAAGCGCTGTCCGGCGTAATCTACGGCGGCATTGGCGAAAGAACAGGTTTTCCGCCGACGGAGAAAGAGCTGGACAATGTCGTGCGCGCGCTGGGTGAAGCGGCATCTTACGCAAAATCGTTAGGCCTTCTGCTCGGTATTGAACCGGTGAACCGGTATGAGTCTCATCTGATTAATACGGGACGTCAGAGTGTGGAAATGATTGAAAAGGTCGGTGCCTCTAACATGTTTGTTCACCTTGACACATACCACATGAATATTGAAGAAAAGGGTGTGGCACAAGGGATTCTGGACGCGCGCGAACACATAAAATATATCCACCTGTCAGAGTCTGACCGAGGGACTCCGGGCGCAGGCACGTGTGACTGGGATGAGATTTTCGCAGTCCTGGCCGCAATTAACTTTAAAGGCGGCTTAGCGATGGAATCGTTCGTGAACATGCCGCCGGAAATTGCCTATGGTCTGTCCATTTGGCGTCCGGTGGCTAAATCGGCTGATGAAGTCATGGAAAACGGCCTGCCGTTTCTACGCAACAAAGCGCGCCAGTATCAGCTGCTGtaa

[0103] SEQ ID NO:5; Protein; Thermoanaerobacter sludgei

[0104] MKYGIFYAYWEKEWKGDFITYIEKVKKLGFDILEVGCGDFHKQPDSYFHTLRDAAREYDIILTGGYGPRAEHNLCSPDTAVVENALAFYSDIFRKMEIAGIRSIGGGLYAYWPVDYSREPDKAGDLERSIKNMRRLADIAERHGITLNMEVLNRFEGYLINDTNEGLAYIRAVDKPNVKLMLDTFHMNIEEDSFTEPILQAGKYLGHVHVGEPNRKPPREGRIPWGEIGKALRQIGYDGPVVMEPFVTMGGQVGKDICVWRDLSQGATEEDLDRDAEKSLAFLKGMFEA

[0105] SEQ ID NO:6; DNA; Clostridium thermosaccharolyticum

[0106] ATGAAATATGGTATATTTTACGCATATTGGGAGAAAGAATGGAAAGGCGACTTTATCACATACATTGAGAAAGTTAAGAAATTAGGCTTTGACATTTTGGAAGTCGGCTGCGGTGATTTTCATAAACAGCCGGATTCATACTTCCACACCCTGCGTGATGCCGCTCGCGAATACGACATTATTCTGACCGGCGGCTATGGGCCGCGCGCCGAACACAACTTGTGTAGCCCGGATACAGCGGTCGTTGAAAATGCCCTGGCATTTTACTCCGATATATTTCGCAAAATGGAAATTGCCGGCATCCGTTCGATCGGCGGCGGTCTATATGCGTATTGGCCAGTTGATTACAGCCGTGAACCCGACAAAGCCGGCGATTTAGAACGTTCCATTAAGAACATGCGTCGCTTAGCCGATATTGCGGAACGGCATGGTATCACGCTGAATATGGAAGTGCTCAATCGCTTCGAAGGTTACCTTATTAATGATACCAATGAAGGTCTGGCCTACATTCGTGCCGTCGATAAACCCAACGTAAAGTTGATGCTGGATACATTTCACATGAACATTGAGGAAGATAGCTTTACCGAACCCATTCTCCAGGCTGGTAAGTACCTGGGCCATGTACATGTCGGCGAACCAAATCGTAAACCACCACGGGAAGGTCGTATTCCGTGGGGCGAGATTGGCAAAGCGCTGCGCCAGATTGGGTACGACGGTCCCGTGGTGATGGAACCGTTTGTTACCATGGGCGGCCAAGTGGGGAAAGACATTTGTGTGTGGCGGGACCTTAGCCAAGGTGCGACCGAAGAAGATCTGGATCGCGATGCTGAGAAAAGCCTGGCCTTTCTGAAAGGAATGTTTGAAGCCtaa

[0107] SEQ ID NO:7; Protein; Deep-sea hydrothermal vent metagenome

[0108] MNLPAKKMVLGVHTFAVAPFWDLEVMRHEARRLKSHGVGLLEIPLLRPEEINIKATRAFAREFGFELVTSLGLPSNIDAVEDPQSALAFLEPAFKVAAEIGSNMLSGVTYAPIGKISGQPVTQREKDGICRFLQQAAALAANHGLKLGVEPCNRYETHLMNTAAQAVEYVEAVAAENLFIHLDTYHMNIEEESFAAGFAKAAPYLGYVHLSESNRGVPGRAMINWDAVMGALADIGYHGALTLESMNYVDPDIATALAVWRPVAKNPDDVIDFGLAFLLKAAADAGLTFG

[0109] SEQ ID NO:8; DNA; Deep-sea hydrothermal vent metagenome

[0110] ATGAACCTGCCGGCTAAGAAGATGGTCCTGGGAGTCCATACGTTCGCAGTCGCCCCGTTTTGGGATCTTGAAGTAATGCGCCACGAGGCCCGCCGTTTAAAGAGCCATGGCGTAGGGTTATTGGAAATCCCACTTCTCCGGCCAGAAGAAATTAATATTAAAGCGACCCGCGCATTTGCTCGTGAATTTGGGTTTGAATTAGTGACCAGTTTAGGCCTGCCTTCGAATATCGACGCTGTAGAAGATCCGCAGTCTGCTTTAGCCTTTCTGGAACCGGCGTTCAAAGTGGCTGCTGAAATTGGTAGCAATATGCTTTCAGGCGTTACCTATGCTCCGATTGGAAAGATAAGTGGTCAGCCGGTGACCCAGCGTGAGAAGGATGGTATTTGTCGGTTCCTACAACAAGCGGCTGCGTTGGCCGCGAACCATGGGTTGAAACTGGGCGTAGAGCCTTGTAATCGCTATGAAACGCATCTGATGAACACAGCAGCCCAAGCAGTTGAATACGTTGAGGCTGTGGCAGCGGAAAACCTTTTCATCCACTTAGATACCTACCACATGAACATCGAAGAAGAAAGTTTTGCAGCAGGGTTTGCGAAAGCGGCGCCTTATCTGGGGTACGTGCATCTGAGCGAGAGCAATCGCGGCGTACCGGGCCGCGCCATGATCAATTGGGATGCAGTGATGGGTGCCCTTGCCGACATCGGGTATCATGGGGCACTGACTCTGGAGTCTATGAATTATGTGGATCCGGATATTGCGACTGCCCTTGCGGTCTGGCGTCCGGTTGCCAAGAACCCGGACGATGTGATAGACTTTGGCCTGGCGTTTCTGCTGAAGGCCGCAGCGGATGCGGGTTTGACATTCGGTtaa

[0111] SEQ ID NO:9; Protein; Thermus subtilis subsp. spizizenii

[0112] MARFGAHAFIWSADWTPQAAEKVAAGAAAGLDFVEIPLLRPEAFDSTLTRRLLEHHGLGCTCSLGLPTEAALPDHPQAAARFLIQALDVAHQMGSPVLSGVTYATLGALSGRPPTEADYETLAKTLKPVAQHAARLGMRLGLEPVNRYETYLINLGSQALDLIQRIGEPNVFVHLDTYHMNIEEKGFKNPIVTVGKHLGYIHLSESDRGTPGSGNVHWDEVFSGLQAIGFQGDLVMESFVALNPDIARATCMWRDVVGDPKTLVHDGLAFLRGKARAYGLL

[0113] SEQ ID NO:10; DNA;

[0114] ATGGCGCGCTTTGGTGCGCATGCTTTTATTTGGAGTGCTGATTGGACCCCTCAAGCCGCCGAAAAGGTCGCAGCGGGAGCAGCGGCGGCGGGACTGGATTTTGTCGAAATACCGCTTCTTCGTCCAGAGGCGTTCGATTCGACACTTACCCGTAGGTTATTAGAGCATCATGGACTGGGATGCACCTGCTCTCTGGGTCTGCCTACCGAGGCAGCACTCCCGGACCATCCTCAAGCAGCAGCCCGTTTTCTCATCCAGGCCTTGGATGTAGCCCATCAAATGGGCAGCCCGGTCCTGAGTGGGGTCACCTACGCAACCCTGGGCGCACTGTCAGGCCGCCCGCCGACCGAGGCCGATTATGAAACCCTGGCGAAAACACTGAAGCCAGTAGCGCAGCACGCCGCCCGCCTGGGTATGCGACTCGGGCTTGAACCGGTTAACCGCTATGAAACATACCTGATAAACTTAGGGTCACAGGCGCTGGATCTCATCCAACGTATCGGCGAACCGAACGTGTTCGTACACCTGGATACTTATCACATGAACATCGAAGAGAAAGGTTTTAAGAACCCGATTGTTACCGTCGGTAAACATCTTGGTTATATTCACCTGTCTGAAAGCGACCGTGGTACCCCAGGTAGCGGTAATGTGCATTGGGATGAAGTGTTCTCCGGCCTGCAAGCAATCGGTTTTCAGGGTGACCTTGTGATGGAGTCGTTTGTGGCACTGAATCCGGATATTGCCCGCGCCACGTGCATGTGGCGGGATGTTGTGGGAGATCCGAAAACACTGGTGCATGATGGCCTAGCTTTCCTGCGCGGTAAAGCTCGCGAATACGGACTGCTGtaa

[0115] SEQ ID NO:11; Protein; Microvirga thermophila

[0116] MQGFGVHTSMWTMHWDRAGAERTIPAAAAYKMDFIEIALLDTAIVDAAHTRALLEKHGLRAVCSLGLPEPVWASVNPEGAIAHLKRALDKTAEMGAEALSGVTYGGIGQRTGVPPTPQEYDNIARALEAAAKHAKALGLAFGIEPVNRYENHLINTGRQAVEMIEKVGADNIFIHLDTYHMNIEEKGVANGILDAREHLRYIHLSESDRGTPGEGTCDWDEIFAALAAIGFKGGLAMESFINMPPQVAYGLAVWRPVAESFEEVMDRGLPFLRNKARQYRLIA

[0117] SEQ ID NO:12; DNA; Microvirga thermophila

[0118] ATGCAAGGTTTCGGCGTACACACTAGCATGTGGACGATGCATTGGGATCGCGCGGGCGCCGAGCGCACCATTCCGGCCGCCGCCGCCTACAAAATGGACTTTATCGAAATTGCGCTGCTGGATACAGCTATAGTCGATGCAGCGCACACCCGTGCGCTGCTGGAGAAGCACGGCCTGCGTGCAGTTTGTTCACTCGGATTACCGGAGCCGGTCTGGGCCTCTGTGAACCCGGAAGGGGCCATTGCTCACCTGAAACGCGCGCTGGACAAGACCGCAGAGATGGGAGCTGAAGCTCTGTCTGGTGTGACTTACGGCGGTATCGGCCAGCGCACAGGCGTACCACCAACGCCCCAAGAATACGATAACATTGCCCGTGCCCTTGAAGCAGCTGCGAAACATGCCAAAGCCTTAGGCCTGGCCTTTGGTATCGAACCGGTTAACAGGTATGAAAACCATTTGATAAACACCGGACGTCAAGCTGTAGAAATGATCGAAAAGGTCGGGGCAGATAACATTTTCATACACCTGGATACGTATCACATGAATATCGAAGAGAAAGGTGTGGCAAACGGTATCCTGGATGCCCGAGAACACCTTCGTTATATTCACCTGTCAGAAAGCGATCGCGGCACTCCTGGCGAAGGGACGTGTGATTGGGACGAAATCTTCGCTGCGTTGGCCGCCATCGGTTTTAAAGGCGGCCTTGCTATGGAATCCTTTATTAACATGCCGCCTCAAGTCGCTTACGGCCTGGCGGTGTGGCGTCCTGTTGCCGAATCTTTTGAAGAAGTCATGGACCGTGGCCTGCCGTTTCTACGTAACAAGGCTCGTCAGTATAGATTGATCGCTtaa

[0119] SEQ ID NO:13; Protein; Tentative Class Thermofonsia branch 1 bacterium

[0120] MPTFGAHAFVWIGDWTTESGNYAIAQAGALGFDFIEIPLLAPQRFDAASHRQALAQAGIQATCSLVLPKGAHMPRYPERARQFLYEALEKVEAVGSQYLGGCIAYELGYLTGQPPTPEERQVVVEVLRDVAAEARRRGIQLALEACNRYETYLYNTLADVRETVLAVGAPNLKLHADTYHMNIEEEGFAQPLIACADVLDYIHMSESHRGLVGSGNVNWAQVWQALAAIRFNGKLVLESFAAINPDLQAATCLWRPPNQPPEVLAREGLRFLREGAAQAQLP

[0121] SEQ ID NO:14; DNA; Tentative class Thermofons ia, branch 1 bacteria

[0122] ATGCCAACCTTCGGCGCCCATGCATTTGTGTGGATTGGAGATTGGACGACAGAATCGGGAAACTATGCGATCGCGCAAGCCGGCGCACTGGGCTTCGACTTCATTGAGATTCCGCTTTTGGCACCGCAGCGTTTTGATGCCGCTTCCCACCGTCAAGCCCTGGCGCAGGCCGGCATTCAGGCGACGTGCAGCCTGGTATTGCCAAAAGGCGCTCACATGCCCAGGTATCCAGAACGTGCGCGTCAGTTCTTGTATGAAGCGTTAGAAAAGGTAGAGGCGGTTGGAAGTCAATACTTGGGCGGTTGCATCGCGTACGAACTTGGGTATCTTACCGGTCAGCCACCGACCCCGGAAGAGCGTCAGGTTGTTGTGGAAGTGCTGCGCGATGTAGCCGCAGAAGCACGCCGTCGCGGCATTCAGTTGGCACTGGAAGCATGCAATCGCTATGAGACTTACCTGTACAACACGCTGGCGGACGTGCGCGAAACGGTTTTAGCGGTGGGCGCGCCAAACTTGAAGCTGCATGCGGATACCTACCACATGAACATCGAAGAAGAAGGCTTTGCCCAGCCGCTTATTGCCTGTGCCGACGTGCTGGATTATATCCACATGTCGGAATCCCATCGCGGCCTGGTCGGAAGCGGCAACGTTAATTGGGCGCAGGTCTGGCAAGCGCTCGCCGCAATTCGCTTCAATGGCAAACTGGTGCTGGAATCGTTTGCCGCGATTAACCCGGACCTTCAGGCCGCTACTTGTCTGTGGCGCCCGCCCAACCAGCCGCCGGAAGTGCTGGCCCGCGAAGGACTGCGATTTCTCCGAGAAGGTGCGGCACAGGCCCAACTACCGtaa

[0123] SEQ ID NO:15; Protein; Deinococcus gobiensis

[0124] MLKFGAHAFCWEGDWTDEIGDRVIEQAARAGLDFIEIPLLHPETFDARRHRRHLEAVGLACVSSLGLPRDAHMPHEPEKAVTFLTGVLDRMEELGARDLTGCTGYSIGVLTGQGPTSQELDRMVDGLARVTEDARSRGIGVGLEAINRYETYMVNTLDDALAVVNRVGSDNLRVHADTYHMNIEETNLREALGRVKGKLNFIHMSESHRGLVGTGTVPWEDVWQGLADIEFSGYLTLESFAAPNAELAAATCIWKPPRHSGQELAQGGLAFLLREGATRHGLM

[0125] SEQ ID NO:16;DNA;

[0126] ATGTTGAAGTTCGGCGCGCACGCCTTTTGCTGGGAAGGCGATTGGACCGATGAAATTGGCGATCGCGTCATCGAGCAAGCGGCGCGGGCCGGATTGGATTTCATTGAAATCCCGCTCTTGCACCCTGAAACCTTCGATGCGCGCCGTCATCGTCGCCATCTTGAGGCTGTGGGCCTGGCGTGCGTTTCCAGCCTGGGCCTTCCGCGTGACGCCCACATGCCACATGAACCGGAGAAAGCGGTAACATTCCTTACCGGCGTACTGGATCGCATGGAAGAACTGGGCGCTCGCGACCTTACCGGTTGTACCGGTTATAGCATTGGCGTACTGACCGGCCAGGGTCCGACCTCTCAAGAGCTCGATCGTATGGTGGATGGTCTGGCTCGTGTGACTGAAGATGCACGGTCGCGCGGGATTGGCGTAGGCCTTGAAGCCATCAATCGTTATGAAACTTATATGGTAAACACGCTGGACGATGCGTTAGCGGTCGTGAATCGTGTCGGGAGTGACAATCTTCGCGTTCATGCCGATACATACCACATGAACATAGAAGAAACCAACCTGCGTGAGGCGCTTGGGCGAGTTAAGGGTAAGCTGAACTTCATTCACATGAGTGAAAGTCATCGTGGTCTGGTTGGTACCGGGACAGTTCCATGGGAAGATGTTTGGCAGGGTCTGGCGGATATTGAGTTCAGCGGGTATCTTACTCTTGAATCGTTTGCAGCTCCGAATGCCGAGTTGGCGGCAGCTACCTGTATCTGGAAACCACCACGGCATAGCGGCCAGGAACTGGCGCAGGGTGGCCTAGCTTTTCTGCGCGAGGGCGCGACTCGCCACGGCTTAATGtaa

[0127] SEQ ID NO:17; Protein; Endosymbiont of Heligmosomoides bakeri

[0128] MKFGAHAFVWEPEWNDTTSRRVISEAARIGLDFVEIPLLRPERFDGAATKVLLDEHAVGATYSLGLPSDKSLPERPYLAEPFLRSAIDAIESAGGDTLTGVLYGTLGELPGRPPNEKDYKVIAQVLRSVADYAKDRGIKLGIEPVNRYETFLVNTAEQAITLLDRIESDNVFIHLDTYHVNIEEDSFGAAIRLAGDRLGYIHLSESHRGTPGKGTVDWDDVFGALSDIGFAGPLVMESFVKLNADIARATCMWRDIVKDPEALIRDGIAFLEGKAKGYGLF

[0129] SEQ ID NO:18; DNA;

[0130] ATGAAATTTGGTGCGCATGCTTTCGTCTGGGAACCAGAATGGAATGATACCACCTCGCGCCGTGTGATTTCAGAGGCCGCGCGCATCGGCCTGGATTTTGTGGAAATACCGCTCCTTCGCCCAGAACGCTTTGACGGCGCCGCTACCAAAGTCTTGCTGGATGAACATGCCGTGGGCGCCACTTATTCATTGGGCCTGCCCAGTGACAAAAGCCTGCCAGAACGCCCGTATCTGGCTGAACCCTTCTTACGGTCTGCAATCGACGCCATTGAATCCGCAGGCGGTGATACACTGACAGGCGTGCTGTATGGCACTCTGGGTGAGCTGCCGGGCCGCCCGCCGAACGAAAAGGACTACAAAGTGATTGCGCAGGTATTACGTAGTGTTGCTGATTACGCGAAAGACCGTGGCATCAAACTGGGCATTGAGCCGGTAAACCGTTATGAAACCTTTCTGGTGAATACCGCAGAGCAGGCGATCACATTGTTAGACCGTATCGAAAGCGACAATGTCTTTATTCATCTGGATACCTATCACGTGAACATCGAAGAAGATAGTTTTGGCGCGGCTATCCGCCTGGCTGGGGATCGTTTAGGCTACATCCACCTGTCCGAAAGCCACCGTGGCACGCCAGGCAAGGGTACTGTGGATTGGGACGACGTTTTCGGAGCGCTTTCCGATATTGGTTTCGCAGGCCCTCTTGTGATGGAAAGCTTTGTCAAGCTGAATGCAGACATTGCACGTGCGACCTGCATGTGGCGTGATATTGTGAAAGACCCAGAAGCCCTGATTCGCGATGGTATTGCGTTTCTCGAGGGGAAAGCGAAAGGCTATGGTCTGTTTtaa

[0131] SEQ ID NO:19; Protein; Acidiphilium

[0132] MKGFGIHTSLWAHDWTEQAARLAIPEAAKHGLAFVEIALIEPDRAETEVTRGLLEQHGLAACCSLGLPEEARPTTNPDKALEFVTLALEKTAAIGASLFTGVTYGSIGERTGQPPTAAELDAVARFLDKAAAVARGFGIIFGIEVVNRYESHLFNTTEQAVALIERVGAPNIVLHLDTYHMNIEATGQANAIRAAGAHLAYIHLSESHRGVPGTGTIAWDEVFAGLAGLGFTGGMALESFIHMPPRLAAALSVWRPVAPSRAAIIDEGLPFLRNKARQYGLI

[0133] SEQ ID NO:20; DNA; Acidiphilium

[0134] ATGAAAGGCTTTGGCATTCATACAAGTCTGTGGGCCCATGACTGGACCGAGCAAGCGGCTCGTTTAGCAATTCCGGAAGCGGCAAAACATGGCCTGGCGTTTGTCGAAATTGCTTTAATTGAACCAGATCGCGCGGAAACTGAAGTTACCCGCGGGCTGCTGGAACAGCACGGCTTGGCTGCCTGCTGCTCTCTGGGCTTACCTGAGGAAGCGCGGCCAACGACGAACCCGGATAAAGCGTTGGAGTTCGTCACTTTAGCTCTGGAAAAGACCGCGGCGATAGGGGCTAGCCTGTTTACCGGTGTGACCTACGGGAGCATTGGCGAACGTACGGGCCAGCCACCTACTGCCGCCGAACTTGATGCAGTAGCAAGGTTCTTGGATAAGGCTGCCGCAGTAGCGCGCGGTTTCGGTATCATTTTCGGTATCGAAGTAGTGAATCGGTACGAGTCTCATCTGTTTAACACCACCGAACAGGCCGTTGCACTGATCGAACGTGTCGGCGCACCGAACATAGTGCTGCACTTAGACACGTATCACATGAACATCGAAGCCACGGGCCAGGCCAACGCAATACGCGCGGCGGGTGCACATCTGGCCTATATTCACCTGAGTGAATCGCACAGGGGTGTTCCGGGCACAGGGACCATCGCCTGGGACGAGGTGTTTGCAGGTCTTGCCGGCCTGGGCTTTACCGGCGGGATGGCCCTCGAATCATTTATTCACATGCCGCCCCGCCTGGCCGCCGCGCTGAGCGTCTGGCGTCCAGTTGCACCGAGTCGCGCGGCCATCATCGATGAAGGCCTTCCATTTCTGCGAAATAAAGCCCGTCAATACGGCCTCATTtaa

[0135] SEQ ID NO:21; Protein; Rhodobacterales bacterium

[0136] MLSLGLHALAAAPEWRPDLWAAILPRMAAHGVSIEPLLRPAELDIAGTRALAAKHDVELVCSLGLPATLNVAERPDEAFDFIRVALEVTASAGATALSGVTFGVIGQTTGAAPTTREIDAMTRHVSRSAALAAKKLGLRFG IEPCNRYETHLLNTGAAARAVIERSGADNAFIHLDTYHMNIEEVSHAQGFADAGDLLGYVHLSESNRGVPGRGTVDWANVFQGLKAAGFDGCAALESFVFVDADLASGLAIWRPVAENPDDVIDVGFPFLRTAGEAAGLRWAR

[0137] SEQ ID NO:22; DNA;

[0138] ATGCTTTCTCTGGGACTGCATGCGCTTGCAGCCGCCCCTGAATGGCGCCCGGATCTGTGGGCGGCGATCCTGCCGCGTATGGCGGCCCACGGTGTTAGCGTTATCGAAATTCCGCTGCTCCGTCCTGCTGAACTGGATATTGCGGGCACGCGGGCTTTAGCGGCTAAACATGATGTTGAACTCGTTTGTTCATTGGGGCTGCCAGCCACCCTCAATGTTGCGGAACGCCCTGATGAGGCGTTTGATTTTATCCGCGTGGCGTTAGAAGTGACCGCGAGCGCGGGTGCGACCGCGCTGTCAGGAGTGACATTTGGCGTGATTGGGCAGACGACGGGCGCCGCCCCAACCACGCGCGAAATCGATGCTATGACCCGGCATGTGAGCCGTAGCGCAGCCCTCGCCAAGAAACTGGGCTTGCGCTTTGGCATTGAGCCGTGCAACCGCTACGAAACCCACTTACTGAATACTGGCGCAGCCGCTCGGGCCGTGATAGAACGCTCAGGCGCTGATAACGCCTTCATCCATTTGGATACATATCACATGAATATCGAAGAAGTGTCTCATGCTCAGGGTTTTGCAGATGCGGGCGACCTGCTGGGATATGTGCACTTGTCAGAAAGCAATCGAGGCGTCCCGGGGCGTGGTACCGTCGATTGGGCGAATGTATTTCAGGGTCTGAAAGCCGCCGGATTCGATGGTTGCGCCGCACTCGAAAGTTTCGTGTTCGTTGACGCGGACTTAGCATCCGGACTTGCCATCTGGCGTCCTGTTGCTGAAAACCCAGATGACGTGATTGACGTAGGTTTCCCGTTCTTGCGGACAGCCGGAGAGGCGGCGGGCTTGCGCTGGGCGCGTtaa

[0139] SEQ ID NO:23; Protein; Chloroflexi bacterium

[0140] MVLFGAHTFIWSAEWNPETAEHVIDGAARAGLDFVEIPLLHPDQMDAGGTRRLLENYGLQCTCSLGLPREAHLPFAPDKATGFLEQAVDVTSDLGSPVLTGCLYTHLGTLTGKPPTDEEIDLVVRVLKRIARYAQDRGISLGIEPVNRYETYLLNLAEQASALLDRIDEANVFVHLDTYHMNIEEKGFYTPIVDTGPRLQYIHLSESDRGIPGTGNVHWDEVFRGLKAVKYEGRLVMESFAAVNEDLMGATAMWRDVVGDPDRLVTEGLAFLRGKAIEYGLL

[0141] SEQ ID NO:24; DNA; Chloroflexi bacterium

[0142] ATGGTACTGTTTGGGGCGCATACCTTTATTTGGAGCGCCGAATGGAACCCAGAAACGGCGGAACATGTAATTGATGGCGCAGCTCGCGCCGGCCTGGATTTCGTTGAGATTCCGTTGCTGCATCCCGATCAGATGGACGCAGGCGGCACGCGCCGGCTTTTGGAAAACTATGGTCTGCAATGCACATGTAGCTTGGGCCTGCCGCGCGAAGCGCATCTGCCATTCGCCCCTGACAAGGCAACAGGCTTTCTGGAACAGGCCGTCGATGTGACAAGTGACCTGGGTAGCCCTGTTTTGACCGGCTGTTTATATACCCACTTAGGAACGCTGACAGGAAAGCCGCCTACCGACGAAGAGATTGATTTGGTCGTTCGTGTCCTGAAGCGAATTGCGCGCTACGCCCAGGACCGAGGGATTAGTCTGGGCATCGAACCGGTCAACCGCTATGAAACCTATCTTCTGAATCTGGCGGAGCAAGCGTCTGCACTGCTCGATCGTATTGACGAAGCCAATGTATTTGTGCATCTGGATACCTATCACATGAACATTGAAGAAAAGGGCTTTTATACTCCGATCGTTGATACCGGGCCGCGTTTACAGTACATTCACCTGTCCGAATCGGACCGCGGTATCCCGGGTACTGGCAACGTTCATTGGGACGAGGTGTTTCGAGGCCTCAAGGCCGTTAAATACGAAGGCCGTCTTGTGATGGAATCTTTCGCCGCAGTCAACGAAGATCTGATGGGTGCAACGGCGATGTGGCGGGATGTGGTGGGTGATCCGGATCGATTAGTCACGGAAGGCCTGGCGTTCTTACGTGGAAAGGCGATTGAGTACGGCCTGCTGtaa

[0143] SEQ ID NO:25; Protein; Helicobacter species E85

[0144] MAEFGAHAFIWESDWNPASARRVIAGAAAAGLDFVEIPLLRPESMDTAGTRRLLAEHRLGVRCSLGLPPAASLPAHPQAAEAFLCRALDVTRALGGPVLTGVIYGTLGQLPGHPPRPGDLDIVAQTLRRVAAYAADQGLALGIEPVNRYETHLVNLTDQALELLDAIGADNVFLHLDTYHMNVEEKGFRGPVEAAGKRLRYIHLSESDRGTPGTGNVHWDEVFDGLAAIGYRGDLVMESFAAVNEDIARATCIWRQVAPDPDTLVREGLAFLRGKATARGLIP

[0145] SEQ ID NO:26; DNA; Helicobacter species E85

[0146] ATGGCGGAGTTCGGTGCACATGCTTTCATTTGGGAGTCTGACTGGAATCCGGCGTCAGCTCGCCGTGTCATTGCCGGCGCCGCTGCCGCCGGCCTAGATTTCGTCGAGATCCCACTGCTCCGCCCGGAATCCATGGACACCGCGGGAACCCGCAGATTACTTGCTGAACATCGCCTCGGCGTGCGCTGTTCACTGGGCCTGCCGCCGGCAGCGAGTCTACCCGCTCATCCGCAGGCCGCAGAAGCATTCCTTTGCCGCGCCCTAGACGTTACGCGTGCGTTGGGCGGACCCGTACTCACTGGTGTCATCTATGGCACTCTGGGCCAGTTACCGGGTCACCCGCCACGCCCTGGCGATCTTGACATTGTCGCACAGACCTTACGGCGCGTGGCAGCGTACGCCGCAGATCAGGGTCTGGCCCTGGGCATTGAACCAGTGAACCGTTATGAAACACATTTAGTGAATCTCACGGATCAAGCCCTAGAACTGTTAGATGCGATTGGCGCCGACAATGTATTCCTGCATCTGGATACCTATCACATGAACGTGGAAGAGAAAGGTTTTCGTGGTCCAGTTGAAGCAGCTGGTAAACGTTTGAGATACATTCATCTTTCGGAGTCAGATCGTGGTACCCCGGGGACAGGCAACGTTCACTGGGATGAAGTATTCGACGGTCTGGCTGCAATCGGATACCGTGGGGATCTGGTGATGGAAAGTTTTGCGGCCGTGAATGAGGACATTGCGCGTGCGACCTGCATCTGGCGTCAAGTTGCCCCAGACCCGGACACTCTGGTTAGAGAAGGCTTGGCGTTTCTTCGCGGCAAAGCGACCGCCCGCGGCCTTATCCCTtaa

[0147] SEQ ID NO:27; Protein; Bartonella species TH137

[0148] MEGFGVHTSMWTMHWDRAGAERTIPAAAAYRMDFIEIALLNTAIVDAAHTRALLERHGMRAVCSLGLPERNWASVNPEGAIAHLCDCIDTAAAMGAEALSGVTYGGIGQRTGLPPTMAEYDNIARALAAVAKHAKARGIAFGIEPVNRYENHLINTAAQAKWMIEKVGADNIFIHLDTYHMNIEEKGAGNGILDARDHLRYIHLSESDRGTPGEGTCDWDEVYATLAAIGFKGGLAMESFINMPPEVAYGLAVWRPVAENFEEVMDKGLPFLRNKARQYRLI

[0149] SEQ ID NO:28; DNA; Bartonella species TH137

[0150] ATGGAAGGATTTGGTGTACATACTTCCATGTGGACAATGCACTGGGATCGCGCGGGCGCTGAACGTACCATTCCTGCCGCAGCTGCGTACCGCATGGATTTCATAGAGATTGCTTTATTAAACACAGCCATTGTTGACGCGGCACACACGAGGGCTCTCCTTGAACGGCATGGCATGCGCGCAGTGTGTTCATTGGGCTTACCTGAACGCAATTGGGCGAGCGTGAACCCGGAAGGCGCCATCGCACATCTGTGCGATTGTATTGACACAGCAGCAGCCATGGGCGCCGAAGCCCTATCCGGCGTCACGTATGGCGGCATCGGGCAACGTACGGGGCTCCCGCCTACGATGGCCGAATACGATAACATTGCCCGCGCTCTGGCAGCTGTGGCGAAACATGCTAAAGCACGCGGCATTGCGTTTGGTATCGAACCCGTGAATCGCTACGAAAATCACTTGATTAATACCGCCGCACAAGCTAAATGGATGATCGAAAAGGTTGGGGCTGATAATATATTTATTCATCTGGACACCTATCACATGAACATAGAGGAAAAGGGAGCCGGCAATGGGATCCTCGATGCGCGTGATCATTTGAGATATATCCACTTATCGGAATCGGATCGCGGCACTCCGGGCGAAGGCACGTGTGACTGGGATGAGGTATATGCTACCCTGGCCGCCATTGGTTTTAAAGGCGGTTTGGCTATGGAATCGTTTATTAATATGCCACCGGAAGTGGCCTATGGTCTTGCCGTCTGGCGTCCGGTGGCGGAAAATTTTGAGGAAGTGATGGATAAAGGACTCCCGTTCTTGCGTAATAAAGCGCGCCAATATCGCCTGATAtaa

[0151] SEQ ID NO:29; Protein; Tropomonas isopenteniformis

[0152] MKGFGVHTSMWTMHWDRDGAERTIPATAAYGMDFVEIALLDTSIVDAAHTRALLEKHELRAVCSLGLPEGSWPSVAPEAAVAHLKDVFETAAAMGAEAVSGVTYGGIGQRSGVPPTEEEYDNVARALQQAAAYAKRLGLAFGIEPVNRYENHLINTAWQARDMIEKVGSDNIFIHLDTYHMNIEEKGVANGILDARDHLRYIHLSESDRGTPGEGCCDWNEVFGTLSAIGFEGGMAMESFINMPPQIAYGLAVWRPVADSFEDVMDRGLPFLRNKARQYRLA

[0153] SEQ ID NO:30; DNA; Tropomonas isoparvula

[0154] ATGAAGGGTTTCGGGGTGCACACGAGTATGTGGACGATGCATTGGGACCGGGATGGCGCGGAACGTACAATTCCAGCGACGGCCGCGTATGGCATGGATTTTGTCGAGATCGCTCTGTTGGATACCTCTATCGTTGATGCTGCGCACACCCGGGCGCTCCTGGAGAAACACGAGCTGCGCGCCGTTTGCTCGTTGGGCCTGCCGGAAGGCTCGTGGCCTTCGGTTGCCCCAGAGGCGGCGGTAGCGCATCTGAAAGACGTATTTGAAACCGCAGCCGCGATGGGCGCCGAAGCGGTTTCGGGCGTCACATACGGCGGTATTGGCCAACGTTCCGGTGTGCCGCCCACCGAGGAAGAATATGACAACGTAGCTCGTGCCCTGCAACAGGCAGCGGCTTACGCCAAACGCTTAGGCTTAGCCTTTGGCATTGAACCTGTGAACCGCTACGAAAACCATTTGATCAACACTGCATGGCAGGCCCGTGACATGATTGAAAAGGTAGGCTCGGACAATATTTTCATCCATTTAGATACTTACCACATGAATATAGAAGAGAAAGGTGTGGCGAACGGTATCTTAGACGCACGTGATCACCTTCGTTATATTCATCTTTCAGAGAGCGACCGTGGCACTCCGGGTGAAGGTTGCTGCGATTGGAACGAAGTTTTCGGGACCTTGAGCGCTATTGGGTTTGAAGGTGGTATGGCGATGGAATCGTTTATTAACATGCCGCCGCAGATAGCTTACGGCCTGGCCGTTTGGCGACCAGTTGCGGACTCCTTCGAAGATGTGATGGATCGCGGGCTGCCGTTTCTGCGTAACAAAGCGCGTCAGTACCGTCTGGCTtaa

[0155] SEQ ID NO:31; Protein; Rhodobacteraceae bacterium

[0156] MQGFGVHTSMWTMHWDRLGAERTIPAAAAYKMDFIEIALLDTSVVDAHHTRDLLAKHEMRAVCSLGLPRESWASVNPDGAIAHLIDAMDYTKEMGGEALSGVTYGGIGERSGVPPTEAEYDNIARALEVAAKYAKTLGIAFGIEPVNRYESHLINTSWQAKEMIDKIGADNIFIHLDTYHMNIEEKGAGNGILAAREHLRYIHLSESDRGTPGEGTCDWDEIFATLAAVEFKGGLAMESFINMPPQVGYGLGIWRPVANSFEEVMDKGLPFLRNKAAQYRLI

[0157] SEQ ID NO:32; DNA; Rhodobacteraceae bacterium

[0158] ATGCAAGGTTTTGGTGTCCATACCAGTATGTGGACAATGCACTGGGATCGCCTGGGCGCGGAACGAACCATTCCGGCCGCGGCTGCGTACAAAATGGACTTTATTGAAATTGCTCTTCTGGACACCAGCGTCGTGGATGCGCATCACACTCGTGATCTTCTGGCGAAACATGAAATGCGTGCGGTTTGCTCGCTGGGCTTGCCTCGCGAATCTTGGGCCAGTGTTAACCCGGATGGTGCCATAGCTCACCTTATAGATGCAATGGACTATACTAAAGAGATGGGCGGCGAGGCACTGTCTGGCGTCACCTATGGCGGTATCGGTGAACGTAGCGGTGTTCCGCCGACGGAAGCTGAATATGATAATATCGCACGTGCCCTAGAAGTTGCCGCCAAATACGCCAAAACGCTGGGCATTGCCTTTGGCATTGAACCCGTGAACCGCTATGAAAGCCACCTGATTAATACCAGTTGGCAGGCTAAAGAAATGATTGATAAAATCGGTGCAGATAACATCTTCATTCACTTAGATACCTACCACATGAACATTGAAGAGAAAGGCGCCGGTAATGGAATCCTGGCAGCTCGGGAACACCTTCGCTACATTCATTTGAGCGAAAGCGACCGGGGTACACCCGGCGAAGGCACCTGTGATTGGGACGAAATATTTGCGACCCTTGCTGCCGTTGAGTTCAAAGGTGGCTTGGCCATGGAGAGTTTCATTAATATGCCGCCGCAAGTGGGATATGGCTTGGGTATCTGGCGCCCGGTCGCCAATTCATTCGAGGAAGTGATGGATAAAGGCCTGCCGTTCCTGCGTAACAAAGCGGCCCAGTATCGTCTTATTtaa

[0159] SEQ ID NO:33; Protein; Rhodobacter sp. SW2

[0160] MEGFGVHTSMWTMQWDRAGAERTIPAAAAYKMDFIEIALLNTAIVDAPHTRALLQKHGLRAVASLGLPQQNWASVNPEGAIEQLCRSLDTAAAMGCEALSGVTYGGIGERSGLPPTMAEYDNVARALAAAAKHAKKLGLAFGIEPVNRYESHLINTGWQAKWMIEKVGADNIFIHLDTYHMNIEEKGAGNGILDAREHLRYVHLSESDRGTPGEGTVDWDEIFATLAAVGFKGGLAMESFINMPPELGYGLAVWRPVAESFQAVMDKGLPFLRNKARQYRLI

[0161] SEQ ID NO:34; DNA; Rhodobacter species SW2

[0162] ATGGAAGGTTTCGGTGTCCATACTTCCATGTGGACCATGCAATGGGACCGTGCTGGTGCCGAACGGACGATACCAGCCGCTGCGGCATATAAGATGGATTTCATCGAAATCGCGCTGCTGAATACGGCCATTGTGGATGCCCCGCATACCCGCGCACTGTTACAGAAACATGGTCTGCGCGCCGTTGCGAGTCTGGGCCTGCCACAACAGAACTGGGCGAGTGTCAATCCTGAGGGTGCCATTGAGCAGTTATGCCGCTCACTGGACACCGCTGCCGCGATGGGTTGCGAAGCCCTGTCAGGGGTTACCTACGGCGGCATCGGAGAGCGCAGCGGTCTGCCGCCAACAATGGCAGAATATGACAACGTGGCCCGTGCGTTGGCGGCGGCAGCAAAACACGCTAAGAAACTGGGCCTGGCGTTTGGAATCGAACCTGTGAACAGATATGAGTCTCACTTGATCAACACCGGATGGCAAGCAAAATGGATGATTGAAAAGGTGGGTGCGGATAACATCTTTATCCACCTGGATACTTATCACATGAATATTGAGGAAAAGGGTGCAGGCAACGGCATTCTCGATGCGCGCGAACACCTGCGTTACGTGCATCTTAGCGAAAGCGACCGTGGGACCCCTGGAGAAGGTACTGTTGACTGGGATGAAATATTCGCGACACTCGCCGCAGTTGGATTTAAAGGCGGATTAGCAATGGAAAGCTTTATCAACATGCCACCGGAACTGGGATATGGCCTGGCAGTATGGCGCCCGGTGGCGGAATCCTTTCAAGCAGTAATGGATAAAGGTTTACCTTTTCTGCGGAACAAAGCGCGTCAGTATCGTCTGATCtaa

[0163] SEQ ID NO:35; Protein; Alphaproteobacterium bacterium HGW-Alphaproteobacteria-8

[0164] MRGFGVHTSMWTMKWDRAGAERAVAAAAHYKMDFIEIALLNAPGVDAPHSRALLEKHGLRAVCSLGLPEAAWPSRNPEAAAVALKVALDKTAEMGCealTGVTYGGIGERTGLPPSATELDNVARALREAAAHAKKLGLLF GIEPVNRYETHLINTGRQAVEMIEKVGAENMFIHLDTYHMNIEEKGAANGILDAREHIKYIHLSESDRGTPGWGTCDWDEIFAVLSAIGFKGGLAMESFINMPPEVAYGLSVWRPVARDEAEVMDNGLPFLRGKARQYRLI

[0165] SEQ ID NO:36; DNA; Alphaproteobacteria HGW-Alphaproteobacterium-8

[0166] ATGCGTGGTTTTGGCGTCCATACATCTATGTGGACTATGAAATGGGACCGTGCAGGTGCCGAGCGTGCCGTGGCGGCGGCAGCCCACTATAAGATGGACTTCATCGAAATTGCGCTGCTGAACGCGCCGGGTGTCGATGCACCGCATAGTCGGGCACTCCTGGAGAAACATGGACTGCGCGCGGTCTGTTCATTGGGGCTTCCGGAAGCCGCTTGGCCGAGCCGCAACCCGGAAGCCGCCGTTGCTCATCTGAAAGTGGCGCTCGATAAAACCGCTGAAATGGGTTGTGAAGCGCTGACCGGAGTCACCTACGGCGGCATCGGGGAACGGACCGGCCTGCCACCGTCTGCCACCGAACTGGATAACGTGGCACGCGCTTTACGCGAAGCCGCAGCACACGCTAAGAAGCTGGGGCTCCTCTTCGGCATCGAACCGGTTAACCGCTACGAAACCCATCTGATTAACACTGGTCGGCAAGCCGTTGAAATGATTGAGAAAGTCGGCGCAGAGAATATGTTTATTCACCTGGACACCTATCACATGAACATCGAGGAAAAGGGCGCAGCGAATGGTATTCTGGATGCACGCGAACACATTAAATATATCCATCTTTCAGAATCTGATCGCGGTACCCCGGGCTGGGGTACCTGCGATTGGGATGAAATTTTCGCAGTGCTGAGCGCAATTGGATTTAAAGGTGGTTTGGCGATGGAAAGTTTTATTAACATGCCGCCGGAAGTTGCGTATGGCCTGTCAGTATGGCGACCAGTCGCACGGGATGAAGCGGAAGTTATGGATAACGGCCTTCCGTTTCTTCGTGGTAAAGCCCGCCAGTACCGCTTAATTtaa

[0167] SEQ ID NO:37; Protein; Rhodomicrobium tepidum

[0168] MQGFGVHTSMWTMNWDRAGAERTIPAASAYGMDFIEIALLNAPAVDAPHTRALLEKHGMRAVCSLGLPERNWASVNPEGAIEHLRQALEVTAALGAEALSGVTYGGIGQRTGVPPTAGEYDNIARALEAAARYARELGIAFGIEPVNRYENHLVNTAAQAKWMIEKVGADNIFIHLDTYHMNIEEKGVGNGILDAREHLRYIHLSESDRGTPGEGTCDWDEVFATLAAIGFKGGLAMESFINMPPEIGYGLAVWRPVAESFEEVMDRGLPFLRNKAKQYRLV

[0169] SEQ ID NO:38; DNA; Rhodomicrobium tepidum

[0170] ATGCAAGGGTTTGGCGTCCACACTTCGATGTGGACCATGAACTGGGATCGTGCCGGCGCCGAACGTACAATCCCGGCCGCGAGCGCATACGGTATGGATTTTATTGAAATTGCGCTGCTGAATGCCCCGGCGGTGGACGCTCCACATACGCGCGCCCTTCTTGAAAAGCACGGGATGCGTGCAGTTTGCAGTTTGGGTTTGCCTGAGCGTAATTGGGCAAGCGTCAACCCTGAAGGCGCGATCGAGCACCTTCGACAAGCCCTCGAAGTGACCGCCGCCCTTGGGGCAGAAGCGTTAAGCGGTGTCACATACGGCGGTATAGGTCAGCGCACAGGGGTGCCACCCACCGCTGGCGAGTATGATAACATCGCCCGTGCACTGGAAGCCGCTGCACGCTATGCACGAGAACTGGGTATCGCCTTTGGTATCGAACCGGTTAACCGGTACGAGAACCATCTGGTGAACACCGCGGCCCAAGCTAAATGGATGATAGAAAAGGTTGGCGCGGATAATATTTTCATCCATTTGGATACATATCACATGAACATTGAGGAGAAAGGTGTCGGCAACGGTATCCTGGATGCGCGTGAGCACCTGCGCTATATTCATCTGTCTGAGAGCGATCGCGGCACACCAGGCGAGGGTACTTGCGATTGGGACGAAGTATTCGCAACCCTGGCAGCAATTGGTTTCAAAGGCGGACTTGCTATGGAATCGTTTATCAACATGCCGCCAGAAATTGGATACGGCTTGGCCGTTTGGCGACCTGTGGCAGAGAGTTTTGAAGAAGTCATGGACCGCGGCCTGCCGTTCCTGCGGAACAAGGCGAAACAGTACCGTCTTGTGtaa

[0171] SEQ ID NO:39; Protein; Rhodobacterales bacterium

[0172] MEGFGVHTSMWTMHWDRAGAERTIPAAAAYKMDFIEIALLNAAMIDPPHTRALLEKHNMRAVASLGLPQRNWASVNPDGAAHLIEAMDVAAAMGAEALSGVTYGGIGERTGLPPTMAEYDNIARALGQAAKHAKKLGIAFGIEPVNRYENHLINTGWQAKWMIEKVGADNIFIHLDTYHMNIEEKGAGNGILDAREHLRYIHLSESDRGTPGEGTCDWDEVYATLAAIGFKGGLAMESFINMPPEVGYGLAVWRPVANSFEEVMDKGLPFLRNKARQYRLI

[0173] SEQ ID NO:40; DNA;

[0174] ATGGAAGGCTTCGGTGTGCACACGAGTATGTGGACGATGCACTGGGATCGTGCTGGTGCTGAACGTACGATTCCGGCCGCCGCCGCGTACAAGATGGACTTCATTGAAATTGCGCTGCTGAACGCAGCGATGATCGATCCGCCACATACTCGCGCGCTTCTGGAGAAACATAACATGCGGGCTGTTGCGAGTCTCGGACTTCCACAACGCAATTGGGCCTCGGTTAATCCAGATGGTGCCGCGGCCCATCTTATCGAGGCGATGGACGTGGCGGCAGCAATGGGTGCCGAAGCGCTTTCCGGAGTGACATACGGCGGAATCGGTGAACGCACTGGCCTGCCGCCAACTATGGCAGAGTACGATAACATTGCACGAGCACTCGGGCAGGCCGCCAAACATGCCAAGAAGCTTGGGATCGCTTTCGGCATAGAGCCGGTGAATCGCTATGAGAATCATCTGATCAACACTGGCTGGCAGGCCAAATGGATGATTGAGAAAGTGGGTGCTGATAACATCTTTATCCATCTGGATACATATCACATGAATATCGAGGAAAAGGGCGCTGGTAATGGTATTTTAGATGCACGTGAACACCTGCGTTATATTCATTTGTCTGAGTCCGATCGCGGCACGCCGGGTGAAGGCACGTGTGACTGGGATGAAGTTTACGCGACACTGGCAGCCATTGGCTTTAAAGGTGGACTCGCGATGGAATCATTCATCAACATGCCGCCGGAAGTGGGTTATGGTCTGGCAGTTTGGCGCCCGGTGGCGAACAGCTTTGAAGAAGTTATGGACAAAGGTCTGCCGTTTCTCCGCAATAAAGCGCGGCAGTACCGCCTGATCtaa

[0175] SEQ ID NO:41; Protein; Rhodobacterales bacterium RIFCSPHIGHO2_02_FULL_62_130>

[0176] MEGFGVHTSMWTMKWDREGTERAVQAAVDYKMDFLEIALLDAPSVDAAHTRKLLEDNDMRAVCSLGLPEAVWPSRDPESAIAFMKGVFDKANEMGAEAVSGVTYGGIGERTDMPPTEAELSNVARALEACASYAKSLGLRFGIEPVNRYETHLLNTGWQARDMIERIGSDNIFIHLDTYHMNIEEKGAASGILDAREHLKYIHLSESDRGTPGEGCCDWDEIFATLAAINFTGGLAMESFINMPPELAHGLSVWRPVAPDFQAVMDKGLPFLRNKAAQYRLV

[0177] SEQ ID NO:42; DNA; Rhodobacterales bacterium RIFCSPHIGHO2_02_FULL_62_130

[0178] ATGGAAGGTTTTGGGGTACACACCAGCATGTGGACGATGAAATGGGATCGTGAGGGCACCGAGCGGGCTGTACAGGCGGCAGTTGATTACAAAATGGATTTTCTGGAAATTGCTCTGCTTGACGCTCCGAGCGTGGACGCGGCACACACGCGCAAACTCCTGGAAGATAACGATATGCGTGCGGTGTGCAGTTTAGGGTTACCAGAAGCGGTGTGGCCGAGTCGCGACCCAGAGAGCGCAATTGCTTTTATGAAAGGAGTATTCGATAAGGCCAACGAAATGGGCGCCGAGGCAGTGAGTGGGGTCACGTATGGCGGCATAGGCGAACGCACAGACATGCCACCGACAGAAGCGGAGCTGAGCAACGTAGCCCGCGCACTGGAAGCGTGCGCGAGTTATGCAAAATCATTAGGTCTGCGATTTGGCATTGAGCCTGTGAATCGTTATGAAACCCATCTGCTGAATACCGGTTGGCAAGCACGCGATATGATCGAACGTATCGGTTCAGATAACATTTTCATTCATTTGGATACGTACCACATGAATATCGAAGAGAAAGGCGCGGCCTCTGGTATTTTAGATGCCCGTGAACACTTGAAATACATCCATTTATCTGAAAGTGACAGAGGCACGCCAGGCGAAGGTTGTTGCGACTGGGATGAAATTTTCGCCACCCTCGCCGCCATTAATTTTACCGGCGGTCTTGCCATGGAAAGTTTCATTAACATGCCGCCAGAGCTGGCGCATGGTCTGAGCGTGTGGCGCCCCGTGGCCCCGGACTTCCAAGCGGTGATGGATAAAGGCCTGCCGTTCCTGCGAAACAAAGCCGCGCAGTACCGCCTGGTCtaa

[0179] SEQ ID NO:43; Protein; Clostridium thermosaccharolyticum, encoded by the codon-optimized DNA sequence SEQ ID NO:44

[0180] KYGIFYAYWEKEWKGDFITYIEKVKKLGFDILEVGCGDFHKQPDSYFHTLRDAAREYDI ILTGGYGPRAEHNLCSPDTAVVENALAFYSDIFRKMEIAGIRSIGGGLYAYWPVDYSREPDKAGDLERSIKNMRRLADIAERHGITLNMEVLNRFEGYLINDTNEGLAYIRAVDKPNVKLMLDTFHMNIEEDSFTEPILQAGKYLGHVHVGEPNRKPPREGRIPWGEIGKALRQIGYDGPVVMEPFVTMGGQVGKDICVWRDLSQGATEEDLDRDAEKSLAFLKGMFEA

[0181] SEQ ID NO:44; DNA; Clostridium thermosaccharolyticum, codon-optimized

[0182] AAGTACGGTATCTTCTATGCTTATTGGGAAAAGGAATGGAAGGGTGACTTCATTACTTACATTGAAAAGGTTAAGAAGCTGGGATTCGATATTCTTGAAGTTGGTTGTGGTGACTTCCATAAACAACCTGATTCTTACTTCCATACTCTTAGAGATGCTGCTAGAGAATATGATATTATTCTTACTGGTGGTTACGGTCCTAGAGCTGAACATAACTTGTGTTCTCCAGATACTGCTGTTGTTGAAAACGCTCTTGCCTTCTATTCTGATATCTTCAGAAAAATGGAGATCGCTGGTATTAGATCCATTGGTGGTGGTCTGTATGCTTACTGGCCTGTTGATTATTCTAGAGAACCTGATAAGGCTGGTGATCTTGAAAGATCCATTAAGAACATGAGAAGATTGGCTGATATTGCTGAAAGACATGGTATTACTCTTAATATGGAAGTTCTTAACAGATTCGAAGGTTATCTTATTAACGATACTAACGAAGGTCTTGCTTATATTAGAGCTGTTGATAAACCAAACGTTAAATTGATGTTGGATACCTTCCACATGAACATTGAAGAAGATTCCTTCACTGAACCTATTCTTCAAGCTGGTAAGTATCTTGGTCATGTTCATGTTGGTGAACCTAATAGAAAGCCACCTAGAGAAGGTAGAATCCCTTGGGGTGAAATTGGTAAAGCTCTTAGACAAATTGGTTACGATGGTCCTGTTGTTATGGAACCATTCGTTACTATGGGTGGTCAAGTTGGTAAGGATATCTGTGTCTGGAGAGACTTATCTCAAGGTGCTACTGAAGAGGATCTTGATAGAGATGCTGAAAAGTCTCTTGCCTTCTTGAAAGGTATGTTCGAAGCT

[0183] Other embodiments

[0184] All features disclosed in this specification can be combined in any combination. Each feature disclosed in this specification can be used for the replacement feature replacement of the same, equivalent or similar purpose. Therefore, unless otherwise clearly stated, each feature disclosed is only an example of a series of equivalent or similar features. From the above description, those skilled in the art can easily determine the essential features of the present disclosure, and without departing from the spirit and scope of the present disclosure, various changes and modifications can be made to the present disclosure to make it suitable for various usages and conditions. Therefore, other embodiments are also within the scope of the claims.

[0185] Equivalents and scope

[0186] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments of the disclosure described herein.The scope of the present disclosure is not intended to be limited to the above description, but rather is set forth in the appended claims.

[0187] As used herein in the specification and claims, the indefinite articles "a" and "an" should be understood to mean "at least one", unless explicitly stated to the contrary.

[0188] The phrase "and / or" as used herein in the specification and claims should be understood to refer to "one or both" of the elements so combined, i.e., elements that are present in combination in some cases and separately in other cases. Multiple elements listed with "and / or" should be interpreted in the same manner, i.e., "one or more / one or more" of the elements so combined. In addition to the elements specifically identified by the "and / or" clause, other elements may optionally be present, whether related or unrelated to those specifically identified elements. Thus, as a non-limiting example, when used in conjunction with open language such as "comprising," a reference to "A and / or B" may refer to A only (optionally including elements other than B) in one embodiment; to B only (optionally including elements other than A) in another embodiment; to both A and B (optionally including other elements) in another embodiment; and so on.

[0189] As used herein in the specification and claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" should be interpreted as being inclusive, i.e. including at least one of a plurality of elements or a list of elements, but also including more than one thereof, and (optionally) other unlisted items. Only when the opposite terms are clearly indicated, such as "only one" or "exactly one", or when used in the claims, "consisting of..." will refer to including exactly one element in a plurality of elements or a list of elements. Generally speaking, when there are exclusive terms in the back, such as "any one", "one of", "only one of" or "exactly one of", the term "or" as used herein should only be interpreted as representing exclusive alternatives (i.e. "one or the other but not both"). When used in the claims, "consisting essentially of..." should have its ordinary meaning used in the field of patent law.

[0190] As used herein in the specification and claims, the phrase "at least one" when referring to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows that elements other than the elements specifically identified in the list of elements referred to by the phrase "at least one" may be optionally present, whether related or unrelated to those specifically identified elements. Thus, as a non-limiting example, "at least one of A and B" (or equivalently, "at least one of A or B", or equivalently "at least one of A and / or B") may mean, in one embodiment, at least one, optionally including more than one A, no B (and optionally including elements other than B); in another embodiment, at least one, optionally including more than one B, no A (and optionally including elements other than A); in another embodiment, at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements); and so on.

[0191] It should also be understood that, in any method claimed herein that includes more than one step or action, the order of the steps or actions of the method are not necessarily limited to the order of the steps or actions of the method as listed unless explicitly stated to the contrary.

[0192] In the claims and the above specification, all conjunctions such as "comprising", "including", "carrying", "having", "containing", "involving", "holding", "consisting of", etc. should be understood to be open-ended, i.e., meaning including but not limited to. As set forth in Section 2111.03 of the Manual of Patent Examining Procedures of the United States Patent Office, only the conjunctions "consisting of" and "consisting essentially of" shall be closed or semi-closed conjunctions, respectively. It should be understood that embodiments described in this document using open conjunctions (e.g., "comprising") are also contemplated in alternative embodiments as "consisting of the features described by the open conjunctions" and "consisting essentially of". For example, if the present disclosure describes "a composition comprising A and B", the present disclosure also contemplates the alternative embodiments "a composition consisting of A and B" and "a composition consisting essentially of A and B".

[0193] In addition, the present disclosure encompasses all variations, combinations and arrangements, wherein one or more limitations, elements, clauses and descriptive terms from one or more listed claims are introduced into another claim. For example, any claim subordinate to another claim may be modified to include one or more limitations present in any other claim subordinate to the same basic claim. In the case where an element is presented as a list, for example, in the format of a Markush group, each subgroup of an element is also disclosed, and any one or more elements may be removed from the group. It should be understood that, in general, in the case where the present disclosure or aspects of the present disclosure are referred to as including specific elements and / or features, certain embodiments of the present disclosure or aspects of the present disclosure are composed of such elements and / or features, or are substantially composed of them. For simplicity, those embodiments are not specifically described in the same words herein. It should also be noted that the terms "comprising" and "containing" are intended to be open and allow the inclusion of additional elements or steps. In the case of giving a range, endpoints are included. In addition, unless otherwise indicated or otherwise apparent from the context and understanding of one of ordinary skill in the art, values ​​expressed as ranges may assume any specific value or sub-range within the stated range, to the tenth of the unit of the lower limit of the range, in different embodiments of the present disclosure, unless the context clearly indicates otherwise.

[0194] This application refers to various granted patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. All references, patents, and patent applications disclosed herein are incorporated by reference with respect to each subject matter to which they are directed, and in some cases, the entire file may be covered. If there is a conflict between any incorporated reference and this specification, this specification shall prevail. In addition, any specific embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more claims. Because such embodiments are considered to be known to those of ordinary skill in the art, they may be excluded even if they are not explicitly set forth herein for exclusion. Any specific embodiment of the present disclosure may be excluded from any claim for any reason, whether or not it is related to the existence of the prior art.

[0195] Those skilled in the art will recognize or be able to determine many equivalents to the specific embodiments described herein using only routine experimentation. The scope of the embodiments of the present invention described herein is not intended to be limited to the above specific embodiments, but is as set forth in the appended claims. However, it will be appreciated by those of ordinary skill in the art that various changes and modifications may be made to this specification without departing from the spirit or scope of the present disclosure as defined in the following claims. Sequence Listing <110> Conagen Inc. <120> D-psicose 3-epimerase for bioconversion of D-fructose to D-psicose <130> C1497.70057WO00 <140> Not yet assigned <141> Submit together <150> US 63 / 022,617 <151> 2020-05-11 <160> 44 <170> PatentIn version 3.5 <210> 1 <211> 283 <212> PRT <213> Unknown <220> <223> Meiothermus silvanus <400> 1 Met Pro Arg Phe Gly Ala His Ala Phe Ile Trp Ala Ala Glu Trp Asn 1 5 10 15 Pro Glu Ala Ala Glu Lys Val Ile Gln Gly Ala Thr Arg Val Gly Leu 20 25 30 Asp Phe Val Glu Ile Pro Leu Leu His Pro Glu Ser Phe Asp Val Glu 35 40 45 Leu Thr Arg Arg Leu Leu Asn Gly Tyr Gly Val Gly Cys Thr Cys Ser 50 55 60 Leu Gly Leu Pro Arg Glu Ala Ser Leu Pro Glu His Pro Glu Ala Ala 65 70 75 80 Thr Arg Phe Leu Ile Gln Ala Leu Asn Val Ala His Gln Ile Gly Ser 85 90 95 Glu Val Leu Thr Gly Val Thr Tyr Ala Thr Leu Gly Thr Leu Ser Gly 100 105 110 Arg Ala Pro Arg Glu Ala Asp Tyr Gln Ala Val Val Lys Ala Leu Lys 115 120 125 Pro Ala Ala Arg His Ala Ala Ala Leu Gly Met Arg Leu Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr Tyr Leu Ile Asn Leu Ala Ala Gln Gly 145 150 155 160 Leu Glu Leu Ile Arg Arg Leu Asp Glu Pro Asn Val Phe Leu His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Phe Arg Gly Pro Ile 180 185 190 Val Glu Ala Gly Ala His Leu Gly Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Thr Gly Asn Val His Trp Asp Ala Val Phe Ala 210 215 220 Gly Leu Arg Glu Ile Gly Phe Ser Gly Asp Leu Val Met Glu Ser Phe 225 230 235 240 Val Ala Leu Asn Pro Asp Ile Ala Arg Ala Thr Cys Met Trp Arg Asp 245 250 255 Val Val Gly Asp Pro Gln Ala Leu Val Gln Glu Gly Leu Ala Phe Leu 260 265 270 Arg Gly Lys Ala Ser Glu Tyr Gly Leu Leu Gly 275 280 <210> 2 <211> 852 <212> DNA <213> Unknown <220> <223> Thermus silvanus <400> 2 atgccacgct tcggagctca tgcgttcatt tgggccgctg agtggaaccc ggaagcggca 60 gagaaagtga ttcagggtgc aacccgtgtt ggcctcgact tcgttgaaat tccgctcttg 120 cacccggaat ccttcgatgt cgagctcaca cgtcgtttac taaacgggta cggcgtcggt 180 tgtacgtgtt ctctgggctt gccgcgtgaa gcgtccctgc cggagcaccc cgaagccgcg 240 actcgttttc tgattcaggc cctgaatgtt gcacatcaga ttggctcaga ggtgctgacc 300 ggggtaacct acgcgacttt aggcacttta tctggccggg cgccgcgcga agctgattat 360 caagcagtcg taaaagcact gaaaccggcg gctagacatg cagcagcact gggtatgcgc 420 ctcggcattg aacccgttaa tcgttatgaa acttatttaa ttaacctggc cgctcagggc 480 ctggagttaa tccgtcgtct ggacgaaccg aatgtgttcc tgcacctgga tacgtaccac 540 atgaatattg aagagaaagg tttccggggc ccgatcgtgg aagcgggcgc ccacctgggg 600 tatattcatc tgagcgagag tgatcgcggc acccctggaa cgggcaacgt ccattgggac 660 gcggtgttcg cggggctgcg tgaaattggc ttctcaggcg atttggtcat ggaaagcttc 720 gtcgcactga accctgatat tgcgcgcgct acatgcatgt ggcgggacgt cgttggcgat 780 ccccaggcgc tggtccagga aggcttagca tttctccgtg gcaaggctag cgagtatggc 840 ctactcggtt aa 852 <210> 3 <211> 282 <212> PRT <213> Unknown <220> <223> hydrothermal vent metagenome <400> 3 Met Glu Gly Phe Gly Val His Thr Ser Met Trp Thr Met Ser Trp Asp 1 5 10 15 Lys Lys Gly Ala Glu Tyr Ala Val Ser Gln Ala Val Arg Tyr Gln Met 20 25 30 Asp Phe Leu Glu Ile Ala Leu Leu Ser Pro Leu Asp Val Asp Ala Gln 35 40 45 His Ser Arg Arg Leu Leu Glu Lys Asn His Met Arg Ala Ile Cys Ser 50 55 60 Leu Gly Leu Pro Glu Gly Ala Trp Leu Ser Asn Asn Pro Gln Ala Gly 65 70 75 80 Val Glu Tyr Leu Lys Ile Ala Leu Glu Lys Thr Ala Glu Met Gly Cys 85 90 95 Glu Ala Leu Ser Gly Val Ile Tyr Gly Gly Ile Gly Glu Arg Thr Gly 100 105 110 Phe Pro Pro Thr Glu Lys Glu Leu Asp Asn Val Val Arg Ala Leu Gly 115 120 125 Glu Ala Ala Ser Tyr Ala Lys Ser Leu Gly Leu Leu Leu Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Ser His Leu Ile Asn Thr Gly Arg Gln Ser 145 150 155 160 Val Glu Met Ile Glu Lys Val Gly Ala Ser Asn Met Phe Val His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Val Ala Gln Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Ile Lys Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Ala Gly Thr Cys Asp Trp Asp Glu Ile Phe Ala 210 215 220 Val Leu Ala Ala Ile Asn Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Val Asn Met Pro Pro Glu Ile Ala Tyr Gly Leu Ser Ile Trp Arg Pro 245 250 255 Val Ala Lys Ser Ala Asp Glu Val Met Glu Asn Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Gln Leu Leu 275 280 <210> 4 <211> 849 <212> DNA <213> Unknown <220> <223> Deep-sea hydrothermal vent metagenome <400> 4 atggaaggtt ttggagtgca cacgagcatg tggactatga gttgggataa gaagggtgcg 60 gagtatgctg taagccaggc ggtgcgctat cagatggatt ttcttgaaat cgctctgctg 120 agtccactgg atgtcgatgc ccagcatagt cgccgtctgc tagagaagaa ccacatgcgc 180 gcgatttgca gccttgggtt accggaaggt gcgtggctca gtaataaccc tcaggccgga 240 gtggaatacc tgaagatcgc cctggaaaag actgcggaga tgggttgcga agcgctgtcc 300 ggcgtaatct acggcggcat tggcgaaaga acaggttttc cgccgacgga gaaagagctg 360 gacaatgtcg tgcgcgcgct gggtgaagcg gcatcttacg caaaatcgtt aggccttctg 420 ctcggtattg aaccggtgaa ccggtatgag tctcatctga ttaatacggg acgtcagagt 480 gtggaaatga ttgaaaaggt cggtgcctct aacatgtttg ttcaccttga cacataccac 540 atgaatattg aagaaaaggg tgtggcacaa gggattctgg acgcgcgcga acacataaaa 600 tatatccacc tgtcagagtc tgaccgaggg actccgggcg caggcacgtg tgactgggat 660 gagattttcg cagtcctggc cgcaattaac tttaaaggcg gcttagcgat ggaatcgttc 720 gtgaacatgc cgccggaaat tgcctatggt ctgtccattt ggcgtccggt ggctaaatcg 780 gctgatgaag tcatggaaaa cggcctgccg tttctacgca acaaagcgcg ccagtatcag 840 ctgctgtaa 849 <210> 5 <211> 289 <212> PRT <213> Unknown <220> <223> Thermoclostridium caenicola <400> 5 Met Lys Tyr Gly Ile Phe Tyr Ala Tyr Trp Glu Lys Glu Trp Lys Gly 1 5 10 15 Asp Phe Ile Thr Tyr Ile Glu Lys Val Lys Lys Leu Gly Phe Asp Ile 20 25 30 Leu Glu Val Gly Cys Gly Asp Phe His Lys Gln Pro Asp Ser Tyr Phe 35 40 45 His Thr Leu Arg Asp Ala Ala Arg Glu Tyr Asp Ile Ile Leu Thr Gly 50 55 60 Gly Tyr Gly Pro Arg Ala Glu His Asn Leu Cys Ser Pro Asp Thr Ala 65 70 75 80 Val Val Glu Asn Ala Leu Ala Phe Tyr Ser Asp Ile Phe Arg Lys Met 85 90 95 Glu Ile Ala Gly Ile Arg Ser Ile Gly Gly Gly Leu Tyr Ala Tyr Trp 100 105 110 Pro Val Asp Tyr Ser Arg Glu Pro Asp Lys Ala Gly Asp Leu Glu Arg 115 120 125 Ser Ile Lys Asn Met Arg Arg Leu Ala Asp Ile Ala Glu Arg His Gly 130 135 140 Ile Thr Leu Asn Met Glu Val Leu Asn Arg Phe Glu Gly Tyr Leu Ile 145 150 155 160 Asn Asp Thr Asn Glu Gly Leu Ala Tyr Ile Arg Ala Val Asp Lys Pro 165 170 175 Asn Val Lys Leu Met Leu Asp Thr Phe His Met Asn Ile Glu Glu Asp 180 185 190 Ser Phe Thr Glu Pro Ile Leu Gln Ala Gly Lys Tyr Leu Gly His Val 195 200 205 His Val Gly Glu Pro Asn Arg Lys Pro Pro Arg Glu Gly Arg Ile Pro 210 215 220 Trp Gly Glu Ile Gly Lys Ala Leu Arg Gln Ile Gly Tyr Asp Gly Pro 225 230 235 240 Val Val Met Glu Pro Phe Val Thr Met Gly Gly Gln Val Gly Lys Asp 245 250 255 Ile Cys Val Trp Arg Asp Leu Ser Gln Gly Ala Thr Glu Glu Asp Leu 260 265 270 Asp Arg Asp Ala Glu Lys Ser Leu Ala Phe Leu Lys Gly Met Phe Glu 275 280 285 Ala <210> 6 <211> 870 <212> PRT <213> Unknown <220> <223> Thermoanaerobacter sludgei <400> 6 Ala Thr Gly Ala Ala Ala Thr Ala Thr Gly Gly Thr Ala Thr Ala Thr 1 5 10 15 Thr Thr Thr Ala Cys Gly Cys Ala Thr Ala Thr Thr Gly Gly Gly Ala 20 25 30 Gly Ala Ala Ala Gly Ala Ala Thr Gly Gly Ala Ala Ala Gly Gly Cys 35 40 45 Gly Ala Cys Thr Thr Thr Ala Thr Cys Ala Cys Ala Thr Ala Cys Ala 50 55 60 Thr Thr Gly Ala Gly Ala Ala Ala Gly Thr Thr Ala Ala Gly Ala Ala 65 70 75 80 Ala Thr Thr Ala Gly Gly Cys Thr Thr Thr Gly Ala Cys Ala Thr Thr 85 90 95 Thr Thr Gly Gly Ala Ala Gly Thr Cys Gly Gly Cys Thr Gly Cys Gly 100 105 110 Gly Thr Gly Ala Thr Thr Thr Thr Cys Ala Thr Ala Ala Ala Cys Ala 115 120 125 Gly Cys Cys Gly Gly Ala Thr Thr Cys Ala Thr Ala Cys Thr Thr Cys 130 135 140 Cys Ala Cys Ala Cys Cys Cys Thr Gly Cys Gly Thr Gly Ala Thr Gly 145 150 155 160 Cys Cys Gly Cys Thr Cys Gly Cys Gly Ala Ala Thr Ala Cys Gly Ala 165 170 175 Cys Ala Thr Thr Ala Thr Thr Cys Thr Gly Ala Cys Cys Gly Gly Cys 180 185 190 Gly Gly Cys Thr Ala Thr Gly Gly Gly Cys Cys Gly Cys Gly Cys Gly 195 200 205 Cys Cys Gly Ala Ala Cys Ala Cys Ala Ala Cys Thr Thr Gly Thr Gly 210 215 220 Thr Ala Gly Cys Cys Cys Gly Gly Ala Thr Ala Cys Ala Gly Cys Gly 225 230 235 240 Gly Thr Cys Gly Thr Thr Gly Ala Ala Ala Ala Thr Gly Cys Cys Cys 245 250 255 Thr Gly Gly Cys Ala Thr Thr Thr Thr Ala Cys Thr Cys Cys Gly Ala 260 265 270 Thr Ala Thr Ala Thr Thr Thr Cys Gly Cys Ala Ala Ala Ala Thr Gly 275 280 285 Gly Ala Ala Ala Thr Thr Gly Cys Cys Gly Gly Cys Ala Thr Cys Cys 290 295 300 Gly Thr Thr Cys Gly Ala Thr Cys Gly Gly Cys Gly Gly Cys Gly Gly 305 310 315 320 Thr Cys Thr Ala Thr Ala Thr Gly Cys Gly Thr Ala Thr Thr Gly Gly 325 330 335 Cys Cys Ala Gly Thr Thr Gly Ala Thr Thr Ala Cys Ala Gly Cys Cys 340 345 350 Gly Thr Gly Ala Ala Cys Cys Cys Gly Ala Cys Ala Ala Ala Gly Cys 355 360 365 Cys Gly Gly Cys Gly Ala Thr Thr Thr Ala Gly Ala Ala Cys Gly Thr 370 375 380 Thr Cys Cys Ala Thr Thr Ala Ala Gly Ala Ala Cys Ala Thr Gly Cys 385 390 395 400 Gly Thr Cys Gly Cys Thr Thr Ala Gly Cys Cys Gly Ala Thr Ala Thr 405 410 415 Thr Gly Cys Gly Gly Ala Ala Cys Gly Gly Cys Ala Thr Gly Gly Thr 420 425 430 Ala Thr Cys Ala Cys Gly Cys Thr Gly Ala Ala Thr Ala Thr Gly Gly 435 440 445 Ala Ala Gly Thr Gly Cys Thr Cys Ala Ala Thr Cys Gly Cys Thr Thr 450 455 460 Cys Gly Ala Ala Gly Gly Thr Thr Ala Cys Cys Thr Thr Ala Thr Thr 465 470 475 480 Ala Ala Thr Gly Ala Thr Ala Cys Cys Ala Ala Thr Gly Ala Ala Gly 485 490 495 Gly Thr Cys Thr Gly Gly Cys Cys Thr Ala Cys Ala Thr Thr Cys Gly 500 505 510 Thr Gly Cys Cys Gly Thr Cys Gly Ala Thr Ala Ala Ala Cys Cys Cys 515 520 525 Ala Ala Cys Gly Thr Ala Ala Ala Gly Thr Thr Gly Ala Thr Gly Cys 530 535 540 Thr Gly Gly Ala Thr Ala Cys Ala Thr Thr Thr Cys Ala Cys Ala Thr 545 550 555 560 Gly Ala Ala Cys Ala Thr Thr Gly Ala Gly Gly Ala Ala Gly Ala Thr 565 570 575 Ala Gly Cys Thr Thr Thr Ala Cys Cys Gly Ala Ala Cys Cys Cys Ala 580 585 590 Thr Thr Cys Thr Cys Cys Ala Gly Gly Cys Thr Gly Gly Thr Ala Ala 595 600 605 Gly Thr Ala Cys Cys Thr Gly Gly Gly Cys Cys Ala Thr Gly Thr Ala 610 615 620 Cys Ala Thr Gly Thr Cys Gly Gly Cys Gly Ala Ala Cys Cys Ala Ala 625 630 635 640 Ala Thr Cys Gly Thr Ala Ala Ala Cys Cys Ala Cys Cys Ala Cys Gly 645 650 655 Gly Gly Ala Ala Gly Gly Thr Cys Gly Thr Ala Thr Thr Cys Cys Gly 660 665 670 Thr Gly Gly Gly Gly Cys Gly Ala Gly Ala Thr Thr Gly Gly Cys Ala 675 680 685 Ala Ala Gly Cys Gly Cys Thr Gly Cys Gly Cys Cys Ala Gly Ala Thr 690 695 700 Thr Gly Gly Gly Thr Ala Cys Gly Ala Cys Gly Gly Thr Cys Cys Cys 705 710 715 720 Gly Thr Gly Gly Thr Gly Ala Thr Gly Gly Ala Ala Cys Cys Gly Thr 725 730 735 Thr Thr Gly Thr Thr Ala Cys Cys Ala Thr Gly Gly Gly Cys Gly Gly 740 745 750 Cys Cys Ala Ala Gly Thr Gly Gly Gly Gly Ala Ala Ala Gly Ala Cys 755 760 765 Ala Thr Thr Thr Gly Thr Gly Thr Gly Thr Gly Gly Cys Gly Gly Gly 770 775 780 Ala Cys Cys Thr Thr Ala Gly Cys Cys Ala Ala Gly Gly Thr Gly Cys 785 790 795 800 Gly Ala Cys Cys Gly Ala Ala Gly Ala Ala Gly Ala Thr Cys Thr Gly 805 810 815 Gly Ala Thr Cys Gly Cys Gly Ala Thr Gly Cys Thr Gly Ala Gly Ala 820 825 830 Ala Ala Ala Gly Cys Cys Thr Gly Gly Cys Cys Thr Thr Thr Cys Thr 835 840 845 Gly Ala Ala Ala Gly Gly Ala Ala Thr Gly Thr Thr Thr Gly Ala Ala 850 855 860 Gly Cys Cys Thr Ala Ala 865 870 <210> 7 <211> 290 <212> PRT <213> Unknown <220> <223> Deep-sea hydrothermal vent metagenome <400> 7 Met Asn Leu Pro Ala Lys Lys Met Val Leu Gly Val His Thr Phe Ala 1 5 10 15 Val Ala Pro Phe Trp Asp Leu Glu Val Met Arg His Glu Ala Arg Arg 20 25 30 Leu Lys Ser His Gly Val Gly Leu Leu Glu Ile Pro Leu Leu Arg Pro 35 40 45 Glu Glu Ile Asn Ile Lys Ala Thr Arg Ala Phe Ala Arg Glu Phe Gly 50 55 60 Phe Glu Leu Val Thr Ser Leu Gly Leu Pro Ser Asn Ile Asp Ala Val 65 70 75 80 Glu Asp Pro Gln Ser Ala Leu Ala Phe Leu Glu Pro Ala Phe Lys Val 85 90 95 Ala Ala Glu Ile Gly Ser Asn Met Leu Ser Gly Val Thr Tyr Ala Pro 100 105 110 Ile Gly Lys Ile Ser Gly Gln Pro Val Thr Gln Arg Glu Lys Asp Gly 115 120 125 Ile Cys Arg Phe Leu Gln Gln Ala Ala Ala Leu Ala Ala Asn His Gly 130 135 140 Leu Lys Leu Gly Val Glu Pro Cys Asn Arg Tyr Glu Thr His Leu Met 145 150 155 160 Asn Thr Ala Ala Gln Ala Val Glu Tyr Val Glu Ala Val Ala Ala Glu 165 170 175 Asn Leu Phe Ile His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Glu 180 185 190 Ser Phe Ala Ala Gly Phe Ala Lys Ala Ala Pro Tyr Leu Gly Tyr Val 195 200 205 His Leu Ser Glu Ser Asn Arg Gly Val Pro Gly Arg Ala Met Ile Asn 210 215 220 Trp Asp Ala Val Met Gly Ala Leu Ala Asp Ile Gly Tyr His Gly Ala 225 230 235 240 Leu Thr Leu Glu Ser Met Asn Tyr Val Asp Pro Asp Ile Ala Thr Ala 245 250 255 Leu Ala Val Trp Arg Pro Val Ala Lys Asn Pro Asp Asp Val Ile Asp 260 265 270 Phe Gly Leu Ala Phe Leu Leu Lys Ala Ala Ala Asp Ala Gly Leu Thr 275 280 285 Phe Gly 290 <210> 8 <211> 873 <212> DNA <213> Unknown <220> <223> Deep-sea hydrothermal vent metagenome <400> 8 atgaacctgc cggctaagaa gatggtcctg ggagtccata cgttcgcagt cgccccgttt 60 tgggatcttg aagtaatgcg ccacgaggcc cgccgtttaa agagccatgg cgtagggtta 120 ttggaaatcc cacttctccg gccagaagaa attaatatta aagcgacccg cgcatttgct 180 cgtgaatttg ggtttgaatt agtgaccagt ttaggcctgc cttcgaatat cgacgctgta 240 gaagatccgc agtctgcttt agcctttctg gaaccggcgt tcaaagtggc tgctgaaatt 300 ggtagcaata tgctttcagg cgttacctat gctccgattg gaaagataag tggtcagccg 360 gtgacccagc gtgagaagga tggtatttgt cggttcctac aacaagcggc tgcgttggcc 420 gcgaaccatg ggttgaaact gggcgtagag ccttgtaatc gctatgaaac gcatctgatg 480 aacacagcag cccaagcagt tgaatacgtt gaggctgtgg cagcggaaaa ccttttcatc 540 cacttagata cctaccacat gaacatcgaa gaagaaagtt ttgcagcagg gtttgcgaaa 600 gcggcgcctt atctggggta cgtgcatctg agcgagagca atcgcggcgt accgggccgc 660 gccatgatca attgggatgc agtgatgggt gcccttgccg acatcgggta tcatggggca 720 ctgactctgg agtctatgaa ttatgtggat ccggatattg cgactgccct tgcggtctgg 780 cgtccggttg ccaagaaccc ggacgatgtg atagactttg gcctggcgtt tctgctgaag 840 gccgcagcgg atgcgggttt gacattcggt taa 873 <210> 9 <211> 282 <212> PRT <213> Unknown <220> <223> Meiothermus granaticius <400> 9 Met Ala Arg Phe Gly Ala His Ala Phe Ile Trp Ser Ala Asp Trp Thr 1 5 10 15 Pro Gln Ala Ala Glu Lys Val Ala Ala Gly Ala Ala Ala Ala Gly Leu 20 25 30 Asp Phe Val Glu Ile Pro Leu Leu Arg Pro Glu Ala Phe Asp Ser Thr 35 40 45 Leu Thr Arg Arg Leu Leu Glu His His Gly Leu Gly Cys Thr Cys Ser 50 55 60 Leu Gly Leu Pro Thr Glu Ala Ala Leu Pro Asp His Pro Gln Ala Ala 65 70 75 80 Ala Arg Phe Leu Ile Gln Ala Leu Asp Val Ala His Gln Met Gly Ser 85 90 95 Pro Val Leu Ser Gly Val Thr Tyr Ala Thr Leu Gly Ala Leu Ser Gly 100 105 110 Arg Pro Pro Thr Glu Ala Asp Tyr Glu Thr Leu Ala Lys Thr Leu Lys 115 120 125 Pro Val Ala Gln His Ala Ala Arg Leu Gly Met Arg Leu Gly Leu Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr Tyr Leu Ile Asn Leu Gly Ser Gln Ala 145 150 155 160 Leu Asp Leu Ile Gln Arg Ile Gly Glu Pro Asn Val Phe Val His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Phe Lys Asn Pro Ile 180 185 190 Val Thr Val Gly Lys His Leu Gly Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Ser Gly Asn Val His Trp Asp Glu Val Phe Ser 210 215 220 Gly Leu Gln Ala Ile Gly Phe Gln Gly Asp Leu Val Met Glu Ser Phe 225 230 235 240 Val Ala Leu Asn Pro Asp Ile Ala Arg Ala Thr Cys Met Trp Arg Asp 245 250 255 Val Val Gly Asp Pro Lys Thr Leu Val His Asp Gly Leu Ala Phe Leu 260 265 270 Arg Gly Lys Ala Arg Glu Tyr Gly Leu Leu 275 280 <210> 10 <211> 849 <212> DNA <213> Unknown <220> <223> Thermoanaerobacter italicus <400> 10 atggcgcgct ttggtgcgca tgcttttatt tggagtgctg attggacccc tcaagccgcc 60 gaaaaggtcg cagcgggagc agcggcggcg ggactggatt ttgtcgaaat accgcttctt 120 cgtccagagg cgttcgattc gacacttacc cgtaggttat tagagcatca tggactggga 180 tgcacctgct ctctgggtct gcctaccgag gcagcactcc cggaccatcc tcaagcagca 240 gccgttttc tcatccaggc cttggatgta gcccatcaaa tgggcagccc ggtcctgagt 300 ggggtcacct acgcaaccct gggcgcactg tcaggccgcc cgccgaccga ggccgattat 360 gaaaccctgg cgaaaacact gaagccagta gcgcagcacg ccgcccgcct gggtatgcga 420 ctcgggcttg aaccggttaa ccgctatgaa acatacctga taaacttagg gtcacaggcg 480 ctggatctca tccaacgtat cggcgaaccg aacgtgttcg tacacctgga tacttatcac 540 atgaacatcg aagagaaagg ttttaagaac ccgattgtta ccgtcggtaa acatcttggt 600 tatattcacc tgtctgaaag cgaccgtggt accccaggta gcggtaatgt gcattgggat 660 gaagtgttct ccggcctgca agcaatcggt tttcagggtg accttgtgat ggagtcgttt 720 gtggcactga atccggatat tgcccgcgcc acgtgcatgt ggcgggatgt tgtgggagat 780 ccgaaaacac tggtgcatga tggcctagct ttctgcgcg gtaaagctcg cgaatacgga 840 ctgctgtaa 849 <210> 11 <211> 283 <212> PRT <213> Unknown <220> <223> Rubellimicrobium thermophilum <400> 11 Met Gln Gly Phe Gly Val His Thr Ser Met Trp Thr Met His Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Thr Ile Pro Ala Ala Ala Ala Tyr Lys Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asp Thr Ala Ile Val Asp Ala Ala 35 40 45 His Thr Arg Ala Leu Leu Glu Lys His Gly Leu Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Pro Val Trp Ala Ser Val Asn Pro Glu Gly Ala 65 70 75 80 Ile Ala His Leu Lys Arg Ala Leu Asp Lys Thr Ala Glu Met Gly Ala 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Gln Arg Thr Gly 100 105 110 Val Pro Pro Thr Pro Gln Glu Tyr Asp Asn Ile Ala Arg Ala Leu Glu 115 120 125 Ala Ala Ala Lys His Ala Lys Ala Leu Gly Leu Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Asn His Leu Ile Asn Thr Gly Arg Gln Ala 145 150 155 160 Val Glu Met Ile Glu Lys Val Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Val Ala Asn Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Cys Asp Trp Asp Glu Ile Phe Ala 210 215 220 Ala Leu Ala Ala Ile Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Gln Val Ala Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Glu Ser Phe Glu Glu Val Met Asp Arg Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Arg Leu Ile Ala 275 280 <210> 12 <211> 852 <212> DNA <213> Unknown <220> <223> Thermomicrobium roseum <400> 12 atgcaaggtt tcggcgtaca cactagcatg tggacgatgc attgggatcg cgcgggcgcc 60 gagcgcacca ttccggccgc cgccgcctac aaaatggact ttatcgaaat tgcgctgctg 120 gatacagcta tagtcgatgc agcgcacacc cgtgcgctgc tggagaagca cggcctgcgt 180 gcagtttgtt cactcggatt accggagccg gtctgggcct ctgtgaaccc ggaaggggcc 240 attgctcacc tgaaacgcgc gctggacaag accgcagaga tgggagctga agctctgtct 300 ggtgtgactt acggcggtat cggccagcgc acaggcgtac caccaacgcc ccaagaatac 360 gataacattg cccgtgccct tgaagcagct gcgaaacatg ccaaagcctt aggcctggcc 420 tttggtatcg aaccggttaa caggtatgaa aaccatttga taaacaccgg acgtcaagct 480 gtagaaatga tcgaaaaggt cggggcagat aacattttca tacacctgga tacgtatcac 540 atgaatatcg aagagaaagg tgtggcaaac ggtatcctgg atgcccgaga acaccttcgt 600 tatattcacc tgtcagaaag cgatcgcggc actcctggcg aagggacgtg tgattgggac 660 gaaatcttcg ctgcgttggc cgccatcggt tttaaaggcg gccttgctat ggaatccttt 720 attaacatgc cgcctcaagt cgcttacggc ctggcggtgt ggcgtcctgt tgccgaatct 780 tttgaagaag tcatggaccg tggcctgccg tttctacgta acaaggctcg tcagtataga 840 ttgatcgctt aa 852 <210> 13 <211> 282 <212> PRT <213> Unknown <220> <223> Candidatus Thermofonsia Clade 1 bacterium <400> 13 Met Pro Thr Phe Gly Ala His Ala Phe Val Trp Ile Gly Asp Trp Thr 1 5 10 15 Thr Glu Ser Gly Asn Tyr Ala Ile Ala Gln Ala Gly Ala Leu Gly Phe 20 25 30 Asp Phe Ile Glu Ile Pro Leu Leu Ala Pro Gln Arg Phe Asp Ala Ala 35 40 45 Ser His Arg Gln Ala Leu Ala Gln Ala Gly Ile Gln Ala Thr Cys Ser 50 55 60 Leu Val Leu Pro Lys Gly Ala His Met Pro Arg Tyr Pro Glu Arg Ala 65 70 75 80 Arg Gln Phe Leu Tyr Glu Ala Leu Glu Lys Val Glu Ala Val Gly Ser 85 90 95 Gln Tyr Leu Gly Gly Cys Ile Ala Tyr Glu Leu Gly Tyr Leu Thr Gly 100 105 110 Gln Pro Pro Thr Pro Glu Glu Arg Gln Val Val Val Glu Val Leu Arg 115 120 125 Asp Val Ala Ala Glu Ala Arg Arg Arg Gly Ile Gln Leu Ala Leu Glu 130 135 140 Ala Cys Asn Arg Tyr Glu Thr Tyr Leu Tyr Asn Thr Leu Ala Asp Val 145 150 155 160 Arg Glu Thr Val Leu Ala Val Gly Ala Pro Asn Leu Lys Leu His Ala 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Glu Gly Phe Ala Gln Pro Leu 180 185 190 Ile Ala Cys Ala Asp Val Leu Asp Tyr Ile His Met Ser Glu Ser His 195 200 205 Arg Gly Leu Val Gly Ser Gly Asn Val Asn Trp Ala Gln Val Trp Gln 210 215 220 Ala Leu Ala Ala Ile Arg Phe Asn Gly Lys Leu Val Leu Glu Ser Phe 225 230 235 240 Ala Ala Ile Asn Pro Asp Leu Gln Ala Ala Thr Cys Leu Trp Arg Pro 245 250 255 Pro Asn Gln Pro Pro Glu Val Leu Ala Arg Glu Gly Leu Arg Phe Leu 260 265 270 Arg Glu Gly Ala Ala Gln Ala Gln Leu Pro 275 280 <210> 14 <211> 849 <212> DNA <213> Unknown <220> <223> Tentative class Thermofonsia branch 1 bacteria <400> 14 atgccaacct tcggcgccca tgcatttgtg tggattggag attggacgac agaatcggga 60 aactatgcga tcgcgcaagc cggcgcactg ggcttcgact tcattgagat tccgcttttg 120 gcaccgcagc gttttgatgc cgcttcccac cgtcaagccc tggcgcaggc cggcattcag 180 gcgacgtgca gcctggtatt gccaaaaggc gctcacatgc ccaggtatcc agaacgtgcg 240 cgtcagttct tgtatgaagc gttagaaaag gtagaggcgg ttggaagtca atacttgggc 300 ggttgcatcg cgtacgaact tgggtatctt accggtcagc caccgacccc ggaagagcgt 360 caggttgttg tggaagtgct gcgcgatgta gccgcagaag cacgccgtcg cggcattcag 420 ttggcactgg aagcatgcaa tcgctatgag acttacctgt acaacacgct ggcggacgtg 480 cgcgaaacgg ttttagcggt gggcgcgcca aacttgaagc tgcatgcgga tacctaccac 540 atgaacatcg aagaagaagg ctttgcccag ccgcttattg cctgtgccga cgtgctggat 600 tatatccaca tgtcggaatc ccatcgcggc ctggtcggaa gcggcaacgt taattgggcg 660 caggtctggc aagcgctcgc cgcaattcgc ttcaatggca aactggtgct ggaatcgttt 720 gccgcgatta acccggacct tcaggccgct acttgtctgt ggcgcccgcc caaccagccg 780 ccggaagtgc tggcccgcga aggactgcga tttctccgag aaggtgcggc acaggcccaa 840 ctaccgtaa 849 <210> 15 <211> 282 <212> PRT <213> Unknown <220> <223> Deinococcus koreensis <400> 15 Met Leu Lys Phe Gly Ala His Ala Phe Cys Trp Glu Gly Asp Trp Thr 1 5 10 15 Asp Glu Ile Gly Asp Arg Val Ile Glu Gln Ala Ala Arg Ala Gly Leu 20 25 30 Asp Phe Ile Glu Ile Pro Leu Leu His Pro Glu Thr Phe Asp Ala Arg 35 40 45 Arg His Arg Arg His Leu Glu Ala Val Gly Leu Ala Cys Val Ser Ser 50 55 60 Leu Gly Leu Pro Arg Asp Ala His Met Pro His Glu Pro Glu Lys Ala 65 70 75 80 Val Thr Phe Leu Thr Gly Val Leu Asp Arg Met Glu Glu Leu Gly Ala 85 90 95 Arg Asp Leu Thr Gly Cys Thr Gly Tyr Ser Ile Gly Val Leu Thr Gly 100 105 110 Gln Gly Pro Thr Ser Gln Glu Leu Asp Arg Met Val Asp Gly Leu Ala 115 120 125 Arg Val Thr Glu Asp Ala Arg Ser Arg Gly Ile Gly Val Gly Leu Glu 130 135 140 Ala Ile Asn Arg Tyr Glu Thr Tyr Met Val Asn Thr Leu Asp Asp Ala 145 150 155 160 Leu Ala Val Val Asn Arg Val Gly Ser Asp Asn Leu Arg Val His Ala 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Thr Asn Leu Arg Glu Ala Leu 180 185 190 Gly Arg Val Lys Gly Lys Leu Asn Phe Ile His Met Ser Glu Ser His 195 200 205 Arg Gly Leu Val Gly Thr Gly Thr Val Pro Trp Glu Asp Val Trp Gln 210 215 220 Gly Leu Ala Asp Ile Glu Phe Ser Gly Tyr Leu Thr Leu Glu Ser Phe 225 230 235 240 Ala Ala Pro Asn Ala Glu Leu Ala Ala Ala Thr Cys Ile Trp Lys Pro 245 250 255 Pro Arg His Ser Gly Gln Glu Leu Ala Gln Gly Gly Leu Ala Phe Leu 260 265 270 Arg Glu Gly Ala Thr Arg His Gly Leu Met 275 280 <210> 16 <211> 849 <212> DNA <213> Unknown <220> <223> Deinococcus koreensis <400> 16 atgttgaagt tcggcgcgca cgccttttgc tgggaaggcg attggaccga tgaaattggc 60 gatcgcgtca tcgagcaagc ggcgcgggcc ggattggatt tcattgaaat cccgctcttg 120 caccctgaaa ccttcgatgc gcgccgtcat cgtcgccatc ttgaggctgt gggcctggcg 180 tgcgtttcca gcctgggcct tccgcgtgac gcccacatgc cacatgaacc ggagaaagcg 240 gtaacattcc ttaccggcgt actggatcgc atggaagaac tgggcgctcg cgaccttacc 300 ggttgtaccg gttatagcat tggcgtactg accggccagg gtccgacctc tcaagagctc 360 gatcgtatgg tggatggtct ggctcgtgtg actgaagatg cacggtcggcg cgggattggc 420 gtaggccttg aagccatcaa tcgttatgaa acttatatgg taaacacgct ggacgatgcg ttagcggtcg tgaatcgtgt cgggagtgac aatcttcgcg ttcatgccga tacataccac 540 atgaacatag aagaaaccaa cctgcgtgag gcgcttgggc gagttaaggg tagctgaac ttcattcaca tgagtgaaag tcatcgtggt ctggttggta ccgggacagt tccatgggaa 660 gatgtttggc agggtctggc ggatattgag ttcagcgggt atcttactct tgaatcgttt 720 gcagctccga atgccgagtt ggcggcagct acctgtatct ggaaaccacc acggcatagc 780 ggccaggaac tggcgcaggg tggcctagct tttctgcgcg aggcgcgac tcgccacggc 840 ttaatgtaa 849 <210> 17 <211> 281 <212> PRT <213> Unknown <220> <223> Endosymbionts of the spirochete Olavius ​​algarvensis <400> 17 Met Lys Phe Gly Ala His Ala Phe Val Trp Glu Pro Glu Trp Asn Asp 1 5 10 15 Thr Thr Ser Arg Arg Val Ile Ser Glu Ala Ala Arg Ile Gly Leu Asp 20 25 30 Phe Val Glu Ile Pro Leu Leu Arg Pro Glu Arg Phe Asp Gly Ala Ala 35 40 45 Thr Lys Val Leu Leu Asp Glu His Ala Val Gly Ala Thr Tyr Ser Leu 50 55 60 Gly Leu Pro Ser Asp Lys Ser Leu Pro Glu Arg Pro Tyr Leu Ala Glu 65 70 75 80 Pro Phe Leu Arg Ser Ala Ile Asp Ala Ile Glu Ser Ala Gly Gly Asp 85 90 95 Thr Leu Thr Gly Val Leu Tyr Gly Thr Leu Gly Glu Leu Pro Gly Arg 100 105 110 Pro Pro Asn Glu Lys Asp Tyr Lys Val Ile Ala Gln Val Leu Arg Ser 115 120 125 Val Ala Asp Tyr Ala Lys Asp Arg Gly Ile Lys Leu Gly Ile Glu Pro 130 135 140 Val Asn Arg Tyr Glu Thr Phe Leu Val Asn Thr Ala Glu Gln Ala Ile 145 150 155 160 Thr Leu Leu Asp Arg Ile Glu Ser Asp Asn Val Phe Ile His Leu Asp 165 170 175 Thr Tyr His Val Asn Ile Glu Glu Asp Ser Phe Gly Ala Ala Ile Arg 180 185 190 Leu Ala Gly Asp Arg Leu Gly Tyr Ile His Leu Ser Glu Ser His Arg 195 200 205 Gly Thr Pro Gly Lys Gly Thr Val Asp Trp Asp Asp Val Phe Gly Ala 210 215 220 Leu Ser Asp Ile Gly Phe Ala Gly Pro Leu Val Met Glu Ser Phe Val 225 230 235 240 Lys Leu Asn Ala Asp Ile Ala Arg Ala Thr Cys Met Trp Arg Asp Ile 245 250 255 Val Lys Asp Pro Glu Ala Leu Ile Arg Asp Gly Ile Ala Phe Leu Glu 260 265 270 Gly Lys Ala Lys Gly Tyr Gly Leu Phe 275 280 <210> 18 <211> 846 <212> DNA <213> Unknown <220> <223> Spiroplasma eriocheiris endosymbiont <400> 18 atgaaatttg gtgcgcatgc tttcgtctgg gaaccagaat ggaatgatac cacctcgcgc 60 cgtgtgattt cagaggccgc gcgcatcggc ctggattttg tggaaatacc gctccttcgc 120 ccagaacgct ttgacggcgc cgctaccaaa gtcttgctgg atgaacatgc cgtgggcgcc 180 acttattcat tgggcctgcc cagtgacaaa agcctgccag aacgcccgta tctggctgaa 240 cccttcttac ggtctgcaat cgacgccatt gaatccgcag gcggtgatac actgacaggc 300 gtgctgtatg gcactctggg tgagctgccg ggccgcccgc cgaacgaaaa ggactacaaa 360 gtgattgcgc aggtattacg tagtgttgct gattacgcga aagaccgtgg catcaaactg 420 ggcattgagc cggtaaaccg ttatgaaacc tttctggtga ataccgcaga gcaggcgatc 480 acattgttag accgtatcga aagcgacaat gtctttattc atctggatac ctatcacgtg 540 aacatcgaag aagatagttt tggcgcggct atccgcctgg ctggggatcg tttaggctac 600 atccacctgt ccgaaagcca ccgtggcacg ccaggcaagg gtactgtgga ttgggacgac 660 gttttcggag cgctttccga tattggtttc gcaggccctc ttgtgatgga aagctttgtc 720 aagctgaatg cagacattgc acgtgcgacc tgcatgtggc gtgatattgt gaaagaccca 780 gaagccctga ttcgcgatgg tattgcgttt ctcgagggga aagcgaaagg ctatggtctg 840 ttttaa 846 <210> 19 <211> 282 <212> PRT <213> Unknown <220> <223> Acidiphilium <400> 19 Met Lys Gly Phe Gly Ile His Thr Ser Leu Trp Ala His Asp Trp Thr 1 5 10 15 Glu Gln Ala Ala Arg Leu Ala Ile Pro Glu Ala Ala Lys His Gly Leu 20 25 30 Ala Phe Val Glu Ile Ala Leu Ile Glu Pro Asp Arg Ala Glu Thr Glu 35 40 45 Val Thr Arg Gly Leu Leu Glu Gln His Gly Leu Ala Ala Cys Cys Ser 50 55 60 Leu Gly Leu Pro Glu Glu Ala Arg Pro Thr Thr Asn Pro Asp Lys Ala 65 70 75 80 Leu Glu Phe Val Thr Leu Ala Leu Glu Lys Thr Ala Ala Ile Gly Ala 85 90 95 Ser Leu Phe Thr Gly Val Thr Tyr Gly Ser Ile Gly Glu Arg Thr Gly 100 105 110 Gln Pro Pro Thr Ala Ala Glu Leu Asp Ala Val Ala Arg Phe Leu Asp 115 120 125 Lys Ala Ala Ala Val Ala Arg Gly Phe Gly Ile Ile Phe Gly Ile Glu 130 135 140 Val Val Asn Arg Tyr Glu Ser His Leu Phe Asn Thr Thr Glu Gln Ala 145 150 155 160 Val Ala Leu Ile Glu Arg Val Gly Ala Pro Asn Ile Val Leu His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Ala Thr Gly Gln Ala Asn Ala Ile 180 185 190 Arg Ala Ala Gly Ala His Leu Ala Tyr Ile His Leu Ser Glu Ser His 195 200 205 Arg Gly Val Pro Gly Thr Gly Thr Ile Ala Trp Asp Glu Val Phe Ala 210 215 220 Gly Leu Ala Gly Leu Gly Phe Thr Gly Gly Met Ala Leu Glu Ser Phe 225 230 235 240 Ile His Met Pro Pro Arg Leu Ala Ala Ala Leu Ser Val Trp Arg Pro 245 250 255 Val Ala Pro Ser Arg Ala Ala Ile Ile Asp Glu Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Gly Leu Ile 275 280 <210> 20 <211> 849 <212> DNA <213> Unknown <220> <223> Acidiphilium <400> 20 atgaaaggct ttggcattca tacaagtctg tgggcccatg actggaccga gcaagcggct 60 cgtttagcaa ttccggaagc ggcaaaacat ggcctggcgt ttgtcgaaat tgctttaatt 120 gaaccagatc gcgcggaaac tgaagttacc cgcgggctgc tggaacagca cggcttggct 180 gcctgctgct ctctgggctt acctgaggaa gcgcggccaa cgacgaaccc ggataaagcg 240 ttggagttcg tcactttagc tctggaaaag accgcggcga taggggctag cctgtttacc 300 ggtgtgacct acgggagcat tggcgaacgt acgggccagc cacctactgc cgccgaactt 360 gatgcagtag caaggttctt ggataaggct gccgcagtag cgcgcggttt cggtatcatt 420 ttcggtatcg aagtagtgaa tcggtacgag tctcatctgt ttaacaccac cgaacaggcc 480 gttgcactga tcgaacgtgt cggcgcaccg aacatagtgc tgcacttaga cacgtatcac 540 atgaacatcg aagccacggg ccaggccaac gcaatacgcg cggcgggtgc acatctggcc 600 tatattcacc tgagtgaatc gcacaggggt gttccgggca cagggaccat cgcctgggac 660 gaggtgtttg caggtcttgc cggcctgggc tttaccggcg ggatggccct cgaatcattt 720 attcacatgc cgccccgcct ggccgccgcg ctgagcgtct ggcgtccagt tgcaccgagt 780 cgcgcggcca tcatcgatga aggccttcca tttctgcgaa ataaagcccg tcaatacggc 840 ctcatttaa 849 <210> 21 <211> 285 <212> PRT <213> Unknown <220> <223> Rhodobacterales bacterium <400> 21 Met Leu Ser Leu Gly Leu His Ala Leu Ala Ala Ala Pro Glu Trp Arg 1 5 10 15 Pro Asp Leu Trp Ala Ala Ile Leu Pro Arg Met Ala Ala His Gly Val 20 25 30 Ser Val Ile Glu Ile Pro Leu Leu Arg Pro Ala Glu Leu Asp Ile Ala 35 40 45 Gly Thr Arg Ala Leu Ala Ala Lys His Asp Val Glu Leu Val Cys Ser 50 55 60 Leu Gly Leu Pro Ala Thr Leu Asn Val Ala Glu Arg Pro Asp Glu Ala 65 70 75 80 Phe Asp Phe Ile Arg Val Ala Leu Glu Val Thr Ala Ser Ala Gly Ala 85 90 95 Thr Ala Leu Ser Gly Val Thr Phe Gly Val Ile Gly Gln Thr Thr Gly 100 105 110 Ala Ala Pro Thr Thr Arg Glu Ile Asp Ala Met Thr Arg His Val Ser 115 120 125 Arg Ser Ala Ala Leu Ala Lys Lys Leu Gly Leu Arg Phe Gly Ile Glu 130 135 140 Pro Cys Asn Arg Tyr Glu Thr His Leu Leu Asn Thr Gly Ala Ala Ala 145 150 155 160 Arg Ala Val Ile Glu Arg Ser Gly Ala Asp Asn Ala Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Val Ser His Ala Gln Gly Phe 180 185 190 Ala Asp Ala Gly Asp Leu Leu Gly Tyr Val His Leu Ser Glu Ser Asn 195 200 205 Arg Gly Val Pro Gly Arg Gly Thr Val Asp Trp Ala Asn Val Phe Gln 210 215 220 Gly Leu Lys Ala Ala Gly Phe Asp Gly Cys Ala Ala Leu Glu Ser Phe 225 230 235 240 Val Phe Val Asp Ala Asp Leu Ala Ser Gly Leu Ala Ile Trp Arg Pro 245 250 255 Val Ala Glu Asn Pro Asp Asp Val Ile Asp Val Gly Phe Pro Phe Leu 260 265 270 Arg Thr Ala Gly Glu Ala Ala Gly Leu Arg Trp Ala Arg 275 280 285 <210> 22 <211> 858 <212> DNA <213> Unknown <220> <223> Rhodobacter bacteria <400> 22 atgctttctc tgggactgca tgcgcttgca gccgcccctg aatggcgccc ggatctgtgg 60 gcggcgatcc tgccgcgtat ggcggcccac ggtgttagcg ttatcgaaat tccgctgctc 120 cgtcctgctg aactggatat tgcgggcacg cgggctttag cggctaaaca tgatgttgaa 180 ctcgtttgtt cattggggct gccagccacc ctcaatgttg cggaacgccc tgatgaggcg 240 tttgatttta tccgcgtggc gttagaagtg accgcgagcg cgggtgcgac cgcgctgtca 300 ggagtgacat ttggcgtgat tgggcagacg acggggcgccg ccccaaccac gcgcgaaatc 360. gatgctatga cccggcatgt gagccgtagc gcagccctcg ccaagaaact gggcttgcgc 420 tttggcattg agccgtgcaa ccgctcgaa acccacttac tgaatactgg cgcagccgct cgggccgtga tagaacgctc aggcgctgat aacgccttca tccatttgga tacatatcac 540 atgaatatcg aagaagtgtc tcatgctcag ggttttgcag atgcggggcga cctgctggga 660. tatgtgcact tgtcagaaag caatcgaggc gtcccggggc gtggtaccgt cgattgggcg aatgtatttc agggtctgaa agccgccgga ttcgatggtt gcgccgcact cgaaagtttc 720 gtgttcgttg acgcggactt agcatccgga cttgccatct ggcgtcctgt tgctgaaaac 780 ccagatgacg tgattgacgt aggtttcccg ttcttgcgga cagccggaga ggcggcgggc 840 ttgcgctggg cgcgttaa 858 <210> 23 <211> 282 <212> PRT <213> Unknown <220> <223> Chloroflexi bacterium <400> 23 Met Val Leu Phe Gly Ala His Thr Phe Ile Trp Ser Ala Glu Trp Asn 1 5 10 15 Pro Glu Thr Ala Glu His Val Ile Asp Gly Ala Ala Arg Ala Gly Leu 20 25 30 Asp Phe Val Glu Ile Pro Leu Leu His Pro Asp Gln Met Asp Ala Gly 35 40 45 Gly Thr Arg Arg Leu Leu Glu Asn Tyr Gly Leu Gln Cys Thr Cys Ser 50 55 60 Leu Gly Leu Pro Arg Glu Ala His Leu Pro Phe Ala Pro Asp Lys Ala 65 70 75 80 Thr Gly Phe Leu Glu Gln Ala Val Asp Val Thr Ser Asp Leu Gly Ser 85 90 95 Pro Val Leu Thr Gly Cys Leu Tyr Thr His Leu Gly Thr Leu Thr Gly 100 105 110 Lys Pro Pro Thr Asp Glu Glu Ile Asp Leu Val Val Arg Val Leu Lys 115 120 125 Arg Ile Ala Arg Tyr Ala Gln Asp Arg Gly Ile Ser Leu Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr Tyr Leu Leu Asn Leu Ala Glu Gln Ala 145 150 155 160 Ser Ala Leu Leu Asp Arg Ile Asp Glu Ala Asn Val Phe Val His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Phe Tyr Thr Pro Ile 180 185 190 Val Asp Thr Gly Pro Arg Leu Gln Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Ile Pro Gly Thr Gly Asn Val His Trp Asp Glu Val Phe Arg 210 215 220 Gly Leu Lys Ala Val Lys Tyr Glu Gly Arg Leu Val Met Glu Ser Phe 225 230 235 240 Ala Ala Val Asn Glu Asp Leu Met Gly Ala Thr Ala Met Trp Arg Asp 245 250 255 Val Val Gly Asp Pro Asp Arg Leu Val Thr Glu Gly Leu Ala Phe Leu 260 265 270 Arg Gly Lys Ala Ile Glu Tyr Gly Leu Leu 275 280 <210> 24 <211> 849 <212> DNA <213> Unknown <220> <223> Chloroflexi bacteria <400> 24 atggtactgt ttggggcgca tacctttatt tggagcgccg aatggaaccc agaaacggcg 60 gaacatgtaa ttgatggcgc agctcgcgcc ggcctggatt tcgttgagat tccgttgctg 120 catcccgatc agatggacgc aggcggcacg cgccggcttt tggaaaacta tggtctgcaa 180 tgcacatgta gcttgggcct gccgcgcgaa gcgcatctgc cattcgcccc tgacaaggca 240 acaggctttc tggaacaggc cgtcgatgtg acaagtgacc tgggtagccc tgttttgacc 300 ggctgtttat atacccactt aggaacgctg acaggaaagc cgcctaccga cgaagagatt 360 gatttggtcg ttcgtgtcct gaagcgaatt gcgcgctacg cccaggaccg agggattagt 420 ctgggcatcg aaccggtcaa ccgctatgaa acctatcttc tgaatctggc ggagcaagcg 480 tctgcactgc tcgatcgtat tgacgaagcc aatgtatttg tgcatctgga tacctatcac 540 atgaacattg aagaaaaggg cttttatact ccgatcgttg ataccgggcc gcgtttacag 600 tacattcacc tgtccgaatc ggaccgcggt atcccgggta ctggcaacgt tcattgggac 660 gaggtgtttc gaggcctcaa ggccgttaaa tacgaaggcc gtcttgtgat ggaatctttc 720 gccgcagtca acgaagatct gatgggtgca acggcgatgt ggcgggatgt ggtgggtgat 780 ccggatcgat tagtcacgga aggcctggcg ttcttacgtg gaaaggcgat tgagtacggc 840 ctgctgtaa 849 <210> 25 <211> 283 <212> PRT <213> Unknown <220> <223> Spiribacter sp. E85 <400> 25 Met Ala Glu Phe Gly Ala His Ala Phe Ile Trp Glu Ser Asp Trp Asn 1 5 10 15 Pro Ala Ser Ala Arg Arg Val Ile Ala Gly Ala Ala Ala Ala Gly Leu 20 25 30 Asp Phe Val Glu Ile Pro Leu Leu Arg Pro Glu Ser Met Asp Thr Ala 35 40 45 Gly Thr Arg Arg Leu Leu Ala Glu His Arg Leu Gly Val Arg Cys Ser 50 55 60 Leu Gly Leu Pro Pro Ala Ala Ser Leu Pro Ala His Pro Gln Ala Ala 65 70 75 80 Glu Ala Phe Leu Cys Arg Ala Leu Asp Val Thr Arg Ala Leu Gly Gly 85 90 95 Pro Val Leu Thr Gly Val Ile Tyr Gly Thr Leu Gly Gln Leu Pro Gly 100 105 110 His Pro Pro Arg Pro Gly Asp Leu Asp Ile Val Ala Gln Thr Leu Arg 115 120 125 Arg Val Ala Ala Tyr Ala Ala Asp Gln Gly Leu Ala Leu Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr His Leu Val Asn Leu Thr Asp Gln Ala 145 150 155 160 Leu Glu Leu Leu Asp Ala Ile Gly Ala Asp Asn Val Phe Leu His Leu 165 170 175 Asp Thr Tyr His Met Asn Val Glu Glu Lys Gly Phe Arg Gly Pro Val 180 185 190 Glu Ala Ala Gly Lys Arg Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Thr Gly Asn Val His Trp Asp Glu Val Phe Asp 210 215 220 Gly Leu Ala Ala Ile Gly Tyr Arg Gly Asp Leu Val Met Glu Ser Phe 225 230 235 240 Ala Ala Val Asn Glu Asp Ile Ala Arg Ala Thr Cys Ile Trp Arg Gln 245 250 255 Val Ala Pro Asp Pro Asp Thr Leu Val Arg Glu Gly Leu Ala Phe Leu 260 265 270 Arg Gly Lys Ala Thr Ala Arg Gly Leu Ile Pro 275 280 <210> 26 <211> 852 <212> DNA <213> Unknown <220> <223> Helicobacter species E85 <400> 26 atggcggagt tcggtgcaca tgctttcatt tgggagtctg actggaatcc ggcgtcagct 60 cgccgtgtca ttgccggcgc cgctgccgcc ggcctagatt tcgtcgagat cccactgctc 120 cgcccggaat ccatggacac cgcgggaacc cgcagattac ttgctgaaca tcgcctcggc 180 gtgcgctgtt cactgggcct gccgccggca gcgagtctac ccgctcatcc gcaggccgca 240 gaagcattcc tttgccgcgc cctagacgtt acgcgtgcgt tgggcggacc cgtactcact 300 ggtgtcatct atggcactct gggccagtta ccgggtcacc cgccacgccc tggcgatctt 360 gacattgtcg cacagacctt acggcgcgtg gcagcgtacg ccgcagatca gggtctggcc 420 ctgggcattg aaccagtgaa ccgttatgaa acacatttag tgaatctcac ggatcaagcc ctagaactgt taggcgat tggcgccgac aatgtattcc tgcatctgga tacctatcac 540 atgaacgtgg aagagaaagg ttttcgtggt ccagttgaag cagctggtaa acgtttgaga tacattcatc tttcggagtc agatcgtggt accccgggga caggcaacgt tcactgggat 660 gaagtattcg acggtctggc tgcaatcgga taccgtgggg atctggtgat ggaaagtttt gcggccgtga atgaggacat tgcgcgtgcg acctgcatct ggcgtcaagt tgccccagac 780 ccggacactc tggttagaga aggcttggcg tttcttcgcg gcaaagcgac cgcccgcggc 840 852 cttatccctt <210> 27 <211> 282 <212> PRT <213> Description(Unknown) <220> <223> Sweet potato (Tabrizicola sp.) TH137 <400> 27 Met Glu Gly Phe Gly Val His Thr Ser Met Trp Thr Met His Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Thr Ile Pro Ala Ala Ala Ala Tyr Arg Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asn Thr Ala Ile Val Asp Ala Ala 35 40 45 His Thr Arg Ala Leu Leu Glu Arg His Gly Met Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Arg Asn Trp Ala Ser Val Asn Pro Glu Gly Ala 65 70 75 80 Ile Ala His Leu Cys Asp Cys Ile Asp Thr Ala Ala Ala Met Gly Ala 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Gln Arg Thr Gly 100 105 110 Leu Pro Pro Thr Met Ala Glu Tyr Asp Asn Ile Ala Arg Ala Leu Ala 115 120 125 Ala Val Ala Lys His Ala Lys Ala Arg Gly Ile Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Asn His Leu Ile Asn Thr Ala Ala Gln Ala 145 150 155 160 Lys Trp Met Ile Glu Lys Val Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Gly Asn Gly Ile 180 185 190 Leu Asp Ala Arg Asp His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Cys Asp Trp Asp Glu Val Tyr Ala 210 215 220 Thr Leu Ala Ala Ile Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Val Ala Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Glu Asn Phe Glu Glu Val Met Asp Lys Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Arg Leu Ile 275 280 <210> 28 <211> 849 <212> DNA <213> Unknown <220> <223> Bartonella species TH137 <400> 28 atggaaggat ttggtgtaca tacttccatg tggacaatgc actgggatcg cgcgggcgct 60 gaacgtacca ttcctgccgc agctgcgtac cgcatggatt tcatagagat tgctttatta 120 aacacagcca ttgttgacgc ggcacacacg agggctctcc ttgaacggca tggcatgcgc 180 gcagtgtgtt cattgggctt acctgaacgc aattgggcga gcgtgaaccc ggaaggcgcc 240 atcgcacatc tgtgcgattg tattgacaca gcagcagcca tgggcgccga agccctatcc 300 ggcgtcacgt atggcggcat cgggcaacgt acggggctcc cgcctacgat ggccgaatac 360 gataacattg cccgcgctct ggcagctgtg gcgaaacatg ctaaagcacg cggcattgcg 420 tttggtatcg aacccgtgaa tcgctacgaa aatcacttga ttaataccgc cgcacaagct 480 aaatggatga tcgaaaaggt tggggctgat aatatattta ttcatctgga cacctatcac 540 atgaacatag aggaaaaggg agccggcaat gggatcctcg atgcgcgtga tcatttgaga 600 tatatccact tatcggaatc ggatcgcggc actccgggcg aaggcacgtg tgactgggat 660 gaggtatatg ctaccctggc cgccattggt tttaaaggcg gtttggctat ggaatcgttt 720 attaatatgc caccggaagt ggcctatggt cttgccgtct ggcgtccggt ggcggaaaat 780 tttgaggaag tgatggataa aggactcccg ttcttgcgta ataaagcgcg ccaatatcgc 840 ctgatataa 849 <210> 29 <211> 282 <212> PRT <213> Unknown <220> <223> Tropicimonas isoalkanivorans <400> 29 Met Lys Gly Phe Gly Val His Thr Ser Met Trp Thr Met His Trp Asp 1 5 10 15 Arg Asp Gly Ala Glu Arg Thr Ile Pro Ala Thr Ala Ala Tyr Gly Met 20 25 30 Asp Phe Val Glu Ile Ala Leu Leu Asp Thr Ser Ile Val Asp Ala Ala 35 40 45 His Thr Arg Ala Leu Leu Glu Lys His Glu Leu Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Gly Ser Trp Pro Ser Val Ala Pro Glu Ala Ala 65 70 75 80 Val Ala His Leu Lys Asp Val Phe Glu Thr Ala Ala Ala Met Gly Ala 85 90 95 Glu Ala Val Ser Gly Val Thr Tyr Gly Gly Ile Gly Gln Arg Ser Gly 100 105 110 Val Pro Pro Thr Glu Glu Glu Tyr Asp Asn Val Ala Arg Ala Leu Gln 115 120 125 Gln Ala Ala Ala Tyr Ala Lys Arg Leu Gly Leu Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Asn His Leu Ile Asn Thr Ala Trp Gln Ala 145 150 155 160 Arg Asp Met Ile Glu Lys Val Gly Ser Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Val Ala Asn Gly Ile 180 185 190 Leu Asp Ala Arg Asp His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Cys Cys Asp Trp Asn Glu Val Phe Gly 210 215 220 Thr Leu Ser Ala Ile Gly Phe Glu Gly Gly Met Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Gln Ile Ala Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Asp Ser Phe Glu Asp Val Met Asp Arg Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Arg Leu Ala 275 280 <210> 30 <211> 849 <212> DNA <213> Unknown <220> <223> Tropicomonas isoparaffinivorans <400> 30 atgaagggtt tcggggtgca cacgagtatg tggacgatgc attgggaccg ggatggcgcg 60 gaacgtacaa ttccagcgac ggccgcgtat ggcatggatt ttgtcgagat cgctctgttg 120 gatacctcta tcgttgatgc tgcgcacacc cgggcgctcc tggagaaaca cgagctgcgc 180 gccgtttgct cgttgggcct gccggaaggc tcgtggcctt cggttgcccc agaggcggcg 240 gtagcgcatc tgaaagacgt atttgaaacc gcagccgcga tgggcgccga agcggtttcg 300 ggcgtcacat acggcggtat tggccaacgt tccggtgtgc cgcccaccga ggaagaatat 360 gacaacgtag ctcgtgccct gcaacaggca gcggcttacg ccaaacgctt aggcttagcc 420 tttggcattg aacctgtgaa ccgctacgaa aaccatttga tcaacactgc atggcaggcc 480 cgtgacatga ttgaaaaggt aggctcggac aatattttca tccatttaga tacttaccac 540 atgaatatag aagagaaagg tgtggcgaac ggtatcttag acgcacgtga tcaccttcgt 600 tatattcatc tttcagagag cgaccgtggc actccgggtg aaggttgctg cgattggaac 660 gaagttttcg ggaccttgag cgctattggg tttgaaggtg gtatggcgat ggaatcgttt 720 attaacatgc cgccgcagat agcttacggc ctggccgttt ggcgaccagt tgcggactcc 780 ttcgaagatg tgatggatcg cgggctgccg tttctgcgta acaaagcgcg tcagtaccgt 840 ctggcttaa 849 <210> 31 <211> 282 <212> PRT <213> Unknown <220> <223> Rhodobacteraceae bacterium <400> 31 Met Gln Gly Phe Gly Val His Thr Ser Met Trp Thr Met His Trp Asp 1 5 10 15 Arg Leu Gly Ala Glu Arg Thr Ile Pro Ala Ala Ala Ala Tyr Lys Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asp Thr Ser Val Val Asp Ala His 35 40 45 His Thr Arg Asp Leu Leu Ala Lys His Glu Met Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Arg Glu Ser Trp Ala Ser Val Asn Pro Asp Gly Ala 65 70 75 80 Ile Ala His Leu Ile Asp Ala Met Asp Tyr Thr Lys Glu Met Gly Gly 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Glu Arg Ser Gly 100 105 110 Val Pro Pro Thr Glu Ala Glu Tyr Asp Asn Ile Ala Arg Ala Leu Glu 115 120 125 Val Ala Ala Lys Tyr Ala Lys Thr Leu Gly Ile Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Ser His Leu Ile Asn Thr Ser Trp Gln Ala 145 150 155 160 Lys Glu Met Ile Asp Lys Ile Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Gly Asn Gly Ile 180 185 190 Leu Ala Ala Arg Glu His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Cys Asp Trp Asp Glu Ile Phe Ala 210 215 220 Thr Leu Ala Ala Val Glu Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Gln Val Gly Tyr Gly Leu Gly Ile Trp Arg Pro 245 250 255 Val Ala Asn Ser Phe Glu Glu Val Met Asp Lys Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Ala Gln Tyr Arg Leu Ile 275 280 <210> 32 <211> 849 <212> DNA <213> Unknown <220> <223> Rhodobacteraceae bacterium <400> 32 atgcaaggtt ttggtgtcca taccagtatg tggacaatgc actgggatcg cctgggcgcg 60 gaacgaacca ttccggccgc ggctgcgtac aaaatggact ttattgaaat tgctcttctg 120 gacaccagcg tcgtggatgc gcatcacact cgtgatcttc tggcgaaaca tgaaatgcgt 180 gcggtttgct cgctgggctt gcctcgcgaa tcttgggcca gtgttaaccc ggatggtgcc 240 atagctcacc ttatagatgc aatggactat actaaagaga tgggcggcga ggcactgtct 300 ggcgtcacct atggcggtat cggtgaacgt agcggtgttc cgccgacgga agctgaatat 360 gataatatcg cacgtgccct agaagttgcc gccaaatacg ccaaaacgct gggcattgcc 420 tttggcattg aacccgtgaa ccgctatgaa agccacctga ttaataccag ttggcaggct 480 aaagaaatga ttgataaaat cggtgcagat aacatcttca ttcacttaga tacctaccac 540 atgaacattg aagagaaagg cgccggtaat ggaatcctgg cagctcggga acaccttcgc 600 tacattcatt tgagcgaaag cgaccggggt acacccggcg aaggcacctg tgattgggac 660 gaaatatttg cgacccttgc tgccgttgag ttcaaaggtg gcttggccat ggagagtttc 720 attaatatgc cgccgcaagt gggatatggc ttgggtatct ggcgcccggt cgccaattca 780 ttcgaggaag tgatggataa aggcctgccg ttcctgcgta acaaagcggc ccagtatcgt 840 cttatttaa 849 <210> 33 <211> 282 <212> PRT <213> Unknown <220> <223> Rhodobacter sp. SW2 <400> 33 Met Glu Gly Phe Gly Val His Thr Ser Met Trp Thr Met Gln Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Thr Ile Pro Ala Ala Ala Ala Tyr Lys Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asn Thr Ala Ile Val Asp Ala Pro 35 40 45 His Thr Arg Ala Leu Leu Gln Lys His Gly Leu Arg Ala Val Ala Ser 50 55 60 Leu Gly Leu Pro Gln Gln Asn Trp Ala Ser Val Asn Pro Glu Gly Ala 65 70 75 80 Ile Glu Gln Leu Cys Arg Ser Leu Asp Thr Ala Ala Ala Met Gly Cys 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Glu Arg Ser Gly 100 105 110 Leu Pro Pro Thr Met Ala Glu Tyr Asp Asn Val Ala Arg Ala Leu Ala 115 120 125 Ala Ala Ala Lys His Ala Lys Lys Leu Gly Leu Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Ser His Leu Ile Asn Thr Gly Trp Gln Ala 145 150 155 160 Lys Trp Met Ile Glu Lys Val Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Gly Asn Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Leu Arg Tyr Val His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Val Asp Trp Asp Glu Ile Phe Ala 210 215 220 Thr Leu Ala Ala Val Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Leu Gly Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Glu Ser Phe Gln Ala Val Met Asp Lys Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Arg Leu Ile 275 280 <210> 34 <211> 849 <212> DNA <213> Unknown <220> <223> Rhodobacter sp. SW2 <400> 34 atggaaggtt tcggtgtcca tacttccatg tggaccatgc aatgggaccg tgctggtgcc 60 gaacggacga taccagccgc tgcggcatat aagatggatt tcatcgaaat cgcgctgctg 120 aatacggcca ttgtggatgc cccgcatacc cgcgcactgt tacagaaaca tggtctgcgc 180 gccgttgcga gtctgggcct gccacaacag aactgggcga gtgtcaatcc tgagggtgcc 240 attgagcagt tatgccgctc actggacacc gctgccgcga tgggttgcga agccctgtca 300 ggggttacct acggcggcat cggagagcgc agcggtctgc cgccaacaat ggcagaatat 360 gacaacgtgg cccgtgcgtt ggcggcggca gcaaaacacg ctaagaaact gggcctggcg 420 tttggaatcg aacctgtgaa cagatatgag tctcacttga tcaacaccgg atggcaagca 480 aaatggatga ttgaaaaggt gggtgcggat aacatcttta tccacctgga tacttatcac 540 atgaatattg aggaaaaggg tgcaggcaac ggcattctcg atgcgcgcga acacctgcgt 600 tacgtgcatc ttagcgaaag cgaccgtggg acccctggag aaggtactgt tgactgggat 660 gaaatattcg cgacactcgc cgcagttgga tttaaaggcg gattagcaat ggaaagcttt 720 atcaacatgc caccggaact gggatatggc ctggcagtat ggcgcccggt ggcggaatcc 780 tttcaagcag taatggataa aggtttacct tttctgcgga acaaagcgcg tcagtatcgt 840 ctgatctaa 849 <210> 35 <211> 282 <212> PRT <213> Unknown <220> <223> Alphaproteobacteria bacteriumHGW-Alphaproteobacteria-8 <400> 35 Met Arg Gly Phe Gly Val His Thr Ser Met Trp Thr Met Lys Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Ala Val Ala Ala Ala Ala Ala His Tyr Lys Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asn Ala Pro Gly Val Asp Ala Pro 35 40 45 His Ser Arg Ala Leu Leu Glu Lys His Gly Leu Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Ala Ala Trp Pro Ser Arg Asn Pro Glu Ala Ala 65 70 75 80 Val Ala His Leu Lys Val Ala Leu Asp Lys Thr Ala Glu Met Gly Cys 85 90 95 Glu Ala Leu Thr Gly Val Thr Tyr Gly Gly Ile Gly Glu Arg Thr Gly 100 105 110 Leu Pro Pro Ser Ala Thr Glu Leu Asp Asn Val Ala Arg Ala Leu Arg 115 120 125 Glu Ala Ala Ala His Ala Lys Lys Leu Gly Leu Leu Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr His Leu Ile Asn Thr Gly Arg Gln Ala 145 150 155 160 Val Glu Met Ile Glu Lys Val Gly Ala Glu Asn Met Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Ala Asn Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Ile Lys Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Trp Gly Thr Cys Asp Trp Asp Glu Ile Phe Ala 210 215 220 Val Leu Ser Ala Ile Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Val Ala Tyr Gly Leu Ser Val Trp Arg Pro 245 250 255 Val Ala Arg Asp Glu Ala Glu Val Met Asp Asn Gly Leu Pro Phe Leu 260 265 270 Arg Gly Lys Ala Arg Gln Tyr Arg Leu Ile 275 280 <210> 36 <211> 849 <212> DNA <213> Unknown <220> <223> Gammaproteobacterium HGW-Gammaproteobacteria-8 <400> 36 atgcgtggtt ttggcgtcca tacatctatg tggactatga aatgggaccg tgcaggtgcc 60 gagcgtgccg tggcggcggc agcccactat aagatggact tcatcgaaat tgcgctgctg 120 aacgcgccgg gtgtcgatgc accgcatagt cgggcactcc tggagaaaca tggactgcgc 180 gcggtctgtt cattggggct tccggaagcc gcttggccga gccgcaaccc ggaagccgcc 240 gttgctcatc tgaaagtggc gctcgataaa accgctgaaa tgggttgtga agcgctgacc 300 ggagtcacct acggcggcat cggggaacgg accggcctgc caccgtctgc caccgaactg 360 gataacgtgg cacgcgcttt acgcgaagcc gcagcacacg ctaagaagct ggggctcctc 420 ttcggcatcg aaccggttaa ccgctacgaa acccatctga ttaacactgg tcggcaagcc 480 gttgaaatga ttgagaaagt cggcgcagag aatatgttta ttcacctgga cacctatcac 540 atgaacatcg aggaaaaggg cgcagcgaat ggtattctgg atgcacgcga acacattaaa 600 tatatccatc tttcagaatc tgatcgcggt accccgggct ggggtacctg cgattgggat 660 gaaattttcg cagtgctgag cgcaattgga tttaaaggtg gtttggcgat ggaaagtttt 720 attaacatgc cgccggaagt tgcgtatggc ctgtcagtat ggcgaccagt cgcacgggat 780 gaagcggaag ttatggataa cggccttccg tttcttcgtg gtaaagcccg ccagtaccgc 840 ttaatttaa 849 <210> 37 <211> 282 <212> PRT <213> Unknown <220> <223> Rubellimicrobium mesophilum <400> 37 Met Gln Gly Phe Gly Val His Thr Ser Met Trp Thr Met Asn Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Thr Ile Pro Ala Ala Ser Ala Tyr Gly Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asn Ala Pro Ala Val Asp Ala Pro 35 40 45 His Thr Arg Ala Leu Leu Glu Lys His Gly Met Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Arg Asn Trp Ala Ser Val Asn Pro Glu Gly Ala 65 70 75 80 Ile Glu His Leu Arg Gln Ala Leu Glu Val Thr Ala Ala Leu Gly Ala 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Gln Arg Thr Gly 100 105 110 Val Pro Pro Thr Ala Gly Glu Tyr Asp Asn Ile Ala Arg Ala Leu Glu 115 120 125 Ala Ala Ala Arg Tyr Ala Arg Glu Leu Gly Ile Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Asn His Leu Val Asn Thr Ala Ala Gln Ala 145 150 155 160 Lys Trp Met Ile Glu Lys Val Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Val Gly Asn Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Cys Asp Trp Asp Glu Val Phe Ala 210 215 220 Thr Leu Ala Ala Ile Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Ile Gly Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Glu Ser Phe Glu Glu Val Met Asp Arg Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Lys Gln Tyr Arg Leu Val 275 280 <210> 38 <211> 849 <212> DNA <213> Unknown <220> <223> Microbacterium thermoxanthum <400> 38 atgcaagggt ttggcgtcca cacttcgatg tggaccatga actgggatcg tgccggcgcc 60 gaacgtacaa tcccggccgc gagcgcatac ggtatggatt ttattgaaat tgcgctgctg 120 aatgccccgg cggtggacgc tccacatacg cgcgcccttc ttgaaaagca cgggatgcgt 180 gcagtttgca gtttgggttt gcctgagcgt aattgggcaa gcgtcaaccc tgaaggcgcg 240 atcgagcacc ttcgacaagc cctcgaagtg accgccgccc ttggggcaga agcgttaagc 300 ggtgtcacat acggcggtat aggtcagcgc acaggggtgc cacccaccgc tggcgagtat 360 gataacatcg cccgtgcact ggaagccgct gcacgctatg cacgagaact gggtatcgcc tttggtatcg aaccggttaa ccggtacgag aaccatctgg tgaacaccgc ggcccaagct aaatggatga tagaaaaggt tggcgcggat aatattttca tccatttgga tacatatcac atgaacattg aggagaaagg tgtcggcaac ggtatcctgg atgcgcgtga gcacctgcgc 600 tatattcatc tgtctgagag cgatcgcggc acaccaggcg agggtacttg cgattgggac 660 gaagtattcg caaccctggc agcaattggt ttcaaaggcg gacttgctat ggaatcgttt atcaacatgc cgccagaaat tggatacggc ttggccgttt ggcgacctgt ggcagagagt 780 840. tttgaagaag tcatggaccg cggcctgccg ttcctgcgga acaaggcgaa acagtaccgt cttgtgtaa 849 <210> 39 <211> 282 <212> PRT <213> Description(Unknown) <220> <223> Antibiotic (Rhodobacterales bacterium) <400> 39 Met Glu Gly Phe Gly Val His Thr Ser Met Trp Thr Met His Trp Asp 1 5 10 15 Arg Ala Gly Ala Glu Arg Thr Ile Pro Ala Ala Ala Ala Tyr Lys Met 20 25 30 Asp Phe Ile Glu Ile Ala Leu Leu Asn Ala Ala Met Ile Asp Pro Pro 35 40 45 His Thr Arg Ala Leu Leu Glu Lys His Asn Met Arg Ala Val Ala Ser 50 55 60 Leu Gly Leu Pro Gln Arg Asn Trp Ala Ser Val Asn Pro Asp Gly Ala 65 70 75 80 Ala Ala His Leu Ile Glu Ala Met Asp Val Ala Ala Ala Met Gly Ala 85 90 95 Glu Ala Leu Ser Gly Val Thr Tyr Gly Gly Ile Gly Glu Arg Thr Gly 100 105 110 Leu Pro Pro Thr Met Ala Glu Tyr Asp Asn Ile Ala Arg Ala Leu Gly 115 120 125 Gln Ala Ala Lys His Ala Lys Lys Leu Gly Ile Ala Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Asn His Leu Ile Asn Thr Gly Trp Gln Ala 145 150 155 160 Lys Trp Met Ile Glu Lys Val Gly Ala Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Gly Asn Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Leu Arg Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Thr Cys Asp Trp Asp Glu Val Tyr Ala 210 215 220 Thr Leu Ala Ala Ile Gly Phe Lys Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Val Gly Tyr Gly Leu Ala Val Trp Arg Pro 245 250 255 Val Ala Asn Ser Phe Glu Glu Val Met Asp Lys Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Arg Gln Tyr Arg Leu Ile 275 280 <210> 40 <211> 849 <212> DNA <213> Unknown <220> <223> Rhodobacterales bacterium <400> 40 atggaaggct tcggtgtgca cacgagtatg tggacgatgc actgggatcg tgctggtgct 60 gaacgtacga ttccggccgc cgccgcgtac aagatggact tcattgaaat tgcgctgctg 120 aacgcagcga tgatcgatcc gccacatact cgcgcgcttc tggagaaaca taacatgcgg gctgttgcga gtctcggact tccacaacgc aattgggcct cggttaatcc agatggtgcc 240 gcggcccatc ttatcgaggc gatggacgtg gcggcagcaa tgggtgccga agcgctttcc 300 ggagtgacat acggcggat cggtgaacgc actggcctgc cgccaactat ggcagagtac 360 420. gataacattg cacgagcact cgggcaggcc gccaaacatg ccaagaagct tgggatcgct ttcggcatag agccggtgaa tcgctatgag aatcatctga tcaacactgg ctggcaggcc aaatggatga ttgagaaagt gggtgctgat aacatcttta tccatctgga tacatatcac atgaatatcg aggaaaaggg cgctggtaat ggtattttag atgcacgtga acacctgcgt tatattcatt tgtctgagtc cgatcgcggc acgccggggtg aaggcacgtg tgactgggat 660 gaagtttacg cgacactggc agccattggc tttaaaggtg gactcgcgat ggaatcattc atcaacatgc cgccggaagt gggttatggt ctggcagttt ggcgcccggt ggcgaacagc 780 tttgaagaag ttatggacaa aggtctgccg tttctccgca ataaagcgcg gcagtaccgc ctgatctaa 849 <210> 41 <211> 282 <212> PRT <213> Unknown <220> <223> Rhodobacterales bacterium RIFCSPHIGHO2_02_FULL_62_130 <400> 41 Met Glu Gly Phe Gly Val His Thr Ser Met Trp Thr Met Lys Trp Asp 1 5 10 15 Arg Glu Gly Thr Glu Arg Ala Val Gln Ala Ala Val Asp Tyr Lys Met 20 25 30 Asp Phe Leu Glu Ile Ala Leu Leu Asp Ala Pro Ser Val Asp Ala Ala 35 40 45 His Thr Arg Lys Leu Leu Glu Asp Asn Asp Met Arg Ala Val Cys Ser 50 55 60 Leu Gly Leu Pro Glu Ala Val Trp Pro Ser Arg Asp Pro Glu Ser Ala 65 70 75 80 Ile Ala Phe Met Lys Gly Val Phe Asp Lys Ala Asn Glu Met Gly Ala 85 90 95 Glu Ala Val Ser Gly Val Thr Tyr Gly Gly Ile Gly Glu Arg Thr Asp 100 105 110 Met Pro Pro Thr Glu Ala Glu Leu Ser Asn Val Ala Arg Ala Leu Glu 115 120 125 Ala Cys Ala Ser Tyr Ala Lys Ser Leu Gly Leu Arg Phe Gly Ile Glu 130 135 140 Pro Val Asn Arg Tyr Glu Thr His Leu Leu Asn Thr Gly Trp Gln Ala 145 150 155 160 Arg Asp Met Ile Glu Arg Ile Gly Ser Asp Asn Ile Phe Ile His Leu 165 170 175 Asp Thr Tyr His Met Asn Ile Glu Glu Lys Gly Ala Ala Ser Gly Ile 180 185 190 Leu Asp Ala Arg Glu His Leu Lys Tyr Ile His Leu Ser Glu Ser Asp 195 200 205 Arg Gly Thr Pro Gly Glu Gly Cys Cys Asp Trp Asp Glu Ile Phe Ala 210 215 220 Thr Leu Ala Ala Ile Asn Phe Thr Gly Gly Leu Ala Met Glu Ser Phe 225 230 235 240 Ile Asn Met Pro Pro Glu Leu Ala His Gly Leu Ser Val Trp Arg Pro 245 250 255 Val Ala Pro Asp Phe Gln Ala Val Met Asp Lys Gly Leu Pro Phe Leu 260 265 270 Arg Asn Lys Ala Ala Gln Tyr Arg Leu Val 275 280 <210> 42 <211> 849 <212> DNA <213> Unknown <220> <223> Rhodobacterales bacterium RIFCSPHIGHO2_02_FULL_62_130 <400> 42 atggaaggtt ttggggtaca caccagcatg tggacgatga aatgggatcg tgagggcacc 60 gagcgggctg tacaggcggc agttgattac aaaatggatt ttctggaaat tgctctgctt 120 gacgctccga gcgtggacgc ggcacacacg cgcaaactcc tggaagataa cgatatgcgt 180 gcggtgtgca gtttagggtt accagaagcg gtgtggccga gtcgcgaccc agagagcgca 240 attgctttta tgaaaggagt attcgataag gccaacgaaa tgggcgccga ggcagtgagt 300 ggggtcacgt atggcggcat aggcgaacgc acagacatgc caccgacaga agcggagctg 360 agcaacgtag cccgcgcact ggaagcgtgc gcgagttatg caaaatcatt aggtctgcga 420 tttggcattg agcctgtgaa tcgttatgaa acccatctgc tgaataccgg ttggcaagca 480 cgcgatatga tcgaacgtat cggttcagat aacattttca ttcatttgga tacgtaccac 540 atgaatatcg aagagaaagg cgcggcctct ggtattttag atgcccgtga acacttgaaa 600 tacatccatt tatctgaaag tgacagaggc acgccaggcg aaggttgttg cgactgggat 660 gaaattttcg ccaccctcgc cgccattaat tttaccggcg gtcttgccat ggaaagtttc 720 attaacatgc cgccagagct ggcgcatggt ctgagcgtgt ggcgccccgt ggccccggac 780 ttccaagcgg tgatggataa aggcctgccg ttcctgcgaa acaaagccgc gcagtaccgc 840 ctggtctaa 849 <210> 43 <211> 288 <212> PRT <213> Unknown <220> <223> Clostridium thermopalmarium <400> 43 Lys Tyr Gly Ile Phe Tyr Ala Tyr Trp Glu Lys Glu Trp Lys Gly Asp 1 5 10 15 Phe Ile Thr Tyr Ile Glu Lys Val Lys Lys Leu Gly Phe Asp Ile Leu 20 25 30 Glu Val Gly Cys Gly Asp Phe His Lys Gln Pro Asp Ser Tyr Phe His 35 40 45 Thr Leu Arg Asp Ala Ala Arg Glu Tyr Asp Ile Ile Leu Thr Gly Gly 50 55 60 Tyr Gly Pro Arg Ala Glu His Asn Leu Cys Ser Pro Asp Thr Ala Val 65 70 75 80 Val Glu Asn Ala Leu Ala Phe Tyr Ser Asp Ile Phe Arg Lys Met Glu 85 90 95 Ile Ala Gly Ile Arg Ser Ile Gly Gly Gly Leu Tyr Ala Tyr Trp Pro 100 105 110 Val Asp Tyr Ser Arg Glu Pro Asp Lys Ala Gly Asp Leu Glu Arg Ser 115 120 125 Ile Lys Asn Met Arg Arg Leu Ala Asp Ile Ala Glu Arg His Gly Ile 130 135 140 Thr Leu Asn Met Glu Val Leu Asn Arg Phe Glu Gly Tyr Leu Ile Asn 145 150 155 160 Asp Thr Asn Glu Gly Leu Ala Tyr Ile Arg Ala Val Asp Lys Pro Asn 165 170 175 Val Lys Leu Met Leu Asp Thr Phe His Met Asn Ile Glu Glu Asp Ser 180 185 190 Phe Thr Glu Pro Ile Leu Gln Ala Gly Lys Tyr Leu Gly His Val His 195 200 205 Val Gly Glu Pro Asn Arg Lys Pro Pro Arg Glu Gly Arg Ile Pro Trp 210 215 220 Gly Glu Ile Gly Lys Ala Leu Arg Gln Ile Gly Tyr Asp Gly Pro Val 225 230 235 240 Val Met Glu Pro Phe Val Thr Met Gly Gly Gln Val Gly Lys Asp Ile 245 250 255 Cys Val Trp Arg Asp Leu Ser Gln Gly Ala Thr Glu Glu Asp Leu Asp 260 265 270 Arg Asp Ala Glu Lys Ser Leu Ala Phe Leu Lys Gly Met Phe Glu Ala 275 280 285 <210> 44 <211> 864 <212> DNA <213> Unknown <220> <223> Clostridium thermopalmarium <400> 44 aagtacggta tcttctatgc ttattgggaa aaggaatgga agggtgactt cattacttac 60 attgaaaagg ttaagaagct gggattcgat attcttgaag ttggttgtgg tgacttccat 120 aaacaacctg attcttactt ccatactctt agagatgctg ctagagaata tgatattatt 180 cttactggtg gttacggtcc tagagctgaa cataacttgt gttctccaga tactgctgtt 240 gttgaaaacg ctcttgcctt ctattctgat atcttcagaa aaatggagat cgctggtatt 300 agatccattg gtggtggtct gtatgcttac tggcctgttg attattctag agaacctgat 360 aaggctggtg atcttgaaag atccattaag aacatgagaa gattggctga tattgctgaa agacatggta ttactcttaa tatggaagtt cttaacagat tcgaaggtta tcttattaac 540. gatactaacg aaggtcttgc ttatattaga gctgttgata aaccaaacgt taaattgatg ttggatacct tccacatgaa cattgaagaa gattccttca ctgaacctat tcttcaagct ggtaagtatc ttggtcatgt tcatgttggt gacctaata gaaagccacc taggaaggt agaatccctt ggggtgaat tggtaaagct cttagacaaa ttggttacga tggtcctgtt gttatggaac cattcgttac tatgggtggt caagttggta aggatatctg tgtctggaga 780 840. gacttatctc aaggtgctc tgaagaggat cttgatagag atgctgaaaa gtctcttgcc ttcttgaaag gtatgttcga agct

Claims

1. A method for producing psicose, the method comprising contacting a fructose substrate with a reaction mixture under conditions such that the fructose substrate is converted into psicose, the reaction mixture comprising: (a) an enzyme system comprising D-psicose 3-epimerase; (b) a host cell transformed with a recombinant vector comprising a nucleic acid molecule encoding D-psicose 3-epimerase; and / or (c) a lysate of the host cell of (b); The D-psicose 3-epimerase comprises the amino acid sequence of SEQ ID NO:

1.

2. The method of claim 1, wherein the conditions comprise maintaining the enzyme system and the fructose substrate at a temperature between 25°C and 75°C.

3. The method of claim 1, wherein the conditions comprise maintaining the enzyme system and the fructose substrate at a pH between 4 and 10. The method according to claim 1 , wherein the D-psicose 3-epimerase is in an isolated form. 5 . The method according to claim 1 , wherein the D-psicose 3-epimerase is immobilized on a solid matrix.

6. The method according to claim 1, wherein the reaction mixture comprises a host cell transformed with a recombinant vector comprising a nucleic acid molecule encoding the D-psicose 3-epimerase or a lysate of the host cell, wherein the nucleic acid molecule encoding the D-psicose 3-epimerase comprises the polynucleotide sequence of SEQ ID NO:

2.

7. The method of claim 6, wherein the host cell is selected from the group consisting of a yeast cell, a filamentous fungal cell, a bacterial cell, a mammalian cell, a plant cell, and a Labyrinthus cell.

8. The method according to claim 7, wherein the host cell is Escherichia coli (E. coli) or Pichia pastoris (P. pastoris).

9. The method of claim 6, wherein the recombinant vector exists as a self-replicating nucleic acid molecule in the host cell.

10. The method according to claim 6, wherein the recombinant vector is integrated into the host cell chromosome.

11. The method according to claim 1, further comprising adding at least one type of metal ion to the reaction system, wherein the at least one type of metal ion is selected from the group consisting of magnesium ion, manganese ion, copper ion, zinc ion, nickel ion, cobalt ion, iron ion, aluminum ion and calcium ion.

12. The method of claim 11, wherein the metal ion is added at a concentration of about 0.01 mM to about 5 mM. 13 . The method of claim 1 , comprising removing a product stream comprising psicose from a reaction system when the fructose substrate is converted into psicose.

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