CN117043341A

CN117043341A - Compositions for degrading trypsin or TMPRSS2

Info

Publication number: CN117043341A
Application number: CN202280010679.5A
Authority: CN
Inventors: 李优先; 渡边荣一郎; 川岛祐介; 王主君; 小原收; 本田贤也; 新幸二
Original assignee: Keio School; RIKEN Institute of Physical and Chemical Research
Current assignee: Keio School; RIKEN Institute of Physical and Chemical Research
Priority date: 2021-01-19
Filing date: 2022-01-18
Publication date: 2023-11-10

Abstract

A composition for degrading trypsin or TMPRSS2, the active ingredient of which comprises a bacterium having 00502 protein or a bacterium having a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding ability, or a bacterium having 00509 protein or a bacterium having a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding ability.

Description

Compositions for degrading trypsin or TMPRSS2

Technical Field

The present application relates to compositions for degrading trypsin or TMPRSS 2. More particularly, the present application relates to compositions for degrading trypsin or TMPRSS2, diagnostic agents for diseases caused by trypsin or TMPRSS2, and quasi-drugs for diseases caused by trypsin or TMPRSS 2. The present application claims priority based on U.S. provisional application No. US 63/138,798 at 2021, month 1, and U.S. provisional application No. US 63/229,077 at 2021, month 8, 4, the contents of which are incorporated herein by reference.

Background

The gastrointestinal tract is a unique organ that is constantly exposed to numerous food and microorganism-derived molecules. In addition to foreign substances, the gastrointestinal tract encounters host-derived molecules, such as digestive enzymes and the like. In the upper intestine, digestive enzymes play an important role in breaking down the large amount of nutrients ingested in the diet into smaller components for physical absorption. On the other hand, the large intestine mainly absorbs water and does not require digestive enzymes, but rather, the imbalance of digestive enzyme activity is related to changes in microbial flora composition, disruption of mucosal barriers, and development of inflammation.

Intestinal tissue has a variety of regulatory and protective mechanisms, such as the production of mucins and enzyme inactivating molecules, to maintain balance and barrier function. It is also known that intestinal microbiota also contributes significantly to maintaining a stable environment by reducing or modifying substances in cavities (for example, refer to non-patent document 1).

Prior art literature

Non-patent literature

Non-patent document 1: j.l. round and s.k. mazmanian, the gut microbiota shapes intestinal immune responses during health and disease, nature reviews immunology,9,313-323,2009.

Disclosure of Invention

Problems to be solved by the invention

However, the details of the regulation of the protein in the cavity by the microorganism are not completely clear. In particular, the properties of microorganisms related to the regulation of digestive enzymes remaining in the large intestine remain largely unknown. The object of the present invention is to elucidate the functions of microorganisms involved in the regulation of residual proteases in the large intestine and to provide a technique for controlling the protease activity.

Means for solving the problems

The present invention includes the following aspects.

[1] A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria: a bacterium having 00502 protein or a bacterium having a protein which has 30% or more of sequence identity with the amino acid sequence of 00502 protein and has trypsin-binding ability; or alternatively

Bacteria having 00509 protein or bacteria having proteins having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding capacity.

[2] A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria: a bacterium having 00502 protein or a bacterium having a protein which has 30% or more of sequence identity with the amino acid sequence of 00502 protein and has trypsin-binding ability; and

[3] A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria: a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 1 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 1 and encodes a protein having trypsin binding ability; or a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 2 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 2 and encodes a protein having trypsin binding ability.

[4] A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria: a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 1 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 1 and encodes a protein having trypsin binding ability; and a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 2 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 2 and encodes a protein having trypsin binding ability.

[5] The composition for degrading trypsin or TMPRSS2 according to any of claims 1-4, wherein the bacteria have a secretion system of type IX (T9 SS).

[6] The composition for degrading trypsin or TMPRSS2 of claim 5, wherein the T9SS comprises PorV protein, porU protein, porN protein, porM protein, porL protein, porK protein, or PorP protein.

[7] The composition for degrading trypsin or TMPRSS2 according to any of the claims 1-6, wherein the bacterium is a bacterium of Paraprevatella, prevoltella, prevoltelamabilia or Bactoidetes genus.

[8] The composition for degrading trypsin or TMPRSS2 according to claim 7, wherein the bacterium of the genus Paraprevatella is Paraprevotella clara and the bacterium of the genus Prevoltella is at least one bacterium selected from the group consisting of Prevoltella rara, prevotella rodentium and Prevoltella muris.

[9] The composition for degrading trypsin or TMPRSS2 as claimed in any of claims 1-8, wherein the bacterium is a bacterium having a 16S rRNA gene comprising a base sequence as shown in SEQ ID NO. 3 or SEQ ID NO. 4 or a bacterium having a 16S rRNA gene comprising a base sequence having 97% or more sequence identity with the base sequence as shown in SEQ ID NO. 3 or SEQ ID NO. 4.

[10] The composition for degrading trypsin or TMPRSS2 according to any of claims 1-6, wherein the bacterium is at least one bacterium selected from the group consisting of Paraprevatella sp.MSP 0303, paraprevatella sp.MSP 0335, prevotellamassilia timonensis, bactoides sp.MSP 0288, bactoides sp.MSP 0410, bactoides sp.MSP 0435 and Porphyromonas gingivalis.

[11] The composition for degrading trypsin or TMPRSS2 of any one of claims 1-10, wherein the bacteria is a living bacteria.

[12] The composition for degrading trypsin or TMPRSS2 of any one of claims 1-10, wherein the bacteria is dead bacteria.

[13] A composition for degrading trypsin or TMPRSS2, comprising as active ingredients the following proteins:

00502 protein or a protein having 30% or more sequence identity to the amino acid sequence of said 00502 protein and having trypsin binding capacity; or alternatively

00509 protein or a protein having 30% or more sequence identity to the amino acid sequence of said 00509 protein and having trypsin binding capacity.

[14] A composition for degrading trypsin or TMPRSS2, comprising as active ingredients the following proteins:

00502 protein or a protein having 30% or more sequence identity to the amino acid sequence of said 00502 protein and having trypsin binding capacity; and

[15] A composition for degrading trypsin or TMPRSS2 according to any one of claims 1-14 for use in treating a disease caused by trypsin or TMPRSS 2.

[16] The composition for degrading trypsin or TMPRSS2 of any one of claims 1-15, wherein the disease caused by trypsin or TMPRSS2 is inflammatory bowel disease (ulcerative colitis, crohn's disease), irritable bowel disease, infection, acute pancreatitis, or chronic pancreatitis.

[17] The composition for degrading trypsin or TMPRSS2 of claim 16, wherein the infection is a viral infection or a bacterial infection.

[18] The composition for degrading trypsin or TMPRSS2 of claim 16 or 17, wherein the inflammatory bowel disease, irritable bowel disease, or infection is a disease involving TMPRSS2 or IgA.

[19] A diagnostic agent for a disease caused by trypsin or TMPRSS2, the diagnostic agent comprising a specific binding substance for detecting: 00502 protein or a protein having 30% or more sequence identity to the amino acid sequence of 00502 protein and having trypsin binding capacity; or alternatively

00509 protein or a protein having 30% or more sequence identity to the amino acid sequence of 00509 protein and having trypsin binding capacity.

[20] A diagnostic agent for a disease caused by trypsin or TMPRSS2, the diagnostic agent comprising a primer set or probe for detecting:

A gene comprising a base sequence shown as SEQ ID NO. 1 or a gene having 30% or more sequence identity with a base sequence shown as SEQ ID NO. 1 and encoding a protein having trypsin binding ability; or alternatively

A gene comprising a base sequence shown as SEQ ID NO. 2 or a gene having 30% or more sequence identity with a base sequence shown as SEQ ID NO. 2 and encoding a protein having trypsin binding ability.

[21] A quasi-drug for diseases caused by trypsin or TMPRSS2, which contains bacteria having the following proteins as active ingredients: 00502 protein or a protein having 30% or more sequence identity to the amino acid sequence of 00502 protein and having trypsin binding capacity; or alternatively

[22] A quasi drug for diseases caused by trypsin or TMPRSS2, which contains as active ingredients the following proteins:

ADVANTAGEOUS EFFECTS OF INVENTION

The present invention can provide a technique for controlling protease activity.

Drawings

FIG. 1 is a graph showing the results of proteomic analysis in example 1;

FIG. 2 is a graph showing the results of trypsin activity detection of feces of Specific Pathogen Free (SPF) mice and sterile (GF) mice in example 1;

FIG. 3 is a photograph showing the result of Western Blot analysis of feces of SPF mice and GF mice in example 1, showing the result of anionic trypsin (PRSS 2);

FIG. 4 is a fluorescence micrograph showing the results of mucus (UEA 1) and anionic trypsin (PRSS 2) detected by immunostaining large intestine sections of SPF mice and GF mice in example 1;

FIG. 5 is a graph showing the results of trypsin activity measured at different locations of the small intestine and the large intestine of GF mice and SPF mice in example 1;

fig. 6 is a graph showing the measurement results of trypsin activity in GF mice feces to which a fecal sample collected from a healthy human donor was applied in example 2;

FIG. 7 is a graph showing the results of measurement of trypsin activity in the feces of each group of mice in example 2;

FIG. 8 is a photograph showing the result of trypsin degradation by Western Blot measurement in example 2;

FIG. 9 is a photograph showing the result of measuring degradation of human trypsin by Western Blot in example 2;

FIG. 10 is a photograph showing the result of measuring trypsin degradation ability of various bacteria by Western Blot in example 2;

FIG. 11 is a photograph showing the results of incubating recombinant mouse PRSS2 (rmPRSS 2) pretreated with a protease inhibitor in combination with P.clara (1C 4) and analyzing rmPRSS2 degradation using Western Blot in example 3;

FIG. 12 is a photograph showing the result of observing the binding of rmPRSS2 to the surface of bacteria by confocal microscopy in example 3;

FIG. 13 is a photograph showing the result of incubating P.clara (1C 4) pretreated with tunicamycin with rmPRSS2 and analyzing the degradation of rmPRSS2 by Western Blot in example 3;

FIG. 14 is a photograph showing the results of incubating P.clara (1C 4) treated or not with tunicamycin with rmPRSS2, respectively, and observing the binding of rmPRSS2 to the bacterial surface with a confocal microscope in example 3;

FIG. 15 is a schematic diagram showing the arrangement of the gene compositions of type IX secretion mechanism (T9 SS) in the genomes of P.clara (JCM 14859), P.xylanipila (JCM 14860) and P.gingivalis (ATCC 33277) of example 3;

FIG. 16 is a schematic diagram showing homologous recombination absent PorU expression in example 3;

FIG. 17 is a photograph showing the result of incubating mutation P.clara (JCM 14859) with rmPRSS2 and analyzing the degradation of rmPRSS2 by Western Blot in example 3;

FIG. 18 is a photograph showing the results of Western Blot analysis of rmPRSS2 degradation mediated by wild-type P.clara (JCM 14859) or a series of mutant P.clara (JCM 14859) in example 3;

FIG. 19 is a photograph P.clara showing the results of incubating P.clara (Δ00502) and P.clara (Δ00509) with rmPRSS2, respectively, and observing the binding of rmPRSS2 to the bacterial surface with a confocal microscope in example 3;

FIG. 20 is a photograph P.clara showing the results of incubating P.clara (Δ00502) and P.clara (Δ00509) with rmPRSS2, respectively, and analyzing the degradation of rmPRSS2 by Western Blot in example 3;

FIG. 21 is a schematic diagram showing the genomic sequence of Parapreviella strain analyzed in example 3;

FIG. 22 is a photograph showing the result of incubating a P.clara mutant obtained by deleting each of 00502 gene to 00509 gene with rmPRSS2 and analyzing the degradation of P.clalarmPRSS2 by Western Blot in example 3;

FIG. 23 is a graph showing the results of measuring the protease activity of recombinant 00502 protein or 00509 protein by cleaving FITC-labeled casein in example 4;

FIG. 24 is a photograph showing the result of incubating recombinant 00502 protein and 00509 protein in free form or recombinant 00502 protein and 00509 protein bound to microbeads with rmPRSS2 and analyzing the degradation of rmPRSS2 by Western Blot in example 4;

FIG. 25 is a photograph showing the results of incubating recombinant 00502 protein, recombinant 00509 protein and Bovine Serum Albumin (BSA) bound to microbeads with rmPRSS2, respectively, and observing the binding of rmPRSS2 and protein-bound microbeads with a confocal microscope in example 4;

FIG. 26 is a photograph showing the result of incubating the content of the cecum of GF mice with recombinant 00502 protein bound to a culture medium control (-) or microbeads and analyzing trypsin degradation by Western Blot in example 4;

FIG. 27 is a graph showing the trypsin degradation model mediated by Parapreviella bacteria proposed by the inventors;

FIG. 28 is a graph showing the results of DNA quantification of the P.clara strain in feces of GF mice inoculated with the P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix in example 5;

FIG. 29 is a graph showing the results of measurement of trypsin activity in feces of GF mice inoculated with the P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix in example 5;

FIG. 30 is a graph showing the results of measurement of trypsin activity in feces of GF mice inoculated with the P.clara strains (wild type (WT), Δ00502) and 34-mix in example 5;

FIG. 31 is a photograph showing the results of Western Blot analysis of the results of the Western Blot analysis of proteins in feces of GF mice inoculated with the P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix in example 5;

FIG. 32 is a photograph showing the results of Western Blot analysis of proteins in the feces of GF mice, the feces of GF mice inoculated with the wild-type P.clara strain and 2-mix, a mixed sample of the two, and a mixed sample of the two plus trypsin inhibitor (TCLK) in example 5;

Fig. 33 is a graph showing the change in body weight of each group of mice after c.rodenium infection in example 6;

fig. 34 the upper half of fig. 34 shows the large intestine images of groups of mice 7 days after c.rodenium infection in example 6; the lower half of fig. 34 is a representative micrograph showing hematoxylin-eosin staining of cecal tissue in example 6;

fig. 35 is a graph showing the cecal histological scores based on hematoxylin-eosin staining in example 6;

FIG. 36 is a photograph showing the results of Western Blot analysis of proteins in feces in example 6;

FIG. 37 is a photograph showing the result of evaluating fecal IgA aggregation effect by in vitro incubation of living C.rodentum and fecal fluid in example 6;

FIG. 38 is a schematic view showing the experimental schedule of example 7;

FIG. 39 is a graph showing the change in body weight of mice after C.rodentum infection in example 7;

fig. 40 is a graph showing CFU measurements of c.rodentum in cecum plaques in example 7;

FIG. 41 is a graph showing CFU measurements of C.rodentium in the lumen contents of example 7;

FIG. 42 is a photograph showing the results of Western Blot analysis of the proteins in the lumen contents of the cecum in example 7;

FIG. 43 is a schematic diagram showing an experiment of infection with Mouse Hepatitis Virus (MHV) in example 8;

FIG. 44 is a graph showing the survival curve of mice after MHV infection in example 8;

FIG. 45 is a graph showing the results of measuring MHV virus titers in the liver, brain and stool in example 8;

fig. 46 is a representative image showing hematoxylin-eosin staining results of liver tissue of each group of mice in example 8; FIG. 47 is a diagram showing homologues of genes related to trypsin degradation (00502 gene and 00509 gene) and species encoded thereby, which were retrieved on a computer in example 9;

FIG. 48 is a graph showing the locus structure of the gene involved in trypsin degradation and the sequence identity (%) with respect to the gene of the P.clara strain in each bacterial species of example 9;

fig. 49 is a graph showing the measurement results of trypsin activity in feces of a non-IBD control group (healthy subjects), ulcerative Colitis (UC) group and Crohn's Disease (CD) group in japanese cohort in example 9;

FIG. 50 is a graph showing Paraprevatela carrier rates in a patient population diagnosed with Ulcerative Colitis (UC) and Crohn's Disease (CD) in PRISM and HMP2 cohorts of example 9;

FIG. 51 is a photograph showing the results of measurement of trypsin degradation mediated by strain Prevotella rodentium, prevolella muris in example 9;

FIG. 52 is a photograph showing the results of measurement of trypsin degradation activity mediated by the Prevotella ara (MSP 0081) strain in example 9;

FIG. 53 is a schematic view showing an example of a fusion protein of 00502 protein or its homolog and/or 00509 protein or its homolog with an antibody constant region.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

[ composition for degrading trypsin or TMPRSS2 ]

In one embodiment, the present invention provides a composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria: a bacterium having 00502 protein or a bacterium having a protein having 30% or more of sequence identity with the amino acid sequence of 00502 protein and having trypsin binding ability, or a bacterium having 00509 protein or a bacterium having a protein having 30% or more of sequence identity with the amino acid sequence of 00509 protein and having trypsin binding ability.

As described in the examples below, the inventors have shown that bacteria having 00502 protein or 00509 protein adsorb trypsin or TMPRSS2 on the surface of the bacteria, and degrade trypsin or TMPRSS2 by autolysis of trypsin or TMPRSS2.

Therefore, a bacterium containing 00502 protein or a composition containing a bacterium containing 00509 protein as an active ingredient can be used for degrading trypsin or TMPRSS2. Thus, the composition of this embodiment can also be said to be a degradation agent for trypsin or TMPRSS2.

Degradation of trypsin or TMPRSS2 may be useful in industrial or pharmaceutical applications, as described below.

The UniProtKB accession number of 00502 protein of strain Paraprevotella clara (YIT 11840) described below (catalog number: JCM 14859) is G5SNC9. In addition, the 00502 protein of Paraprevotella clara (YIT 11840) strain is encoded by the HMPREF41_ 00858 gene (UniProtKB), the NCBI accession number of the HMPREF9441_00858 gene being NZ_JH376591 REGION:complex (87340..91131). The amino acid sequence of 00502 protein of Paraprevotella clara (YIT 11840) strain is shown as SEQ ID NO. 5, and the cDNA base sequence of HMPREF9441_00858 gene is shown as SEQ ID NO. 1.

The amino acid sequence of 00502 protein of Paraprevotella clara (1C 4) strain is shown in SEQ ID NO. 6, and the cDNA base sequence of gene encoding 00502 protein of Paraprevotella clara (1C 4) strain is shown in SEQ ID NO. 7.

The amino acid sequence of 00502 protein of Paraprevotella xylaniphila (82A 6) strain is shown as SEQ ID NO. 8, and the cDNA base sequence of gene encoding Paraprevotella xylaniphila (82A 6) 00502 protein of strain is shown as SEQ ID NO. 9.

The amino acid sequence of 00502 protein of Paraprevotella xylaniphila (YIT 11841) strain (catalog number: JCM 14860) is shown in SEQ ID NO. 10, and the cDNA base sequence of the gene encoding 00502 protein of Paraprevotella xylaniphila (YIT 11841) strain is shown in SEQ ID NO. 11.

The amino acid sequence of 00502 protein of Prevolella ra (109) strain (catalog number: DSM 105141) is shown in SEQ ID NO. 12, and the cDNA base sequence of the gene encoding 00502 protein of Prevolella ra (109) strain is shown in SEQ ID NO. 13.

The amino acid sequence of 00502 protein of Prevotella rodentium (PJ 1A) strain (catalog number: DSM 105243) is shown in SEQ ID NO. 14, and the cDNA base sequence of the gene encoding 00502 protein of Prevotella rodentium (PJ 1A) strain is shown in SEQ ID NO. 15.

The amino acid sequence of 00502 protein of Prevolella Muris (PMUR) strain (catalog number: DSM 103722) is shown in SEQ ID NO. 16, and the cDNA base sequence of the gene encoding 00502 protein of Prevolella Muris (PMUR) strain is shown in SEQ ID NO. 17.

Paraprevotella clara (YIT 11840) strain (catalog number: JCM 14859) 00509 protein UniProtKB accession number G5SNC1. In addition, the Paraprevotella clara (YIT 11840) strain 00509 protein is encoded by the HMPREF9441_00850 gene (UniProtKB). The NCBI accession number of the HMPREF9441_00850 gene is NZ_JH376591REGION 73848.76931. The amino acid sequence of 00509 protein of Paraprevotella clara (YIT 11840) strain is shown as SEQ ID NO. 18, and the cDNA base sequence of HMPREF9441_00850 gene is shown as SEQ ID NO. 2.

The amino acid sequence of 00509 protein of Paraprevotella clara (1C 4) strain is shown as SEQ ID NO. 19, and the cDNA base sequence of gene encoding Paraprevotella clara (1C 4) 00509 protein of strain is shown as SEQ ID NO. 20.

The amino acid sequence of 00509 protein of Parapprevotella xvlaniphila (82A 6) strain is shown as SEQ ID NO. 21, and the cDNA base sequence of the gene encoding 00509 protein of Parapprevotella xvlaniphila (82A 6) strain is shown as SEQ ID NO. 22.

The amino acid sequence of 00509 protein of Paraprevotella xylaniphila (YIT 11841) strain (catalog number: JCM 14860) is shown in SEQ ID NO. 23, and the cDNA base sequence of the gene encoding 00509 protein of Paraprevotella xylaniphila (YIT 11841) strain is shown in SEQ ID NO. 24.

The amino acid sequence of 00509 protein of Prevolella ra (109) strain (catalog number: DSM 105141) is shown in SEQ ID NO. 25, and the cDNA base sequence of the gene encoding 00509 protein of Prevolella ra (109) strain is shown in SEQ ID NO. 26.

The amino acid sequence of 00509 protein of Prevotella rodentium (PJ 1A) strain (catalog number: DSM 105243) is shown in SEQ ID NO. 27, and the cDNA base sequence of the gene encoding 00509 protein of Prevotella rodentium (PJ 1A) strain is shown in SEQ ID NO. 28.

As discussed in the examples below, the composition for degrading trypsin or TMPRSS2 according to the present embodiment may include bacteria having 00502 protein homolog or bacteria having 00509 protein homolog as an active ingredient.

00502 protein homologues include proteins having 30%, 40%, 50%, 60%, 70%, 80%, 90% or more than 95% sequence identity with the amino acid sequence shown in SEQ ID NO. 5 and having trypsin binding capacity. Bacteria with 00502 protein homologues should have trypsin or TMPRSS2 binding capacity and further have trypsin or TMPRSS2 degrading activity.

00509 protein homologues include proteins having 30%, 40%, 50%, 60%, 70%, 80%, 90% or more than 95% sequence identity with the amino acid sequence shown in SEQ ID NO. 18 and having trypsin binding capacity. Bacteria with 00509 protein homolog should have trypsin or TMPRSS2 binding capacity and further have trypsin or TMPRSS2 degrading activity.

The "containing as an active ingredient" of the composition for degrading trypsin or TMPRSS2 according to the present embodiment means containing a sufficient amount of a bacterium having 00502 protein or 00502 protein homolog, or containing a sufficient amount of a bacterium having 00509 protein or 00509 protein homolog, to degrade trypsin or TMPRSS2. Alternatively, "containing as an active ingredient" means containing a bacterium having 00502 protein or a 00502 protein homolog, or containing a bacterium having 00509 protein or a 00509 protein homolog as a main active ingredient.

The composition for degrading trypsin or TMPRSS2 according to the present embodiment, the bacterium having 00502 protein or 00502 protein homolog refers to a bacterium expressing 00502 protein or 00502 protein homolog.

The bacterium expressing 00502 protein may be a bacterium having hmpref9441_00858 gene (SEQ ID NO: 1) encoding 00502 protein. The bacterium having the 00502 protein homolog may also be a bacterium having a gene which has 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% or more sequence identity with the base sequence shown in SEQ ID NO. 1 and has cDNA encoding a protein having trypsin binding ability. 00502 protein may be derived from an endogenous gene or an exogenous gene of the bacterium.

Likewise, a bacterium having 00509 protein or a 00509 protein homolog refers to a bacterium expressing 00509 protein or a 00509 protein homolog.

The bacterium expressing 00509 protein may be a bacterium having hmpref9441_00850 gene (SEQ ID NO: 2) encoding 00509 protein. The bacterium having the 00509 protein homolog may also be a bacterium having a gene which has 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% or more sequence identity with the base sequence shown in SEQ ID NO. 1 and has cDNA encoding a protein having trypsin binding ability. 00509 protein may be derived from an endogenous gene or an exogenous gene of the bacterium.

The composition for degrading trypsin or TMPRSS2 according to the present embodiment may contain only bacteria having 00502 protein or 00502 protein homolog, only bacteria having 00509 protein or 00509 protein homolog, or both bacteria having 00502 protein or 00502 protein homolog and bacteria having 00509 protein or 00509 protein homolog, as long as it has an activity of degrading trypsin or TMPRSS 2. Alternatively, the composition for degrading trypsin or TMPRSS2 according to the present embodiment may comprise a bacterium having 00502 protein or 00502 protein homolog and a bacterium having 00509 protein or 00509 protein homolog. Such bacteria may be prepared by genetic engineering or other means.

In the composition for degrading trypsin or TMPRSS2 according to the present embodiment, bacteria having 00502 protein or 00502 protein homolog, or bacteria having 00509 protein or 00509 protein homolog should have IX type secretion system (IX type secretion mechanism, T9 SS).

For example, T9SS contains PorV protein, porU protein, porN protein, porM protein, porL protein, porK protein, or PorP protein.

As discussed in the examples below, the inventors have demonstrated that 00502 protein or 00502 protein homolog, or 00509 protein homolog, is transported across the outer membrane to and bound to the bacterial surface by T9 SS.

For example, in the Paraprevotella clara strain described below, the NCBI of PorV protein is WP_008622445.1, the NCBI of PorU protein is WP_008622443.1, the NCBI of PorN protein is WP_008623210.1, the NCBI of PorM protein is WP_008623211.1, the NCBI of PorL protein is WP_008623213.1, the NCBI of PorK protein is WP_008623215.1, and the NCBI of PorP protein is WP_008623217.1. In bacteria other than the Paraprevotella clara strain, homologs of these proteins in the bacteria constitute T9SS.

The bacterium having 00502 protein or 00502 protein homolog, or the bacterium having 00509 protein or 00509 protein homolog in the composition for degrading trypsin or TMPRSS2 according to the present embodiment may be a bacterium belonging to the genus paraprevatella, prevotella or bacterioides.

As discussed in the examples below, the inventors have demonstrated that bacteria belonging to the genera Paraprefotella, prevoltellella, prevoltellamabilia or Bactoidetes can degrade trypsin or TMPRSS2. Bacteria of the genus Parapreviella include Paraprevotella clara, paraprevotella xylaniphila, parapreviella sp.MSP 0303, parapreviella sp.MSP 0335, and the like. Bacteria of the genus Prevolella include Prevolella rara, prevotella rodentium and Prevolella muris, etc.

The bacterium of Paraprevatella has a 16S rRNA gene composed of a base sequence shown as SEQ ID NO. 3 or SEQ ID NO. 4, or a 16S rRNA gene composed of a base sequence having 97% or more sequence identity with the base sequence shown as SEQ ID NO. 3 or SEQ ID NO. 4. It should be noted that these bacteria are considered to belong to the same species if the sequence identity of the 16S rRNA gene is 97% or more.

The base sequence shown in SEQ ID NO. 3 is the base sequence of the 16S rRNA gene of the Paraprevotella clara (YIT 11840) strain (catalog number: JCM 14859) described below. The base sequence shown in SEQ ID NO. 29 is the base sequence of the 16S rRNA gene of the Paraprevotella clara (1C 4) strain. The base sequence shown in SEQ ID NO. 4 is the base sequence of the 16S rRNA gene of the Paraprevotella xylaniphila (82A 6) strain described below. The base sequence shown in SEQ ID NO. 30 is the base sequence of the 16S rRNA gene of strain Paraprevotella xylaniphila (YIT 11841) (catalog number: JCM 14860).

The bacterium having 00502 protein or 00502 protein homolog, or bacterium having 00509 protein or 00509 protein homolog in the composition for degrading trypsin or TMPRSS2 according to the present embodiment may be at least one bacterium of paraprevatella sp.msp 0303, paraprevatella sp.msp 0335, prevotellamassilia timonensis, bactericides sp.msp 0288, bactericides sp.msp 0410, bactericides sp.msp 0435, and Porphyromonas gingivalis.

Prevolella ra includes the Prevolella ra (MSP 0081) strain and the Prevolella ra (109) strain. Prevotella rodentium includes the Prevotella rodentium (PJ 1A) strain (catalog number: DSM 105243). Prevolella muris included the Prevolella Muris (PMUR) strain (catalog number: DSM 103722). Prevotellamassilia timonensis includes Prevotellamassilia timonensis (MSP 0224) strain. Porphyromonas gingivalis includes the Porphyromonas gingivalis (ATCC 33277) strain.

As discussed in the examples below, the inventors have demonstrated that these bacteria can degrade trypsin or TMPRSS2.

The base sequence shown in SEQ ID NO. 31 is the base sequence of the 16S rRNA gene of the Prevolella ra (109) strain (catalog number: DSM 105141). The base sequence shown in SEQ ID NO. 32 is the base sequence of the 16S rRNA gene of the Prevotella rodentium (PJ 1A) strain (catalog number: DSM 105243). The base sequence shown in SEQ ID NO. 33 is the base sequence of the 16S rRNA gene of the Prevolella Muris (PMUR) strain (catalog number: DSM 103722).

In the composition for degrading trypsin or TMPRSS2 according to the present embodiment, the above bacteria may be living bacteria or dead bacteria as long as they have the degrading activity of trypsin or TMPRSS2.

The composition for degrading trypsin or TMPRSS2 according to the present embodiment may be in an aqueous form or a semi-solid form such as a solution or a suspension, may contain the above bacteria in a powder form or a lyophilized form, etc. In one embodiment, the composition or bacteria are lyophilized. In one embodiment, a subset of bacteria in the composition is lyophilized. Methods of lyophilizing a composition comprising bacteria are well known in the art. For example, reference may be made to the specification of U.S. Pat. No.3261761, the specification of U.S. Pat. No.4205132 and International publication No.2012/098358. These references are incorporated herein by reference.

The bacteria may be lyophilized as a composition or may be separately lyophilized and then combined. The bacteria may be combined with a pharmaceutically acceptable carrier before being combined with other bacteria. A variety of lyophilized bacteria can be combined in lyophilized form. The combined bacterial mixture may then be combined with a pharmaceutically acceptable carrier. In one embodiment, the bacteria are lyophilized solids. In one embodiment, the composition is a lyophilized solid.

In one embodiment, the present invention provides a composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following proteins: 00502 protein or a protein having 30% or more sequence identity with the amino acid sequence of said 00502 protein and having trypsin binding capacity, or 00509 protein or a protein having 30% or more sequence identity with the amino acid sequence of said 00509 protein and having trypsin binding capacity.

As discussed in the examples below, the inventors have demonstrated that 00502 protein or 00509 protein can degrade trypsin or TMPRSS2. Here, the 00502 protein or 00509 protein is preferably immobilized on a solid-phase surface.

The solid phase may be any material including particles made of resin, glass or metal, the surface of a container (e.g., a plate or tube, etc.), a film made of resin, etc. The particles may be magnetic particles.

The solid phase may be a pharmaceutically acceptable solid phase. Pharmaceutically acceptable solid phases include liposomes, polymer nanoparticles (e.g., protein nanoparticles, lipid nanoparticles, etc.), nanoemulsions (e.g., iron nanoparticles, lipid microspheres, etc.), micelles, vaccine adjuvants, nanocrystals, etc. The pharmaceutically acceptable solid phase is preferably approved by the national institutes of medicine and medical devices (Pharmaceutical and Medical Devices Agency, PMDA), the united states food and drug administration (Food and Drug Administration, FDA), the european medicines administration (European Medicines Agency, EMA).

The method of immobilizing 00502 or 00509 protein on a solid phase is not limited, and includes methods of binding using a chemical linker, binding using avidin-biotin, and physical adsorption.

As discussed in the examples below, the compositions for degrading trypsin or TMPRSS2 may comprise 00502 protein homolog or 00509 protein homolog as an active ingredient.

00502 protein homologues may have 30%, 40%, 50%, 60%, 70%, 80%, 90% or more than 95% sequence identity with the amino acid sequence shown in SEQ ID NO. 5, and have trypsin binding capacity, as described above. Preferably, the 00502 protein homologue has trypsin or TMPRSS2 binding ability.

00509 protein homologue may have 30%, 40%, 50%, 60%, 70%, 80%, 90% or more than 95% sequence identity with the amino acid sequence shown in SEQ ID NO. 18 and have trypsin binding capacity as described above. Preferably, the 00509 protein homologue has trypsin or TMPRSS2 binding ability.

In the composition for degrading trypsin or TMPRSS2 according to the present embodiment, "containing as an active ingredient" means containing a sufficient amount of 00502 protein or 00502 protein homolog, or containing a sufficient amount of 00509 protein or 00509 protein homolog, to degrade trypsin or TMPRSS2. Alternatively, "containing as an active ingredient" means containing 00502 protein or 00502 protein homolog, or containing 00509 protein or 00509 protein homolog as a main active ingredient.

00502 protein or 00502 protein homologue, or 00509 protein homologue, may be added with a peptide tag for protein purification, protein detection or binding to a solid phase. Peptide tags include, but are not limited to, histidine tags, FLAG tags, MYC tags, and the like.

The composition for degrading trypsin or TMPRSS2 according to the present embodiment may contain only 00502 protein or 00502 protein homolog, only 00509 protein or 00509 protein homolog, or both 00502 protein or 00502 protein homolog and 00509 protein or 00509 protein homolog, as long as it has an activity of degrading trypsin or TMPRSS 2.

When the composition for degrading trypsin or TMPRSS2 according to the present embodiment contains both 00502 protein or its homolog and 00509 protein or its homolog, 00502 protein or its homolog and 00509 protein or its homolog may be combined.

Here, 00502 protein or its homolog and 00509 protein or its homolog may be combined linearly or circularly. The 00502 protein or its homolog and 00509 protein or its homolog may be directly bound or bound via a linker. The linker is not limited to, for example, a peptide comprising an amino acid sequence in which GGGS (SEQ ID NO: 34) is repeated 1 to 4 times, or the like.

00502 protein or a homolog thereof and/or 00509 protein or a homolog thereof may be present in the form of a protein fused to the constant region of an antibody. The antibody constant region may be a constant region derived from a human antibody or a constant region derived from a human IgG type antibody.

FIG. 53 is a schematic view showing an example of a fusion protein of 00502 protein or its homolog and/or 00509 protein or its homolog with an antibody constant region. In FIG. 53, "protein" represents 00502 protein or its homolog, or 00509 protein or its homolog, "CH2" represents the CH2 domain of the antibody constant region, and "CH3" represents the CH3 domain of the antibody constant region.

As shown in FIG. 53, 00502 protein or its homolog and/or 00509 protein or its homolog and antibody constant region may be linked by a linker. The linker is not limited to, for example, a peptide comprising an amino acid sequence in which GGGS (SEQ ID NO: 34) is repeated 1 to 4 times, or the like.

[ pharmaceutical composition for treating diseases caused by trypsin or TMPRSS2 ]

In one embodiment, the above-described compositions for degrading trypsin or TMPRSS2 are useful for treating diseases caused by trypsin or TMPRSS 2. In other words, in one embodiment, the present invention provides a pharmaceutical composition for treating a disease caused by trypsin or TMPRSS 2. The pharmaceutical composition of the present embodiment contains the following components as active ingredients: a bacterium having 00502 protein or a homolog of 00502 protein, a bacterium having 00509 protein or a homolog of 00509 protein, a bacterium having 00502 protein or a homolog of 00502 protein, a homolog of 00502 protein or a homolog of 00502 protein, or a homolog of 00509 protein.

In the pharmaceutical composition of this embodiment, 00502 protein homolog, 00509 protein homolog, and bacteria having these proteins are the same as those described above.

In the pharmaceutical combination according to the present embodiment, diseases caused by trypsin or TMPRSS2 include inflammatory bowel disease (ulcerative colitis, crohn's disease), irritable bowel disease, acute pancreatitis and chronic pancreatitis.

In addition, in the pharmaceutical composition of the present embodiment, diseases caused by trypsin or TMPRSS2 include infection. Infections include viral or bacterial infections. Such inflammatory bowel disease, irritable bowel disease or infection includes diseases involving TMPRSS2 or IgA. Infections involving TMPRSS2 include coronavirus infections and the like, and infections involving IgA include Salmonella infections and the like.

The pharmaceutical composition according to this embodiment may be formulated with a pharmaceutically acceptable carrier. As pharmaceutically acceptable carriers, those commonly used in formulating pharmaceutical compositions may be used without limitation. More specifically, for example, binders (such as gelatin, corn starch, tragacanth, acacia, and the like), excipients (such as starch, crystalline cellulose and the like), and bulking agents (such as alginic acid and the like).

The pharmaceutical composition according to the present embodiment may contain additives. Additives include lubricants (e.g., calcium stearate, magnesium stearate, and the like), sweeteners (e.g., sucrose, lactose, saccharin, maltitol, and the like), flavoring agents (e.g., peppermint, red monomer oil, and the like), stabilizers (e.g., benzyl alcohol, phenol, and the like), and buffering agents (e.g., phosphate, sodium acetate, and the like).

The pharmaceutical composition according to the present embodiment may be formulated by mixing appropriately combining the above-mentioned carriers and additives in a unit dosage form conforming to the general needs.

The administration method of the pharmaceutical composition according to the present embodiment is not particularly limited, and may be determined according to the symptoms, weight, age, sex, and the like of the patient. Such as tablets, dispersions, capsules, liquid formulations, intravenous formulations, suppositories, and the like. Tablets, dispersions, capsules and liquid formulations are orally administered. Intravenous formulations and suppositories are enterally administered. The pharmaceutical composition is preferably in a form capable of delivering the active ingredient (a bacterium having 00502 protein or a homolog of 00502 protein, a bacterium having 00509 protein or a homolog of 00509 protein, a 00502 protein or a homolog of 00502 protein, or a homolog of 00509 protein) to the intestinal tract.

The dosage of the pharmaceutical composition depends on the symptoms, weight, age, sex, and other factors of the patient, and cannot be approximated. If the active ingredient is a living bacterium, it is considered that the active ingredient is administered 1 or more times in a unit dosage form of 0.1 to 100mg/kg body weight per administration. If the active ingredient is dead bacteria or protein, it is contemplated that the active ingredient may be administered in unit dosage form of 0.1-100mg/kg body weight 1 or more times per day.

The pharmaceutical composition according to the present embodiment may be a quasi drug. Quasi-drugs are products with specific effects and effects, and have light effects on human bodies. Quasi-drugs of this embodiment include, but are not limited to, beverages, gastric drugs, enteral formulations, and the like.

[ diagnostic agent for diseases caused by trypsin or TMPRSS2 ]

In one embodiment, the invention provides a diagnostic agent for a disease caused by trypsin or TMPRSS2 comprising a specific binding substance for detecting: 00502 protein or a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding ability, or 00509 protein or a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding ability.

In other words, the diagnostic agent of the present embodiment contains a specific binding substance for detecting 00502 protein, 00502 protein homolog, 00509 protein, and 00509 protein homolog at the protein level.

Specific binding substances include antibodies, antibody fragments, and aptamers, among others. Antibody fragments include Fab, F (ab ') 2, fab' and single chain antibodies (scFv), and the like. The antibody may be a monoclonal or polyclonal antibody. In addition, commercially available antibodies can be used.

In the diagnostic agent of the present embodiment, 00502 protein homolog, 00509 protein homolog, and bacteria having these proteins are the same as those described above. Diseases caused by trypsin or TMPRSS2 are also the same as previously described.

The diagnostic agents of this embodiment can be used to detect the presence of 00502 protein, 00502 protein homolog, 00509 protein homolog, or bacteria having these in a biological sample from a subject.

The detection principle of the diagnostic agent of the present embodiment is not limited, and for example, methods such as enzyme-linked immunosorbent assay (ELISA), lateral flow immunoassay, western Blot, and flow cytometry (FACS) can be used.

Biological samples derived from subjects include fecal samples and the like. If the presence of 00502 protein, 00502 protein homolog, 00509 protein homolog, or bacteria having these proteins is detected in a biological sample derived from a subject using the diagnostic agent of the present embodiment, it can be determined that the subject is not suffering from a disease caused by trypsin or TMPRSS 2.

If the presence of 00502 protein, 00502 protein homolog, 00509 protein homolog, or bacteria having these proteins is not detected in the biological sample from which the subject originated, it can be judged that the subject is likely to suffer from a disease caused by trypsin or TMPRSS 2. In this case, the symptoms of the disease caused by trypsin or TMPRSS2 can be treated or alleviated by administering the above pharmaceutical composition or quasi-drug to the subject.

In one embodiment, the diagnostic agent of this embodiment may be used to detect 00502 protein, 00502 protein homolog, 00509 protein, and 00509 protein homolog at the gene level.

In other words, in one embodiment, the present invention provides a diagnostic agent for a disease caused by trypsin or TMPRSS2 comprising primers or probes for detecting: the HMPREF41_ 00858 gene encoding 00502 protein or the gene having 30% or more sequence identity with the base sequence of the HMPREF9441_00858 gene and encoding a protein having trypsin binding ability, the HMPREF41_ 00850 gene encoding 00509 protein or the gene having 30% or more sequence identity with the base sequence of the HMPREF9441_00850 gene and encoding a protein having trypsin binding ability. In other words, the diagnostic agent of the present embodiment includes primers or probes for detecting the following genes: a gene comprising a base sequence shown in SEQ ID NO. 1 or a gene having 30% or more of sequence identity to a base sequence shown in SEQ ID NO. 1 and encoding a protein having trypsin binding ability, or a gene comprising a base sequence shown in SEQ ID NO. 2 or a gene having 30% or more of sequence identity to a base sequence shown in SEQ ID NO. 2 and encoding a protein having trypsin binding ability.

The HMPREF41_ 00858 and HMPREF9441_00850 genes are the same as those described above.

The gene which has 30% or more sequence identity with the hmpref9441_00858 gene sequence and which encodes a protein having trypsin binding capacity is a gene encoding a homolog of the 00502 protein described above.

The gene which has 30% or more sequence identity with the hmpref9441_00850 gene sequence and which encodes a protein having trypsin binding capacity is a gene encoding a homolog of the 00509 protein described above.

The diagnostic agent of the present embodiment can be used to detect the presence or absence of a bacterium having 00502 protein, a bacterium having 00502 protein homolog, a bacterium having 00509 protein, and a bacterium having 00509 protein homolog in a biological sample derived from a subject.

The detection principle of the diagnostic agent of the present embodiment is not limited, and for example, methods such as PCR, RNA sequencing (RNA-Seq), genomic analysis, and DNA microarray analysis can be used.

The biological sample derived from the subject is the same as the above sample, and for example, a fecal sample or the like can be used. If the presence of a bacterium having 00502 protein, a bacterium having 00502 protein homolog, a bacterium having 00509 protein, or a bacterium having 00509 protein homolog is detected in a biological sample derived from a subject using the diagnostic agent of the present embodiment, it can be judged that the subject has not suffered from a disease caused by trypsin or TMPRSS 2.

If the presence of a bacterium having 00502 protein, a bacterium having a homolog of 00502 protein, a bacterium having 00509 protein, or a bacterium having a homolog of 00509 protein is not detected in a biological sample derived from the subject, it can be judged that the subject is likely to suffer from a disease caused by trypsin or TMPRSS 2.

In this case, the disease symptoms caused by trypsin or TMPRSS2 may be treated or alleviated by administering the aforementioned pharmaceutical composition or quasi-drug to the subject.

Other embodiments

In one embodiment, the invention provides a method of treating a disease caused by trypsin or TMPRSS2 comprising administering to a patient in need of treatment an effective amount of bacteria: a bacterium having 00502 protein, a bacterium having a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding ability, a bacterium having 00509 protein, or a bacterium having a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding ability.

In one embodiment, the invention provides a method of treating a disease caused by trypsin or TMPRSS2, the method comprising administering to a patient in need of treatment an effective amount of bacteria: a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 1, a bacterium having a gene which has 30% or more of sequence identity to a base sequence shown in SEQ ID NO. 1 and encodes a protein having trypsin binding ability, a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 2, or a bacterium having a gene which has 30% or more of sequence identity to a base sequence shown in SEQ ID NO. 2 and encodes a protein having trypsin binding ability.

In one embodiment, the invention provides a method of treating a disease caused by trypsin or TMPRSS2, the method comprising administering to a patient in need of treatment an effective amount of a protein of: 00502 protein, a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding capacity, 00509 protein, or a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding capacity.

In one embodiment, the invention provides a method of diagnosing and treating a disease caused by trypsin or TMPRSS2, the method comprising detecting the presence or absence of 00502 protein, a homolog of 00509 protein, or a homolog of 00509 protein in a biological sample from a subject; when the presence of the above protein or homolog is not detected, then the subject is indicated to have a disease caused by trypsin or TMPRSS 2; while the presence of the protein or homolog is not detected, administering to the subject an effective amount of: a bacterium having 00502 protein, a bacterium having a protein which has 30% or more of sequence identity to the amino acid sequence of 00502 protein and has trypsin binding ability, a bacterium having 00509 protein, a bacterium having a protein which has 30% or more of sequence identity to the amino acid sequence of 00509 protein and has trypsin binding ability, 00502 protein, a protein which has 30% or more of sequence identity to the amino acid sequence of 00502 protein and has trypsin binding ability, 00509 protein, or a protein which has 30% or more of sequence identity to the amino acid sequence of 00509 protein and has trypsin binding ability.

In one embodiment, the present invention provides a method for diagnosing and treating a disease caused by trypsin or TMPRSS2, the method comprising detecting in a biological sample derived from a subject the presence or absence of a bacterium having 00502 protein, a bacterium having 00502 protein homolog, a bacterium having a gene comprising a base sequence as shown in SEQ ID NO:1, a bacterium having a gene having 30% or more sequence identity to a base sequence as shown in SEQ ID NO:1 and encoding a protein having trypsin binding capacity, a bacterium having 00509 protein homolog, a bacterium having a gene comprising a base sequence as shown in SEQ ID NO:2, or a bacterium having a gene having 30% or more sequence identity to a base sequence as shown in SEQ ID NO:2 and encoding a protein having trypsin binding capacity; when the presence of bacteria having the above proteins or homologs or genes is not detected, then the subject is indicated to have a disease caused by trypsin or TMPRSS 2; while the presence of bacteria having the above proteins or homologs or genes is not detected, an effective amount of the following is administered to the subject: a bacterium having 00502 protein, a bacterium having a protein having 30% or more of sequence identity to the amino acid sequence of 00502 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 1, a bacterium having a gene having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 1 and encoding a protein having trypsin binding ability, a bacterium having 00509 protein, a bacterium having a protein having 30% or more of sequence identity to the amino acid sequence of 00509 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 2, a bacterium having a gene having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 2 and encoding a protein having trypsin binding ability, 00502 protein, a protein having 30% or more of sequence identity to the amino acid sequence of 00502 protein and having trypsin binding ability, 00509 protein, or a protein having 30% or more of sequence identity to the amino acid sequence of 00509 protein and having trypsin binding ability.

In one embodiment, the present invention provides a composition for treating diseases caused by trypsin or TMPRSS2, comprising as active ingredients the following bacteria: a bacterium having 00502 protein, a bacterium having a protein having 30% or more of sequence identity to the amino acid sequence of 00502 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 1, a bacterium having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 1 and encoding a protein having trypsin binding ability, a bacterium having 00509 protein, a bacterium having 30% or more of sequence identity to the amino acid sequence of 00509 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 2, or a bacterium having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 2 and encoding a protein having trypsin binding ability.

In one embodiment, the present invention provides a pharmaceutical composition for treating diseases caused by trypsin or TMPRSS2, comprising as active ingredients the following proteins: 00502 protein, a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding capacity, 00509 protein, or a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding capacity.

In one embodiment, the invention provides a method for preparing a pharmaceutical composition for treating a disease caused by trypsin or TMPRSS2, the method comprising using: a bacterium having 00502 protein, a bacterium having a protein having 30% or more of sequence identity to the amino acid sequence of 00502 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 1, a bacterium having a gene having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 1 and encoding a protein having trypsin binding ability, a bacterium having 00509 protein, a bacterium having a protein having 30% or more of sequence identity to the amino acid sequence of 00509 protein and having trypsin binding ability, a bacterium having a gene comprising the base sequence shown in SEQ ID NO. 2, or a bacterium having 30% or more of sequence identity to the base sequence shown in SEQ ID NO. 2 and encoding a protein having trypsin binding ability.

In one embodiment, the invention provides a method for preparing a pharmaceutical composition for treating a disease caused by trypsin or TMPRSS2, the method comprising using: 00502 protein, a protein having 30% or more sequence identity with the amino acid sequence of 00502 protein and having trypsin binding capacity, 00509 protein, or a protein having 30% or more sequence identity with the amino acid sequence of 00509 protein and having trypsin binding capacity.

[ example ]

The present invention will be described in more detail by way of examples, but the present invention is not limited to the following examples.

[ materials and methods ]

(mice)

C57BL/6 mice were kept under Specific Pathogen Free (SPF) or sterile (GF) conditions, purchased from Sankyo Laboratories Japan, SLC Japan, charles River Japan or CLEA Japan. GF mice and sydney (gnotobiotic) mice are bred and maintained in sterile facilities at the center of life sciences and medical biosciences of the japan physical and chemical institute. All animal experiments were approved by the animal experiment committee of the institute of physical chemistry, the lazy research institute.

(bacterial Strain)

Paraprevotella clara (JCM 14859), paraprevotella xylaniphila (JCM 14860), paraprevotella copri (JCM 13464), paraprevotella denticola (JCM 13449), paraprevotella stercorea (JCM 13469) and Paraprevotella oulorum (JCM 14966) are obtained from Japan Collection of Microorganisms (JCM). Paraprevotella clara (P237E 3B) and (P322B 5) are provided by Vedanta Biosciences. Paraprevotella xylaniphila (82A 6) was isolated by the inventors.

(proteomic analysis of cecal content)

Proteins in cecal content were extracted by pipetting and mixing with Tris-buffered saline (TBST) containing protease inhibitors. The supernatant was transferred to a fresh tube, 25% trichloroacetic acid (final concentration 12.5% v/v) was added and incubated for 1 hour at 4 ℃. The supernatant was subsequently removed by centrifugation at 15,000Xg for 15 minutes at 4℃after which the precipitate was washed twice with acetone, uncovered and dried. The dried sample was then redissolved in 0.5% sodium laurate and 100mM Tris-HCl (pH 8.5) using a water bath sonicator (biosoprotor UCD-200,SonicBio Corp). The protein concentration of the redissolved sample was determined by biuret acid (BCA) and adjusted to 1 μg/μl. Pretreatment of proteomic analysis was performed according to the reported method (Kawashima y., et al Optimisation of Data-Independent Acquisition Mass Spectrometry for Deep and Highly Sensitive Proteomic Analysis, int J Mol sci.20 (23), 5932,2019).

The peptide was injected directly into a 75 μm by 15cm PicoFrit emitter (New Objective), which was internally filled with 2.7 μm CORE-shell C18 particles (CAPCELL CORE MP 2.7 μm,materialOsaka thred limited) and was isolated using a Eksigent ekspert nanoLC HPLC system (Sciex) with a gradient flow rate of 300nL/min for 180 minutes. />

Peptide fragments eluted from the column were analyzed by Shotgun MS and SWATH (Sequential Window Acquisition of All Theoretical Mass Spectra) -MS using a TripleTOF 5600+ mass spectrometer (Sciex). In experiments using Shotgun MS, MS1 spectra were collected at 250 MS intervals in the range of 400-1000 m/z. The first 25 precursor ions with charge states from 2+ to 5+ and counts greater than 150 times/second were selected, fragmented using rolling collision energy, and MS2 spectra were collected at 100 MS intervals in the range of 100-1500 m/z. The dynamic offset time is set to 24 seconds.

In the SWATH-MS experiment, the mass spectrometer was run in a continuous data independent acquisition mode, with a 12m/z increase in the precursor separation window each time. The separation width was 13m/z (window overlap was 1 m/z), and 50 window sets covering the precursor mass range of 400-1000m/z were constructed. SWATH MS2 spectra were run in the range of 100-1500m/z, with each MS2 experiment being run at 60MS intervals in the range of 100-1500 m/z. In each MS2 experiment, precursor ions were fragmented using rolling collision energy.

All Shotgun MS files were matched to the mouse UniProt reference proteome (UniProt id UP000000589, reviewed, cannical) using the proteo software v.4.5 and the Paragon algorithm (Sciex) for protein identification.

The reliability threshold for the protein was set to an unused fraction of 1.3 for ProteinPilot and a 95% confidence level of at least one peptide was required. In this study, the overall false discovery rate for both peptides and proteins was less than 1%. The identified proteins were quantified from SWATH-MS data using PeakView v.2.2 (Sciex).

(P.clara culture supernatant proteomic analysis).

25% trichloroacetic acid (final concentration 12.5% v/v) was added to the P.clara culture supernatant and incubated at 4℃for 1 hour. The supernatant was subsequently removed by centrifugation at 15,000Xg for 15 minutes at 4 ℃. The precipitate was washed twice with acetone, uncovered and dried. The dried sample was then redissolved in 0.5% sodium laurate and 100mM Tris-HCl (pH 8.5) using a water bath sonicator (Biorupter UCD-200), and the protein concentration of the redissolved sample was measured by BCA and adjusted to 1. Mu.g/. Mu.L. Pretreatment of Shotgun proteomic analysis was performed as described above.

The peptide was injected directly into a 75 μm by 20cm PicoFrit emitter filled with 2.7 μm core-shell C18 particles. The separation was then carried out at 50℃using an UltiMate 3000RSLCnano LC system (Thermo Fisher Scientific) with a gradient flow rate of 100nL/min for 80 minutes. The peptide fragments eluted from the column were analyzed by overlap window DIA-MS using Q exact HF-X (Thermo Fisher Scientific). MS1 spectra were collected in the 495-785m/z range with a resolution of 30,000 and the target value for automatic gain control set to 3 Xe ⁶ The maximum sample injection time was set to 55.

MS2 spectra were collected in a range of 200m/z with a resolution of 30,000 and the target value for automatic gain control was set to 3 Xe ⁶ The maximum sample injection time was set to "auto", the stepwise normalized collision energy was set to 22, 26 and 30%, and the isolation window width of MS2 was set to 4m/z. An overlapping window pattern of 500-780m/z is used. Window configuration was optimized using Skyline software.

MS files were matched to the P.clara spectral library using Scaffold DIA (Proteome Software). The spectral library was generated using Prosit software from the protein sequence database of P.clara (Unit Prot id UP000000589, reviewed, cannical).

The clara protein sequence database was self-created by metagenomic analysis. The search parameters of the Scaffold DIA are shown below. Experimental data retrieval enzyme: trypsin, the most wrong cleavage site: precursor mass tolerance range: 8ppm, fragment mass tolerance range: 8ppm, static modification: aminomethylation of cysteines.

The threshold for protein identification was set to give false discovery rates of peptides and proteins below 1%. The quantification of peptides was calculated by the EncyclopeDIA algorithm of Scaffold DIA. The 4 highest mass fragment ions were selected for each peptide fragment and quantified. The quantitative value of the protein is estimated from the sum of the quantitative values of the peptides.

(Peptidomimetic analysis)

Acetonitrile (ACN) containing 0.1% trifluoroacetic acid (TFA) was added to the cecal content and dried in a centrifugal evaporator. Acetone was added to the dried sample, and the low molecular weight compounds in the lipid were dissolved by a water bath sonicator, and then centrifuged at 15,000Xg for 15 minutes at 4 ℃.

After removal of the supernatant, 70% ACN-HCl was added to the precipitate, the peptide was redissolved in a water bath sonicator and centrifuged at 15,000Xg for 15 min at 4 ℃. The supernatant was then transferred to a new tube and dried in a centrifugal evaporator. The dried samples were then redissolved in 100mM Tris-HCl containing protease inhibitors and treated with 10mM dithiothreitol at 50℃for 30 minutes. Alkylation treatment with 30mM iodoacetamide was then carried out for 30 minutes at room temperature in the dark and acidified with 0.5% trifluoroacetic acid (final concentration). The acidified samples were desalted with Monospin C18 (GL Sciences).

The peptide was injected directly into a 75 μm x 25cm PicoFrit emitter (New Objective), which was filled with C18 CORE-shell particles (CAPCELL CORE MP 2.7 μm,osaka threda). The separation was then carried out at 50℃using an UltraMate 3000RSLCnano LC system (Thermo Fisher Scientific) with a gradient flow rate of 100nL/min for 90 minutes.

The peptide fragment eluted from the column was analyzed by DDA-MS using Q exact HF-X (Thermo Fisher Scientific). MS1 spectra were collected in the 350-1500m/z range with a resolution of 120000 to achieve an Automatic Gain Control (AGC) target value of 3×10 ⁶ 。

Using a data dependent mode to normalize collision induced dissociation at a collision energy of 26% for charge states from 2+ to 5+ and intensities exceeding 4.4X10 ³ The 30 strongest ions of (2) were crushed and tandem mass spectrum was obtained by Orbitrap mass spectrometer at 200m/z with a mass resolution of 30000. The AGC target value is set to 1×10 ⁵ 。

MS files were retrieved in the mouse UniProt reference proteome (UniProt id UP000000589, review, canonical) PEAKS Studio.

The search conditions are as follows: the precursor mass tolerance range was 8ppm, the fragment ion mass tolerance range was 0.01Da, the enzyme was present, the immobilized modification was aminomethylated, and the variable modification was oxidized (M). Peptides were filtered and identified to ensure that the false discovery rate of peptides was below 1%.

(Western Blot)

The mouse fecal samples were suspended in Phosphate Buffered Saline (PBS) for 50-fold dilution, supplemented with protease inhibitor cocktail (roche). The suspension samples were then centrifuged at 15,000Xg for 10 minutes at 4℃and the supernatant was subjected to Western Blot. Mouse pancreatic tissue was snap frozen in liquid nitrogen and protein was extracted with TRIzol reagent (Thermo Fisher Scientific) at a final protein concentration of 4. Mu.g/. Mu.L. Novex (R) NuPAGE (R) SDS-PAGE Gel System (Thermo Fisher Scientific) and iBlot were used ^TM 2Dry Blotting System (Thermo Fisher Scientific) was subjected to SDS-PAGE and blotting, and used according to the manufacturer's instructions.

In some experiments, SDS-PAGE and transfer of PVDF membrane (0.2. Mu. mTransfer Membranes Immobilon-PSQ, merck Millipore) was performed according to the manufacturer's instructions (XV PANTERA SYSTEM (DRC)).

By iBind ^TM Western Systems (Thermo Fisher Scientific) staining.

The antibodies used are shown below. Rabbit anti-mouse PRSS2 antibody (cosmo bio), anti-mouse HSP90 antibody (# 4877,Cell Signaling Technology), rabbit anti-human PRSS2 antibody (LS-B15726, LSBio), rabbit anti-human PRSS1 antibody (LS-331381, LSBio), rabbit anti-mouse TMPRSS2 antibody (LS-C373022, LSBio, sequence directed against protease domain), rabbit anti-6-His antibody (a 190-214A,Bethyl laboratories), goat anti-mouse IgA alpha chain antibody (HRP) (ab 97235, abcam), rat anti-mouse kappa chain antibody (HRP) (ab 99632, abcam), rabbit anti-mouse CELA3B antibody (OACD 03205, aviva sybio), anti-rabbit IgG antibody (HRP) (# 7074,Cell Signalling Technology), rabbit anti-mouse Reg3 antibody (51153-R005, sino Biological).

Chemi-Lumi One (Nacalai Tesque) for chemiluminescent enzyme-linked immunosorbent assay, molecular imaging (R) Chemidoc ^TM XRS+System (Bio-Rad) or iBright ^TM FL1500 is used for imaging.

(RT-qPCR)

RNA from mouse pancreas was extracted with TRIzol reagent (Thermo Fisher Scientific). The extracted RNA was transcribed into cDNA using ReverTra Ace (R) qPCR RT Master Mix with gDNA Remover (Toyobo).

RT-qPCR analysis was performed using Thunderbird SYBR qPCR Mix (Toyobo) and Lightcycler 480 (Roche), and analysis was performed using the ΔΔCt method. GAPDH was used as an endogenous control. The base sequences of the primers used are shown below. GAPDH Forward primer:5'-GTCGTGGAGTCTACTGGTCTTC-3' (SEQ ID NO: 35)

GAPDH Reverse primer：5'-GTCATATTTCTCGTGGTTCACACC-3'(SEQ ID NO:36)

PRSS2 Forward primer：5'-TGTGACCCCCTCAATGCCAGAG-3'(SEQ ID NO:37)

PRSS2 Reverse primer：5'-AGCACTGGGGCATCAACAC-3'(SEQ ID NO:38)。

(immunofluorescent staining)

The rat intestinal tissue (including feces) was collected and fixed overnight with Kornoi solution (60% methanol, 30% chloroform, 10% glacial acetic acid) at 4 ℃. Paraffin embedding was performed using a tissue processor (Leica Microsystems). Paraffin blocks were then cut into thin slices (5.0 μm) using a microtome, paraffin removed and immunostained.

Antibodies used in immunofluorescence are as follows: rabbit anti-PRSS 2 antibody (LSBio), alexa 488-labeled goat anti-rabbit IgG antibody (Thermo Fisher Scientific), 4' -6-diamidino-2-phenylindole (DAPI) (college of chemistry), rhodamine-labeled UEA1 (Ulex Europaeus Agglutinin 1,Vector Laboratories). Immunofluorescence imaging was performed using Leica AF600 and confocal Leica TCS SP5.

(measurement of trypsin Activity in mouse and human faecal samples)

The intestinal contents or stool samples of mice were diluted 500-fold (w/v) in 0.9% NaCl solution. Human feces were diluted 200-fold (w/v) in 0.9% sodium chloride solution. The diluted solution was vortexed in a small shaker at 2000rmp for 20 minutes, homogenized by a pipette, and centrifuged at 10,000Xg for 15 minutes at 4 ℃. The supernatant was then collected and trypsin activity was measured using Trypsin Activity Assay Kit (colorimeter) 100tests (ab 102531) according to the manufacturer's protocol. Absorbance measurements were performed in kinetic mode at 405nm wavelength using PerkinElmer 2030Multilabel Reader.

(colonization of human microbiota in GF mice)

Human stool samples were collected at the institute of chemical and university of Qing according to the institutional review board approved study protocol. Informed consent was obtained for each subject. Human fecal samples (stored in 20% (v/v) glycerol) were transferred to an anaerobic chamber, thawed, filtered through a 100 μm mesh screen, transferred to GF isolator, and orally administered to GF mice at a dose of 200 μl/mouse.

Antibiotic treatment was prepared using the following solutions: a solution of 0.5g/L ampicillin (Nacalai Tesque), 0.5g/L metronidazole (Nacalai Tesque) and 1.0g/L tylosin (Sigma) was prepared using autoclaved tap water. Mice orally administered the fecal content of donor C were continuously given antibiotic solution for 12 days. The antibiotic solution was changed once a week.

(isolation and identification of colony forming species from the fecal content of mice)

The intestinal contents of the mice were mixed with PBS containing glycerol (20%), placed in an anaerobic chamber and stored at-80 ℃. In the anaerobic chamber, the stock was mixed with an equal volume of TS solution (BD) and smeared on different agar plates as shown below: EG. ES, M10, NBGT, VS, TS (BD), BL (Rongsheng chemical), BBE (Jidong pharmaceutical), oxoid CM0619 (Thermo Fisher Scientific), CM 0619-supported SR0107 (Thermo Fisher Scientific), CM 0619-supported SR0108 (Thermo Fisher Scientific), mGAM (Niday Water pharmaceutical), schaedler (BD). After 2 days of incubation, colonies with different appearances were transferred to new EG plates. The colonies were then grown overnight in EGEF broth, mixed with glycerol (final concentration 20% (v/v)) and stored at-80 ℃.

The formulation of EG (Eggerth Gagnon) agar medium is shown below. Peptone No. 3 (10.0 g), yeast extract (5.0 g), na ₂ HPO ₄ (4.0 g), glucose (1.5 g), soluble starch (0.5 g), L-cysteine hydrochloride (0.5 g), L-cysteine (0.2 g), tween 80 (0.5 g), agar (4.8 g), horse meat extract (500 mL), up to 1000mL water+defibrinated horse blood (50 mL); in EGEF medium, defibrinated horse blood (50 mL) was replaced with Fildes solution (40 mL) in addition to agar.

The bacterial DNA genome was then extracted from the isolate using the same protocol as DNA was isolated from feces. 16S rRNA was amplified by PCR using KOD plus Neo Kit (Toyobo) according to the manufacturer' S protocol. Sanger sequencing work was performed by the Eurofins company. The nucleotide sequence was matched to the NCBI database. Primer sequences used for Sanger sequencing are shown below. F27 primer:5'-AGRGTTTGATYMTGGCTCAG-3' (SEQ ID NO: 39) R1492 primer:5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID NO: 40)

(16S rRNA sequencing)

Frozen mouse faecal samples were thawed and 100. Mu.L of the suspension was mixed with 900. Mu.L of TE10 (10 mM Tris-HCl,10mM EDTA) buffer containing RNase A (final concentration 100. Mu.g/mL, thermo Fisher Scientific) and lysozyme (final concentration 3.0mg/mL, sigma). The suspension was incubated at 37℃for 1 hour with gentle agitation. Purified aromatic peptidase (and light) was added to a final concentration of 2,000 units/mL and incubated for a further 30 minutes at 37 ℃. Sodium dodecyl sulfate (1% final concentration) and proteinase K (1 mg/mL final concentration, nacalai) were then added to the suspension and incubated for 1 hour at 55 ℃.

Then phenol is used for: chloroform: the polymerized DNA was extracted with isoamyl alcohol (25:24:1), precipitated with isopropanol, washed with 70% ethanol and resuspended in 100. Mu.L of TE.

PCR analysis was performed using Ex taq (Takara). Primer sequences for amplifying the V1-V2 region of the 16S rRNA gene are as follows. 27Fmod primer:5'-AATGATACGGCGACCACCGAGATCTACACxxxxxxxxACACTCTTTCC CTACACGACGCTCTTCCGATCTAGRGTTTGATYMTGGCTCAG-3' (SEQ ID NO:41, "xxxxxxxx" means the Miseq (Illumina) index sequence).

338R primer:5'-CAAGCAGAAGACGGCATACGAGATxxxxxxxxGTGACTGGAGTTCAGAC GTGTGCTCTTCCGATCTTGCTGCCTCCCGTAGGAGT-3' (SEQ ID NO:42, "xxxxxxxxxxxx" means the Miseq (Illumina) index sequence).

The PCR products were purified on a Agencourt AMPure XP (Beckman Coulter) according to the manufacturer's protocol. The 16S rRNA library was prepared according to the protocol using Kapa library quantification Kit (Kapa Biosystems). 16S rRNA sequencing was performed using the standard protocol of MiSeq Reagent kit ver 3.

(Gnetobiotic experiment)

The isolated strain was cultured in an anaerobic chamber at 37℃for 1-2 days except Phasolactobacterium faecium (3G 4). Phasolactobacterium faecium was cultured on an Oxoid CM0619 agar plate supplemented with 80mM sodium succinate for 2-3 days, colonies were collected and resuspended in EGEF medium. According to OD ₆₀₀ Values bacterial density was adjusted and the mixture of cultured strains was orally administered to GF mice (150 μl/mouse, total bacteria number about 1-2×10) ⁸ CFU)。

To quantify p.clara DNA in feces, mouse fecal DNA was purified and the base sequence of the 16s rRNA gene of the p.clara strain was amplified in the LightCycler 480 system (roche) using THUNDERBIRD SYBR qPCR Mix (eastern ocean). Standard curves were prepared from serial dilutions of genomic DNA of the p.clara (JCM 14859) strain. The primer sequences used are shown below.

5'-CGTAGGAGTTTGGACCGTGT-3'(SEQ ID NO:43)

5'-GCATGGGAGCGACAAATAAA-3'(SEQ ID NO:44)

(Citrobacter rodentium (C. Condensing) infection)

GF mice were orally administered 200. Mu.L of 2-mix (B.uniformis and P.merdae) +P.clara (WT), 2-mix+P.clara (Δ00502) or only 2-mix. After 14 days, mice were infected with overnight cultured c.rodentum (150 μl/mouse) orally and euthanized on day 7.

For histological analysis, mice necks were fixed with 4% paraformaldehyde and paraffin embedded. Paraffin blocks were cut and stained with hematoxylin and eosin.

The extent of colitis was assessed by gastroenterologists according to the following criteria: inflammatory cell infiltration (score, 0-4), mucosal thickening (score, 0-4), goblet cell depletion (score, 0-4), crypt abscess (score, 0-4), and tissue structure destruction (score, 0-4). The final histological score is defined as the sum of the scores of these parameters.

For Colony Forming Unit (CFU) detection, cecal plaque or cecal lumen contents were collected, homogenized in PBS, and gradually diluted homogenates were placed on LB agar plates for colony streaking. CFU were counted after overnight incubation at 37 ℃ under aerobic conditions.

For in vitro evaluation of C.rodenium-specific IgA, the cecal content was resuspended in 5-fold (w/v) LB medium, centrifuged and the supernatant was filtered through a PVDF sterile filter device with a pore size of 0.22 μm and then mixed with an equal volume of in vitro overnight C.rodenium medium. Mix with an equal volume of in vitro overnight c. The mixture was incubated at room temperature with gentle shaking for 1 hour, and the agglutination effect was examined under a confocal microscope (Leica TCS SP 8). Alternatively, after incubation, the mixture was centrifuged, washed once with PBS, and the bacterial particles were lysed in 1% SDS solution (diluted in 50mM Tris-HCl buffer with 5mM EDTA). Lysates were Western Blot stained with goat anti-mouse IgA alpha chain antibody (HRP) (ab 97235) to assess the relative amount of IgA in the fecal content that bound to c.

For vaccination experiments, GF mice were pre-dosed with 200. Mu.L of 2-mix (B.uniformis and P.merdae) +P.clara (WT) or 2-mix+P.clara (Δ00502). After 4 days, peracetic acid inactivated c.rodentum (10) ¹⁰ Individual cells/mouse) 1 time per week for 3 weeks. After 3 weeks of immunization, mice were infected with overnight-cultured c.rodenium (150 μl/mouse) and euthanized on day 14.

The preparation of C.rodenium inactivated by oxyacetic acid is shown below. C.rodentium cultured overnight was collected by centrifugation (16,000Xg, 10 min) and at 10 per ml ¹⁰ The density of individual cells was resuspended in sterile PBS. Peracetic acid (240990, sigma) was added to a final concentration of 0.4% and incubated for 1 hour at room temperature. After three washes with sterile PBS, the final pellet was resuspended in PBS at a final concentration of 10 ¹¹ particles/mL and stored at 4 ℃. 100. Mu.L of the solution was quenched before useLive vaccine was inoculated into 200mL of LB medium and incubated overnight at 37 ℃ to ensure complete inactivation.

(mouse hepatitis Virus (MHV) infection)

MHV-2 is supplied by Makoto Ujiie (Japanese university of veterinary and Life sciences). GF C57BL/6N male mice at 5 weeks of age were obtained from CLEA Japan or Sankyo Lab Services and housed in separate stainless steel isolators. GF mice were orally administered 200. Mu.L of 2-mix (B.uniformis and P.merdae) +P.clara (WT) or 2-mix+P.clara (Δ00502). Two weeks after inoculation, the inoculation was performed with 4.5X10 ⁶ Mice were infected orally with MHV-2 of PFU, and survival was observed daily for 10 days.

To determine viral titers, livers and brains were collected on day 4 or 5 post-infection and homogenized with DNA/RNA Shield (Zymo Research). Viral RNA was extracted using the Quick-RNA visual Kit (Zymo Research) and cDNA was synthesized using the ReverTra Ace (Toyobo) and random primers (Toyobo) according to the manufacturer's instructions.

Quantitative real-time PCR was performed on the LightCycler 480 system (roche) using THUNDERBIRD SYBR qPCR Mix (eastern spun) to amplify the orf1a gene. The primer sequences used are shown below.

5'-AAGAGTGATTGGCGTCCGTAC-3'(SEQ ID NO:45)

5'-ATGGACACGTCACTGGCAGAG-3'(SEQ ID NO:46)

(degradation of trypsin in vitro)

An overnight culture of bacteria was incubated with recombinant mouse trypsin (final concentration 1. Mu.g/mL) for 1 hour or with human trypsin (final concentration 20. Mu.g/mL) for 4 hours. The recombinant trypsin isoforms used were recombinant mouse PRSS2 (50383-M08H, both messenger and organism), recombinant human PRSS1 (LS-G135640), recombinant human PRSS2 (LS-G20167) and recombinant human PRSS3 (His-tag) (NBP 2-52220).

In some experiments, recombinant mouse PRSS2 was treated with one of the following trypsin inhibitors for 30 minutes prior to incubation with p.clara cultures: AEBSF (Sigma, final concentration 2 mM), leupeptin (Sigma, final concentration 100. Mu.M), TLCK (Abcam, final concentration 100. Mu.M).

In some experiments, p.clara was incubated overnight in the presence of: tunicamycin (Sigma, final concentration 10. Mu.g/mL), 2-fluoro-L-fucose (Cayman Chemical, final concentration 250. Mu.M), or corresponding DMSO controls.

In the evaluation of Ca ²⁺ In the experiments affected, P.clara was cultured with or without 1mM Ca prior to incubation with recombinant mouse PRSS2 ²⁺ Low Ca of (2) ²⁺ In mGAM medium. In experiments using p.clara supernatant, overnight cultured p.clara was filtered in a sterile filtration device with a PVDF membrane having a pore size of 0.22 μm.

(confocal microscope)

Using Alexa Fluor ^TM 488 antibody labelling kit (A20181, thermoFisher) recombinant mouse PRSS2 was labelled and pretreated with AEBSF inhibitor (150. Mu.g/mL rmPRSS2 with 20 mMAEBSF).

Alexa Fluor is used ^TM 488-labeled mice PRSS2 were incubated with bacteria at a final concentration of 5 μg/mL in an anaerobic chamber for 20 minutes. The mixture was centrifuged, washed once with PBS and resuspended in PBS. Confocal images were captured using a Leica TCS SP8 confocal microscope.

(crosslinked disuccinimidyl sulfone (DSSO))

DSSO (a 33545) was purchased from Thermo Fisher Scientific. The overnight cultured p.clara (1C 4) strain was incubated with AEBSF treated mouse recombinant PRSS2 for 20 min, washed once with PBS and resuspended in 10mM DSSO. After incubation for 10 min at room temperature, the reaction was stopped by adding Tris-HCl buffer (final concentration 20 mM).

After washing with PBS, the particles were dissolved in 1% SDS solution (diluted in 50mM Tris-HCl buffer containing 5mM EDTA). Only p.clara (1C 4) strain (not hatched with PRSS 2) was similarly treated and served as a negative control. The supernatant was stained with rabbit anti-6-His antibody (A190-214A,Bethyl laboratories) and anti-rabbit IgG antibody (HRP) (# 7074,Cell Signalling Technology) and analyzed by Western Blot.

(Whole cell lysate, supernatant, glycoprotein-containing protein staining)

The P.clara (1C 4) strain was isolated in the presence of tunicamycin (Sigma, final concentration 10. Mu.g/mL), 2-fluoro-L-fucose (Cayman Chemical, final concentration)Degree 250 μm) or the corresponding DMSO control. The cultured bacteria were then precipitated, washed once with PBS, and dissolved in 1% SDS solution (diluted in 50mM Tris-HCl buffer containing 5mM EDTA) and subjected to SDS-PAGE using Novex (R) NuPAGE (R) SDS-PAGE Gel System (Thermo Fisher Scientific). Use of Pro-Q according to manufacturer's protocol ^TM Emerald 300Glycoprotein Gel and Blot Stain Kit (Thermo Fisher Scientific) stained for glycoproteins. Proteins from whole cell lysates were stained with Colloidal Blue Staining Kit (Thermo Fisher Scientific). The proteins in the supernatant were concentrated in Amicon Ultra Centrifugal Filters (10 kD NMWL) and stained with Colloidal Blue Staining Kit (ThermoFisher Scientific).

(mutant preparation)

The preparation of deletion mutants (Delta03049-03053, delta 000502 and Delta 000509) of the P.clara (JCM 14859) strain is shown below. First, a sequence of about 1kb between the coding regions was amplified by PCR and incorporated into suicide vector pLGB30 using HiFi DNA Assembly (NEB) according to the manufacturer's protocol. Then, 1. Mu.L of each reaction solution was transformed into inductive E.coli S17-1. Lambda. Pir.

The transformants were then combined with the p.clara (JCM 14859) strain as follows. Culturing donor strain and recipient strain in LB and EGEF medium respectively to OD ₆₀₀ 0.5 and mixed in a ratio of 1:1. The mixture was dropped onto an EGEF agar plate and aerobically incubated at 37℃for 16 hours. The ligation completed bodies were then selected on EGEF agar plates containing tetracycline (10. Mu.g/mL). The ligation completed body was partially sensitive to rhamnose-induced ss-bfe1 toxin expression and growth was inhibited in the presence of 10mM rhamnose (overnight OD ₆₀₀ About 0.3). Subsequently, for the disappearance of the plasmid from the genome due to the second hybridization, the ligation completed body was cultured at least three times in EGEF medium containing 10mM rhamnose until the ligation completed body was defeated by the revertants (overnight OD ₆₀₀ Reaching-1.0). Then, bacterial culture was performed, and individual colonies were collected, and normal deletion was confirmed by PCR.

A homologous sequence of about 0.5-1kb in the coding region was inserted into suicide vector pLGB30 and transformed into inductive E.coli S17-1 strain, and an insertional mutant was prepared by the same procedure.

Transformants were ligated with the P.clara (JCM 14859) strain using the same protocol, and the ligation completed was selected on EGEF agar plates containing tetracycline (10. Mu.g/mL), confirmed by PCR, and maintained in EGEF medium containing tetracycline (10. Mu.g/mL). The sequences of all the primers used to prepare the mutants are listed in tables 1-5 below.

[ Table 1 ]

/>

[ Table 2 ]

/>

[ Table 3 ]

/>

[ Table 4 ]

[ Table 5 ]

(Transmission electron microscope (TEM))

The Wild Type (WT) or.DELTA.00502 P.clara (JCM 14589) strain was incubated with recombinant mouse PRSS2 (50383-M08H, sino Biological, final concentration 5. Mu.g/mL) for 20 min, washed with PBS, 4% paraformaldehyde-1% glutaraldehyde solution, and fixed at room temperature for 2 hours. After washing with 0.05M PBS, the mixture was dehydrated stepwise with ethanol (50%, 70%, 80%, 90%, 95%, 100%). The dehydrated pellets were infiltrated with LRW resin (100% ethanol and LRW 1:1 for 1 hour, then 100% ethanol and LRW 1:2 for overnight, 5 hours with 100% LRW). After infiltration, the samples were cured in gelatin capsules (53 ℃,24 hours). The polymerized LRW blocks were sectioned with Leica Ultracut UCT to obtain 80nm sections.

For immunostaining, sections were blocked with 0.05M PBS plus 1% BSA, and then stained with rabbit anti-6-His antibody (A190-214A,Bethyl laboratories) for 60 min. After washing with 0.05M PBS, the sections were stained with 12nm colloidal gold labeled goat anti-rabbit IgG antibody for 60 minutes. After washing again with 0.05M PBS, the sections were fixed with 1% glutaraldehyde in 0.05M PBS, washed with water and stained with uranium acetate for 5 minutes. All images were taken with a JEOL JEM-1400 transmission electron microscope.

(expression of recombinant proteins and binding to magnetic microbeads)

To prepare recombinant 00502 and recombinant 00509 proteins, the gene coding regions of both proteins (excluding the N-terminal sequence encoding the signal peptide) were cloned into the expression vector pET-28b (+) (# 69865, novagen) and a His-tag was introduced at the C-terminus according to the supplier's protocol.

The expression vector was then transformed into Rosetta-gami B (DE 3) component Cells (# 71136, novagen). The transformants were then grown exponentially and protein expression was induced by addition of 0.4mM IPTG (I6758, sigma). After incubation at 25℃overnight, B-PER was used ^TM Bacterial Protein Extraction Reagent (# 78243,Thermo Fisher Scientific) lyse cells using Pierce ^TM Ni-NTA magnetic agarose beads (# 78605,Thermo Fisher Scient)ific) lyse cells and use Pierce ^TM Polyacrylamide Spin Desalting Columns (# 89849,Thermo Fisher Scientific) recombinant 00502 and 00509 proteins were prepared.

Dynabeads is then used ^TM Antibody Coupling Kit (14311D, thermo Fisher Scientific), purified recombinant 00502 protein, recombinant 00509 protein or bovine serum albumin (# 23209,Thermo Fisher Scientific) was combined with microbeads (Dynabeads) according to the manufacturer's protocol ^TM ) And (5) combining. To each mg of magnetic beads 15. Mu.g of protein was added.

1mg of protein-bound Dynabeads were then used ^TM Resuspended in 200. Mu.L of EGEF medium and combined with recombinant mouse PRSS2 (final concentration 3. Mu.g/mL), AEBSF-pretreated Alexa Fluor ^TM 488-labeled mice PRSS2 (final concentration 5. Mu.g/mL) or 50. Mu.L GF cecal content (50-fold diluted in PBS) were mixed. The sequences of all primers used to prepare recombinants are shown in Table 6 below.

[ Table 6 ]

(protease Activity measurement)

Using Pierce ^TM Fluorescent Protease Assay Kit the protease activity of the P.clara broth, P.clara broth supernatant, recombinant 00502 protein and recombinant 00509 protein was determined according to the manufacturer's protocol (# 23266,Thermo Fisher Scientific).

FITC-casein substrate was digested into smaller fluorescein-labeled fragments using a Perkinelmer 2030Multilabel Reader with fluorescein excitation and emission filters (485/538 nm). An increase in total fluorescence was detected. Protease activity is measured as a change in Relative Fluorescence Units (RFU).

(Meta-genome analysis of human fecal samples in public queues)

The metagenome of human stool samples from the PRISM, HMP2, FHS, 500FG, CVON, and Jie cohorts were assembled from scratch into non-redundant gene catalogs and assigned to metagenomic species using MSPminer to quantify relative abundance.

To retrieve homologues of trypsin-related genes from p.clara and p.xylanphilia strains, including genes encoding 00502 protein and 00509 protein, and six other adjacent genes, in the gene catalog, userchublast (at the protein level) was used and the matching result with a minimum e value of 0.1 was retained. The results confirm that all 8 genes are present in the gene list for each species.

To identify other putative homologs and species encoding the locus, we first assessed the similarity between the corresponding homologs of the p.clara and p.xylanphilia strains and set the minimum identity (Id) and coverage (Cov) thresholds shown below for each gene at the locus. 00502: id=25%, cov=90%; 00503: id=70%, cov=90%; 00504: id=60%, cov=90%; 00505: id=60%, cov=90%; 00506: id=50%, cov=90%; 00507: id=25%, cov=90%; 0.0508: id=45%, cov=80%; 00509: id=20%, cov=30%.

Then, we assessed which other metagenomic species (MPS) encoded homologs of the 00502-00509 genes of the p.clara and p.xylanphilia strains, and found MSP 0355 and MSP 0305, which had 8 and 7 homologs, respectively. MSP 0355 and MSP 0305 were previously only annotated as the doors of bacterioides, but this study compared these proteomes with the Unified Human Gastrointestinal Genome (UHGG) using ublast. The results show that MSP 0355 and MSP 0305 are annotated as GUT_GENOME 140082 and GUT_GENOME 016875, respectively, most of which genes [ ] >90%) at high confidence (median amino acid identity>99, e value<1×e ^-184 ) Down-mapped onto a single species representing UHGG. In UHGG, both are phylogenetically classified as Paraprevatella species.

(statistics)

All statistical analyses were performed using GraphPad Prism software (GraphPad Software, inc). Multiple comparisons employ one-way analysis of variance and Tukey's test; the comparison between the two groups used either a Welch corrected Mann-Whitney test (non-parametric) or a paired t-test (parametric). The Spearman rank correlation coefficient was used to study the correlation between the two variables. The survival analysis used a log rank (Mantel-Cox) test.

Example 1

(control of trypsin in the large intestine by microbiota)

To investigate the effect of intestinal microbiota on protein distribution in the large intestine, we collected the cecal content of sterile (GF) and pathogen-free (SPF) mice and performed proteomic analysis by mass spectrometry. Among 713 host-derived proteins detected, 324 proteins were found to be higher in SPF mice than GF mice (> 2-fold, p < 0.05). They include immune related molecules such as alpha-defensin 21 (Defa 21) and peptidoglycan recognition protein 1 (Pglyrp 1). On the other hand, 45 proteins were higher in GF mice (> 2-fold, p < 0.05) than SPF mice.

FIG. 1 is a graph showing the results of proteomic analysis. FIG. 1 shows proteins with increased expression levels in the cecum of GF mice compared to SPF mice. Among these, we focused on anionic trypsin (encoded by the Prss2 gene). The level of anionic trypsin (PRSS 2) was significantly elevated in the cecal content of GF mice.

FIG. 2 is a graph showing the results of trypsin activity assay of feces of SPF mice and GF mice. FIG. 3 is a photograph showing the result of Western Blot analysis of feces from SPF mice and GF mice with anionic trypsin (PRSS 2). Fig. 4 is a fluorescence micrograph showing the results of immunostaining of mucus (UEA 1) and anionic trypsin (PRSS 2) detected in large intestine sections of SPF mice and GF mice. Nuclei were also stained with DAPI.

The results of trypsin activity detection of SPF mice and GF mice stool, western Blot analysis of SPF mice and GF mice stool, and immunostaining of large intestine sections were similar to those of proteomic analysis, indicating that the content of anionic trypsin was higher in the large intestine content and stool of GF mice than SPF mice. The anionic trypsin may be referred to below simply as trypsin.

Trypsin is produced in the pancreas in the form of an inactive precursor (trypsinogen) and is then secreted into the duodenum where it is activated by intestinal peptidases. Thus, we studied the expression of trypsinogen in the pancreas of GF mice and SPF mice. The results show that mRNA and protein levels of PRSS2 in the pancreas of GF mice and SPF mice are comparable. These results rule out the possibility that the difference in trypsin level observed in the large intestine is due to the difference in pancreas production.

Intraluminal trypsin activity was then assayed at different locations in the small and large intestines of GF mice and SPF mice. FIG. 5 is a graph showing the results of measurements of trypsin activity in jejunum, ileum, cecum, colon and stool. In fig. 5, "n.s." means no significant difference, "×" means significant difference at P < 0.05.

The results showed similar trypsin levels in SPF mice and GF mice at the distal end of the small intestine. On the other hand, in the large intestine, the trypsin activity of SPF mice was significantly reduced compared to GF mice, suggesting that microbiota play an important role in trypsin regulation in the large intestine.

Example 2

(identification of a strain of Paraprevatela capable of promoting trypsin degradation)

Although the large intestine microbiota has been previously reported to inactivate trypsin, bacteria involved in this process have not been identified. Attempts have therefore been made to isolate and identify trypsin-reducing bacteria from the human microbiota.

First, stool samples collected from 6 healthy Japanese donors (donors A-F) were administered to GF mice. Trypsin activity in GF mice feces was then measured. FIG. 6 is a graph showing the measurement of trypsin activity in the stool of GF mice administered with stool samples collected from each healthy human donor (A-F). In fig. 6, "n.s." means no significant difference, ", means significant difference at P < 0.001.

The results show that the human fecal samples examined differ in their ability to reduce the trypsin activity of the mouse feces. Specifically, the fecal trypsin activity of mice treated with the microbial flora of donor B was not reduced, whereas the fecal trypsin activity of mice treated with the microbial flora of donors C, D, E and F was significantly reduced.

Mice receiving donor C microbiota (mouse c#5) were then selected and their cecal content was collected. In addition, cecal content was orally administered to new GF mice (gf+c5 mice). Here, in order to perfect the microbiota, gf+c5 mice were divided into four groups, each group was administered ampicillin (Amp), metronidazole (MNZ), tylosin (Tyl) or a control group (No antibiotic, no Abx) by drinking water, and trypsin activity in each group of feces was measured over a period of time. The results of the study are as follows.

FIG. 7 is a graph showing the results of measurement of trypsin activity in the feces of each group of mice. In fig. 7, "n.s." means no significant difference, ", means significant difference at P < 0.001. Thus, fecal trypsin activity of gf+c5 mice not treated with antibiotics decreased within a few days. Notably, the fecal trypsin activity was reduced more in the Amp-treated group, while the fecal trypsin activity was not reduced in either the MNZ-treated group or the Tyl-treated group. These results indicate that the microbiota of C #5 contains trypsin-reducing bacteria that are enriched in Amp-treated groups and reduced in MNZ or Tyl-treated groups.

One of the Amp-treated mice (C5-Amp #5 mice) was followed, and its cecal content was collected and cultured in vitro with various media under anaerobic conditions. 432 different colonies were then isolated and subjected to 16S rRNA gene sequencing, which was subdivided into 35 unique strains. These 35 strains cover bacterial species that colonize extensively in C5-amp#5 mice; when a mixture of 35 isolated strains (35-mix) was administered to GF mice (gf+35-mix), trypsin activity in the feces was significantly reduced, and this reduction was comparable to that of mice colonized with the original donor C microbiota.

Next, in order to narrow the range of effector bacteria, the 16S rRNA gene sequences of fecal samples obtained in the above-described antibiotic administration study were analyzed, and Spearman rank correlation test was performed to evaluate the correlation between the relative abundance of each of 35 strains and the decrease in trypsin activity. The results showed that 1 of 35 strains was inversely related to trypsin activity (ρ. Ltoreq. -0.3).

The effects of these 14 bacteria and the remaining 21 bacteria were then compared by preparing Sydney mice. The results showed that trypsin activity in the feces of GF+14-mix mice was significantly reduced, comparable to GF+35-mix mice, while trypsin activity in the feces of GF+21-mix mice was not reduced. From these 14-mix, 9 strains (p.ltoreq.0.5, p.ltoreq.0.05) were subsequently further selected which were markedly associated with a reduction in trypsin activity.

The colonization of 9-mix in GF mice showed a significant decrease in trypsin activity in feces, comparable to the colonization of 14-mix mice. Finally, 9-mix is classified into 3-mix including Bacteroides species and 6-mix including non-Bacteroides species.

3-mix consisting of Paraprevotella clara (P.clara, strain number: 1C 4), bacteroides uniformis (B.uniformis, strain number: 3H 3) and Parabacteroides merdae (P.merdae, strain number: 1D 4) was sufficient to reduce fecal trypsin activity in vivo, whereas 6 non-bacteroidal mixtures were found to be completely ineffective in reducing fecal trypsin activity.

To identify trypsin-degrading bacteria, each strain in 9-mix was incubated with recombinant mouse trypsin (rmPRSS 2, added with C-terminal His tag) and trypsin degradation was measured by Western Blot.

FIG. 8 is a photograph showing the result of Western Blot. The results showed that only P.clara (strain number: 1C 4) was able to degrade trypsin. In agreement therewith, the mixture of B.unitoris 3H3 and P.merdae 1D4 (3-mix excluding P.clara (1C 4)) or 34-mix did not show the ability to reduce fecal trypsin activity in vivo.

Based on the above results, contrary to the original assumption that the community of microbiota plays a key role in the decrease of trypsin activity, we conclude that a single p.clara (1C 4) is necessary for the decrease of trypsin activity.

Recombinant human trypsin isomers PRSS1, PRSS2 and PRSS3 (rhPRSS 1-3) were then mixed and incubated with P.clara (1C 4), and then analyzed for human trypsin degradation by Western Blot.

FIG. 9 is a photograph showing the result of Western Blot. In fig. 9, "-" indicates that p.clara was not added, and "+" indicates that p.clara was added. The results show that p.clara can promote the reduction of three known human trypsin (PRSS 1 and PRSS2, and to a lesser extent PRSS 3) in addition to mouse trypsin.

It was then investigated whether the effect of p.clara on trypsin activity was strain specific. Specifically, recombinant mouse PRSS2 (rmPRSS 2) was hatched in vitro with Paraprefortella strain or Prevoltellella strain, and the degradation of rmPRSS2 was analyzed by Western Blot. The Paraprresolvotela strains used were P.clara (JCM 14859, P237E3B, P322B 5) and P.xylanipila (JCM 14860, 82A 6). The Prevoltella strains used were Paraprevotella copri, paraprevotella denticola, paraprevotella stercorea, prevotella oulorum. FIG. 10 is a photograph showing the result of Western Blot. In fig. 10, "×" indicates the position of the cleaved fragment of PRSS2 detected by the anti-PRSS 2 antibody.

Paraprefotella is a recently discovered genus of Prevoltellaceae, comprising only two species, namely P.clara and Paraprevotella xylaniphila. Thus, the P.xylaniella (JCM 14860) strain and the P.xylaniella (82A 6) strain isolated from human healthy stool samples were studied.

The results show that the other three p.clara strains (JCM 14859, P237E3B and P322B 5) isolated from fecal samples of healthy people (including non-japanese donors) have a common trypsin reduction. It was also found that both the P.xylanipila (JCM 14860) strain and the P.xylanipila (82A 6) strain exhibited strong trypsin reducing ability similar to that of P.clara.

On the other hand, bacteria of the genus Prevotella (Prevotella copri, prevotella denticola, prevotella stercorea and Prevotella oulorum) which are phylogenetically similar to the genus Paraprefotella are unable to reduce trypsin activity.

These results indicate that Parapreviella is a representative component of the trypsin-degrading human microbiota.

Example 3

(type IX secretion mechanism dependent autolysis of trypsin by Polysacchoride binding molecules)

The substrate specificity of p.clara was studied. Specifically, GF mice cecal content containing large amounts of trypsin and various proteins were incubated in vitro with p.clara (1C 4). Subsequently, the abundance of each protein was studied by peptide group analysis of LC-MS. The results showed that of the 7614 peptides from 276 proteins, trypsin was the only one that showed a pattern of significant increases in peptide concentration over time in the presence of p.clara.

These results indicate that p.clara has a narrower substrate specificity and higher activity towards trypsin. In addition, the rate of p.clara degrading trypsin was studied and found to occur gradually and proportionally.

Furthermore, P.clara mediated trypsin degradation occurs only in the presence of a sufficient number of divalent cations (e.g., ca2+). Thus, this degradation is obviously mediated by enzymes (proteases). However, when trypsin was incubated with p.clara culture supernatant, no such degradation was observed. Furthermore, no activity of protease degradation was detected when the p.clara live bacteria or the filtered p.clara supernatant was incubated with protease substrate (fluorescein-labeled casein).

FIG. 11 is a photograph showing the results of incubating protease inhibitor-pretreated recombinant mouse PRSS2 (rmPRSS 2) in combination with P.clara (1C 4) and analyzing rmPRSS2 degradation using Western Blot. The results show that pretreatment with the serine protease inhibitor AEBSF or Leupeptin and the specific trypsin inhibitor TLCK inhibited the trypsin degradation capacity of P.clara. This result suggests that p.clara effects degradation of trypsin by mediating trypsin-dependent autolysis.

Subsequently, trypsin was labeled with Alexa Fluor 488 to observe the interaction between trypsin and p.clara, in order to elucidate the mechanism by which p.clara promotes trypsin autolysis. FIG. 12 is a photograph showing the results of incubating Alexa Fluor 488-labeled rmPRSS2 with P.clara (1C 4), P.dentritica and P.olorum, respectively, and observing the binding of rmPRSS2 to bacterial surfaces with a confocal microscope. In fig. 12, a black square shows an enlarged image of p.clara (1C 4).

The results show that the fluorescently labeled trypsin was able to aggregate to the surface of p.clara in a few minutes. In contrast, no trypsin aggregation was observed in p.dentate and p.olorum. This suggests that degradation of trypsin occurs at the surface of p.clara and that trypsin-bound surface molecules promote aggregation and autolysis of p.clara trypsin.

Next, to identify trypsin-bound surface molecules of p.clara, trypsin (His-tag rmPRSS 2) and p.clara-derived molecules were crosslinked to form complexes using disuccinimide sulfone (DSSO) as a chemical crosslinking agent. The crosslinked complexes of rmPRSS2 and p.clara source molecules were then analyzed by Western Blot.

The results show that DSSO treatment resulted in a new band of high molecular weight (about 250 kDa) recognized by anti-His-tag antibodies. The band indicates the presence of a high molecular weight complex containing trypsin.

Although the mass spectrometer at hand was not sensitive enough to detect the cross-linked peptide fragment of the complex, the band around 250kDa was smeared, indicating that trypsin is interacting with a heterogeneous molecule.

It is well known that the cell surface of Bacteroides (including Paraprevitelella) has modified glycan complexes. This suggests that sugar-containing molecules on the p.clara surface are involved in trypsin binding and degradation.

To investigate this hypothesis, experiments were performed using inhibitors against p.clara saccharide synthesis. First, p.clara is treated with tunicamycin, an inhibitor against the WecA class of transferases, which mediate the first step of bacterial Lipopolysaccharide (LPS) O-glycan formation.

FIG. 13 is a photograph showing the results of incubating rmPRSS2 with P.clara (1C 4) pretreated with tunicamycin (Tuni.) or a control and analyzing rmPRSS2 degradation by Western Blot. The results showed no degradation of trypsin in clara treated with tunicamycin.

FIG. 14 is a photograph showing the results of incubating P.clara (1C 4) treated or not with tunicamycin with Alexa Fluor 488-labeled rmPRSS2, respectively, and observing the binding of rmPRSS2 to bacterial surfaces with confocal microscopy. In FIG. 14, "Tunicamycin (+)" represents the results of treatment with Tunicamycin and "Tunicamycin (-)" represents the results of treatment without Tunicamycin. The results showed that in the tunicamycin treated p.clara, trypsin did not aggregate at the surface of p.clara. Also, when p.clara is treated with the glycosyltransferase inhibitor 2-fluoro-L-fucose (2F-Fuc), which widely inhibits glycosyl-rich synthesis, both surface binding of p.clara to trypsin and degradation of trypsin are inhibited.

T9SS (type IX secretion mechanism) is a bacterial mechanism that works in conjunction with the Sec system to transport proteins with conserved C-terminal domains across the outer membrane to the surface. T9SS has a transpeptidase-like protease activity, can remove the C-terminal domain, and can bind the extracellular transported protein to surface polysaccharides.

The inventors speculate that cell surface proteins secreted by T9SS may be responsible for the recruitment and degradation of trypsin, and conducted the following study. First, the gene sequences that were supposed to be included in T9SS were identified in the genomes of P.clara and P.xylanishila.

FIG. 15 is a schematic diagram showing the arrangement of the T9SS gene composition in the P.clara (JCM 14859), P.xylanipila (JCM 14860) and P.gingivalis (ATCC 33277) genomes.

A mutant P.clara (JCM 14859) strain was then prepared by homologous recombination of the plasmid sequences, which had deleted expression of the T9SS essential factor PorU. FIG. 16 is a schematic diagram showing homologous recombination absent PorU expression.

FIG. 17 is a photograph showing the result of incubating mutation P.clara (JCM 14859) with rmPRSS2 and analyzing the degradation of rmPRSS2 by Western Blot. In FIG. 17, "PorU variant" represents the result of the mutation P.clara (JCM 14859), and "WT" represents the result of the wild-type P.clara (JCM 14859). The results showed that the mutation p.clara (JCM 14859) was completely absent for trypsin degradation, indicating that T9 SS-dependent surface proteins are involved in trypsin degradation.

The p.clara culture supernatants were then proteomic analyzed in the presence or absence of tunicamycin to identify cell surface proteins with trypsin degradation. As a result, 20 bacteria-derived proteins were found in the P.clara culture supernatant treated with tunicamycin.

Thus, by introducing a plasmid sequence into each of the 20 target loci, or by removing the gene cluster (Δ03048-03053), a series of mutant p.clara strains inhibiting synthesis of the tunicamycin-sensitive protein were prepared.

FIG. 18 is a photograph showing the results of Western Blot analysis of rmPRSS2 degradation mediated by wild-type (WT) P.clara (JCM 14859) or a series of mutants P.clara (JCM 14859). In fig. 18, "Δ03048-03053" represents the results of removing mutant p.clara of gene clusters, "00029", "00890", "00822", "00342", "00104", "03191", "00502", "02199", "00472", "01041", "00823", "00729", "00935", "01686", "03166", "00509", "00002", "PorU" and "WecA" represent the results of deleting mutant p.clara of the described genes, respectively, after insertion into a plasmid.

The results showed that when the gene encoding 00502 protein (UniProtKB ID: G5SNC9, omp 28-associated extracellular protein) or 00509 protein (UniProtKB ID: G5SNC1, unknown protein) was disrupted, trypsin degradation in vitro disappeared, as was the case for mutants lacking PorU or WecA (target factor for tunicamycin).

Next, the same study was performed by preparing p.clara strain (Δ00502 and Δ00509) lacking 00502 protein or 00509 protein instead of the insertion mutant.

FIG. 19 is a photograph showing the results of incubating Δ00502 and Δ00509 with Alexa Fluor 488-labeled rmPRSS2, respectively, and observing the binding of rmPRSS2 to the bacterial surface with a confocal microscope. The results showed that there was no recruitment of trypsin in both the Δ00502 strain and the Δ00509 strain.

FIG. 20 is a photograph showing the results of incubating the strain Δ00502 and the strain Δ00509 with rmPRSS2, respectively, and analyzing the degradation of rmPRSS2 by Western Blot. In fig. 20, "WT" represents the result of wild-type p.clara (JCM 14859). The results showed no degradation of trypsin in both the Δ00502 strain and the Δ00509 strain.

The genomic sequence of the Paraprevatella strain was then analyzed. FIG. 21 is a schematic diagram showing the genomic sequence of Paraprevatella strain. The results showed that all strains with trypsin degradation ability possess 00502 gene and 00509 gene. However, none of the species of Prevotella investigated in the study had homologs of 00502 gene and 00509 gene.

In addition, the 00503 to 00508 genes are conserved among the Paraprevatella strains, the loci of which are far apart from those of the 00502 gene and 00509 gene. FIG. 22 is a photograph showing the result of incubating a P.clara mutant strain lacking 00502 to 00509 genes with rmPRSS2 and analyzing the degradation of rmPRSS2 by Western Blot. The results showed that p.clara mutants deleted from 00503 to 00508 gene retained trypsin degradation activity, excluding their possibility of participating in these trypsin degradation processes.

Example 4

(study of 00502 protein and 00509 protein)

Recombinant 00502 protein and 00509 protein were prepared. The expression vector of 00502 protein or 00509 protein was introduced into E.coli, and treated with isopropyl-beta-thiogalactoside (IPTG) to induce recombinant protein expression, and then the expressed recombinant 00502 or 00509 protein was purified from the cell lysate using magnetic agarose beads.

FIG. 23 is a graph showing the results of measuring the protease activity of recombinant 00502 protein or 00509 protein by cleaving FITC-labeled casein. Trypsin (1 ng/. Mu.L) was used as positive control. In FIG. 23, (-) indicates that no protein was added. Protease activity is expressed as a change in relative fluorescent units. The results showed that neither the recombinant 00502 protein nor 00509 protein had protease activity.

FIG. 24 is a photograph showing the result of incubating free recombinant 00502 protein and 00509 protein or recombinant 00502 protein and 00509 protein bound to microbeads with rmPRSS2 and degrading rmPRSS2 by Western Blot analysis.

FIG. 25 is a photograph showing the results of incubating microbead-bound recombinant 00502 protein, recombinant 00509 protein, and Bovine Serum Albumin (BSA) with rmPRSS2, respectively, and observing the binding of rmPRSS2 to protein-bound microbeads with a confocal microscope. The scale bar is 5 μm. BSA was used as a negative control.

The results showed that neither the free 00502 protein nor 00509 protein showed trypsin degradation activity. In contrast, recombinant 00502 protein bound to microbeads showed efficient recruitment and degradation of trypsin.

FIG. 26 is a photograph showing the results of in vitro degradation of trypsin by Western Blot analysis after incubation of GF mouse cecal content with recombinant 00502 protein conjugated to medium control (-) or microbeads for 24 or 48 hours. In fig. 26, "×" indicates the position of the cleaved fragment of PRSS2 detected by the anti-PRSS 2 antibody. Thus, bead-bound recombinant 00502 protein detected significant trypsin degradation.

These results support the model proposed by the inventors, namely 00502 protein as a scaffold for trypsin binding and promote trypsin autolysis. In addition, trypsin binds efficiently to recombinant 00509 protein bound to the microbeads, but degradation of trypsin does not occur.

These results indicate that 00502 protein is the main component that exerts trypsin recruitment and autolysis, while 00509 protein acts to assist trypsin.

FIG. 27 is a graph showing the model of trypsin degradation mediated by Paraprefotella. In FIG. 27, "Sec" represents a Sec system in which proteins are excreted through the cytoplasmic membrane.

As shown in FIG. 27, 00502 and 00509 proteins were transported across Paraprevatella cell outer membrane by a type IX secretion mechanism (T9 SS). PorU is a basic building block for T9SS, which degrades the C-terminal domain (CTD) of T9 SS-dependent proteins, linking LPS molecules of ParaPrevoltella bacteria and these proteins. WecA mediates an initial step in LPS O-glycan synthesis, e.g., disrupting the function of WecA (e.g., by tunicamycin treatment) results in the release of T9 SS-dependent proteins. 00502 protein is involved in trypsin recruitment and autolysis as a major effector. Whereas 00509 protein aids in trypsin recruitment.

The exact mechanism by which the 00502 protein, but not 00509 protein, promotes autolysis of trypsin is still unclear, but it is speculated that binding of trypsin to 00502 protein may alter the structure of trypsin, making it more susceptible to autolysis.

Example 5

(colonisation P.clara helps to maintain IgA)

To confirm the contribution of 00502 and 00509 proteins to trypsin degradation in vivo, GF mice colonized with one of three p.clara (JCM 14859) strains (wild-type (WT), Δ00502 or Δ00509) were analyzed.

Since a single P.clara strain was unable to colonize mice, the P.clara strain was inoculated with two strains that were not trypsin-degrading (2-mix: bacteroides uniformis H3 and Parabacteroides merdae 1D 4). All three p.clara strains were effectively colonized in the intestinal tract of mice by inoculation with 2-mix.

FIG. 28 is a graph showing the results of DNA quantification of P.clara strain in feces of GF mice inoculated with P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix. In fig. 28, "n.s." means no significant difference. The results showed that in vivo, the deletion of 00502 gene or 00509 gene did not affect the number of p.clara strains.

FIG. 29 is a graph showing measurement results of trypsin activity in feces of GF mice inoculated with the P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix. In fig. 29, "and" "indicate significant differences between P <0.05 and P <0.001, respectively. The results showed that, in agreement with the in vitro results, mice colonized with the p.clara strain of Δ00502 maintained higher trypsin activity in the faeces, whereas mice colonized with the p.clara strain of Δ00509 showed a partial decrease in trypsin activity.

Subsequently, the importance of Δ00502 was investigated in the presence of more complex microbiota. FIG. 30 is a graph showing the results of measurement of trypsin activity in feces of GF mice inoculated with the P.clara strain (wild type (WT), Δ00502) and the 34-mix described above (P.clara (1C 4) excluded from 35-mix). In fig. 30, "×" indicates that there is a significant difference at P < 0.01. The results showed that mice colonized with the p.clara strain of Δ00502 showed higher trypsin activity in faeces than mice colonized with the wild-type (WT) p.clara strain.

These results demonstrate that the basic role of 00502 protein is to promote trypsin degradation in vivo.

The effect of Wild Type (WT) and mutant p.clara on the regulation of intestinal trypsin levels on important intestinal defenses factors such as IgA and antibacterial peptides were subsequently studied.

FIG. 31 is a photograph showing the results obtained by Western Blot analysis of trypsin (PRSS 2), type II transmembrane serine protease (TMPRSS 2), igA heavy chain, kappa light chain, gram negative antibacterial peptide Reg 3. Beta. And Chymotrypsin Like Elastase B (CELA 3B) in the feces of GF mice inoculated with P.clara strain (wild type (WT), Δ00502 or Δ00509) and 2-mix.

According to the results, more IgA heavy chains (alpha chains) were detected in the feces of mice colonized with wild-type p.clara strains compared to mice colonized with p.clara strains of Δ00502 or Δ00509. In contrast, kappa light chains are resistant to trypsin and therefore the kappa light chain levels in the mouse feces of the various p.clara strains were similar. In addition, the gram-negative antibacterial peptide Reg3 beta is similar to kappa light chain and also resistant to trypsin.

These results indicate that colonization of the p.clara strain protects IgA from trypsin degradation in vivo.

FIG. 32 is a photograph showing the results obtained by incubating feces of GF mice, feces of GF mice inoculated with the wild-type P.clara strain and 2-mix, a mixed sample of the two, and a mixed sample of the two plus trypsin inhibitor (TCLK) at 37℃for 48 hours, and then analyzing trypsin (PRSS 2), igA heavy chain, kappa light chain, and Chymotrypsin Like Elastase B (CELA 3B) with Western Blot.

The results in lane 1 show that trypsin was present in the feces of GF mice, indicating that the IgA heavy chain was degraded. The results in lane 2 show that trypsin is reduced in the feces of GF mice vaccinated with the wild-type P.clara strain and 2-mix, and IgA heavy chain is still present. The results in lane 3 also show that residual trypsin in GF mouse faeces degrades the IgA heavy chain remaining in GF mouse faeces vaccinated with wild type p.clara strain and 2-mix. The results in lane 4 also show that the addition of trypsin inhibitor can inhibit degradation of IgA heavy chains in lane 3. The results in lanes 3 and 4 also indicate that IgA heavy chains are more susceptible to trypsin degradation than kappa light chains.

Example 6

(study of the effect of colonization of the P.clara Strain on infection with an enteric pathogen)

It was investigated whether trypsin degradation mediated by the p.clara strain can maintain IgA levels under conditions of enteric pathogen infection.

GF mice were inoculated with 2-mix or with 2-mix in combination with wild-type (WT) or with p.clara strain Δ00502 and infected with mouse pathogen Citrobacter rodentium (c.tridentatum) 14 days after inoculation, c.tridentatum mainly infecting the large intestine.

Figure 33 shows the change in body weight of each group of mice after c. In fig. 33, ", and", respectively, indicate that there is a significant difference (2-mix+Δ00502vs.2-mix) between p < 0.01 and p < 0.001. "#" also indicates a significant difference at P <0.05 (2-mix+WT vs.2-mix).

The upper half of fig. 34 is an image showing the large intestine of each group of mice 7 days after infection with c. In the 2-mix group, the cecum color whitened and contracted. The bottom half of fig. 34 is a representative micrograph showing hematoxylin-eosin staining of cecal tissue.

Fig. 35 is a graph showing histological scores of cecal tissue based on hematoxylin-eosin staining. In fig. 35, "n.s." means no significant difference, ", means significant difference at P < 0.001.

The results show that the 2-mix group showed a rapid weight loss and severe cecal inflammation after infection with c. In contrast, mice colonized with p.clara showed milder weight loss and milder cecal inflammation. This is surprisingly similar in both the 2-mix+WT and 2-mix+Δ00502 groups. These results indicate that the p.clara strain protects mice from c.tridentatum infection by a mechanism independent of trypsin degradation.

FIG. 36 is a photograph showing the results of analyzing total IgA, C.condensing specific IgA and Chymotrypsin Like Elastase B (CELA 3B) in feces by Western Blot.

The results show that in the case of similar cecal inflammation levels, mice in the 2-mix+wt group maintained higher total IgA levels, starting 7 days earlier after c.rodenium infection, compared to the 2-mix+Δ00502 group, which was found to have considerable c.rodenium-specific IgA.

FIG. 37 is a photograph showing the results of evaluating fecal IgA aggregation effect by in vitro incubation of living C.rodenium and fecal fluid. The results show that incubating the fecal microbiota of the 2-mix+wt group with live c. This result indicates the presence of C.condensing specific IgA.

In summary, the p.clara strain, in addition to having a defensive effect on c.rodenium infection through an unknown mechanism, enhances the adaptive immune response to pathogens by protecting pathogen-specific IgA.

Example 7

(enhancing the efficacy of the vaccine against C.condensing infection by trypsin degradation of the P.clara strain)

The p.clara strain mediated trypsin degradation and the consequent pathogen-specific IgA protection may enhance the efficacy of oral vaccines against enteric pathogens and provide greater resistance when re-exposed to the same pathogen.

To confirm this hypothesis, we conducted the following study. Fig. 38 is a schematic view showing the schedule of the present embodiment. GF mice colonized with WT or Δ00502 p.clara strain were orally vaccinated with acetic acid inactivated c.tridentium vaccine 1 time per week for 3 weeks on a 2-mix basis. Oral infection was then performed with c.

Fig. 39 is a graph showing the change in body weight of mice after c. In fig. 39, "×" indicates that there is a significant difference at P < 0.05. The results showed that mice pre-inoculated with the wild type p.clara strain had a lighter weight loss.

The cecal plaque and lumen contents were then collected from mice 14 days after c.rodenium infection, and CFU of c.rodenium was measured. Figure 40 is a graph showing CFU measurements of c. Fig. 41 is a graph showing CFU measurements of c.

The results show that although the amount of C.rodents present in the cecum was similar in the 2-mix+WT and 2-mix+Δ00502 groups, there were more C.rodents present in the cecum tissue in the 2-mix+WT groups. In the 2-mix+wt group, infiltration of cecum tissue by c.condensing is inhibited.

FIG. 42 is a photograph showing the results of analysis of trypsin (PRSS 2), total IgA, C.condensing specific IgA and Chymotrypsin Like Elastase B (CELA 3B) in the lumen contents of the blind intestine by Western Blot.

The results show that significantly higher total IgA levels and significantly higher C.rodentum-specific IgA levels were detected in the cecum of mice colonized with the wild-type (WT) P.clara strain. From these results, it can be speculated that the vaccine is very effective in preventing invasion of C.rodenium into the 2-mix+WT group.

These results support the inventors' opinion that the delivery of the p.clara strain allows the host to more effectively cope with previously encountered enteropathogens.

Example 8

(P.clara strain protects mice from coronavirus infection)

Trypsin and trypsin-like proteases such as type II transmembrane serine protease (TMPRSS 2) are known to be involved in the proteolytic activation of coronavirus spike proteins and fusion of viral and host cell membranes.

TMPRSS2 is expressed as a transmembrane protein in lung and intestinal epithelial cells, but can undergo self-cleavage to release its protease domain. Interestingly, the inventors found in example 5 above that the content of TMPRSS2 was reduced in the feces of GF mice colonized with the wild type p.clara strain. This suggests that the p.clara strain has an effect similar to the release of TMPRSS2 active form in vivo. This suggests that in the gut, episomal TMPRSS2 may promote coronavirus infection along with trypsin.

The possibility that the p.clara strain may protect the gut from coronavirus infection by degradation of trypsin and TMPRSS2 was investigated. Specifically, the effect of colonization of the p.clara strain on infection with the Mouse Hepatitis Virus (MHV) (Maustropic coronavirus) was studied.

Fig. 43 is a schematic diagram showing an outline of an MHV infection experiment. GF mice were inoculated with WT or Δ00502 p.clara (JCM 14859) strain, simultaneously with 2-mix, and after 14 days were subjected to oral infection with MHV.

FIG. 44 is a graph showing survival curves of mice after MHV infection. The results show that colonization of the wild-type p.clara strain prolonged survival of mice after MHV lethal infection.

FIG. 45 is a graph showing the results of measuring MHV virus titers in the liver, brain and stool. In fig. 45, ", and", respectively, indicate that there is a significant difference between p <0.05, p <0.01, and p < 0.01.

The results showed that colonization of the wild-type p.clara strain successfully defended against viral transmission in the liver and brain. In addition, the number of viral particles excreted in the feces of mice from group 2-mix+WT was significantly lower than that of mice from group 2-mix+Δ00502.

Fig. 46 is a representative image showing hematoxylin-eosin staining results of liver tissue of each group of mice. Histological analysis showed that mice colonized with the wild-type p.clara strain were protected from liver necrosis lesions caused by MHV.

These results indicate that the colonization of the p.clara strain protects the host during coronavirus infection.

Example 9

(detection of Paraprevatellella and related genes in the human microbiota)

First, about 600 ten thousand non-redundant complete genes were collected from a six-region cohort, a new enterobacteria gene list was established, and abundance and prevalence of the paraprevatela, trypsin-related 00502 gene, and 00509 gene were analyzed.

Homology searches were performed on 5,929,528 genes in the non-redundant enterobacteriaceae gene catalog using USEARCH ublast (protein level), and the results were clustered on a match with a minimum e-value of 0.1.

FIG. 47 is a diagram showing computer-retrieved homologs of genes related to trypsin degradation (00502 gene and 00509 gene) and species encoded thereby.

FIG. 48 is a graph showing the locus structure of genes involved in trypsin degradation and sequence identity (%) with respect to the P.clara strain gene in each bacterial species.

The results showed that the 00502 gene of the P.clara strain and the homolog of the 00502 gene of the P.xylanphilia strain were the same or almost the same. Similarly, it was confirmed that the 00509 gene of the P.clara strain and the homolog of the 00509 gene of the P.xylanphilia strain were identical or nearly identical. In addition, it was confirmed that the homologs of 00503 to 00508 genes located between 00502 gene and 00509 gene were identical or nearly identical with the genes of the p.clara strain and the p.xylanphilia strain.

In addition, two metagenomic species (MSP 0303 and MSP 0335) with 00502 to 00509 gene homologs were identified and identified as likely members of Paraprevatella.

Two metagenomic species (MSP 0303 and MSP 0335) annotated as parapreviella encode homologs of all or nearly all 00502 to 00509 genes of the p.clara strain; five MSPs (MSP 0081, MSP 0224, MSP 0288, MSP 0410 and MSP 0435) annotated as bacterioides encode homologs of the 00502 gene and 00509 gene, but lack homologs of the 00503 to 00508 genes.

Prevolella muris is also believed to lack homologs of the 00509 gene.

In metagenomic analysis, the average maximum relative abundance of Paraprevatella was 3%, with abundance varying widely between the queues (7-50% of the samples), with P.clara strain being the most common species, followed by P.xylanigilla strain.

These data indicate that Parapreviella forms an important part of the human microbiota, which may be related to individual differences in susceptibility to infection by enteric pathogens.

Like mice, the concentration of human trypsin is thought to be regulated by bacterial species that degrade trypsin like Paraprevatella. Patients with Inflammatory Bowel Disease (IBD), such as Ulcerative Colitis (UC) and Crohn's Disease (CD), have been reported to have high trypsin concentrations in their feces. Thus, we measured trypsin activity in the feces of non-IBD control group (healthy), ulcerative Colitis (UC) and Crohn's Disease (CD) patients in the japanese cohort.

FIG. 49 is a graph showing the measurement results of trypsin activity. In fig. 49, "×" indicates that there is a significant difference at p < 0.05. The results showed that trypsin activity was higher in faeces of UC and CD patients compared to the non-IBD control group.

Subsequently, we analyzed Paraprevotella carryover rates for the non-IBD control group, UC and CD subgroups of the two IBD queues (PRISM and HMP 2). FIG. 50 is a graph showing Parapreviella carrier rates in patient populations diagnosed with PRISM and HMP2 cohorts as Ulcerative Colitis (UC) and Crohn's Disease (CD). The results show that in both studies, paraprevatela was present in the non-IBD samples at a greater rate than in the UC and CD patients. This trend is also statistically significant in the larger scale HMP2 queues.

These results indicate that the Parapreviella carry-over is related to intestinal health.

FIG. 51 is a photograph showing the results obtained by incubating Prevotella rodentium strain, prevolella Muris strain, wild-type P.clara strain (P.clara (WT) P.clara, P.clara strain (P.clara (00502 KO)) of the 00502 gene knocked out as a negative control) in combination with recombinant mouse trypsin (rmPRSS 2, C-terminal His tag added) and measuring trypsin degradation with Western Blot.

FIG. 52 is a photograph showing the results obtained by incubating a Prevolella ra (MSP 0081) strain, a wild-type P.clara strain (P.clara (WT)) as a positive control, and a P.clara strain (P.clara (00502 KO)) with a recombinant mouse trypsin (rmPRSS 2, C-terminal His tag added) as a negative control in a mixed manner, and measuring trypsin degradation by Western Blot.

[ Industrial availability ]

The present invention provides a technique for controlling protease activity.

SEQUENCE LISTING

<110> national institute of research and development method for human and physical chemistry

Celebration Ying Yishu

<120> composition for degrading trypsin or TMPRSS2

<130> PC-33307

<160> 178

<170> PatentIn version 3.5

<210> 1

<211> 3792

<212> DNA

<213> Paraprevotella clara

<400> 1

atgaaaaaaa aatcttgttt ggtgttcctc tttctattgg gaatactatg gaatgtacaa 60

gtccacgctg acggcgtgag ccaacctacg ttccatacct tcaagttcac agaccaagct 120

acactccaaa acatgtccga caatgggaaa tgggctgtag ctttcggtac gaacggtaac 180

agtgtggaag attttccgaa gctcatagac ttggctacgg acaaggctac cgaacttttg 240

agcgaaagcg gaaccgcatt aggcaacggc gcctacgatg tcaataacga gggaactctc 300

gtagtaggac gatatgaagg caatccggcc gtttggacca aaaccaataa caaatggaca 360

gtacttcctg ttcctacggg atgggacggt ggtttagtta acgccattac tcccgacgga 420

aagtgggcca taggtagggc caccaaaggt cagtatgacg aaactcccgt actgtgggat 480

atgagtaaag gcggtataat cacagaaacg cctaatatcc cggtaaaaga catgaccgga 540

ctagaccagc accaaagccg ctttgtaggc atttccgctg acggacgtta tatcgtaggt 600

tgcctgtctt tcagctatat ccagcctatc gcttgttgtt attacgtcta tgatcgggat 660

aagcaaacat ataaattcat cggattcgat gtagatgaga attacaaatg gactccgttg 720

gctcagaatc tggcctttat cgatgatgca cgtatcagca acaacggaaa gtatataacg 780

ggtactgctt atatggtaaa agaaggaaat gccgaataca aaactccttt tctttatgac 840

gtcgaagccg gttctttctc tatttacgac cagaatcagg atcaaggtct catcggacta 900

tgttgtgata acgagggtca tgtgttcggt gcttcacctt cagacagtcc ggcacgtgaa 960

tggagcatac gcgtaggcca atactggtac agcctaaccc aaatcctgaa acagcattac 1020

aatatagact tcaatacagc ttcggggttt gaaaatagcg gcactccgat tgcccttaat 1080

gtagaaggga ctaaaatcgc cgttttcgtc tataaaggtg aaagttatat actggagatg 1140

cctcaatcgc tcgttgaact gaccaatgaa attgatttat tggccaacta ttccgtcagc 1200

ccggaggaag gttcacaatt ctctgccctg aaatcactaa gcctcacttt tgaccgcgac 1260

attgaagtga caggaaagag cagtgccgta atcttaaaag acgaaaacgg gaacaaagta 1320

acaagttcag taacgttcaa acgtaacgcc aataacagca aaatcgtcga tattagtttc 1380

cgtggcgcaa acctcgaaga cggtaaaaaa tatactgttt ctgtcccggc tggttccatc 1440

gttattaatg gtgatgccga gaaggcctgc aaggaaatta acattagcta caccggacgg 1500

gccaataagc cagtcgcacc tactagcatt gcaccggttg acaactcaac cctatcattg 1560

ttgaacttct ctacgaaccc tatcatcatt aattttgatg caggaatcgc cttgaccgac 1620

acggcttacg ccgcagttta ccgcaataac gaaacagagc cgatgtgcga actgaaaatg 1680

gctgtgtcct ctctcgacag taaacaatta ggcgtgtacc cgtctgccgg acaatatttg 1740

tataagggta atgagtatta cgtaaaaatc aaggcaggat cagtcacgga cgttttaggt 1800

agcaatccga acgaagccat caacctgcac tacacgggta actatgaacg tgaaatcaac 1860

tacgacgatg taacattgtt caaagacgat tttaacaacg gtttgggcca attcctcttc 1920

tatgaaggag ataaacgcga accggtggaa agcatggccc aatggggatt cactgcgact 1980

acaactcctt ggagcatcgt atgggatgag gacaacacca gcgatttggc agcagcttca 2040

cattccatgt actctccggc aggaaagtct gacgactgga tggtaacaac gcaaattttc 2100

attccgtcta accaatgcta tttaagatgg gagtcacaga gttatctcaa gagtaaagga 2160

gaccgtttga aaataatggt ttgggaatac gaccccgtat tgaatgctct gaacgacgat 2220

cttatcgcta aattcaagaa tgaaggaaag gttatttacg acgaatttga aaaacctggt 2280

gaagacgaaa acaaattagc aggagaatgg acttcccaca tcgtaaaact ggaagagttt 2340

aaaggcaaaa acgtatatat tgctttcgtg aacgagaacg aagatcaaag tgccatcttt 2400

atagataatg tggaagtaac caacgaccag aagtttttgg tgggtttgac caacgaaacc 2460

tccgtggtaa accaaaaaga aattaaaatc agtggccgta tcagtatcaa tgctttggaa 2520

gatacttatc agagcgtaca cattatcatg aaagacgcta atggtaacgc catagatgaa 2580

atcagcgaat ctggtctctc tttaaagaat ggtgacaagt atgactttgc tttccaaaaa 2640

gctctcccgc taagcgtagg catcgcaaac aagttcacat tggatatcac tttagatgat 2700

gaagaaaaaa caacaggata ttctatcaag aatctggctt tcgcaccgac caagcgtttg 2760

gtgatagaag aatttacagg tacggattgc ccgaactgtc cgctcggaat tctggctttg 2820

ggcaacatgg agaaaatgtt tggcgaccaa atcatcccca tggctatcca cacttacgat 2880

ggcgacatct actcgaccaa agaacttgaa gagtattccg cgttcctgaa cttcagtggt 2940

gctcccagtg gcgtggttaa ccgtcagggc ggagctaccc ccgttccttc ttatccaatg 3000

gccagcgtta aaaacacaga aggaaaagtg aattatatat tcacaaacgg ttcggattta 3060

tggttagacc aggccgagaa agaattcaaa gtggctgcag atgccgaact gaacatcacg 3120

gcgaagtatc aggacggcaa aatcgtagtg ccttgtacct ataaatatgc actgaatgcg 3180

acagatctaa acgtaagttt gttcatggct attctggaag ataacttaac ccgttatcaa 3240

tgtaacaact tgggaggtac ttctgacccg aatctcggag aatggggatt gggcggccag 3300

tacgctgcca gtgttgtggc accttacacg ttcaacgatg tagttcgcag catacccagt 3360

gcttactatg gtgtaagcgg tctgattcct tcttccgtaa cagccggtga ggagaataca 3420

actagcctgg acttaaatat cccggaaaat gtcattgacc tcacgaactg taaagttgta 3480

tgtatgatga tcgatgccaa cacgggaaac atcatcaatg cagccagagc tgacatcaat 3540

acagacgatt ataattccat cgaacaaaca aaagcagacg aaagtatctc cgtgaatgct 3600

gacaacaaca ccatccacat taatgccggc acaacggcac aagttacagt gtacagtgta 3660

gacggaagtg ttctcgacca agttgttatc gaaggagaag gcagcctgaa gctgcaaggc 3720

cataagggta tcgtcttggt tgaggtaacg accgagaata ctcgtgttgt gaagaaagta 3780

ttcgtaaaat aa 3792

<210> 2

<211> 3084

<212> DNA

<213> Paraprevotella clara

<400> 2

atgagagaga aactttacac gaaatggaag ggaggcctga ataggctttg ttttcttctt 60

atgtgttttt gttggactac ggttcagtca tgggccgttg gtgaagattt acatttaacc 120

attgaaaatg gtaagacgta tgaatttgaa gctttcaaca gttattatct tacttatgta 180

gccacggcta acggacaatt atcattgtat caaaccggtg gagacttttg tcgtcagtat 240

acagataaca cttttgaaac ggaattgcct tctacccctc agtatgttaa tgagggaaaa 300

cttgtagaag tgaaggttga gtcaggtaag acttattatt tcttgactcg aggtttaagt 360

aaaggagaac tgaccgttac tttcggagaa aaagcaactc cacttgaatt actttcgctc 420

tccaaagaag agggaacgac attgaatctt tctattgata ccctcttagg cttcactttc 480

aatagaatgg taaaagtagg aaattgcaca ttgtcttccg ggtcggtaat cggaaacttg 540

accgcttcga cgcatgatta tggtttcacc gtttccatta aagatgtgtt gtataaatgg 600

ttgaaggagg gtaatgtaaa agccggagat gaagtcgttt tgactgtaac tggattatgt 660

aatgccaatg acgagagcga caaatacaat ggtaatggcg tgctgacggt gaaatatatt 720

gcaggagcgt tgcctgccga gctggttagc gttgcgaact cacccgaaga gatgaatttc 780

ctgtcttatt acctgcctac tgatgaaagg gggttggtta ctttgacttt caaccgttcg 840

atgggcgaag atgctacggc aaaattattc tatggcgata tagaaggttc aaatgtctat 900

acggaaagtc ttccggtgaa ggtgaaagat actcagttga ttgtagacct ccgtggaaaa 960

cgtcgtttgc catctgatat gttaccgaat gccagtgctg atgtgtggga acagtataaa 1020

accatttcct taaaggttag caatgtgagg gacgcggaag gaaattatgc catgtctacc 1080

tctcagggta ccacaggttc gtatgctttt aattatacaa tagaagctgt tgagtttgat 1140

atcgctcgtc agtttacacc agatgaaggc acatctttgg atgattatcc gacaatagaa 1200

ctttggattc gggaagacga ttcgtttact tatgagggtg tagatttcac ttatagcgtg 1260

aatggaaaac ctgagacggt atttgtaccc atcagccaga ttacgaaaaa agactctcca 1320

gacggagatg gtgtactatt gacgattccg gtgccgaata aagcggcgga tgagaattca 1380

gatgtcgtag tttctttgaa taatttgaaa gcggccgatg gtgcggatta ttcagaagat 1440

tttacgataa tatataaaac gtcaggaaag agtatggcag ggcttgagat agaatctgtg 1500

tctcctgctg atggagcagc tatcgcggct ttagaagctg gttcgtttct ggaattgtct 1560

accaatatga atgatcgtgt aggatatgta tggtttagaa tagatgacca gaacccgaag 1620

gatcctgatc aggcttgtgt taagacgatg acatcgatga agaaagagac ggttgaggga 1680

aaagtttctt tccggacaga aataattcgt agtgtgactt tctttgaagg gcatacttat 1740

aaagttactt tcaatgctta tgcttcggag agcgattatc agcacggtgc agatgctctt 1800

ggtgttatga ctgtgactta cactggtaca actccggaat ttaagtttag tccggttaag 1860

tttgtaggaa taacgccgga tcctgattat actgtaattt cagacgtaag ccagaatgaa 1920

tttacgttga cttttgacgg tcctgtagta ttgaatcatg aaaatgcttt tgtagtatac 1980

ggtcagggaa tgaattatcc atttgaaagt atcgagtcga atgaagacgg tacggaatgg 2040

acggtaaagg catccttgga aaaattaatg gaaatgggta tgatgcctcg attgtctatg 2100

agctttatgc cggtggataa agacggaatg ttagttgaag gtaatgccgg taaagatgcg 2160

acaagttatt tgaattttgc ctatgattgt acgattggta taccagattt gcaagtgagt 2220

ccggaaagcg gttctaaagt ggaaagtttg aaagagatca tagtaggatg taaagatatc 2280

atggacgaca atggtgaggt gatatttcaa ggtggtatca gcgaatctta tatggcggcc 2340

gaaaagatca ttttgtataa gaacggacgt gagccggttg caacggtaac gagtatcgaa 2400

cctattatac cggaggacca agaggataat tacttctatg ttcctgtgga aatgaaatta 2460

acattggata ctgaaattac agataacggt aattatcgtt tgcacattcc tgccaactac 2520

tttatattgg gaacaggtat gtctaatgta aacagtaaag aaacaagtgt gctgtatact 2580

atcgaccagc cgattaagat aacggttacg ccagagaata actctacggt agaaagcctg 2640

aaagaaatta ctatcgaatg tgaatcaggt atagatgtgc cgagtatggg tacaattcaa 2700

ttattgaatg ctcagaatga ggtggtagct tcggcaacgg gtgaggattg cgaattgttg 2760

ttacccgaag gtgccggtcc ttgggatcct tatactggtg tgacaattaa attggatcaa 2820

gaggtgacag agaaaggtac atataagttg gttattcctg aaggattctt ctatttgggt 2880

gaaaactatg aaaattcgga cgaaatgacg tttacttatt cgattaatgc cagtggtatt 2940

cattcgattg gtactgagtc taagggagta gtcgtttata ctgtagacgg taaatttatc 3000

ttaaagtcga gcgatgccaa agacgtgaag aatctgaaga aaggtttgta cattgtgaac 3060

ggtaagaaga tgatggtaaa ataa 3084

<210> 3

<211> 1483

<212> DNA

<213> Paraprevotella clara

<400> 3

gatgaacgct agctacaggc ttaacacatg caagtcgagg ggcagcatgr rcycagcttt 60

gctgggtttg atggcgaccg gcgcacgggt gagtaacgcg tatccaacct gccctttact 120

ccgggatagt ctcctgaaag ggagtttaat accggatgtg tttgtctttc cgcatgggag 180

cgacaaataa agattgattg gtaaaggatg gggatgcgtc ccattagctr gttggcgggg 240

taacggccca ccaaggcrac gatgggtagg ggttctgaga ggaaggtccc ccacattgga 300

actgagacac ggtccaaact cctacgggag gcagcagtga ggaatattgg tcaatgggcg 360

agagcctgaa ccagccaagt agcgtgaagg acgacggccc tacgggttgt aaacttcttt 420

tataagggaa taaagttcgc cacgcgtggt gttttgtatg taccttatga ataagcatcg 480

gctaattccg tgccagcagc cgcggtaata cggaagatgc gagcgttatc cggatttatt 540

gggtttaaag ggagcgtagg cgggctttta agtcagcggt caaatgtcac ggctcaaccg 600

tggccagccg ttgaaactgt aagccttgag tctgcacagg gcacatggaa ttcgtggtgt 660

agcggtgaaa tgcttagata tcacgaagaa ctccgatcgc gaaggcattg tgccggggca 720

gcactgacgc tgaggctcga aagtgcgggt atcaaacagg attagatacc ctggtagtcc 780

gcacggtaaa cgatgaatgc tcgctatggg cgatayawtg tccgtggcca agcgaaagcg 840

ttaagcattc cacctgggga gtacgccggc aacggtgaaa ctcaaaggaa ttgacggggg 900

cccgcacaag cggaggaaca tgtggtttaa ttcgatgata cgcgaggaac cttacccggg 960

cttgaattgc aggtgcatga gtcagagatg attctttcct tcgggactcc tgtgaaggtg 1020

ctgcatggtt gtcgtcagct cgtgccgtga ggtgtcggct taagtgccat aacgagcgca 1080

acccttctcc ccagttgcca tcgggtaatg ccgggccctc tggggacact gccatcgtaa 1140

gatgcgagga aggtggggat gacgtcaaat cagcacggcc cttacgtccg gggctacaca 1200

cgtgttacaa tggggggtac agagggccgc tgtccggtga cggttggcca atccctaaaa 1260

cccctctcag ttcggactgg agtctgcaac ccgactccac gaagctggat tcgctagtaa 1320

tcgcgcatca gccatggcgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcaa 1380

gccatgaaag ccgggggtgc ctgaagtccg tgaccgcgag ggtcggccta gggtaaaact 1440

ggtgattggg gctaagtcgt aacaaggtag ccgtaccgga agg 1483

<210> 4

<211> 1482

<212> DNA

<213> Paraprevotella xylaniphila

<400> 4

gatgaacgct agctacaggc ttaacacatg caagtcgagg ggcagcatga gctcagttat 60

tctgggtttg atggcgaccg gcgcacgggt gagtaacgcg tatccaacct gccctttacc 120

cggggatagc cttctgaaag gaagtttaat acccgatgat ttcgtttagt cgcatgactt 180

gatgaataaa gattaattgg taaaggatgg ggatgcgtcc cattagcttg ttggcggggt 240

aacggcccac caaggcgacg atgggtaggg gttctgagag gaaggtcccc cacattggaa 300

ctgagacacg gtccaaactc ctacgggagg cagcagtgag gaatattggt caatgggcgc 360

gagcctgaac cagccaagta gcgtggagga cgacggccct acgggttgta aactcctttt 420

ataaggggat aaagttggcc atgtatggcc atttgcaggt accttatgaa taagcatcgg 480

ctaattccgt gccagcagcc gcggtaatac ggaagatgcg agcgttatcc ggatttattg 540

ggtttaaagg gagcgtaggc gggcagtcaa gtcagcggtc aaatggcgcg gctcaaccgc 600

gttccgccgt tgaaactggc agccttgagt atgcacaggg tacatggaat tcgtggtgta 660

gcggtgaaat gcttagatat cacgaggaac tccgatcgcg caggcattgt accggggcat 720

tactgacgct gaggctcgaa ggtgcgggta tcaaacagga ttagataccc tggtagtccg 780

cacagtaaac gatgaatgcc cgctgtcggc gacatagtgt cggcggccaa gcgaaagcgt 840

taagcattcc acctggggag tacgccggca acggtgaaac tcaaaggaat tgacgggggc 900

ccgcacaagc ggaggaacat gtggtttaat tcgatgatac gcgaggaacc ttacccgggc 960

ttgaatcgca ggtgcatggg ccggagacgg ccctttcctt cgggactcct gcgaaggtgc 1020

tgcatggttg tcgtcagctc gtgccgtgag gtgtcggctt aagtgccata acgagcgcaa 1080

cccccctccc cagttgccac cgggtaatgc cgggcacttt ggggacactg ccaccgcaag 1140

gtgcgaggaa ggtggggatg acgtcaaatc agcacggccc ttacgtccgg ggcgacacac 1200

gtgttacaat ggggggtaca gagggccgct gcccggtgac ggttggccaa tccctaaaac 1260

ccctctcagt tcggactgga gtctgcaacc cgactccacg aagctggatt cgctagtaat 1320

cgcgcatcag ccatggcgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaag 1380

ccatgaaagc cgggggtgcc tgaagtccgt gaccgcgagg gtcggcctag ggtaaaaccg 1440

gtgattgggg ctaagtcgta acaaggtagc cgtaccggaa gg 1482

<210> 5

<211> 1263

<212> PRT

<213> Paraprevotella clara

<400> 5

Met Lys Lys Lys Ser Cys Leu Val Phe Leu Phe Leu Leu Gly Ile Leu

1 5 10 15

Trp Asn Val Gln Val His Ala Asp Gly Val Ser Gln Pro Thr Phe His

20 25 30

Thr Phe Lys Phe Thr Asp Gln Ala Thr Leu Gln Asn Met Ser Asp Asn

35 40 45

Gly Lys Trp Ala Val Ala Phe Gly Thr Asn Gly Asn Ser Val Glu Asp

50 55 60

Phe Pro Lys Leu Ile Asp Leu Ala Thr Asp Lys Ala Thr Glu Leu Leu

65 70 75 80

Ser Glu Ser Gly Thr Ala Leu Gly Asn Gly Ala Tyr Asp Val Asn Asn

85 90 95

Glu Gly Thr Leu Val Val Gly Arg Tyr Glu Gly Asn Pro Ala Val Trp

100 105 110

Thr Lys Thr Asn Asn Lys Trp Thr Val Leu Pro Val Pro Thr Gly Trp

115 120 125

Asp Gly Gly Leu Val Asn Ala Ile Thr Pro Asp Gly Lys Trp Ala Ile

130 135 140

Gly Arg Ala Thr Lys Gly Gln Tyr Asp Glu Thr Pro Val Leu Trp Asp

145 150 155 160

Met Ser Lys Gly Gly Ile Ile Thr Glu Thr Pro Asn Ile Pro Val Lys

165 170 175

Asp Met Thr Gly Leu Asp Gln His Gln Ser Arg Phe Val Gly Ile Ser

180 185 190

Ala Asp Gly Arg Tyr Ile Val Gly Cys Leu Ser Phe Ser Tyr Ile Gln

195 200 205

Pro Ile Ala Cys Cys Tyr Tyr Val Tyr Asp Arg Asp Lys Gln Thr Tyr

210 215 220

Lys Phe Ile Gly Phe Asp Val Asp Glu Asn Tyr Lys Trp Thr Pro Leu

225 230 235 240

Ala Gln Asn Leu Ala Phe Ile Asp Asp Ala Arg Ile Ser Asn Asn Gly

245 250 255

Lys Tyr Ile Thr Gly Thr Ala Tyr Met Val Lys Glu Gly Asn Ala Glu

260 265 270

Tyr Lys Thr Pro Phe Leu Tyr Asp Val Glu Ala Gly Ser Phe Ser Ile

275 280 285

Tyr Asp Gln Asn Gln Asp Gln Gly Leu Ile Gly Leu Cys Cys Asp Asn

290 295 300

Glu Gly His Val Phe Gly Ala Ser Pro Ser Asp Ser Pro Ala Arg Glu

305 310 315 320

Trp Ser Ile Arg Val Gly Gln Tyr Trp Tyr Ser Leu Thr Gln Ile Leu

325 330 335

Lys Gln His Tyr Asn Ile Asp Phe Asn Thr Ala Ser Gly Phe Glu Asn

340 345 350

Ser Gly Thr Pro Ile Ala Leu Asn Val Glu Gly Thr Lys Ile Ala Val

355 360 365

Phe Val Tyr Lys Gly Glu Ser Tyr Ile Leu Glu Met Pro Gln Ser Leu

370 375 380

Val Glu Leu Thr Asn Glu Ile Asp Leu Leu Ala Asn Tyr Ser Val Ser

385 390 395 400

Pro Glu Glu Gly Ser Gln Phe Ser Ala Leu Lys Ser Leu Ser Leu Thr

405 410 415

Phe Asp Arg Asp Ile Glu Val Thr Gly Lys Ser Ser Ala Val Ile Leu

420 425 430

Lys Asp Glu Asn Gly Asn Lys Val Thr Ser Ser Val Thr Phe Lys Arg

435 440 445

Asn Ala Asn Asn Ser Lys Ile Val Asp Ile Ser Phe Arg Gly Ala Asn

450 455 460

Leu Glu Asp Gly Lys Lys Tyr Thr Val Ser Val Pro Ala Gly Ser Ile

465 470 475 480

Val Ile Asn Gly Asp Ala Glu Lys Ala Cys Lys Glu Ile Asn Ile Ser

485 490 495

Tyr Thr Gly Arg Ala Asn Lys Pro Val Ala Pro Thr Ser Ile Ala Pro

500 505 510

Val Asp Asn Ser Thr Leu Ser Leu Leu Asn Phe Ser Thr Asn Pro Ile

515 520 525

Ile Ile Asn Phe Asp Ala Gly Ile Ala Leu Thr Asp Thr Ala Tyr Ala

530 535 540

Ala Val Tyr Arg Asn Asn Glu Thr Glu Pro Met Cys Glu Leu Lys Met

545 550 555 560

Ala Val Ser Ser Leu Asp Ser Lys Gln Leu Gly Val Tyr Pro Ser Ala

565 570 575

Gly Gln Tyr Leu Tyr Lys Gly Asn Glu Tyr Tyr Val Lys Ile Lys Ala

580 585 590

Gly Ser Val Thr Asp Val Leu Gly Ser Asn Pro Asn Glu Ala Ile Asn

595 600 605

Leu His Tyr Thr Gly Asn Tyr Glu Arg Glu Ile Asn Tyr Asp Asp Val

610 615 620

Thr Leu Phe Lys Asp Asp Phe Asn Asn Gly Leu Gly Gln Phe Leu Phe

625 630 635 640

Tyr Glu Gly Asp Lys Arg Glu Pro Val Glu Ser Met Ala Gln Trp Gly

645 650 655

Phe Thr Ala Thr Thr Thr Pro Trp Ser Ile Val Trp Asp Glu Asp Asn

660 665 670

Thr Ser Asp Leu Ala Ala Ala Ser His Ser Met Tyr Ser Pro Ala Gly

675 680 685

Lys Ser Asp Asp Trp Met Val Thr Thr Gln Ile Phe Ile Pro Ser Asn

690 695 700

Gln Cys Tyr Leu Arg Trp Glu Ser Gln Ser Tyr Leu Lys Ser Lys Gly

705 710 715 720

Asp Arg Leu Lys Ile Met Val Trp Glu Tyr Asp Pro Val Leu Asn Ala

725 730 735

Leu Asn Asp Asp Leu Ile Ala Lys Phe Lys Asn Glu Gly Lys Val Ile

740 745 750

Tyr Asp Glu Phe Glu Lys Pro Gly Glu Asp Glu Asn Lys Leu Ala Gly

755 760 765

Glu Trp Thr Ser His Ile Val Lys Leu Glu Glu Phe Lys Gly Lys Asn

770 775 780

Val Tyr Ile Ala Phe Val Asn Glu Asn Glu Asp Gln Ser Ala Ile Phe

785 790 795 800

Ile Asp Asn Val Glu Val Thr Asn Asp Gln Lys Phe Leu Val Gly Leu

805 810 815

Thr Asn Glu Thr Ser Val Val Asn Gln Lys Glu Ile Lys Ile Ser Gly

820 825 830

Arg Ile Ser Ile Asn Ala Leu Glu Asp Thr Tyr Gln Ser Val His Ile

835 840 845

Ile Met Lys Asp Ala Asn Gly Asn Ala Ile Asp Glu Ile Ser Glu Ser

850 855 860

Gly Leu Ser Leu Lys Asn Gly Asp Lys Tyr Asp Phe Ala Phe Gln Lys

865 870 875 880

Ala Leu Pro Leu Ser Val Gly Ile Ala Asn Lys Phe Thr Leu Asp Ile

885 890 895

Thr Leu Asp Asp Glu Glu Lys Thr Thr Gly Tyr Ser Ile Lys Asn Leu

900 905 910

Ala Phe Ala Pro Thr Lys Arg Leu Val Ile Glu Glu Phe Thr Gly Thr

915 920 925

Asp Cys Pro Asn Cys Pro Leu Gly Ile Leu Ala Leu Gly Asn Met Glu

930 935 940

Lys Met Phe Gly Asp Gln Ile Ile Pro Met Ala Ile His Thr Tyr Asp

945 950 955 960

Gly Asp Ile Tyr Ser Thr Lys Glu Leu Glu Glu Tyr Ser Ala Phe Leu

965 970 975

Asn Phe Ser Gly Ala Pro Ser Gly Val Val Asn Arg Gln Gly Gly Ala

980 985 990

Thr Pro Val Pro Ser Tyr Pro Met Ala Ser Val Lys Asn Thr Glu Gly

995 1000 1005

Lys Val Asn Tyr Ile Phe Thr Asn Gly Ser Asp Leu Trp Leu Asp

1010 1015 1020

Gln Ala Glu Lys Glu Phe Lys Val Ala Ala Asp Ala Glu Leu Asn

1025 1030 1035

Ile Thr Ala Lys Tyr Gln Asp Gly Lys Ile Val Val Pro Cys Thr

1040 1045 1050

Tyr Lys Tyr Ala Leu Asn Ala Thr Asp Leu Asn Val Ser Leu Phe

1055 1060 1065

Met Ala Ile Leu Glu Asp Asn Leu Thr Arg Tyr Gln Cys Asn Asn

1070 1075 1080

Leu Gly Gly Thr Ser Asp Pro Asn Leu Gly Glu Trp Gly Leu Gly

1085 1090 1095

Gly Gln Tyr Ala Ala Ser Val Val Ala Pro Tyr Thr Phe Asn Asp

1100 1105 1110

Val Val Arg Ser Ile Pro Ser Ala Tyr Tyr Gly Val Ser Gly Leu

1115 1120 1125

Ile Pro Ser Ser Val Thr Ala Gly Glu Glu Asn Thr Thr Ser Leu

1130 1135 1140

Asp Leu Asn Ile Pro Glu Asn Val Ile Asp Leu Thr Asn Cys Lys

1145 1150 1155

Val Val Cys Met Met Ile Asp Ala Asn Thr Gly Asn Ile Ile Asn

1160 1165 1170

Ala Ala Arg Ala Asp Ile Asn Thr Asp Asp Tyr Asn Ser Ile Glu

1175 1180 1185

Gln Thr Lys Ala Asp Glu Ser Ile Ser Val Asn Ala Asp Asn Asn

1190 1195 1200

Thr Ile His Ile Asn Ala Gly Thr Thr Ala Gln Val Thr Val Tyr

1205 1210 1215

Ser Val Asp Gly Ser Val Leu Asp Gln Val Val Ile Glu Gly Glu

1220 1225 1230

Gly Ser Leu Lys Leu Gln Gly His Lys Gly Ile Val Leu Val Glu

1235 1240 1245

Val Thr Thr Glu Asn Thr Arg Val Val Lys Lys Val Phe Val Lys

1250 1255 1260

<210> 6

<211> 1263

<212> PRT

<213> Paraprevotella clara

<400> 6

Met Lys Lys Lys Ser Cys Leu Val Phe Leu Phe Leu Leu Gly Ile Leu

1 5 10 15

Trp Asn Val Gln Val His Ala Asp Gly Val Ser Gln Pro Thr Phe His

20 25 30

Thr Phe Lys Phe Thr Asp Gln Ala Thr Leu Gln Asn Met Ser Asp Asn

35 40 45

Gly Lys Trp Ala Val Ala Phe Gly Thr Asn Gly Asn Ser Val Glu Asp

50 55 60

Phe Pro Lys Leu Ile Asp Leu Ala Thr Asp Lys Ala Thr Glu Leu Leu

65 70 75 80

Ser Glu Ser Gly Thr Ala Leu Gly Asn Gly Ala Tyr Asp Val Asn Asn

85 90 95

Glu Gly Thr Leu Val Val Gly Arg Tyr Glu Gly Asn Pro Ala Val Trp

100 105 110

Thr Lys Thr Asn Asn Lys Trp Thr Val Leu Pro Val Pro Thr Gly Trp

115 120 125

Asp Gly Gly Leu Val Asn Ala Ile Thr Pro Asp Gly Lys Trp Ala Ile

130 135 140

Gly Arg Ala Thr Lys Gly Gln Tyr Asp Glu Thr Pro Val Leu Trp Asp

145 150 155 160

Met Ser Lys Gly Gly Ile Ile Thr Glu Thr Pro Asn Ile Pro Val Lys

165 170 175

Asp Met Thr Gly Leu Asp Gln His Gln Ser Arg Phe Val Gly Ile Ser

180 185 190

Ala Asp Gly Arg Tyr Ile Val Gly Cys Leu Ser Phe Ser Tyr Ile Gln

195 200 205

Pro Ile Ala Cys Cys Tyr Tyr Val Tyr Asp Arg Asp Lys Gln Thr Tyr

210 215 220

Lys Phe Ile Gly Phe Asp Val Asp Glu Asn Tyr Lys Trp Thr Pro Leu

225 230 235 240

Ala Gln Asn Leu Ala Phe Ile Asp Asp Ala Arg Ile Ser Asn Asn Gly

245 250 255

Lys Tyr Ile Thr Gly Thr Ala Tyr Met Val Lys Glu Gly Asn Ala Glu

260 265 270

Tyr Lys Thr Pro Phe Leu Tyr Asp Val Glu Ala Gly Ser Phe Ser Ile

275 280 285

Tyr Asp Gln Asn Gln Asp Gln Gly Leu Ile Gly Leu Cys Cys Asp Asn

290 295 300

Glu Gly His Val Phe Gly Ala Ser Pro Ser Asp Ser Pro Ala Arg Glu

305 310 315 320

Trp Ser Ile Arg Val Gly Gln Tyr Trp Tyr Ser Leu Thr Gln Ile Leu

325 330 335

Lys Gln His Tyr Asn Ile Asp Phe Asn Thr Val Ser Gly Phe Glu Asn

340 345 350

Ser Gly Thr Pro Ile Ala Leu Asn Val Glu Gly Thr Lys Ile Ala Val

355 360 365

Phe Val Tyr Lys Gly Glu Ser Tyr Ile Leu Glu Met Pro Gln Ser Leu

370 375 380

Val Glu Leu Thr Asn Glu Ile Asp Leu Leu Ala Asn Tyr Ser Val Ser

385 390 395 400

Pro Glu Glu Gly Ser Gln Phe Ser Ala Leu Lys Ser Leu Ser Leu Thr

405 410 415

Phe Asp Arg Asp Ile Glu Val Thr Gly Lys Ser Ser Ala Val Ile Leu

420 425 430

Lys Asp Glu Asn Gly Asn Lys Val Thr Ser Ser Val Thr Phe Lys Arg

435 440 445

Asn Ala Asn Asn Ser Lys Ile Val Asp Ile Ser Phe Arg Gly Ala Asn

450 455 460

Leu Glu Asp Gly Lys Lys Tyr Thr Val Ser Val Pro Ala Gly Ser Ile

465 470 475 480

Val Ile Asn Gly Asp Ala Glu Lys Ala Cys Lys Glu Ile Asn Ile Ser

485 490 495

Tyr Thr Gly Arg Ala Asn Lys Pro Val Ala Pro Thr Ser Ile Ala Pro

500 505 510

Val Asp Asn Ser Thr Leu Ser Leu Leu Asn Phe Ser Thr Asn Pro Ile

515 520 525

Ile Ile Asn Phe Asp Ala Gly Ile Ala Leu Thr Asp Thr Ala Tyr Ala

530 535 540

Ala Val Tyr Arg Asn Asn Glu Thr Glu Pro Met Cys Glu Leu Lys Met

545 550 555 560

Ala Val Ser Ser Leu Asp Ser Lys Gln Leu Gly Val Tyr Pro Ser Ala

565 570 575

Gly Gln Tyr Leu Tyr Lys Gly Asn Glu Tyr Tyr Val Lys Ile Lys Ala

580 585 590

Gly Ser Val Thr Asp Val Leu Gly Ser Asn Pro Asn Glu Ala Ile Asn

595 600 605

Leu His Tyr Thr Gly Asn Tyr Glu Arg Glu Ile Asn Tyr Asp Asp Val

610 615 620

Thr Leu Phe Lys Asp Asp Phe Asn Asn Gly Leu Gly Gln Phe Leu Phe

625 630 635 640

Tyr Glu Gly Asp Lys Arg Glu Pro Val Glu Ser Met Val Gln Trp Gly

645 650 655

Phe Thr Ala Thr Thr Thr Pro Trp Ser Ile Val Trp Asp Glu Asp Asn

660 665 670

Thr Ser Asp Leu Ala Ala Ala Ser His Ser Met Tyr Ser Pro Ala Gly

675 680 685

Lys Ser Asp Asp Trp Met Val Thr Thr Gln Ile Phe Ile Pro Ser Asn

690 695 700

Gln Cys Tyr Leu Arg Trp Glu Ser Gln Ser Tyr Leu Lys Ser Lys Gly

705 710 715 720

Asp Arg Leu Lys Ile Met Val Trp Glu Tyr Asp Pro Val Leu Asn Ala

725 730 735

Leu Asn Asp Asp Leu Ile Ala Lys Phe Lys Asn Glu Gly Lys Val Ile

740 745 750

Tyr Asp Glu Phe Glu Lys Pro Gly Glu Asp Glu Asn Lys Leu Ala Gly

755 760 765

Glu Trp Thr Ser His Ile Val Lys Leu Glu Glu Phe Lys Gly Lys Asn

770 775 780

Val Tyr Ile Ala Phe Val Asn Glu Asn Glu Asp Gln Ser Ala Ile Phe

785 790 795 800

Ile Asp Asn Val Glu Val Thr Asn Asp Gln Lys Phe Leu Val Gly Leu

805 810 815

Thr Asn Glu Thr Ser Val Val Asn Gln Lys Glu Ile Lys Ile Ser Gly

820 825 830

Arg Ile Ser Ile Asn Ala Leu Glu Asp Thr Tyr Gln Ser Val His Ile

835 840 845

Ile Met Lys Asp Ala Asn Gly Asn Ala Ile Asp Glu Ile Ser Glu Ser

850 855 860

Gly Leu Ser Leu Lys Asn Gly Asp Lys Tyr Asp Phe Ala Phe Gln Lys

865 870 875 880

Ala Leu Pro Leu Ser Val Gly Ile Ala Asn Lys Phe Thr Leu Asp Ile

885 890 895

Thr Leu Asp Asp Glu Glu Lys Thr Thr Gly Tyr Ser Ile Lys Asn Leu

900 905 910

Ala Phe Ala Pro Thr Lys Arg Leu Val Ile Glu Glu Phe Thr Gly Thr

915 920 925

Asp Cys Pro Asn Cys Pro Leu Gly Ile Leu Ala Leu Gly Asn Met Glu

930 935 940

Lys Met Phe Gly Asp Gln Ile Ile Pro Met Ala Ile His Thr Tyr Asp

945 950 955 960

Gly Asp Ile Tyr Ser Thr Lys Glu Leu Glu Glu Tyr Ser Ala Phe Leu

965 970 975

Asn Phe Ser Gly Ala Pro Ser Gly Val Val Asn Arg Gln Gly Gly Ala

980 985 990

Thr Pro Val Pro Ser Tyr Pro Met Ala Ser Val Lys Asn Thr Glu Gly

995 1000 1005

Lys Val Asn Tyr Ile Phe Thr Asn Gly Ser Asp Leu Trp Leu Asp

1010 1015 1020

Gln Ala Glu Lys Glu Phe Lys Val Ala Ala Asp Ala Glu Leu Asn

1025 1030 1035

Ile Thr Ala Lys Tyr Gln Asp Gly Lys Ile Val Val Pro Cys Thr

1040 1045 1050

Tyr Lys Tyr Ala Leu Asn Ala Thr Asp Leu Asn Val Ser Leu Phe

1055 1060 1065

Met Ala Ile Leu Glu Asp Asn Leu Thr Arg Tyr Gln Cys Asn Asn

1070 1075 1080

Leu Gly Gly Thr Ser Asp Pro Asn Leu Gly Glu Trp Gly Leu Gly

1085 1090 1095

Gly Gln Tyr Ala Ala Ser Val Val Ala Pro Tyr Thr Phe Asn Asp

1100 1105 1110

Val Val Arg Ser Ile Pro Ser Ala Tyr Tyr Gly Val Ser Gly Leu

1115 1120 1125

Ile Pro Ser Ser Val Thr Ala Gly Glu Glu Asn Thr Thr Ser Leu

1130 1135 1140

Asp Leu Asn Ile Pro Glu Asn Ile Ile Asp Leu Thr Asn Cys Lys

1145 1150 1155

Val Val Cys Met Met Ile Asp Ala Asn Thr Gly Asn Ile Ile Asn

1160 1165 1170

Ala Ala Arg Ala Asp Ile Asn Thr Asp Asp Tyr Asn Ser Ile Glu

1175 1180 1185

Gln Thr Lys Ala Asp Glu Ser Ile Ser Val Asn Ala Asp Asn Asn

1190 1195 1200

Thr Ile His Ile Asn Ala Gly Thr Thr Ala Gln Val Thr Val Tyr

1205 1210 1215

Ser Val Asp Gly Ser Val Leu Asp Gln Val Val Ile Glu Gly Glu

1220 1225 1230

Gly Ser Leu Lys Leu Gln Gly His Lys Gly Ile Val Leu Val Glu

1235 1240 1245

Val Thr Thr Glu Asn Thr Arg Val Val Lys Lys Val Phe Val Lys

1250 1255 1260

<210> 7

<211> 3792

<212> DNA

<213> Paraprevotella clara

<400> 7

atgaaaaaaa aatcttgttt ggtgttcctc tttctattgg gaatactatg gaatgtacaa 60

gtccacgctg acggtgtgag ccaacctacg ttccatacct tcaagttcac agaccaagct 120

acactccaga acatgtccga caatgggaaa tgggctgtag ctttcggtac gaacggtaac 180

agtgtggaag attttccgaa gctcatagac ttggctacgg acaaggctac cgaacttttg 240

agcgaaagcg gaaccgcatt aggcaacggc gcctacgatg tcaataacga gggaactctc 300

gtagtaggac gatatgaagg aaatccggcc gtttggacca aaaccaataa caaatggaca 360

gtacttcctg ttcctacggg atgggacggt ggtttagtta acgccattac tcccgacgga 420

aagtgggcca taggtagggc caccaaaggt cagtatgacg aaactcccgt actgtgggat 480

atgagtaaag gcggtataat cacagaaacg cctaatatcc cggtaaaaga catgaccgga 540

ctagaccagc accaaagccg ctttgtaggc atttccgctg acggacgtta tatcgtaggt 600

tgcctgtctt tcagctatat ccagcctatc gcttgttgtt attacgtcta tgatcgggat 660

aagcaaacat ataaattcat cggattcgat gtagatgaga attacaaatg gactccgttg 720

gctcagaatc tggcctttat cgatgatgca cgtatcagca acaacggaaa gtatataacg 780

ggtactgctt atatggtaaa agaaggaaat gccgaataca aaactccttt tctttatgac 840

gttgaagccg gttccttctc tatttacgac cagaaccagg accaaggtct catcggacta 900

tgttgtgata acgagggtca tgtgttcggt gcttcacctt cagacagtcc ggcacgtgaa 960

tggagcatac gcgtaggcca atactggtac agccttaccc aaatcctgaa acagcattac 1020

aatatagact ttaatacagt ttcggggttt gaaaatagcg gaactccgat tgcccttaat 1080

gtagaaggta ctaaaatcgc cgtcttcgtc tataaaggtg aaagttatat actggagatg 1140

cctcagtcgc tcgttgaact gaccaatgaa attgatttat tggccaacta ttccgtcagc 1200

ccggaggaag gttcacaatt ctctgccctg aaatcactaa gcctcacttt tgaccgcgac 1260

attgaagtga caggaaagag cagtgccgta atcttaaaag acgaaaacgg gaacaaagta 1320

acaagttcag taacgttcaa acgtaacgcc aataacagca aaatcgtcga tattagtttc 1380

cgtggcgcaa acctcgaaga cggtaaaaaa tatactgttt ctgtcccggc tggttccatc 1440

gttattaatg gtgatgccga gaaggcctgc aaggaaatta acattagcta caccggacgg 1500

gccaataagc cagtcgcacc tactagcatt gcaccggttg acaactcaac cctatcattg 1560

ttgaacttct ctacgaaccc tatcatcatt aattttgatg caggaatcgc cttgaccgac 1620

acggcttacg ccgcagttta ccgcaataac gaaacagagc cgatgtgcga actgaaaatg 1680

gctgtgtcct ctctcgacag taaacaatta ggcgtgtacc cgtctgccgg acaatatttg 1740

tataagggta atgagtatta cgtaaaaatc aaggcaggat cagtcacgga cgttttaggt 1800

agcaatccga acgaagccat caacctgcac tacacgggta actatgaacg tgaaatcaac 1860

tacgacgatg taacattgtt caaagacgat tttaacaacg gtttgggcca attcctcttc 1920

tatgaaggag ataaacgcga accggtggaa agcatggtcc aatggggatt cactgcgact 1980

acaactcctt ggagcatcgt atgggatgag gacaacacca gcgatttggc agcagcttca 2040

cattccatgt actctccggc aggaaagtct gacgactgga tggtaacaac gcaaattttc 2100

attccgtcta accaatgcta tttaagatgg gagtcacaga gttatcttaa gagtaaagga 2160

gaccgtttga aaataatggt ttgggaatac gaccccgtat tgaatgctct gaacgacgat 2220

cttatcgcta aattcaagaa tgaaggaaag gttatttacg acgaatttga aaaacctggt 2280

gaagacgaaa acaaattagc aggagaatgg acttcccaca tcgtaaaact ggaagagttt 2340

aaaggcaaaa acgtatatat tgctttcgtg aacgagaacg aagatcaaag tgccatcttt 2400

atagataatg tggaagtaac caacgaccag aagtttttgg tgggtttgac caacgaaacc 2460

tccgtggtaa accaaaaaga aattaaaatc agtggccgta tcagtatcaa tgctttggaa 2520

gatacttatc agagcgtaca cattatcatg aaagacgcta atggtaacgc catagatgaa 2580

atcagcgaat ctggtctctc tttaaagaat ggtgacaagt atgactttgc tttccaaaaa 2640

gctctcccgc taagcgtagg catcgcaaac aagttcacat tggatatcac tttagatgat 2700

gaagaaaaaa caacaggata ttctatcaag aatctggctt tcgcaccgac caagcgtttg 2760

gtgatagaag aatttacagg tacggattgc ccgaactgtc cgctcggaat tctggctttg 2820

ggcaacatgg agaaaatgtt tggcgaccaa atcatcccca tggctatcca cacttacgat 2880

ggcgacatct actcgaccaa agaacttgaa gagtattccg cgttcctgaa cttcagtggt 2940

gctcccagcg gagtagtcaa tcgtcagggc ggagctaccc ccgttccttc ttatccaatg 3000

gccagcgtta aaaacacaga aggaaaagtg aattatatat tcacaaacgg ttcggattta 3060

tggttagacc aggccgagaa agaattcaaa gtggctgcag atgccgaact gaacatcacg 3120

gcgaagtatc aggacggcaa aatcgtagtg ccttgtacct ataaatatgc actgaatgcc 3180

acggatctaa atgtaagttt gttcatggca attctggaag ataacttaac ccgttatcaa 3240

tgtaacaact tgggaggtac ttctgacccg aatctcggag aatggggatt gggcggccag 3300

tacgctgcca gtgtagtggc accttacacg ttcaacgatg tagttcgcag catacccagt 3360

gcttactatg gtgtaagcgg tctgattcct tcttccgtaa cagccggtga ggagaatacg 3420

actagtctgg acttaaatat cccggaaaat atcattgatc tcacaaactg taaagtagta 3480

tgtatgatga tcgatgccaa cacgggaaac atcatcaatg cagccagagc tgatatcaat 3540

acagacgatt ataattccat cgaacaaaca aaagcagacg aaagtatctc cgtgaatgct 3600

gacaacaaca ccatccacat taatgccggc acaacggcac aagttacggt gtacagtgta 3660

gacggaagtg ttctcgacca agttgttatc gaaggagaag gtagcctgaa gctgcaaggc 3720

cataagggta tcgtcttggt tgaggtaacg accgagaata ctcgtgttgt gaagaaagta 3780

ttcgtaaaat aa 3792

<210> 8

<211> 1198

<212> PRT

<213> Paraprevotella xylaniphila

<400> 8

Met Phe Pro Lys Leu Ile Asp Leu Ser Ser Asn Gln Val Thr Glu Leu

1 5 10 15

Leu Thr Gly Glu Asp Tyr Ser Asn Leu Thr Ser Ala Ala Tyr Asp Val

20 25 30

Thr Asp Asp Gly Ser Met Val Ala Gly Ser Tyr Asp Gly Gln Pro Ala

35 40 45

Ile Tyr Ile Thr Ala Gln Lys Arg Trp Ser Met Leu Ala Val Pro Asp

50 55 60

Ser Cys Val Gly Gly Glu Val Thr Ala Ile Thr Pro Asp Gly His Tyr

65 70 75 80

Ala Val Gly Arg Gly Leu Tyr Ala Asn Thr Tyr Met Glu Ala Val Val

85 90 95

Met Trp Asp Leu Gln Gln Asp Gly Leu Leu Leu Asp Leu Ser Gly Ile

100 105 110

Pro Phe Glu Asp Met Val Gly Glu Asn Asn Asn Gln Arg Arg Leu Ile

115 120 125

Gly Val Thr Pro Asp Gly Asn Lys Val Leu Gly Cys Ile Ser Tyr Ser

130 135 140

Tyr Val Ser Pro Ala Gly Ile Phe Phe Phe Val Tyr Asp Arg Gln Thr

145 150 155 160

Lys Lys Cys Gln Pro Ile Gly Phe Asp Thr Thr Thr Glu Thr Thr Glu

165 170 175

Glu Gly Pro Gln Thr Val Trp Thr Pro Arg Ala Glu Gly Leu Tyr Phe

180 185 190

Ile Asn Ser Ala Asp Leu Ser Ala Asn Gly Arg Tyr Ile Thr Gly Glu

195 200 205

Ala Tyr Met Glu Asn Asp Ala Ala Lys Gly Tyr Val Tyr Asp Ile Glu

210 215 220

Thr Asp Lys Phe Thr Leu Tyr Ser Asp Ala Gln Asp Asp Met Pro Gly

225 230 235 240

Phe Thr Ala Cys Ser Asn Gly Ile Val Leu Gly Ala Thr Pro Pro Thr

245 250 255

Asn Pro Tyr Arg Asp Trp Ser Val Arg Val Gly Glu Tyr Trp Tyr Pro

260 265 270

Ile Ser Leu Ile Leu Ser Gln Arg Tyr Gly Ile Asn Phe Ser Thr Glu

275 280 285

Thr Asn Phe Ser Asn Thr Gly Thr Pro Ile Ala Thr Ser Leu Asp Gly

290 295 300

Lys Lys Ile Val Val Phe Val Ala Pro Gln Lys Glu Asn Tyr Ile Leu

305 310 315 320

Glu Leu Pro Glu Thr Leu Asp Ala Ala Thr Thr Asp Ile Asp Leu Leu

325 330 335

Gly Asn Tyr Ile Val Asn Pro Lys Ala Gly Ser Glu Phe Ser Ser Leu

340 345 350

Lys Thr Val Thr Phe Thr Phe Pro Tyr Glu Ile Glu Leu Leu Gly Asp

355 360 365

Lys Ser Asn Val Thr Phe Thr Asp Glu Asn Gly Lys Lys Thr Gly Thr

370 375 380

Ile Leu Ser Leu Lys Thr Asp Asp Lys Asp Leu Asn Ile Ser Phe Arg

385 390 395 400

Ser Thr Asn Leu Gln Ala Gly Val Lys Tyr Thr Leu Thr Leu Ser Ala

405 410 415

Gly Thr Ile Ala Ile Lys Gly Asp Thr Glu Arg Lys Asn Lys Glu Leu

420 425 430

Ser Val Thr Tyr Thr Gly Arg Asp Asn Val Pro Val Lys Met Val Ser

435 440 445

Ala Thr Pro Tyr Glu Asn Thr Glu Val Lys Gln Leu Asn Tyr Thr Asn

450 455 460

Asn Pro Ile Thr Ile Thr Phe Asp Thr Gln Leu Ser Leu Asn Glu Asp

465 470 475 480

Ala Ile Gly Tyr Leu Tyr Lys Asp Asp Glu Thr Thr Pro Asp Gly Thr

485 490 495

Leu Arg Leu Gly Val Thr Asp Asn Lys Leu Tyr Val Phe Pro Val Thr

500 505 510

Ala Arg Asn Leu Phe Lys Gly His Asn Tyr Lys Val Thr Val Pro Arg

515 520 525

Asn Ala Val Tyr Asp Ile Met Gly Asn Asn Gly Asn Asp Ser Ile Thr

530 535 540

Leu His Tyr Ile Gly Gly Phe Glu Arg Glu Val Ser Tyr Asp Asn Asp

545 550 555 560

Thr Leu Phe Val Glu Asp Phe Asn Asn Gly Phe Thr Lys Thr Met Leu

565 570 575

Tyr Glu Gly Asp His Asn Lys Pro Thr Glu Glu Met Gln Lys Met Asp

580 585 590

Phe Asn Asp Ser Asp Asn Tyr Pro Trp Thr Met Val Arg Asp Glu Asp

595 600 605

Asp Pro Asn Asn Tyr Ala Ala Ala Ser Thr Ser Leu Tyr Asp Pro Val

610 615 620

Gly Arg Ser Asp Asp Trp Met Val Thr Ala His Ile Tyr Ile Pro Asp

625 630 635 640

Asp Arg Cys Tyr Leu Gln Phe Asp Ala Gln Ser Phe Arg Glu Asn Lys

645 650 655

Lys Asp Ser Leu Tyr Val Leu Val Trp Val Thr Glu Asp Glu Phe Ser

660 665 670

Ser Met Asn Ser Glu Arg Ile Ala Lys Phe Lys Ala Glu Cys Asp Thr

675 680 685

Ile Tyr Ala Gly Ile Glu Thr Pro Gly Asp Tyr Glu Asp Phe Leu Ala

690 695 700

Gly Asp Trp Thr Glu His Thr Ala Ser Leu Gln Pro Tyr Ala Gly Lys

705 710 715 720

Asn Ile Tyr Ile Ala Phe Val Asn Ser Asn Glu Asn Gln Ser Cys Val

725 730 735

Phe Val Asp Asn Leu Leu Val Arg Gln Asp Gln Gln Phe Gln Ile Ser

740 745 750

Val Thr Thr Asp Gln Thr Val Val Lys Ala Lys Glu Ile Lys Ile Ser

755 760 765

Gly Gln Ile Arg Ile Thr Thr Pro Asp His Thr Tyr Ser Thr Leu Glu

770 775 780

Leu Thr Leu Lys Asp Ser Gln Gly Asn Met Val Asp Gln Ile Ser Glu

785 790 795 800

Ser Asn Leu Glu Leu Lys Glu Asn Asp Lys Tyr Asp Phe Ser Phe Asp

805 810 815

Lys Pro Leu Pro Leu Val Ile Gly Gln Glu Asn Glu Tyr Thr Ile Leu

820 825 830

Val Gln Leu Asp Glu Glu Gln Ser Glu Pro His Tyr Ser Val Lys Asn

835 840 845

Leu Ser Phe Gln Thr Val Lys Arg Val Val Leu Glu Glu Gly Thr Gly

850 855 860

Gln Asp Cys Pro Asn Cys Pro Gln Gly Ile Leu Ala Ile Glu Asn Leu

865 870 875 880

Glu Asp Val Tyr Gly Asp Leu Phe Ile Pro Val Ser Leu His Thr Tyr

885 890 895

Ser Gly Asp Pro Tyr Gly Thr Gly Phe Glu Ser Tyr Ala Ala Phe Leu

900 905 910

Asn Ile Thr Gly Tyr Pro Ser Gly Thr Val Asn Arg Thr Asn Gln Val

915 920 925

Pro Val Gly Ala Met Thr Ser Asp Glu Asn Gly Asn Ala Thr Phe Thr

930 935 940

Ser Thr Ala His Pro Cys Trp Leu Asp Tyr Val Ala Ala Glu Met Glu

945 950 955 960

Ile Ser Ala Asp Ala Asn Phe Asp Ile Ala Lys Ala Lys Val Glu Ala

965 970 975

Asp Ser Thr Ile Asn Met Thr Tyr Lys Tyr Gln Tyr Ala Leu Asn Val

980 985 990

Lys Gly Gln Asn Val Asn Leu Phe Thr Val Ile Leu Glu Asp Ser Leu

995 1000 1005

Ile Gly Lys Gln Arg Asn Ser Tyr Gly Ser Asn Ser Asp Pro Leu

1010 1015 1020

Leu Gly Glu Trp Gly Gln Gly Gly Lys Tyr Ser Ala Ala Thr Val

1025 1030 1035

Arg Asn Tyr Pro Phe Val Asp Val Val Arg Gly Val Val Gly Asp

1040 1045 1050

Thr Phe Tyr Gly Thr Ala Gly Tyr Ile Gln Pro Asn Val Glu Ala

1055 1060 1065

Gly Lys Glu Tyr Thr Ala Glu Ala Thr Phe Lys Leu Pro Ala Lys

1070 1075 1080

Val Leu Lys Ala Lys Asn Cys Lys Ile Val Thr Met Met Ile Asp

1085 1090 1095

Ala Asn Ser Gly Lys Val Ile Asn Ser Ala Arg Ala Lys Leu Asp

1100 1105 1110

Val Ser Asp Phe Ala Asn Ala Ile Ala Asp Ile Gln Ala Glu Lys

1115 1120 1125

Pro Asn Val Glu Ile Ala Val Val Asn Gly Gly Val Glu Ile Thr

1130 1135 1140

Thr Asp Ala Pro Ala Gln Ile Asn Leu Tyr Ser Leu Asn Gly Thr

1145 1150 1155

Leu Ile Gly Thr Ala Lys Ser Gln Gly Phe Val Ser Leu Ser Thr

1160 1165 1170

Asn Gly Tyr Arg Gly Leu Thr Leu Val Lys Val Thr Thr Gly His

1175 1180 1185

Gln Thr Leu Val Lys Lys Val Ile Val Lys

1190 1195

<210> 9

<211> 3597

<212> DNA

<213> Paraprevotella xylaniphila

<400> 9

atgtttccca aacttatcga cttaagctct aaccaagtca cggaactgct taccggcgaa 60

gactactcca acttgacaag cgcggcctat gacgtgacag acgacggtag catggtggca 120

ggaagctatg acggccaacc ggctatctat atcacagccc aaaaacgttg gagcatgttg 180

gccgtacccg acagctgtgt aggtggagaa gtcacagcca tcacccccga tggccattac 240

gccgtgggaa ggggcctata cgcaaatact tacatggaag ctgtcgtcat gtgggattta 300

caacaggacg gcctgcttct cgatttatcc ggcattccat ttgaagacat ggtaggcgaa 360

aataacaacc aacgccgtct gataggtgta acccctgatg gaaataaagt attaggctgc 420

atatcataca gctacgtaag ccctgccggc atcttctttt ttgtatatga ccgccaaacc 480

aagaagtgcc agcctatcgg tttcgatacc acaacagaaa caacagaaga aggccctcaa 540

acggtttgga caccgcgtgc cgaaggactt tattttatca actcagccga tctcagtgcc 600

aacggacgat atatcacagg cgaagcgtac atggaaaatg atgcagccaa aggttacgtt 660

tacgacatag aaacggataa attcacccta tacagcgatg cacaagatga catgcccggt 720

tttacagcat gcagtaatgg aatcgtgttg ggtgcaaccc ctccgaccaa tccttatcgt 780

gactggagcg tgcgcgtggg agaatattgg tatcccattt ctttgattct ctctcaacgt 840

tatggcatca acttctctac ggagactaat ttctccaata caggaacacc gatagccact 900

agcctggatg gcaaaaaaat cgttgtattc gtagctccac aaaaagaaaa ttatattttg 960

gaattaccgg agaccttgga tgcagcaacc acagatattg acttattagg taactacatc 1020

gtcaatccga aggccggaag tgaattctcg tctcttaaaa ccgtcacgtt tacattcccc 1080

tatgaaatag aactattagg agacaaaagc aatgtgacct ttacggacga aaacgggaaa 1140

aaaacgggta cgatcttaag tcttaaaacg gatgacaagg atttgaatat cagcttccgc 1200

agcacaaacc tgcaagcagg agtgaaatat accctgaccc tttctgccgg tactatcgcc 1260

atcaaaggcg acacggaacg taaaaacaaa gagctctccg tcacttatac aggacgtgat 1320

aatgttccgg taaaaatggt atctgccact ccgtatgaaa acacggaagt aaaacagctg 1380

aattacacca acaatcctat aaccataacg ttcgacaccc aattatccct caatgaggac 1440

gctatcggtt acctttacaa agacgacgaa acgaccccgg acggcacctt acgtttagga 1500

gtcacagaca ataaattata tgtgttccct gtcacagccc gcaatttgtt taagggacac 1560

aactacaaag tgacggttcc tcgcaacgcg gtatatgaca tcatgggcaa taatggtaat 1620

gactccatca ccttgcacta catcggcgga ttcgaacgcg aagtaagtta tgacaacgat 1680

acactcttcg tagaggattt caacaacggg ttcactaaga ccatgttgta tgaaggagac 1740

cataataaac ccacggagga aatgcaaaaa atggatttca acgatagcga taactatcct 1800

tggaccatgg tacgcgatga agatgatccc aataactatg ccgctgcttc cacttcgttg 1860

tatgaccccg tcgggaggtc tgacgactgg atggtcacag cgcatatcta tatcccggac 1920

gacagatgtt atttgcaatt tgatgcccaa agtttccggg agaataaaaa agatagcctt 1980

tatgtattgg tttgggtaac tgaagatgag ttttcttcaa tgaacagcga acgtatcgcc 2040

aagttcaaag ccgaatgcga caccatatat gccggcatcg aaactccggg agactatgaa 2100

gacttccttg ccggagactg gacggaacac acggcaagcc tacaacctta tgccggcaaa 2160

aacatctaca tcgcttttgt caacagcaat gaaaaccaaa gttgtgtatt cgtagacaac 2220

cttttggtga gacaagacca acagttccaa atcagcgtga ccacagacca aacggtggta 2280

aaagccaaag agataaaaat cagtggacaa atcagaatca cgacaccaga ccacacttac 2340

tcgaccttgg aactgacctt gaaagattct cagggaaaca tggtagacca aattagcgaa 2400

agcaacctgg aactgaaaga aaatgataag tatgacttct ccttcgacaa gccattaccc 2460

ttggttatcg gccaagaaaa cgaatatact atccttgtcc aattggatga agaacaaagt 2520

gaacctcact attcagtaaa gaacctgtct ttccagaccg ttaaacgggt tgtattggaa 2580

gaaggtaccg gacaggattg tccgaactgt ccgcaaggca ttctggccat tgaaaatctg 2640

gaagatgttt acggcgactt gttcatccct gtaagccttc atacttattc cggtgaccct 2700

tatggaactg gctttgaaag ctatgccgca ttcctgaaca tcacaggata cccttcgggc 2760

accgtcaatc gtaccaatca agtccctgtg ggagccatga cttcggatga gaatggaaat 2820

gccaccttca cctctacggc acatccttgt tggttagact atgtagctgc agaaatggaa 2880

ataagcgccg atgcaaactt cgacatcgca aaggctaagg tagaggcaga cagtacaatt 2940

aacatgactt ataaatacca atatgcccta aacgtgaaag gacaaaacgt aaacctcttt 3000

actgtaatat tagaagatag cctgattggc aagcaaagaa actcttatgg ttccaacagc 3060

gacccgcttt taggcgaatg gggccaggga ggtaaatact ctgccgccac cgtacgtaat 3120

tatcctttcg tagatgtagt gagaggcgta gtcggtgata cattctacgg cactgccggt 3180

tatatccaac cgaatgtcga ggccggtaag gaatatacgg ccgaagccac tttcaagctt 3240

cctgcaaaag tcctgaaagc taaaaactgt aagatcgtaa ccatgatgat agacgccaac 3300

agcggcaaag tcatcaattc ggcgcgcgcc aaattggatg tatcggattt tgctaatgcc 3360

atagctgaca tacaagcaga aaaaccgaat gtcgaaattg cagttgtaaa cggtggcgta 3420

gagattacca cagatgcccc ggcacaaatt aatttgtata gcctgaacgg cactttaatc 3480

ggaacagcca aaagccaagg ctttgtctca ttaagcacaa acggataccg tggccttact 3540

ttggtgaaag tgactaccgg acatcaaact ttagtgaaaa aagtcatcgt gaagtaa 3597

<210> 10

<211> 1260

<212> PRT

<213> Paraprevotella xylaniphila

<400> 10

Met Lys Lys Gly Leu Leu Lys Ile Ala Leu Ser Val Leu Gly Ala Ala

1 5 10 15

Phe Phe Leu Cys Ala Lys Ala Gln Gly Val Pro Gly Ala Ser Ile His

20 25 30

Pro Phe Val Phe Glu Asp Val Ser Ile Leu Thr Asn Ile Ser Asn Asn

35 40 45

Gly Lys Trp Ala Thr Ala His Gly Thr Glu Glu Ser Lys Thr Met Phe

50 55 60

Pro Lys Leu Ile Asp Leu Ser Ser Asn Gln Val Thr Glu Leu Leu Thr

65 70 75 80

Gly Glu Asp Tyr Ser Asn Leu Thr Ser Ala Ala Tyr Asp Val Thr Asp

85 90 95

Asp Gly Ser Met Val Ala Gly Ser Tyr Asp Gly Gln Pro Ala Ile Tyr

100 105 110

Ile Thr Ala Gln Lys Arg Trp Ser Met Leu Ala Val Pro Asp Ser Cys

115 120 125

Val Gly Gly Glu Val Thr Ala Ile Thr Pro Asp Gly His Tyr Ala Val

130 135 140

Gly Arg Gly Leu Tyr Ala Asp Thr Tyr Met Glu Ala Val Val Met Trp

145 150 155 160

Asp Leu Gln Gln Asp Gly Leu Leu Leu Asp Leu Ser Gly Ile Pro Phe

165 170 175

Glu Asp Met Val Gly Glu Asn Asn Asn Gln Arg Arg Leu Ile Gly Val

180 185 190

Thr Pro Asp Gly Asn Lys Val Leu Gly Cys Ile Ser Tyr Ser Tyr Val

195 200 205

Ser Pro Ala Gly Ile Phe Phe Phe Val Tyr Asp Arg Gln Thr Lys Lys

210 215 220

Cys Gln Pro Ile Gly Phe Asp Thr Thr Thr Glu Thr Thr Glu Glu Gly

225 230 235 240

Pro Gln Thr Val Trp Thr Pro Arg Ala Glu Gly Leu Tyr Phe Ile Asn

245 250 255

Ser Ala Asn Leu Ser Ala Asn Gly Arg Tyr Ile Thr Gly Glu Ala Tyr

260 265 270

Met Glu Asn Asp Ala Ala Lys Gly Tyr Val Tyr Asp Ile Glu Thr Asp

275 280 285

Lys Phe Thr Leu Tyr Ser Asp Ala Gln Asp Asp Met Pro Gly Phe Thr

290 295 300

Ala Cys Ser Asn Gly Ile Val Leu Gly Ala Thr Pro Pro Thr Asn Pro

305 310 315 320

Tyr Arg Asp Trp Ser Val Arg Val Gly Glu Tyr Trp Tyr Pro Ile Ser

325 330 335

Leu Ile Leu Ser Gln Arg Tyr Gly Ile Asn Phe Ser Thr Glu Thr Asn

340 345 350

Phe Ser Asn Thr Gly Thr Pro Ile Ala Thr Ser Leu Asp Gly Lys Lys

355 360 365

Ile Val Val Phe Val Ala Pro Gln Lys Glu Asn Tyr Ile Leu Glu Leu

370 375 380

Pro Glu Thr Leu Asp Ala Ala Thr Thr Asp Ile Asp Leu Leu Gly Asn

385 390 395 400

Tyr Ile Val Asn Pro Lys Ala Gly Ser Glu Phe Ser Ser Leu Lys Thr

405 410 415

Val Thr Phe Thr Phe Pro Tyr Glu Ile Glu Leu Leu Gly Asp Lys Ser

420 425 430

Asn Val Thr Phe Thr Asp Glu Asn Gly Lys Lys Thr Gly Thr Ile Leu

435 440 445

Ser Leu Lys Thr Asp Asp Lys Asp Leu Asn Ile Ser Phe Arg Ser Thr

450 455 460

Asn Leu Gln Ala Gly Val Lys Tyr Thr Leu Thr Leu Ser Ala Gly Thr

465 470 475 480

Ile Ala Ile Lys Gly Asp Thr Glu Arg Lys Asn Lys Glu Leu Ser Val

485 490 495

Thr Tyr Thr Gly Arg Asp Asn Val Pro Val Lys Met Val Ser Ala Thr

500 505 510

Pro Tyr Glu Asn Thr Glu Val Lys Gln Leu Asn Tyr Thr Asn Asn Pro

515 520 525

Ile Thr Ile Thr Phe Asp Thr Gln Leu Ser Leu Asn Glu Asp Ala Ile

530 535 540

Gly Tyr Leu Tyr Lys Asp Asp Glu Thr Thr Pro Asp Gly Thr Leu Arg

545 550 555 560

Leu Gly Val Thr Asp Asn Lys Leu Tyr Val Phe Pro Val Thr Ala Arg

565 570 575

Asn Leu Phe Lys Gly His Asn Tyr Lys Val Thr Val Pro Arg Asn Ala

580 585 590

Val Tyr Asp Ile Met Gly Asn Asn Gly Asn Asp Ser Ile Thr Leu His

595 600 605

Tyr Ile Gly Gly Phe Glu Arg Glu Val Ser Tyr Asp Asn Asp Thr Leu

610 615 620

Phe Val Glu Asp Phe Asn Asn Gly Phe Thr Lys Thr Met Leu Tyr Glu

625 630 635 640

Gly Asp His Asn Lys Pro Thr Glu Glu Met Gln Lys Met Asp Phe Asn

645 650 655

Asp Ser Asp Asn Tyr Pro Trp Thr Met Val Arg Asp Glu Asp Asp Pro

660 665 670

Asn Asn Tyr Ala Ala Ala Ser Thr Ser Leu Tyr Asp Pro Val Gly Arg

675 680 685

Ser Asp Asp Trp Met Val Thr Ala His Ile Tyr Ile Pro Asp Asp Arg

690 695 700

Cys Tyr Leu Gln Phe Asp Ala Gln Ser Phe Arg Glu Asn Lys Lys Asp

705 710 715 720

Ser Leu Tyr Val Leu Val Trp Val Thr Glu Asp Glu Phe Ser Ser Met

725 730 735

Asn Ser Glu Arg Ile Ala Lys Phe Lys Ala Glu Cys Asp Thr Ile Tyr

740 745 750

Ala Gly Ile Glu Thr Pro Gly Asp Tyr Glu Asp Phe Leu Ala Gly Asp

755 760 765

Trp Thr Glu His Thr Ala Ser Leu Gln Pro Tyr Ala Gly Lys Asn Ile

770 775 780

Tyr Ile Ala Phe Val Asn Ser Asn Glu Asn Gln Ser Cys Val Phe Val

785 790 795 800

Asp Asn Leu Leu Val Arg Gln Asp Gln Gln Phe Gln Ile Ser Val Thr

805 810 815

Thr Asp Gln Thr Val Val Lys Ala Lys Glu Ile Lys Ile Ser Gly Gln

820 825 830

Ile Arg Ile Thr Thr Pro Asp His Thr Tyr Ser Thr Leu Glu Leu Thr

835 840 845

Leu Lys Asp Ser Gln Gly Asn Met Val Asp Gln Ile Ser Glu Ser Asn

850 855 860

Leu Glu Leu Lys Glu Asn Asp Lys Tyr Asp Phe Ser Phe Asp Lys Pro

865 870 875 880

Leu Pro Leu Val Ile Gly Gln Glu Asn Glu Tyr Thr Ile Leu Val Gln

885 890 895

Leu Asp Glu Glu Gln Ser Glu Pro His Tyr Ser Val Lys Asn Leu Ser

900 905 910

Phe Gln Thr Val Lys Arg Val Val Leu Glu Glu Gly Thr Gly Gln Asp

915 920 925

Cys Pro Asn Cys Pro Gln Gly Ile Leu Ala Ile Glu Asn Leu Glu Asp

930 935 940

Val Tyr Gly Asp Leu Phe Ile Pro Val Ser Leu His Thr Tyr Ser Gly

945 950 955 960

Asp Pro Tyr Gly Thr Gly Phe Glu Ser Tyr Ala Ala Phe Leu Asn Ile

965 970 975

Thr Gly Tyr Pro Ser Gly Thr Val Asn Arg Thr Asn Gln Val Pro Val

980 985 990

Gly Ala Met Thr Ser Asp Glu Asn Gly Asn Ala Thr Phe Thr Ser Thr

995 1000 1005

Ala His Pro Cys Trp Leu Asp Tyr Val Ala Ala Glu Met Glu Ile

1010 1015 1020

Ser Ala Asp Ala Asn Phe Asp Ile Ala Lys Ala Lys Val Glu Ala

1025 1030 1035

Asp Ser Thr Ile Asn Met Thr Tyr Lys Tyr Gln Tyr Ala Leu Asn

1040 1045 1050

Val Lys Gly Gln Asn Val Asn Leu Phe Thr Val Ile Leu Glu Asp

1055 1060 1065

Ser Leu Ile Gly Lys Gln Arg Asn Ser Tyr Gly Ser Asn Ser Asp

1070 1075 1080

Pro Leu Leu Gly Glu Trp Gly Gln Gly Gly Lys Tyr Ser Ala Ala

1085 1090 1095

Thr Val Arg Asn Tyr Pro Phe Val Asp Val Val Arg Gly Val Val

1100 1105 1110

Gly Asp Thr Phe Tyr Gly Thr Ala Gly Tyr Ile Gln Pro Asn Val

1115 1120 1125

Glu Ala Gly Lys Glu Tyr Thr Ala Glu Ala Thr Phe Lys Leu Pro

1130 1135 1140

Ala Lys Val Leu Lys Ala Lys Asn Cys Lys Ile Val Thr Met Met

1145 1150 1155

Ile Asp Ala Asn Ser Gly Lys Val Ile Asn Ser Ala Arg Ala Lys

1160 1165 1170

Leu Asp Val Ser Asp Phe Ala Asn Ala Ile Ala Asp Ile Gln Ala

1175 1180 1185

Glu Lys Pro Asn Val Glu Ile Ala Val Val Asn Gly Gly Val Glu

1190 1195 1200

Ile Thr Thr Asp Ala Pro Ala Gln Ile Asn Leu Tyr Ser Leu Asn

1205 1210 1215

Gly Thr Leu Ile Gly Thr Ala Lys Ser Gln Gly Phe Val Ser Leu

1220 1225 1230

Ser Thr Asn Gly Tyr Arg Gly Leu Thr Leu Val Lys Val Thr Thr

1235 1240 1245

Gly His Gln Thr Leu Val Lys Lys Val Ile Val Lys

1250 1255 1260

<210> 11

<211> 3783

<212> DNA

<213> Paraprevotella xylaniphila

<400> 11

atgaagaaag gtctattaaa aattgccctc tcagtattag gagcggcgtt ttttctttgc 60

gcgaaagcgc aaggggttcc aggagcctcc attcatccgt ttgtatttga agatgtatcc 120

atccttacta acatttcaaa caacgggaaa tgggctaccg ctcatggtac agaggaaagc 180

aaaacgatgt ttcccaaact tatcgactta agctctaacc aagtcacgga actgcttacc 240

ggcgaagact actccaactt gacaagcgcg gcctatgacg tgacagacga cggtagcatg 300

gtggcaggaa gctatgacgg ccaaccggct atctatatca cagcccaaaa acgttggagc 360

atgttggccg tacccgacag ctgtgtaggt ggagaagtca cagccatcac ccccgatggc 420

cattacgccg tgggaagggg cctatacgca gatacttaca tggaagctgt cgtcatgtgg 480

gatttacaac aggacggcct gcttctcgat ttatccggca ttccatttga agacatggta 540

ggcgaaaata acaaccaacg ccgtctgata ggtgtaaccc ctgatggaaa taaagtatta 600

ggctgcatat catacagcta cgtaagccct gccggcatct tcttttttgt atatgaccgc 660

caaaccaaga agtgccagcc tatcggtttc gataccacaa cagaaacaac agaagaaggc 720

cctcaaacgg tttggacacc gcgtgccgaa ggactttatt ttatcaactc agccaatctc 780

agtgctaacg gacgatatat cacaggcgaa gcgtacatgg aaaatgatgc agccaaaggt 840

tacgtttacg acatagaaac ggataaattc accctataca gcgatgcaca agatgacatg 900

cccggtttta cagcatgcag taatggaatc gtgttgggtg caacccctcc gaccaatcct 960

tatcgtgact ggagcgtgcg cgtgggagaa tattggtatc ccatttcttt gattctctct 1020

caacgttatg gcatcaactt ctctacggag actaattttt ctaatacagg aacaccgata 1080

gccactagcc tggatggcaa aaaaatcgtt gtattcgtag ctccacaaaa agaaaattat 1140

attttggaat taccggagac cttggatgca gcaaccacag atattgactt attaggtaac 1200

tacatcgtca atccgaaggc cggaagtgaa ttctcgtctc ttaaaaccgt cacgtttaca 1260

ttcccctatg aaatagaact attaggagac aaaagcaatg tgacctttac ggacgaaaac 1320

gggaaaaaaa cgggtacgat cttaagtctt aaaacggatg acaaggattt gaatatcagc 1380

ttccgcagca caaacctgca agcaggagtg aaatataccc tgaccctttc tgccggtact 1440

atcgccatca aaggcgacac ggaacgtaaa aacaaagagc tctccgtcac ttatacagga 1500

cgtgataatg ttccggtaaa aatggtatct gccactccgt atgaaaacac ggaagtaaaa 1560

cagctgaatt acaccaacaa tcctataacc ataacgttcg acacccaatt atccctcaat 1620

gaggacgcta tcggttacct ttacaaagac gacgaaacga ccccggacgg caccttacgt 1680

ttaggagtca cagacaataa attatatgtg ttccctgtca cagcccgcaa tttgtttaag 1740

ggacacaact acaaagtgac ggttcctcgc aacgcggtat atgacatcat gggcaataat 1800

ggtaatgact ccatcacctt gcactacatc ggcggattcg aacgcgaagt aagttatgac 1860

aacgatacac tcttcgtaga ggatttcaac aacgggttca ctaagaccat gttgtatgaa 1920

ggagaccata ataaacccac ggaggaaatg caaaaaatgg atttcaacga tagcgataac 1980

tatccttgga ccatggtacg cgatgaagat gatcccaata actatgccgc tgcttccact 2040

tcgttgtatg accccgtcgg gaggtctgac gactggatgg tcacagcgca tatctatatc 2100

ccggacgaca gatgttattt gcaatttgat gcccaaagtt tccgggagaa taaaaaagat 2160

agcctttatg tattggtttg ggtaactgaa gatgagtttt cttcaatgaa cagcgaacgt 2220

atcgccaagt tcaaagccga atgcgacacc atatatgccg gcatcgaaac tccgggagac 2280

tatgaagact tccttgccgg agactggacg gaacacacgg caagcctaca accttatgcc 2340

ggcaaaaaca tctacatcgc ttttgtcaac agcaatgaaa accaaagttg tgtattcgta 2400

gacaaccttt tggtgagaca agaccaacag ttccaaatca gcgtgaccac agaccaaacg 2460

gtggtaaaag ccaaagagat aaaaatcagt ggacaaatca gaatcacgac accagaccac 2520

acttactcga ccttggaact aaccttgaaa gattctcagg gaaacatggt agaccaaatt 2580

agcgaaagca acctggaact gaaagaaaat gataagtatg acttctcctt cgacaagcca 2640

ttacccttgg ttatcggcca agaaaacgaa tatactatcc ttgtccaatt ggatgaagaa 2700

caaagtgaac ctcactattc agtaaagaac ctgtctttcc agaccgttaa acgggttgta 2760

ttggaagaag gtaccggaca ggattgtccg aactgtccgc aaggcattct ggccattgaa 2820

aatctggaag atgtttacgg cgacttgttc atccctgtaa gccttcatac ttattccggt 2880

gacccttatg gaactggctt tgaaagctat gccgcattcc tgaacatcac aggataccct 2940

tcgggcaccg tcaatcgtac caatcaagtc cctgtgggag ccatgacttc ggatgagaat 3000

ggaaatgcca ccttcacctc tacggcacat ccttgttggt tagactatgt agctgcagaa 3060

atggaaataa gcgccgatgc aaacttcgac atcgcaaagg ctaaggtaga ggcagacagt 3120

acgattaaca tgacttataa ataccaatat gccctaaacg tgaaaggaca aaacgtaaac 3180

ctctttactg taatattaga agatagcctg attggcaagc aaagaaactc ttatggttcc 3240

aacagcgacc cacttttagg cgaatggggc cagggaggta aatactctgc cgccaccgta 3300

cgtaattatc ctttcgtaga tgtagtgaga ggcgtagtcg gtgatacatt ctacggcact 3360

gccggttata tccaaccgaa tgtcgaggcc ggtaaggaat atacggccga agccactttc 3420

aagcttcctg caaaagtcct gaaagctaaa aactgtaaga tcgtaaccat gatgatagac 3480

gccaacagcg gcaaagtcat caattcggcg cgcgccaaat tggatgtatc ggattttgct 3540

aatgccatag ctgacataca agcagaaaaa ccgaatgtcg aaattgcagt tgtaaacggt 3600

ggcgtagaga ttaccacaga tgccccggca caaattaatt tgtatagcct gaacggcact 3660

ttaatcggaa cagccaaaag ccaaggcttt gtctcattaa gcacaaacgg ataccgtggc 3720

cttactttgg tgaaagtgac taccggacat caaactttag tgaaaaaagt catcgtgaag 3780

taa 3783

<210> 12

<211> 1242

<212> PRT

<213> Prevotella rara

<400> 12

Met Gly Leu Leu Trp Ala Ile Pro Ser Ser Ala Gln Glu Ala Gln Leu

1 5 10 15

Thr Thr Val Ser Phe Pro Lys Ala Ala Ala Phe Thr Ser Leu Ser Asp

20 25 30

Asn Gly Leu Trp Ala Thr Ala Ala Gly Val Asn Asp Asp Asp Gln Ser

35 40 45

Lys Tyr Ala Tyr Pro Tyr Leu Ile Asn Val Glu Thr Gly Ala Leu Thr

50 55 60

Glu Leu Trp Val Glu Ala Asp Leu Ile Lys Ser Leu Glu Ala Thr Asp

65 70 75 80

Val Thr Asn Asp Gly Lys Ile Ile Val Gly Thr Tyr Asp Ser Lys Pro

85 90 95

Ala Tyr Tyr Asp Met Asn Gln Gly Lys Trp Ile Thr Leu Gln Ser Glu

100 105 110

Asn Pro Gly Lys Ala Thr Ser Val Thr Pro Asp Gly Lys Tyr Ile Ser

115 120 125

Gly Trp Ser Asn Ser Gly Ser Phe Ser Gly Asp Ala Tyr Val Glu Thr

130 135 140

Pro Leu Leu Trp Glu Lys Gln Ser Asp Gly Thr Tyr Arg Ala Ile Asp

145 150 155 160

Val Tyr Ala Glu Leu Pro Asn Phe Pro Lys Lys Thr Lys Leu Gly Thr

165 170 175

Asn Thr Gln Gln Val Arg Ile Asp Asn Val Ser Pro Asp Gly Asn Ile

180 185 190

Leu Ser Gly Ile Ile Asn Phe Val Thr Pro Ala Thr Val Cys Tyr Tyr

195 200 205

Val Tyr Asn Lys Thr Thr Gln Glu Cys Lys Tyr Val Asp Asn Ala Leu

210 215 220

Gly Glu Val Pro Asp Glu Thr Phe Val Asp Glu Ser Thr Met Ser Asn

225 230 235 240

Asn Gly Lys Tyr Leu Thr Gly Ile Val Gln Val Ala Gly Gly Ser Tyr

245 250 255

Ile Ser Ser Tyr Leu Tyr Asn Thr Ser Asp Asn Ser Cys Ile Leu Tyr

260 265 270

Asn Thr Glu Ser Glu Glu Gln Asp Arg Ala Gly Ser Ala Val Ser Asn

275 280 285

Thr Gly Val Val Phe Ala Cys Ser Pro Ala Val Asn Pro Val Arg Ser

290 295 300

Ala Tyr Val Arg Val Gly Ser Leu Trp Tyr Gly Ile Asp Glu Ile Leu

305 310 315 320

Ser Gly Arg Tyr Asp Met Asn Phe Tyr Glu Arg Thr Gly Tyr Asp Phe

325 330 335

Thr Gly Thr Ile Gln Gly Val Ser Asp Asp Glu Lys Thr Ile Ile Gly

340 345 350

Met Ser Glu Thr Lys Thr Lys Gly Tyr Ile Ile Arg Leu Pro Glu Thr

355 360 365

Ile Ser Glu Ala Ala Ser Ser Val Asn Pro Leu Gly Thr Tyr Ala Val

370 375 380

Ser Pro Ala Tyr Gly Ser Lys Phe Ala Lys Phe Ser Gln Met Lys Leu

385 390 395 400

Ala Phe Ser Lys Ser Ala Ala Val Thr Ser Gly Val Lys Ala Gln Phe

405 410 415

Met Asp Ala Ser Gly Lys Val Leu Arg Glu Tyr Asn Ile Thr Ala Gln

420 425 430

Ser Gly Asn Lys Thr Phe Thr Ile Gly Gly Ile Pro Gln Ala Leu Asn

435 440 445

Ala Gly Glu Glu Tyr Thr Met Lys Ile Pro Ala Gly Ala Phe Tyr Leu

450 455 460

Met Ala Asp Asn Ser Ile Lys Ser Asp Glu Ile Thr Ile Lys Tyr Ile

465 470 475 480

Gly Arg Ala Glu Ala Pro Ala Ala Val Gln Gln Val Ser Pro Ala Asp

485 490 495

Gln Ala Asn Val Ser Glu Ile Ser Ser Glu His Pro Val Gln Ile Leu

500 505 510

Phe Asp Leu Ser Ile Ala Ile Ser Glu Asp Ala Lys Ala Tyr Leu Tyr

515 520 525

Arg Asp Gly Gln Thr Ser Pro Val Cys Glu Leu Asn Phe Val Gln Gly

530 535 540

Ala Glu Ser Thr Ile Leu Leu Tyr Pro Ser Leu Lys Arg Tyr Leu Glu

545 550 555 560

Lys Asp Glu Lys Tyr Lys Val Val Val Glu Ala Gly Ser Val Thr Asp

565 570 575

Ile Met Gly Phe Cys Pro Asn Asn Glu Ile Thr Ile Asn Tyr Thr Gly

580 585 590

Ala Tyr Glu Pro Glu Ile Ser Ser Asp Gly Ser Leu Phe Ser Asp Asp

595 600 605

Phe Asn Asp Pro Ser Asn Ser Met Val Lys Tyr Leu Leu Tyr Glu Gly

610 615 620

Asp His Asn Lys Pro Ser Ser Ala Met Gln Asp Leu Gly Phe Asp Ala

625 630 635 640

Asp Asn Ser Pro Trp Leu Phe Val Ile Arg Glu Ser Glu Ser Ser Ser

645 650 655

Asp Tyr Cys Ala Ala Ser Thr Ser Ile Tyr Asn Pro Thr Gly Lys Ser

660 665 670

Asp Asp Trp Met Ala Ile Pro Arg Leu Ser Ile Glu Asn Ala Asp Tyr

675 680 685

Tyr Leu Ser Phe Asp Val Gln Ser Tyr Tyr Lys Ser Lys Ala Asp Arg

690 695 700

Leu Lys Val Leu Val Leu Glu Asp Asp Ala Val Tyr Ser Asn Phe Thr

705 710 715 720

Thr Glu Leu Tyr Glu Lys Phe Lys Ala Asn Gly Lys Val Leu Tyr Asp

725 730 735

Glu Gln Leu Ser Pro Gly Glu Ser Glu Glu Asn Leu Thr Gly Asp Trp

740 745 750

Thr His Val Glu Lys Ser Leu Ala Glu Tyr Ala Gly Lys Asn Ile Tyr

755 760 765

Ile Ala Phe Val Asn Glu Asn Glu Asn Gln Ser Met Ile Phe Leu Asp

770 775 780

Asn Leu Lys Val Tyr Tyr Lys Gly Asp Phe Asn Phe Val Pro Asn Val

785 790 795 800

Glu Ser Thr Gln Val Asn Lys Glu Ser Thr Asn Val Gly Val Ile Val

805 810 815

Lys Val Thr Ser Asp Lys Thr Tyr Asn Thr Ile Asn Ala Thr Leu Thr

820 825 830

Ser Glu Asp Gly Ser Phe Lys Ser Thr Tyr Thr Asp Thr Pro Ser Lys

835 840 845

Pro Ile Thr Ser Ala Glu Asn Tyr Ser Phe Thr Phe Pro Asp Lys Leu

850 855 860

Pro Leu Thr Val Gly Ala Gln Asn Lys Tyr Thr Ile Ser Ile Asp Leu

865 870 875 880

Gly Gly Thr Val Tyr Ser Gln Thr Asn Thr Ile Ser Asn Leu Ala Phe

885 890 895

Glu Thr Thr Lys His Val Val Ile Glu Glu Ser Thr Gly Gln Gln Cys

900 905 910

Lys Asn Cys Pro Gln Gly Ile Leu Ala Met Glu Asn Leu Glu Lys Leu

915 920 925

Tyr Gly Glu Gln Val Ile Pro Ile Ala Ile His Ser Ser Ile Ile Gly

930 935 940

Thr Asp Gln Phe Ala Tyr Glu Asn Tyr Asn Thr Tyr Phe Gly Ile Thr

945 950 955 960

Ala Gln Pro Met Gly Leu Val Asn Arg Ile Asp Thr Leu Tyr Ala Pro

965 970 975

Met Tyr Val Asp Gly Ser Glu Gln Tyr His Phe Asp Ser Pro Glu Gly

980 985 990

Asn Asn Thr Phe Tyr Asp Val Ala Gln Arg Glu Phe Glu Asn Tyr Ala

995 1000 1005

Ile Ala Asp Val Ser Ile Asp Lys Ala Ile Tyr Asp Ala Ser Ser

1010 1015 1020

Lys Asn Ile Gln Ile Thr Gly Asn Val Asn Tyr Ala Leu Thr Met

1025 1030 1035

Asn Ser Leu Asn His Asn Leu Ala Phe Val Val Leu Glu Asp Gly

1040 1045 1050

Leu Glu Gly Pro Gln Leu Asn Gly Phe Tyr Val Leu Asp Gln Pro

1055 1060 1065

Ile Phe Gly Glu Phe Gly Lys Gly Gly Lys Tyr Gly Ser Ser Val

1070 1075 1080

Ala Val Val Thr Phe Asn Asp Val Ala Arg Lys Val Pro Asn Asp

1085 1090 1095

Asn Phe Ala Gly Glu Ser Gly Phe Ile Pro Val Ser Val Thr Ala

1100 1105 1110

Gly Gln Pro Val Ala Phe Asn Lys Val Phe Ser Leu Pro Glu Asn

1115 1120 1125

Val Lys Asn Trp Asp Asn Thr Lys Val Val Val Met Leu Leu Asp

1130 1135 1140

Ala Asn Thr Glu Arg Val Leu Asn Ala Ala Arg Leu Lys Met Ser

1145 1150 1155

Ala Gly Thr Ala Gly Ile Asn Asp Ala Thr Val Ser Glu Asn Gly

1160 1165 1170

Ile Thr Ile Ser Gly Glu Asn Gly Ser Ile Ser Val Asn Gly Gly

1175 1180 1185

Ser Asp Leu Asn Val Thr Val Tyr Asp Val Ser Gly Ser Val Ile

1190 1195 1200

Ser Asn Val Asn Ser Thr Ser Gly Ala Val Lys Val Ser Thr Gly

1205 1210 1215

Gly Lys Asn Gly Leu Phe Ile Val Lys Ala Thr Ser Asp Gly Val

1220 1225 1230

Ser Val Val Lys Lys Val Ile Val Lys

1235 1240

<210> 13

<211> 3729

<212> DNA

<213> Prevotella rara

<400> 13

ttgggcctgc tgtgggcaat tccgtcttca gcacaggaag cacagcttac cactgtgagt 60

ttccccaagg ctgcagcttt cacatctttg tcagacaatg gcttgtgggc tactgctgct 120

ggtgtgaatg atgatgatca aagtaagtat gcctatcctt atcttattaa tgtagagaca 180

ggtgctctta ctgaactttg ggttgaagca gacttgataa agtcactcga agcaacagac 240

gttactaatg atggaaaaat catcgttggt acctatgaca gcaagcctgc ttattatgac 300

atgaatcagg gtaagtggat cactcttcag tctgagaatc ctggaaaagc tacatctgta 360

acacctgatg gcaaatacat cagcggatgg agcaattctg gtagcttctc aggtgatgct 420

tatgttgaaa caccgctttt gtgggagaag cagtctgatg gtacatatcg tgcaattgat 480

gtgtatgctg aacttcctaa cttcccaaag aaaacaaagc ttggaacaaa cactcagcag 540

gtacgtatag acaatgttag tcctgatggt aacatccttt ctggtattat taactttgta 600

actcctgcaa ctgtttgcta ttatgtttac aacaagacaa ctcaggagtg caaatatgta 660

gacaatgctc tcggagaagt gcctgacgaa acatttgttg acgagtcaac aatgagtaac 720

aatggtaaat atcttaccgg tatcgtacag gtggctggtg gttcatacat ttcaagctat 780

ctttacaata caagtgacaa ctcttgtata ttatataata cagaaagcga agagcaagac 840

cgtgccggaa gtgctgtttc aaatacaggc gttgtatttg cttgcagtcc tgccgtaaac 900

cctgtccgtt ctgcttacgt acgtgtgggt agtctttggt acggaataga cgaaattctt 960

tctggccgtt atgatatgaa cttctatgag cgtactggct atgatttcac aggaacaatt 1020

caaggagttt ctgatgatga aaagaccata atcggtatgt ctgaaacaaa gacaaagggt 1080

tatatcattc gtcttcctga aactatcagc gaagctgcaa gcagcgtgaa tccattaggc 1140

acttatgccg tttctcctgc ttatggctct aagtttgcta aattcagtca gatgaagttg 1200

gctttctcta agtctgccgc tgtaacatct ggtgttaagg ctcagttcat ggacgcttct 1260

ggtaaggtgt tgagagagta taacattacg gctcagtctg gtaacaagac attcactatt 1320

ggcggtatac ctcaggctct gaatgcaggc gaggaatata caatgaagat tcctgctgga 1380

gcattctatt tgatggcaga caactctatc aagagtgatg aaatcacaat taagtatatc 1440

ggccgtgccg aggctccggc tgctgtacag caggtttctc ctgcagatca ggcaaatgta 1500

tctgaaataa gttcagagca tcctgtacag attctcttcg atttgtctat cgctatatct 1560

gaagatgcta aggcttatct gtatagagat ggtcagacat caccggtttg tgaacttaac 1620

tttgtgcagg gagcagaaag cacaatcctt ctgtatccta gcctcaagcg ttatcttgaa 1680

aaagatgaga aatataaggt tgttgtagag gctggttctg ttacagatat tatgggattc 1740

tgtccgaaca atgagattac aattaattac acaggtgcat acgagcctga aatcagttct 1800

gacggaagcc tcttctctga tgacttcaat gatcctagca actctatggt taaatacctc 1860

ctttatgaag gtgaccacaa caagccgtca agtgcaatgc aggatttagg ctttgatgcc 1920

gacaattctc cttggctttt cgtaatccgt gaatcagaaa gcagctctga ctattgtgca 1980

gcttcaacat ctatctataa tcctacagga aaatctgacg actggatggc aataccccgt 2040

ctgtcaatag aaaatgccga ctattatctc agctttgacg ttcagtcata ctataagtca 2100

aaggcagaca gacttaaggt tctcgttctt gaggatgatg ctgtttattc taacttcaca 2160

acagaacttt atgagaagtt caaggctaat ggtaaggttc tttatgacga gcagctttct 2220

cccggtgagt cagaggaaaa tctgacaggt gactggacac atgttgagaa atctctcgca 2280

gaatatgcag gtaagaatat ctatatcgct ttcgttaatg agaacgaaaa tcagagtatg 2340

atcttcttgg ataatctgaa ggtttactac aagggcgact tcaacttcgt tcctaatgta 2400

gaaagcacgc aggttaacaa agagtctaca aatgttggtg taatagttaa ggttacaagt 2460

gataagactt ataacactat taatgcaaca cttacttctg aagacggctc attcaagagc 2520

acttatacag atacacctag caagcctata acaagtgcag agaattattc tttcacattc 2580

cctgacaagt tgcctctgac tgtaggtgca cagaataagt atactataag cattgatctc 2640

ggcggtactg tttacagcca gacaaacact atcagcaacc ttgcttttga gacaacaaag 2700

catgtagtta tagaagaaag cactggtcag cagtgtaaga actgtcctca gggaatcctt 2760

gcaatggaaa atcttgagaa actttacggt gagcaggtta tacctatcgc tattcatagc 2820

tctataatcg gaacagacca gtttgcttat gagaactata atacatattt cggtataaca 2880

gcacagccta tgggtctggt taatcgtata gacactcttt atgcgccgat gtatgttgat 2940

ggaagcgagc agtatcactt cgattcacca gaaggcaaca atacattcta tgatgtagct 3000

cagcgtgagt tcgaaaacta tgcaattgca gatgtcagca tcgacaaggc tatttacgat 3060

gcttcaagca agaacataca gattacaggt aatgtaaact atgctctaac catgaactct 3120

ctgaaccaca atcttgcatt tgttgttctt gaggatggtt tggaaggacc tcagcttaat 3180

ggtttctacg ttctcgacca gccgatattc ggtgagttcg gtaagggcgg caagtatggt 3240

tcaagcgttg ctgttgtaac attcaacgat gttgcacgta aagtgccaaa cgacaacttc 3300

gcaggtgaat caggtttcat ccctgtaagc gtaactgcag gtcagcctgt tgcattcaat 3360

aaggtattca gcttgcctga aaacgttaag aactgggata acactaaggt tgttgttatg 3420

ttgctcgatg caaatacaga gcgcgttctg aatgctgcaa gacttaagat gtcagccgga 3480

acagctggaa tcaacgatgc aacagtatct gaaaacggta tcacaatttc tggtgagaac 3540

ggttcaataa gcgttaacgg cggaagcgat ctcaacgtta cagtttatga tgtaagcgga 3600

agcgtaataa gcaatgtcaa ctctacttct ggtgctgtta aggtaagcac aggcggaaag 3660

aacggcttgt ttatcgttaa ggctacaagc gacggcgtaa gcgttgttaa gaaagtaatc 3720

gttaaataa 3729

<210> 14

<211> 1230

<212> PRT

<213> Prevotella rodentium

<400> 14

Met Arg Lys Asn Lys Leu Lys Thr Phe Leu Leu Ala Val Ile Gly Ile

1 5 10 15

Ser Ser Phe Ala Cys Val Thr Leu His Ala Gln Asn Val Thr Glu Pro

20 25 30

Lys Met Thr Leu Tyr Asp Phe Lys Asp Gln Ser Met Ile Tyr Ser Leu

35 40 45

Ser Asp Asn Gly Gln Trp Ala Val Ser Tyr Gly Thr Ser Pro Thr Asp

50 55 60

Ala Ser Arg Tyr Thr Asn Ala Arg Arg Thr Asn Val Lys Thr Lys Glu

65 70 75 80

Ser Asp Ile Leu Gly Leu Asp Gly Asp Glu Thr Ile Pro Leu Gln Cys

85 90 95

Gln Ala Asn Asp Val Ala Asp Asp Gly Thr Val Val Gly Ala Tyr His

100 105 110

Asp Gln Pro Ala Ile Trp Thr Lys Ala Gly Gly Trp Lys Tyr Leu Pro

115 120 125

Ile Pro Lys Gly Trp Thr Thr Gly Phe Ala Ser Ala Val Thr Pro Asp

130 135 140

Gly His Tyr Ala Val Gly Arg Met Phe Ser Tyr Ser Gly Asn Ala Glu

145 150 155 160

Asn Tyr Gly Glu Tyr Pro Met Leu Trp Asp Leu Thr Thr Met Gln Ile

165 170 175

Thr Glu Thr Pro Gly Tyr Pro Thr Val Gly Ser Ala Gly Glu Lys Ala

180 185 190

Arg Met Ile Arg Tyr Asp Ala Ile Ser Ser Asp Ser Arg Tyr Ile Thr

195 200 205

Gly Ile Val Asp Phe Ser Tyr Thr Trp Asn Thr Leu His Phe Ile Tyr

210 215 220

Asp Arg Gln Asn Glu Ser Tyr Thr Thr Met Gly Phe Asn Thr Asp Gly

225 230 235 240

Thr Pro Trp Thr Glu Gly Leu Leu Gly Val Glu Gly Thr Leu Ser Pro

245 250 255

Asn Gly Lys Trp Phe Gly Gly Thr Ala Phe Ile Gln Asn Ala Thr Asn

260 265 270

Pro Asn Asp Glu Tyr Ser Val Pro Cys Arg Tyr Asn Met Glu Thr Asn

275 280 285

Glu Phe Glu Met Phe Asn Glu Leu Glu Ala Arg Asp Tyr Gly Ser Ile

290 295 300

Thr Ile Asp Asn Thr Gly Thr Ile Tyr Ser Ala Thr Pro Ser Ser Thr

305 310 315 320

Pro Ile Arg Ser Val Tyr Ile Arg Ala Gly Lys Phe Trp Tyr Ala Leu

325 330 335

Asp Glu Leu Met Ser Gln Ser Tyr Gly Ile Asp Phe Tyr Gly Lys Thr

340 345 350

Gly Leu Asp Asn Thr Gly Thr Ile Met Ser Val Ser Ala Asp Gly Lys

355 360 365

Val Ile Thr Ala Phe Pro Asp Pro Tyr Lys Ser Tyr Ile Leu Glu Leu

370 375 380

Asp Glu Thr Leu Thr Asp Ala Ala Gly Arg Val Asn Leu Leu Asn Asn

385 390 395 400

Tyr Thr Val Thr Pro Ala Asn Gly Ala Ser Phe Ser Gln Met Lys Asp

405 410 415

Val Ser Ile Lys Phe Ser Arg Asp Ile Lys Ile Leu Gly Lys Thr Ser

420 425 430

Asp Ile Lys Phe Thr Asp Asp Ser Gly Ala Ser Val Gly Arg Ile Ile

435 440 445

Thr Phe Ala Val Ser Pro Ser Ser Ser Lys Thr Ala Arg Ile Ala Phe

450 455 460

Arg Thr Leu Asn Leu Thr Ala Gly Lys Lys Tyr Thr Leu Thr Ile Pro

465 470 475 480

Ala Gly Thr Ile Ala Leu Ser Ala Asp Glu Thr Arg Leu Asn Asp Glu

485 490 495

Ile Val Ile Thr Tyr Thr Gly Arg Gly Thr Glu Pro Val Lys Val Val

500 505 510

Thr Ala Val Pro Glu Ser Gly Ser Ala Leu Ser Gln Leu Asn Val Thr

515 520 525

Thr Asn Pro Ile Leu Leu Thr Phe Asp Thr Asn Ile Ser Leu Thr Glu

530 535 540

Asn Ala Ala Ala Thr Leu Tyr Arg Asp Gly Ser Asp Asp Ile Val Ser

545 550 555 560

Pro Leu Ser Val Ala Val Lys Asp Asn Gln Met Leu Ile Tyr Pro Glu

565 570 575

Thr Thr Gln Tyr Leu Tyr Leu Asn Thr Asn Tyr Lys Val Val Leu Asn

580 585 590

Ala Gly Ser Val Thr Asp Val Asn Gly Gly Asn Ala Asn Glu Arg Tyr

595 600 605

Glu Ile Met Tyr Glu Gly Ile Tyr Glu Arg Ile Val Val Ala Asp Asp

610 615 620

Thr Leu Ile Tyr Lys Glu Asp Phe Ala Asn Gly Val Gly Gly Met Met

625 630 635 640

Leu Tyr Asp Gly Asp Gly Asn Ile Pro Asn Glu Glu Met Lys Asp Tyr

645 650 655

Asp Phe Tyr Tyr Asn Asn Thr Pro Gln Pro Trp Val Pro Val Arg Glu

660 665 670

Ser Arg Glu Ser Asp Asp Tyr Ser Ala Ala Ser Thr Ser Ala Tyr Ser

675 680 685

Pro Ala Gly Lys Ser Asp Asp Trp Met Val Thr Pro Gln Ile Tyr Ile

690 695 700

Pro Asp Ala Lys Cys Arg Leu Glu Phe Asn Gly Gln Gly Phe Arg Lys

705 710 715 720

Tyr Lys Gln Asp Lys Leu Lys Val Ile Val Tyr Ala Ser Asp Lys Val

725 730 735

Leu Asn Tyr Phe Ser Lys Asp Asn Ala Asp Glu Phe Arg Ala Asp Gly

740 745 750

Asn Val Ile Met Asp Glu Ile Leu Ser Pro Gly Asn Ser Glu Asp Asn

755 760 765

Leu Ser Asn Glu Trp Thr Thr Tyr Ser Phe Lys Leu Asp Lys Tyr Ala

770 775 780

Gly Lys Asn Ile Tyr Val Ala Phe Ile Asn Glu Asn Glu Asp Gln Ser

785 790 795 800

Ile Val Phe Val Asp Asn Ile Lys Val Val Arg Asp Asn Gly Phe Leu

805 810 815

Thr Ala Leu Thr Ser Ala Thr Thr Val Val Gly Gln Asn Ser His Lys

820 825 830

Ile Glu Gly Arg Val Thr Ala Asn Ser Glu Thr Glu Thr Tyr Thr Thr

835 840 845

Ala Asn Ile Lys Leu Leu Asp Ser Glu Lys Asn Val Val Asp Glu Ile

850 855 860

Ser Glu Asn Gly Leu Ser Leu Lys Lys Gly Asp Arg Tyr Asp Phe Ala

865 870 875 880

Phe Ala Lys Asn Leu Pro Leu Thr Ile Gly Glu Ile Asn Val Phe Tyr

885 890 895

Ile Arg Val Gln Leu Asp Glu Lys Phe Asp Thr Ile Ser Tyr Ala Ile

900 905 910

Lys Asp Leu Ala Phe Gln Pro Thr Lys Arg Val Ile Val Glu Glu Met

915 920 925

Thr Gly Gln Asp Cys Gly Asn Cys Pro Arg Gly His Leu Ala Trp Glu

930 935 940

Asn Leu Glu Arg Val Tyr Gly Asp Arg Val Ile Leu Ala Gly Tyr His

945 950 955 960

Val Tyr Thr Gly Asp Ile Tyr Glu Ser Gly Met Ser Ser Tyr Val Asn

965 970 975

Gln Phe Leu Gly Leu Ala Gly Ala Pro Ser Ala Lys Val Gln Arg Gly

980 985 990

Glu Thr Ile Gly Ser Pro Thr Tyr Ala Ser Ile Thr Ala Gly Arg Thr

995 1000 1005

Ser Tyr Ser Phe Thr Ser Pro Leu Gly Asp Cys Trp Phe Asp Leu

1010 1015 1020

Val Gln Lys Glu Phe Asp Thr Asp Ala Asp Ala Asn Leu Asp Val

1025 1030 1035

Ile Ala Tyr Phe Asp Glu Ala Thr Gln Lys Val Lys Ala Thr Ala

1040 1045 1050

Ser Ala Lys Phe Ala Met Asn Ile Ser Lys Gln Asn Ile Gly Leu

1055 1060 1065

Phe Ile Ile Val Thr Glu Asp Gly Leu Pro Gly Phe Gln His Asn

1070 1075 1080

Tyr His Tyr Asn Asp Asp Ala Asp Gly Leu Gly Glu Trp Gly Lys

1085 1090 1095

Gly Gly Thr Leu Gly Gln Glu Tyr Val Val Tyr Thr His Asn Asp

1100 1105 1110

Val Ala Arg Ala Gln Val Gly Ser Tyr Tyr Gly Thr Thr Gly Tyr

1115 1120 1125

Ile Pro Ser Thr Ile Ala Ser Asn Glu Thr Tyr Thr Ala Asn Ile

1130 1135 1140

Glu Phe Ala Lys Pro Ala Val Asn Glu Ile Tyr Asn Ser Asn Val

1145 1150 1155

Ile Cys Met Met Ile Asn Ala Asn Thr Gly Ala Val Ile Asn Val

1160 1165 1170

Ala Lys Ser Lys Ile Ala Lys Ala Ser Gly Ile Glu Gly Ile Thr

1175 1180 1185

Thr Ser Gly Thr Asp Ala Thr Glu Lys Ile Arg Tyr Asn Ala Ala

1190 1195 1200

Gly Gln Ile Ile Thr Ala Pro Val Lys Gly Leu Asn Ile Ile Arg

1205 1210 1215

Met Gly Asp Gly Ser Ile Arg Lys Val Val Val Lys

1220 1225 1230

<210> 15

<211> 3084

<212> DNA

<213> Prevotella rodentium

<400> 15

atgttttcat ttatgggcat cggacaggcc atagccgatg ttgtgggttc gggatcaaaa 60

gaagatccgt acgtgcttga aaacggcaca acactgacat tgaaagcata ccagtctttc 120

tatgccaaat tcacggcccc ggctgacggg atcttctcat tatatacaaa agacagctat 180

gccctatata ccgacgagag tttcagtcag attgacgagt cggcaaacat tacattcaac 240

ggaagctaca gtgagacggc atactacttt aactgtaaag caggtgttac ctattatata 300

ggaaacagtt ttgtaatggt cggaggtacg gttacagcca aattcaacac agaggctgaa 360

ccattggttc tgcgtgaaat atcacctgag gccggttctg ttttcaatgc aggcaaagga 420

agcgtagaac tgacattcaa ccaaaaagtc caaatagctt ctgttactgt caatgctggc 480

acatcctcaa agacaatgtc agccaatgta aatggagtat atgtatctgt agacgtaaaa 540

gactggctga accaaatgta tgatgaaggt aaggtcaaag aaggaaatga gatcagcttc 600

aagttcaaag gcgtggcccc aaccatttct ccaagcaaat tatacaacgg caatggagag 660

ttggacatta catacacagc aggcaagaaa cccctgcaac ttgtaagcag taccaacacc 720

ccaatgagca ctcctgccgt aaccacattc aagtcattct atacatccga tgacaaccaa 780

ggcatcgtaa cactggaatt cagtgacgaa gtgaactttt cggaaggcaa caaaccaacc 840

gcaaccctta catatggaaa tatggaccaa gatgccgacg gcgaatacta cacagaatca 900

ctccccatac ttccacttgg aagcaacatt ttaatggtta atctcaagga caagctgcgc 960

cgtgcacagg acatggtgac ttccggcact gtttacgacg gaataacact atccatatca 1020

cacgtaaagg atatggacgg aaactatgcc tatggctcgg gctccggtgt actcggatcg 1080

ttcgcattca actacaaact atctgaaata aactataccg ttgatacaga ctgggcacta 1140

cttgacgcaa ccggcaatcc gacaaacaaa gacgtcatag attcagacac caaaagcata 1200

gaactatgga tgagcgaaaa cgacggtcag ataacattca acggagtaga attcaaatat 1260

accgaaagcg gtactgaaaa ggtcaagaca atgaccctca gcgagattaa agttgacaac 1320

aatggaaacg aaacaacaat aacaatcccc gttcccaaca tcgcagccga tgccggtagc 1380

gacatcgcca tatctctcaa agacgtggaa cgccccgacg gtattaccac cagtatcgat 1440

ccggccgcac taagctattt cacaaaaaca ttcacaacaa caggcataac tgaaagcaaa 1500

ttcgacatta caagtgccat atggatgtat gaagagaatg aaggcaatat tgtcgaagtc 1560

aatatgataa acggtaatat cggtgtactg acaagaggca gcaaatctgt aatcaaaacc 1620

aacaaagaca atgagattgg ttatgtagaa tgggaaatca gaggcgttga caatcctgat 1680

atggaatata tcagagctgg atatgtagat acaaccatga caggcaagtt ggtagacggc 1740

tttaccatag aatggagagg agaaggcttg acagccggca aagactacac tttcacgctt 1800

caggcatgga agaatgaagc cgacaaaaac agtggtgccg agcctaatgt aggtgaagca 1860

atgttcatca tacacggtac aaaacaagcc tacatttaca gcgatgttgt gatgaagact 1920

gatataagcc agcctatcag acttgcttct gccaatgaca attaccgtac catcgagttc 1980

agcgctcctg taacgttaaa tgccgtggta aaccttggca tggggacatc ggcagattgc 2040

accgttgaac cctcggccga ccgtactgca tggacagtca caatacctga atatgtaatg 2100

tcacaatatg gagagttcag cgtgaatgtc tttgcaaaag atgacgaagg gcgtgcagta 2160

aacaagactg agaacggact gggtataata atggggacag aagacaatac gtggttccag 2220

attgattttg tcagtgagag ccttgccccg gactttacag taacaccggc aaacgaatca 2280

gtgctcgaga gtctcagcac cgtcactttc ggttatgaag gcagtatcag catcaactgg 2340

aacaacagtg agaaaatcac catatataac agaacaacac gtgagaagat agcagagttc 2400

agcggtgatg acgtcgtgtt ggatgaagat ccggatgact attgggctcc aatactctca 2460

tgccacatca cacttcccga gcctgtcact gctattggag tctacgatgt aaatgttcct 2520

gccggtttct tcgttcttgg tgagcagttc gacagtaact taagtaaggc aacaagcata 2580

gtatacgaaa taaaggaacc ggttgagccg ttcggaatag aaataagccc tactgccgga 2640

ttggtaagcg agattccgtc aaaactcata gtaacggtaa cggacagaag cagttgtaac 2700

tttactgcca accccacatt gaccgacaat accggtaaca gctatcccgt acacaacgat 2760

ttcgactggg gtatcccgga aatgaacaag ttcgtcatca ttcttgacaa cggagccatc 2820

acggctgacg gaatatacac actcactatt cctgccggct cgattatagg tgacgacgag 2880

acagacctca acaaggaaga ctttgtattc atatatacaa tcaatctggc cggaatcaca 2940

gaactcgtca acaatgaggg cggaaaggtt gacgtataca ctctcaaagg aacattgttg 3000

atgaaagacg ccgatgcatc agcagtaaac aagctggcaa aaggcatgta cataatcaat 3060

ggtaaaaaag tgttcataag ataa 3084

<210> 16

<211> 1212

<212> PRT

<213> Prevotella muris

<400> 16

Met Lys Lys Thr Ile Ala Leu Phe Val Trp Ser Phe Ile Phe Ile Leu

1 5 10 15

Gly Leu Ser Ala Gln Ser Glu Ser Pro Val Leu Lys Thr Leu Glu Ile

20 25 30

Pro Phe Gly Thr Ile Cys Gly Met Ser Asp Asn Gly Arg Trp Ala Val

35 40 45

Tyr Asp Asp Gly Asn Asp Gly Val Glu Arg Gly Cys Ser Val Asp Val

50 55 60

Tyr Asp Leu Ala Thr Gly Thr Ile Val Thr Leu Ser Glu Gly Thr Glu

65 70 75 80

Gly Asp Asn Ile His Leu Asn Asp Ile Ser Asp Asp Gly Lys Ile Val

85 90 95

Ala Gly Ser Tyr Asn Asn Val Pro Ala Tyr Phe Lys Asp Gly Lys Trp

100 105 110

Thr Val Leu Pro Leu Pro Ala Gly Thr Arg Gly Tyr Gly Gly Arg Val

115 120 125

Ser Ser Met Thr Pro Asp Gly Ser Val Met Val Gly Met Ile Tyr Asn

130 135 140

Asn Ser Phe Asn Phe Thr Ser Cys Tyr Trp Lys Asp Gly Asn Leu Ile

145 150 155 160

Thr Leu Glu Gly Leu Pro Thr Thr Asp Ile Ser Gly Lys Glu Asn Ala

165 170 175

Glu Leu Asn Ile Val Ala Val Ser Ala Asp Gly Asn Ile Val Leu Gly

180 185 190

Gly Leu Ser Thr Asn His Pro Gly Trp Gly Cys Cys Tyr Phe Val Tyr

195 200 205

Asp Val Arg Thr Lys Thr Tyr Glu Met Leu Gly Gln Asn Leu Thr Ser

210 215 220

Glu Ser Phe Ile Asp Asn Val Val Met Ser Asn Asn Gly Ser Phe Val

225 230 235 240

Gly Gly Asn Ala Tyr Ile Val Arg Glu Val Glu Gly Ser Glu Trp Pro

245 250 255

Glu Glu Leu Ile Val Pro Phe Leu Tyr Asp Val Asn Lys Lys Thr Phe

260 265 270

Glu Ile Tyr Glu Ser Thr Ala Asp Asn Asp Gln Phe Val Ser Ala Val

275 280 285

Ala Asn Asp Gly Leu Met Phe Cys Ala Thr Pro Tyr Gly Asn Pro Val

290 295 300

Arg Thr Met Ser Val Arg Val Asp Gly Lys Tyr Ile Asp Leu Gly Met

305 310 315 320

Ile Leu Lys Gln Arg Tyr Ser Ile Asp Phe Thr Lys Ala Met Gly Val

325 330 335

Glu Tyr Thr Gly Thr Ala Val Ala Val Ser Asp Asp Gly Lys Thr Ile

340 345 350

Val Ala Phe Pro Ser Pro Gln Asp Asp Asn Tyr Ala Leu Thr Leu Pro

355 360 365

Val Ser Phe Ser Glu Ala Ala Gln Gly Val Asn Ile Leu Ala Glu Tyr

370 375 380

Glu Leu Ser Pro Val Ser Gly Ser Ser Ile Ala Lys Ile Arg Gln Val

385 390 395 400

Ala Leu Ser Phe Thr Arg Glu Ala Thr Val Ala Glu Gly Ala Glu Ala

405 410 415

Tyr Ile Tyr Lys Gly Glu Thr Lys Leu Ala Val Thr Asn Ser Ile Ser

420 425 430

Ala Ala Ser Val Lys Asp Asn Arg Ile Phe Ile Leu Asp Phe Pro Glu

435 440 445

Thr Ala Phe Ser Glu Asp Glu Glu Tyr Val Leu Lys Ile Pro Ala Gly

450 455 460

Ile Phe Val Asn Gly Asn Met Arg Asn Asn Glu Ile Thr Ala Val Tyr

465 470 475 480

Lys Gly Arg Ala Asp Arg Pro Val Ala Pro Thr Arg Ile Ala Pro Leu

485 490 495

Glu Gly Ser Val Ile Thr Glu Leu Ser Tyr Asn Asn Pro Val Met Ile

500 505 510

Thr Phe Asp Thr Lys Ile Ala Val Ala Gln Ala Val Thr Gly Ile Leu

515 520 525

Phe Gln Ser Gly Ser Glu Gln Pro Leu Ser Ser Leu Val Leu Val Ala

530 535 540

Asp Gly Asn Lys Leu Tyr Val Tyr Pro Pro Thr Thr Arg Arg Leu Met

545 550 555 560

Lys Gly Ser Asp Tyr Val Val Lys Ile Pro Ala Gly Ala Val Ser Asp

565 570 575

Ile Val Gly Phe Cys Pro Ser Asn Glu Ile Thr Val Asn Tyr His Gly

580 585 590

Ala Phe Val Val Thr Pro Pro Glu Gln Gly Ser Ser Phe Leu Phe Val

595 600 605

Asp Asp Phe Asn Asp Pro Gly Thr Ser Leu Ser Lys Trp Leu Met Tyr

610 615 620

Glu Gly Asp His Asn Thr Pro Thr Ser Glu Met Val Gly Trp Gly Phe

625 630 635 640

Asp Ala Asp Asn Thr Pro Trp Asn Phe Ser Val His Asp Asn Gly Gln

645 650 655

Tyr Asp Tyr Cys Ala Ala Ser Thr Ser Met Tyr Ser Pro Ala Gly Lys

660 665 670

Ala Asn Asp Trp Met Ile Thr Pro Gln Leu Ser Ile Gly Asn Glu Tyr

675 680 685

Tyr Arg Leu Asn Phe Gln Thr Gln Ser Tyr Lys Ala Gly Lys Thr Asp

690 695 700

Lys Leu Lys Val Tyr Val Trp Glu Asn Asn Glu Leu Ile Glu Gly Thr

705 710 715 720

Val Leu Lys Glu His Ile Asp Ala Ile Arg Ala Asn Gly Lys Leu Val

725 730 735

Phe Asp Gly Ile Leu Thr Pro Gly Ala Asn Glu Ala Thr Leu Thr Gly

740 745 750

Glu Trp Glu Ser His Ser Ile Ser Leu Ala Glu Tyr Ala Gly Lys Lys

755 760 765

Val Tyr Ile Ala Phe Leu Asn Glu Asn Glu Asp Gln Ser Val Val Phe

770 775 780

Val Asp Asn Val Asn Val Ile Tyr Lys Gly Asn Phe Val Ala Gly Ser

785 790 795 800

Met Thr Gln Glu Asn Val Val Ser Gln Glu Glu Val Glu Ile Lys Gly

805 810 815

Tyr Val Met Val Thr Asn Asn Ile Ser Tyr Asp Ala Ile Glu Ala Thr

820 825 830

Tyr Lys Thr Val Asp Gly Thr Gln Ile Gly Ile Tyr Lys Ala Glu Gly

835 840 845

Ile Gly Leu Lys Glu Gly Ser Val Tyr Glu Phe Thr Phe Pro Glu Lys

850 855 860

Leu Lys Leu Thr Lys Gly Gln Glu Thr Gln Phe Thr Ile Val Val Lys

865 870 875 880

Met Gly Thr Glu Glu Gln Glu Ile Asn Ala Ser Val Lys Asn Leu Ala

885 890 895

Phe Asn Pro Lys Arg Arg Val Val Leu Glu Glu Gly Thr Gly Ala Trp

900 905 910

Cys Gly Asn Cys Pro Gly Gly Ile Leu Ala Val Glu Tyr Ile Glu Ser

915 920 925

Gln Phe Pro Gly Gln Leu Ile Pro Val Cys Ile His Asn Asp Asp Ala

930 935 940

Tyr Ala Tyr Gly Ala Tyr Glu Asn Phe Leu Gly Phe Arg Ser Phe Pro

945 950 955 960

Thr Gly Arg Val Asn Arg Ile Glu Asn Tyr Leu Ser Tyr Met Asp Ser

965 970 975

Asp Glu Glu Gly Asn Pro Ser Phe Phe Ser Val Asp Gly Asp Val Thr

980 985 990

Phe Ala Asp Tyr Val Lys Ser Glu Ile Asp Lys Leu Thr Asp Val Glu

995 1000 1005

Ile Glu Thr Gly Glu Ala Val Tyr Asn Ser Ser Thr Asp Glu Ile

1010 1015 1020

Thr Val Pro Val Asp Val Arg Phe Ala Leu Asp Lys Asn Ser Val

1025 1030 1035

Asn Tyr Asn Ile Leu Thr Val Val Val Glu Asp Asn Leu Tyr Ala

1040 1045 1050

Asn Gln Arg Asn Tyr Phe Ala Ser Arg Thr Asp Pro Ile Tyr Gly

1055 1060 1065

Asp Trp Gly Leu Gly Gly Lys Tyr Gly Arg Gly Ser Val Trp Tyr

1070 1075 1080

Ala Tyr Lys Asp Val Ala Arg Ala Ile Val Gly Gln Ser Tyr Tyr

1085 1090 1095

Gly Glu Asn Gly Tyr Ile Pro Thr Ser Val Arg Gly Gly Glu Thr

1100 1105 1110

Ile Thr Gly Asn Val Ser Phe Glu Val Pro Gly Lys Ser Ile Thr

1115 1120 1125

Ser Leu Gln Asn Cys Lys Ile Val Cys Met Leu Ile Asp Ala Ala

1130 1135 1140

Asn Gly Tyr Val Leu Asn Ala Ala Arg Cys Asp Ala Ile Ser Asn

1145 1150 1155

Ala Leu Glu Trp Asp Leu Ala Asn Gly Ile Glu Asp Ala Val Val

1160 1165 1170

Asp Thr Asp Thr Ala Gly Val Glu Asn Tyr Asn Leu Ala Gly Gln

1175 1180 1185

Lys Met Asn Ala Gln Arg Lys Gly Leu Asn Ile Val Arg Leu Ser

1190 1195 1200

Asn Gly Arg Thr Val Lys Val Val Lys

1205 1210

<210> 17

<211> 3639

<212> DNA

<213> Prevotella muris

<400> 17

atgaaaaaaa ctattgcatt atttgtatgg agtttcattt tcatattagg gctttccgcc 60

caaagtgaat ctcctgtatt gaaaactctt gaaattccat tcggtactat ttgcgggatg 120

tcagacaacg gccgttgggc tgtttatgac gatggcaatg acggtgttga gcggggctgt 180

tctgtggatg tctatgactt ggctacagga accattgtaa cactctctga aggaacggaa 240

ggggataata ttcatttgaa tgatatttcg gatgacggta agatagtggc aggttcatac 300

aacaatgttc ccgcttactt taaggatgga aagtggacgg tacttccttt accggccggc 360

acacgtggat atggcggaag agtttcgagt atgactccgg atggttcggt aatggtcgga 420

atgatttata ataactcctt taattttacg tcatgctatt ggaaagacgg aaatcttatc 480

actcttgaag gattgcctac aaccgatata tccggtaagg aaaatgcaga attgaatatt 540

gttgctgttt cggctgacgg caatattgtt ctcggaggat tgtcaaccaa tcatccggga 600

tggggatgct gttattttgt atatgatgtc aggacaaaga cttacgagat gcttggtcag 660

aatcttacct cagagtcatt cattgataat gttgtgatga gcaataacgg aagttttgtg 720

ggtggtaatg catatatagt gcgtgaggtt gaaggttcgg aatggccgga ggagcttatt 780

gttcctttcc tttatgatgt gaataaaaaa acgtttgaaa tatatgaaag tacagctgat 840

aatgaccagt ttgtttcagc tgtggcaaat gacggtttga tgttctgtgc cactccgtac 900

ggtaatcctg tgcgtacgat gagtgtgcgt gtggatggaa aatatattga tttgggaatg 960

atactcaaac aacgctattc gatagatttt acaaaggcaa tgggtgttga gtatacggga 1020

actgccgtag cggtgagtga cgatggtaag acaatagtgg ctttcccttc tccacaagat 1080

gataattatg ccttgactct ccctgtttca ttctctgagg ccgcacaagg tgttaatatt 1140

cttgcggaat atgagctttc tcctgtgtca ggaagcagta tcgcaaaaat aaggcaggtc 1200

gctttatctt ttacgcgtga ggccaccgta gctgaaggtg ccgaggcata tatttataaa 1260

ggtgaaacaa agttggcagt cacaaattcc ataagtgcgg cgtctgtaaa agataatcgt 1320

atatttatac ttgattttcc tgagacagcc ttttcggaag atgaagagta tgtgcttaag 1380

atacctgcag gtattttcgt aaatggaaat atgagaaata atgaaatcac ggctgtatat 1440

aaaggacgtg cagacagacc ggtagctcca acacgcatag ctccgttgga aggaagcgta 1500

ataacagagt taagctacaa caatcctgtc atgattactt tcgacacgaa gattgctgtg 1560

gctcaggcag tcacaggcat tcttttccag agtggttcgg aacagccgtt gagcagtctt 1620

gtacttgttg ccgatggaaa caaactatac gtatatcctc ctacaacgcg tcgtttgatg 1680

aaaggttccg actatgtggt gaagatacct gccggtgctg tttccgatat agtagggttc 1740

tgccccagta atgagattac tgtcaattat catggggctt ttgttgtaac acctcccgag 1800

cagggttcaa gcttcctgtt tgttgatgac ttcaatgatc cgggaacgtc actgagcaaa 1860

tggcttatgt acgagggtga ccataacaca ccgacctcag aaatggtagg atggggattt 1920

gatgcagaca atacgccatg gaatttctct gtgcatgaca acggacagta tgattattgt 1980

gccgcatcta cttccatgta ttctccggcc ggcaaggcaa acgactggat gataacccca 2040

cagctttcta taggaaacga atattacaga ctgaatttcc agacgcagtc atataaggcc 2100

ggtaagactg ataaactgaa agtttatgta tgggagaaca atgagcttat agaaggtact 2160

gtgctgaaag agcacattga tgcaatacgt gcaaacggaa agctggtatt tgacggtata 2220

ctgactccgg gagcaaatga agctacgctt accggtgaat gggaaagcca cagcatttca 2280

cttgcagaat atgcaggaaa gaaagtgtat atagcattcc tcaacgaaaa cgaagaccaa 2340

agtgtagtgt ttgttgataa cgtaaatgta atatacaaag gcaactttgt tgccggaagc 2400

atgacacagg agaatgtcgt ctcacaggaa gaggtggaaa tcaagggata tgtaatggtg 2460

accaacaaca tatcatacga tgccattgag gccacatata agacggtaga cggaacccaa 2520

ataggaattt ataaggctga gggtatcggt ttgaaggagg gcagtgttta tgaatttacg 2580

ttccctgaaa aactgaagtt gaccaaaggg caggaaacgc agtttactat agtggtgaag 2640

atgggtacgg aagaacagga gataaatgct tcggttaaga atcttgcgtt caatcccaag 2700

cgtcgtgtcg tacttgaaga aggcaccggc gcatggtgtg gaaattgtcc tggaggtata 2760

cttgctgtgg aatatattga aagtcagttc cccggtcagt tgatacctgt atgcatacat 2820

aatgacgatg cgtatgctta tggtgcgtat gagaatttcc ttggtttccg ctcgttccct 2880

acaggtcgtg taaaccgtat tgagaattat ctttcatata tggactcaga tgaggaaggc 2940

aatccgtctt tcttctctgt tgacggagat gtaacgtttg ctgactatgt gaagagtgag 3000

atagacaaac ttacagatgt tgaaatagag acaggcgagg ctgtctataa ttcgtctacg 3060

gatgagataa ctgtgccggt cgatgtaagg tttgcccttg acaagaattc tgtaaattat 3120

aatattctta ctgttgttgt cgaggataat ctatatgcta atcagcgtaa ttattttgca 3180

agccgtacgg atcctattta tggagactgg gggcttggcg gtaagtatgg ccgtggttcc 3240

gtatggtatg cttataagga tgtcgcacgt gcaatcgtgg gtcagtctta ttatggtgag 3300

aacggatata ttcccacttc tgtcagaggc ggtgagacaa taactggtaa tgtttctttt 3360

gaagttcccg gcaaatcgat tacttcactg cagaattgca agattgtatg tatgctgatt 3420

gatgcggcta acggatacgt tcttaatgca gcacggtgtg atgcaatatc taacgcattg 3480

gaatgggatc ttgcgaatgg aatagaggat gctgttgttg atacagatac agcaggtgtg 3540

gagaattaca atctggcagg ccagaaaatg aatgctcagc gcaaaggtct taatatcgta 3600

aggctttcca acggacgtac ggtgaaagtc gtaaagtga 3639

<210> 18

<211> 1027

<212> PRT

<213> Paraprevotella clara

<400> 18

Met Arg Glu Lys Leu Tyr Thr Lys Trp Lys Gly Gly Leu Asn Arg Leu

1 5 10 15

Cys Phe Leu Leu Met Cys Phe Cys Trp Thr Thr Val Gln Ser Trp Ala

20 25 30

Val Gly Glu Asp Leu His Leu Thr Ile Glu Asn Gly Lys Thr Tyr Glu

35 40 45

Phe Glu Ala Phe Asn Ser Tyr Tyr Leu Thr Tyr Val Ala Thr Ala Asn

50 55 60

Gly Gln Leu Ser Leu Tyr Gln Thr Gly Gly Asp Phe Cys Arg Gln Tyr

65 70 75 80

Thr Asp Asn Thr Phe Glu Thr Glu Leu Pro Ser Thr Pro Gln Tyr Val

85 90 95

Asn Glu Gly Lys Leu Val Glu Val Lys Val Glu Ser Gly Lys Thr Tyr

100 105 110

Tyr Phe Leu Thr Arg Gly Leu Ser Lys Gly Glu Leu Thr Val Thr Phe

115 120 125

Gly Glu Lys Ala Thr Pro Leu Glu Leu Leu Ser Leu Ser Lys Glu Glu

130 135 140

Gly Thr Thr Leu Asn Leu Ser Ile Asp Thr Leu Leu Gly Phe Thr Phe

145 150 155 160

Asn Arg Met Val Lys Val Gly Asn Cys Thr Leu Ser Ser Gly Ser Val

165 170 175

Ile Gly Asn Leu Thr Ala Ser Thr His Asp Tyr Gly Phe Thr Val Ser

180 185 190

Ile Lys Asp Val Leu Tyr Lys Trp Leu Lys Glu Gly Asn Val Lys Ala

195 200 205

Gly Asp Glu Val Val Leu Thr Val Thr Gly Leu Cys Asn Ala Asn Asp

210 215 220

Glu Ser Asp Lys Tyr Asn Gly Asn Gly Val Leu Thr Val Lys Tyr Ile

225 230 235 240

Ala Gly Ala Leu Pro Ala Glu Leu Val Ser Val Ala Asn Ser Pro Glu

245 250 255

Glu Met Asn Phe Leu Ser Tyr Tyr Leu Pro Thr Asp Glu Arg Gly Leu

260 265 270

Val Thr Leu Thr Phe Asn Arg Ser Met Gly Glu Asp Ala Thr Ala Lys

275 280 285

Leu Phe Tyr Gly Asp Ile Glu Gly Ser Asn Val Tyr Thr Glu Ser Leu

290 295 300

Pro Val Lys Val Lys Asp Thr Gln Leu Ile Val Asp Leu Arg Gly Lys

305 310 315 320

Arg Arg Leu Pro Ser Asp Met Leu Pro Asn Ala Ser Ala Asp Val Trp

325 330 335

Glu Gln Tyr Lys Thr Ile Ser Leu Lys Val Ser Asn Val Arg Asp Ala

340 345 350

Glu Gly Asn Tyr Ala Met Ser Thr Ser Gln Gly Thr Thr Gly Ser Tyr

355 360 365

Ala Phe Asn Tyr Thr Ile Glu Ala Val Glu Phe Asp Ile Ala Arg Gln

370 375 380

Phe Thr Pro Asp Glu Gly Thr Ser Leu Asp Asp Tyr Pro Thr Ile Glu

385 390 395 400

Leu Trp Ile Arg Glu Asp Asp Ser Phe Thr Tyr Glu Gly Val Asp Phe

405 410 415

Thr Tyr Ser Val Asn Gly Lys Pro Glu Thr Val Phe Val Pro Ile Ser

420 425 430

Gln Ile Thr Lys Lys Asp Ser Pro Asp Gly Asp Gly Val Leu Leu Thr

435 440 445

Ile Pro Val Pro Asn Lys Ala Ala Asp Glu Asn Ser Asp Val Val Val

450 455 460

Ser Leu Asn Asn Leu Lys Ala Ala Asp Gly Ala Asp Tyr Ser Glu Asp

465 470 475 480

Phe Thr Ile Ile Tyr Lys Thr Ser Gly Lys Ser Met Ala Gly Leu Glu

485 490 495

Ile Glu Ser Val Ser Pro Ala Asp Gly Ala Ala Ile Ala Ala Leu Glu

500 505 510

Ala Gly Ser Phe Leu Glu Leu Ser Thr Asn Met Asn Asp Arg Val Gly

515 520 525

Tyr Val Trp Phe Arg Ile Asp Asp Gln Asn Pro Lys Asp Pro Asp Gln

530 535 540

Ala Cys Val Lys Thr Met Thr Ser Met Lys Lys Glu Thr Val Glu Gly

545 550 555 560

Lys Val Ser Phe Arg Thr Glu Ile Ile Arg Ser Val Thr Phe Phe Glu

565 570 575

Gly His Thr Tyr Lys Val Thr Phe Asn Ala Tyr Ala Ser Glu Ser Asp

580 585 590

Tyr Gln His Gly Ala Asp Ala Leu Gly Val Met Thr Val Thr Tyr Thr

595 600 605

Gly Thr Thr Pro Glu Phe Lys Phe Ser Pro Val Lys Phe Val Gly Ile

610 615 620

Thr Pro Asp Pro Asp Tyr Thr Val Ile Ser Asp Val Ser Gln Asn Glu

625 630 635 640

Phe Thr Leu Thr Phe Asp Gly Pro Val Val Leu Asn His Glu Asn Ala

645 650 655

Phe Val Val Tyr Gly Gln Gly Met Asn Tyr Pro Phe Glu Ser Ile Glu

660 665 670

Ser Asn Glu Asp Gly Thr Glu Trp Thr Val Lys Ala Ser Leu Glu Lys

675 680 685

Leu Met Glu Met Gly Met Met Pro Arg Leu Ser Met Ser Phe Met Pro

690 695 700

Val Asp Lys Asp Gly Met Leu Val Glu Gly Asn Ala Gly Lys Asp Ala

705 710 715 720

Thr Ser Tyr Leu Asn Phe Ala Tyr Asp Cys Thr Ile Gly Ile Pro Asp

725 730 735

Leu Gln Val Ser Pro Glu Ser Gly Ser Lys Val Glu Ser Leu Lys Glu

740 745 750

Ile Ile Val Gly Cys Lys Asp Ile Met Asp Asp Asn Gly Glu Val Ile

755 760 765

Phe Gln Gly Gly Ile Ser Glu Ser Tyr Met Ala Ala Glu Lys Ile Ile

770 775 780

Leu Tyr Lys Asn Gly Arg Glu Pro Val Ala Thr Val Thr Ser Ile Glu

785 790 795 800

Pro Ile Ile Pro Glu Asp Gln Glu Asp Asn Tyr Phe Tyr Val Pro Val

805 810 815

Glu Met Lys Leu Thr Leu Asp Thr Glu Ile Thr Asp Asn Gly Asn Tyr

820 825 830

Arg Leu His Ile Pro Ala Asn Tyr Phe Ile Leu Gly Thr Gly Met Ser

835 840 845

Asn Val Asn Ser Lys Glu Thr Ser Val Leu Tyr Thr Ile Asp Gln Pro

850 855 860

Ile Lys Ile Thr Val Thr Pro Glu Asn Asn Ser Thr Val Glu Ser Leu

865 870 875 880

Lys Glu Ile Thr Ile Glu Cys Glu Ser Gly Ile Asp Val Pro Ser Met

885 890 895

Gly Thr Ile Gln Leu Leu Asn Ala Gln Asn Glu Val Val Ala Ser Ala

900 905 910

Thr Gly Glu Asp Cys Glu Leu Leu Leu Pro Glu Gly Ala Gly Pro Trp

915 920 925

Asp Pro Tyr Thr Gly Val Thr Ile Lys Leu Asp Gln Glu Val Thr Glu

930 935 940

Lys Gly Thr Tyr Lys Leu Val Ile Pro Glu Gly Phe Phe Tyr Leu Gly

945 950 955 960

Glu Asn Tyr Glu Asn Ser Asp Glu Met Thr Phe Thr Tyr Ser Ile Asn

965 970 975

Ala Ser Gly Ile His Ser Ile Gly Thr Glu Ser Lys Gly Val Val Val

980 985 990

Tyr Thr Val Asp Gly Lys Phe Ile Leu Lys Ser Ser Asp Ala Lys Asp

995 1000 1005

Val Lys Asn Leu Lys Lys Gly Leu Tyr Ile Val Asn Gly Lys Lys

1010 1015 1020

Met Met Val Lys

1025

<210> 19

<211> 1027

<212> PRT

<213> Paraprevotella clara

<400> 19

Met Arg Glu Lys Pro Tyr Thr Lys Trp Lys Gly Gly Leu Asn Arg Leu

1 5 10 15

Cys Phe Leu Leu Met Cys Phe Cys Trp Thr Thr Val Gln Ser Trp Ala

20 25 30

Val Gly Glu Asp Val His Leu Thr Ile Glu Asn Gly Lys Thr Tyr Glu

35 40 45

Phe Glu Ala Phe Asn Ser Tyr Tyr Leu Thr Tyr Val Ala Thr Ala Asn

50 55 60

Gly Gln Leu Ser Leu Tyr Gln Thr Gly Gly Asp Phe Cys Arg Gln Tyr

65 70 75 80

Thr Asp Asn Thr Phe Glu Thr Glu Leu Pro Ser Thr Pro Gln Tyr Val

85 90 95

Asn Glu Gly Lys Leu Val Glu Val Lys Val Glu Ser Gly Lys Thr Tyr

100 105 110

Tyr Phe Leu Thr Arg Gly Leu Ser Lys Gly Glu Leu Thr Val Thr Phe

115 120 125

Gly Glu Lys Ala Thr Pro Leu Glu Leu Leu Ser Leu Ser Lys Glu Glu

130 135 140

Gly Thr Thr Leu Asn Leu Ser Ile Asp Thr Leu Leu Gly Phe Thr Phe

145 150 155 160

Asn Arg Met Val Lys Val Gly Asn Cys Thr Leu Ser Ser Gly Ser Val

165 170 175

Ile Gly Asn Leu Thr Ala Ser Thr His Asp Tyr Gly Phe Thr Val Ser

180 185 190

Ile Lys Asp Val Leu Tyr Lys Trp Leu Lys Glu Gly Asn Val Lys Ala

195 200 205

Gly Asp Glu Val Val Leu Thr Val Thr Gly Leu Cys Asn Ala Asn Asp

210 215 220

Glu Asn Asp Lys Tyr Asn Gly Asn Gly Val Leu Thr Val Lys Tyr Ile

225 230 235 240

Ala Gly Ala Leu Pro Ala Glu Leu Val Ser Val Thr Asn Ser Pro Glu

245 250 255

Asp Met Asn Phe Leu Ser Tyr Tyr Leu Pro Thr Asp Glu Arg Gly Leu

260 265 270

Val Thr Leu Thr Phe Asn Arg Ser Met Gly Glu Asp Ala Thr Ala Lys

275 280 285

Leu Phe Tyr Gly Asp Ile Glu Gly Ser Asn Val Tyr Thr Glu Ser Leu

290 295 300

Pro Val Lys Val Lys Asp Thr Gln Leu Ile Val Asp Leu Arg Gly Lys

305 310 315 320

Arg Arg Leu Pro Ser Asp Met Leu Pro Asn Ala Ser Ala Asp Val Trp

325 330 335

Glu Gln Tyr Lys Thr Ile Ser Leu Lys Val Ser Asn Val Lys Asp Val

340 345 350

Glu Gly Asn Tyr Ala Met Ser Thr Ser Gln Gly Thr Thr Gly Ser Tyr

355 360 365

Ala Phe Asn Tyr Thr Ile Glu Ala Val Glu Phe Asp Ile Ala Arg Gln

370 375 380

Phe Thr Pro Asp Glu Gly Thr Ser Leu Asp Asp Tyr Pro Thr Ile Glu

385 390 395 400

Leu Trp Ile Arg Glu Asp Asp Ser Phe Thr Tyr Glu Gly Val Asp Phe

405 410 415

Thr Tyr Ser Val Asn Gly Lys Pro Glu Thr Val Phe Val Pro Ile Ser

420 425 430

Gln Ile Thr Lys Lys Asp Ser Pro Asp Gly Asp Gly Val Leu Leu Thr

435 440 445

Ile Pro Val Pro Asn Lys Ala Ala Asp Glu Asn Ser Asp Val Val Val

450 455 460

Ser Leu Asn Asn Leu Lys Ala Ala Asp Gly Ala Asp Tyr Ser Glu Asp

465 470 475 480

Phe Thr Ile Ile Tyr Lys Thr Ser Gly Lys Ser Met Ala Gly Leu Glu

485 490 495

Ile Glu Ser Val Ser Pro Ala Asp Gly Ala Ala Ile Ala Ala Leu Glu

500 505 510

Ala Gly Ser Phe Leu Glu Leu Ser Thr Asn Met Asn Asp Arg Val Gly

515 520 525

Tyr Val Trp Phe Arg Ile Asp Asp Gln Asn Pro Lys Asp Pro Asp Gln

530 535 540

Ala Cys Val Lys Thr Met Thr Ser Met Lys Lys Glu Thr Val Glu Gly

545 550 555 560

Lys Val Ser Phe Arg Thr Glu Ile Ile Arg Ser Val Thr Phe Phe Glu

565 570 575

Gly His Thr Tyr Lys Val Thr Phe Asn Ala Tyr Ala Ser Glu Ser Asp

580 585 590

Tyr Gln His Gly Ala Asp Ala Leu Gly Val Met Thr Val Thr Tyr Thr

595 600 605

Gly Thr Thr Pro Glu Phe Lys Phe Ser Pro Val Lys Phe Val Gly Ile

610 615 620

Thr Pro Asp Pro Asp Tyr Thr Val Ile Ser Asp Val Ser Gln Asn Glu

625 630 635 640

Phe Thr Leu Thr Phe Asp Gly Pro Val Val Leu Asn His Glu Asn Ala

645 650 655

Phe Val Val Tyr Gly Gln Gly Met Asn Tyr Pro Phe Glu Ser Ile Glu

660 665 670

Ser Asn Glu Asp Gly Thr Glu Trp Thr Val Lys Ala Ser Leu Glu Lys

675 680 685

Leu Met Glu Met Gly Met Met Pro Arg Leu Ser Met Ser Phe Met Pro

690 695 700

Val Asp Lys Asp Gly Met Leu Val Glu Gly Asn Ala Gly Lys Asp Ala

705 710 715 720

Thr Ser Tyr Leu Asn Phe Ala Tyr Asp Cys Thr Ile Gly Ile Pro Asp

725 730 735

Leu Gln Val Ser Pro Glu Ser Gly Ser Lys Val Glu Ser Leu Lys Glu

740 745 750

Ile Ile Val Gly Cys Lys Asp Ile Met Asp Asp Asn Gly Glu Val Ile

755 760 765

Phe Gln Gly Gly Ile Ser Glu Ser Tyr Met Ala Ala Glu Lys Ile Ile

770 775 780

Leu Tyr Lys Asn Gly Arg Glu Pro Val Ala Thr Val Thr Ser Ile Glu

785 790 795 800

Pro Ile Ile Pro Glu Asp Gln Glu Asp Asn Tyr Phe Tyr Val Pro Val

805 810 815

Glu Met Lys Leu Thr Leu Asp Thr Glu Ile Thr Asp Asn Gly Asn Tyr

820 825 830

Arg Leu His Ile Pro Ala Asn Tyr Phe Ile Leu Gly Thr Gly Met Ser

835 840 845

Asn Val Asn Ser Lys Glu Thr Ser Val Leu Tyr Thr Ile Asp Gln Pro

850 855 860

Ile Lys Ile Thr Val Thr Pro Glu Asn Asn Ser Thr Val Glu Ser Leu

865 870 875 880

Lys Glu Ile Thr Ile Glu Cys Glu Ser Gly Ile Asp Val Pro Ser Met

885 890 895

Gly Thr Ile Gln Leu Leu Asn Ala Gln Asn Glu Val Val Ala Ser Ala

900 905 910

Thr Gly Glu Asp Cys Glu Leu Leu Leu Pro Glu Gly Ala Gly Pro Trp

915 920 925

Asp Pro Tyr Thr Gly Val Thr Ile Lys Leu Asp Gln Glu Val Thr Glu

930 935 940

Lys Gly Thr Tyr Lys Leu Val Ile Pro Glu Gly Phe Phe Tyr Leu Gly

945 950 955 960

Glu Asn Tyr Glu Asn Ser Asp Glu Met Thr Phe Thr Tyr Ser Ile Asn

965 970 975

Ala Ser Gly Ile His Ser Ile Gly Thr Glu Ser Lys Gly Val Val Val

980 985 990

Tyr Thr Val Asp Gly Lys Phe Ile Leu Lys Ser Ser Asp Ala Lys Asp

995 1000 1005

Val Lys Asn Leu Lys Lys Gly Leu Tyr Ile Val Asn Gly Lys Lys

1010 1015 1020

Met Met Val Lys

1025

<210> 20

<211> 3084

<212> DNA

<213> Paraprevotella clara

<400> 20

atgagagaga aaccttacac gaaatggaag ggaggcctga ataggctttg ttttcttctt 60

atgtgttttt gttggactac ggttcagtca tgggccgttg gtgaagatgt acatttaacc 120

attgaaaatg gtaagacgta tgaatttgaa gctttcaaca gttattatct tacttatgta 180

gccacggcta acggacaatt atcattgtat caaaccggtg gagacttttg tcgtcagtat 240

acagataaca cttttgaaac ggaattgcct tctacccctc agtatgttaa tgagggaaaa 300

cttgtagaag tgaaggttga gtcaggtaag acttattatt tcttgactcg aggtttaagt 360

aaaggagaac tgaccgttac tttcggagaa aaagcaactc cacttgaatt actttcgctc 420

tccaaggaag agggaacgac attgaatctt tctattgata ccctcttagg cttcactttc 480

aatagaatgg taaaagtagg aaattgcaca ttgtcttccg ggtcggtaat cggaaacttg 540

accgcttcga cgcatgatta tggtttcacc gtttccatta aagatgtgtt gtataaatgg 600

ttgaaggagg gtaatgtaaa agccggagat gaagtcgttt tgactgtaac gggattatgt 660

aatgccaatg atgagaacga caaatacaat ggtaatggcg tgctgacggt gaaatatatt 720

gcaggagcgt tgcctgccga gttggttagc gttacgaact cacccgaaga tatgaacttc 780

ctgtcttatt acctgcctac tgatgaaagg gggttggtta ctttgacttt caaccgttcg 840

atgggcgaag atgctacggc aaaattattc tatggcgata tagaaggttc gaatgtctat 900

acggaaagtc ttccggtgaa ggtgaaagat actcagttga ttgtagacct ccgtggaaaa 960

cgtcgtttgc catctgatat gttgccgaat gccagtgctg atgtttggga acagtataaa 1020

accatttcct tgaaggttag caacgtgaag gatgtggaag gaaattatgc tatgtctacc 1080

tctcagggta ccacaggttc gtatgctttt aattatacaa tagaagctgt tgagtttgat 1140

atcgctcgtc agtttacacc agatgaaggc acatctttgg atgattatcc gacaatagaa 1200

ctttggattc gggaagacga ttcgtttact tatgagggtg tagatttcac ttatagcgtg 1260

aatggaaaac ctgagacggt atttgtaccc atcagccaga ttacgaaaaa agactctcca 1320

gacggagatg gtgtactatt gacgattccg gtgccgaata aagcagcgga tgagaattca 1380

gatgtcgtag tttctttgaa taatttgaaa gcggccgatg gtgcggatta ttcagaagat 1440

tttacgataa tatataaaac gtcaggaaag agtatggcag ggcttgagat agaatctgtg 1500

tctcctgctg atggagcagc tatcgcggct ttagaagctg gttcgtttct ggaattgtct 1560

accaatatga atgatcgtgt aggatatgta tggtttagaa tagatgacca gaacccgaag 1620

gatcctgatc aggcttgtgt taagacgatg acatcgatga agaaagagac ggttgaggga 1680

aaagtttctt tccggacaga aataattcgt agtgtgactt tctttgaagg gcatacttat 1740

aaagttactt tcaatgctta tgcttcggag agcgattatc agcacggtgc agatgctctt 1800

ggtgttatga ctgtgactta cacaggtaca actccggaat ttaagtttag tccggttaag 1860

tttgtaggaa taacgccgga tcctgattat actgtaattt cagacgtaag ccagaatgaa 1920

tttacgttga cttttgacgg tcctgtagta ttgaatcatg aaaatgcttt tgttgtatac 1980

ggtcagggaa tgaattatcc atttgaaagt atcgagtcga atgaagacgg tacggaatgg 2040

acggtaaagg catccttgga aaaattaatg gaaatgggta tgatgcctcg attgtctatg 2100

agctttatgc cggtggataa agacggaatg ttagttgaag gtaatgccgg taaagatgcg 2160

acaagttatt tgaattttgc ctatgattgt acgattggta taccagattt gcaagtgagt 2220

ccggaaagcg gttctaaagt ggaaagtttg aaagagatca tagtaggatg taaagatatc 2280

atggacgaca atggtgaggt gatatttcaa ggtggtatca gcgaatctta tatggcggcc 2340

gaaaagatca ttttgtataa gaacggacgt gagccggttg caacggtaac gagtatcgaa 2400

cctattatac cggaggacca agaggataat tacttctacg ttcctgtgga aatgaaatta 2460

acattggata ctgaaattac agataacggt aattatcgtt tgcacattcc tgccaactac 2520

tttatattgg gaacaggtat gtctaatgta aacagtaaag aaacaagtgt gctgtatact 2580

atcgaccagc cgattaagat aacggttacg ccagagaata actctacggt agaaagcctg 2640

aaagaaatta ctatcgaatg tgaatcaggt atagatgtgc cgagtatggg tacaattcaa 2700

ttattgaatg ctcagaatga ggtggtagct tcggcaacgg gtgaggattg cgaattgttg 2760

ttacctgaag gtgccggtcc ttgggatcct tatactggtg tgacaattaa attggatcaa 2820

gaggtgacag agaaaggtac atataagttg gttattcctg aaggattctt ctatttgggt 2880

gaaaactatg aaaattcgga cgagatgacg tttacttatt cgattaatgc cagtggtatt 2940

cattcgattg gtactgagtc taagggagta gtcgtttata ctgtagacgg taaatttatc 3000

ttaaagtcga gcgatgccaa agacgtgaag aatctgaaga aaggtttgta cattgtgaac 3060

ggtaagaaga tgatggtaaa ataa 3084

<210> 21

<211> 787

<212> PRT

<213> Paraprevotella xylaniphila

<400> 21

Met Arg Lys Met Phe Thr Leu Glu Met Lys Asp Tyr Leu Met Arg Met

1 5 10 15

Val Leu Phe Val Ala Val Cys Met Val Gly Val Gly Gly Leu Lys Ala

20 25 30

Ala Pro Gly Leu Ser Ala Asp Asp Pro Leu Ile Phe Glu Gly Gly Glu

35 40 45

Glu Tyr Asp Val Ser Gly Ser Tyr Phe Lys Asp Leu Tyr Ala Thr Phe

50 55 60

Thr Ala Pro Ser Asp Gly Val Leu Thr Leu Thr Phe Asp Ser Thr Asp

65 70 75 80

Pro Leu Asp Leu Tyr Thr Asp Asn Thr Tyr Gln Thr Met Val Glu Pro

85 90 95

Arg Leu Val Tyr Asn Asn Gly Val Cys Glu Leu Glu Val Lys Ala Gly

100 105 110

Ile Thr Tyr Tyr Phe His Arg Ser Phe Met Leu Ser Ala Arg Thr Ile

115 120 125

Ser Val Glu Phe Gly Lys Glu Ala Val Ala Leu Lys Leu Gln Arg Val

130 135 140

Ser Pro Thr Glu Gly Glu Thr Leu Phe Val Ser Asn Ala Thr Val Ser

145 150 155 160

Phe Thr Phe Asn Arg Asp Val Lys Met Asp Gly Ala Thr Leu Thr Ile

165 170 175

Gly Gly Lys Glu Lys Ser Ile Asp Ala Val Ser Thr Ser Ser Ser Val

180 185 190

Gln Phe Asp Leu Ser Ala Ile Met Met Glu Ala Tyr Asn Asp Gly Ile

195 200 205

Leu Lys Glu Gly Asp Asp Met Leu Leu Thr Leu Lys Gly Val Arg Asn

210 215 220

Ala Asn Lys Glu Ser Asp Ile Tyr Gly Glu Asp Gly Thr Cys Ser Leu

225 230 235 240

Ala Leu Lys Ala Ala Ala Lys Pro Val Val Leu Asn Ser Ser Val Asn

245 250 255

Thr Pro Gly Asn Gly Met Asp Ile Phe His Ser Tyr Ile Met Ser Gly

260 265 270

Asp Glu Asn Val Val Gln Leu Asn Phe Asp Gly Ala Leu Asp Thr Glu

275 280 285

Lys Lys Pro Val Ala Thr Leu Ile Tyr Gly Asp Ile Glu Ala Glu Asn

290 295 300

Gly Phe Tyr Gln Glu Thr Leu Asp Ala Gln Phe Leu Gly Glu Asn Thr

305 310 315 320

Val Thr Val Asp Leu Ser Gly Lys Ser Arg Arg His Lys Asp Met Leu

325 330 335

Pro Asn Tyr Ser Gly Ala Ala Phe Gly Thr Ile Ser Leu Lys Ile Ala

340 345 350

Gly Val Tyr Ala Ala Asp Gly Gln Pro Val Tyr Ser Gly Ala Gln Ser

355 360 365

Ser Val Gly Ser Cys Thr Phe Gln Tyr Ala Tyr Glu Glu Val Lys Val

370 375 380

Asp Ile Ala Val Ala Phe Asp Asp Leu Ala Glu Ser Asn Ser Ile Asp

385 390 395 400

Gly Leu Asp Ser Leu Thr Ile Trp Val Arg Gly Asp Glu Tyr Leu His

405 410 415

Tyr Thr Gly Val Gln Phe Asp Phe Met Val Asn Gly Gln Val Asn Ser

420 425 430

Ile Val Glu Lys Asp Ile Lys Lys Lys Ala Asp Ser Glu Glu Glu Gly

435 440 445

Ala Cys Ile Leu Arg Val Cys Val Pro Lys Ser Asp Ala Asp Ala Asn

450 455 460

Ser Glu Val Lys Val Ser Phe Val Gly Leu Glu Ala Ala Asp Gly Gly

465 470 475 480

Asp Tyr Ser Ala Asp Phe Ser Ala Val Phe Thr Thr Val Gly Asn Leu

485 490 495

Val Asn Val Pro Asp Leu Ile Leu Thr Pro Val Ser Gly Ser Thr Val

500 505 510

Glu Asn Ile Lys Glu Val Leu Val Gly Cys Ser Glu Gly Ile Arg Val

515 520 525

Asn Ala Asp Asn Lys Glu Lys Ile Gln Ile Trp Asp Arg Met Arg Asn

530 535 540

Val Val Ala Thr Ala Val Ser Met Glu Pro Val Ile Pro Glu Asp Glu

545 550 555 560

Lys Glu Asn Pro Asp Tyr Ile Pro Thr Glu Ile Lys Val Val Phe Asp

565 570 575

Asn Glu Val Lys Leu Pro Gly Ala Tyr Met Val Asn Leu Pro Ala Glu

580 585 590

Ala Phe Val Leu Gly Asn Gln Thr Ser Ile Leu Ser Lys Ala Thr Leu

595 600 605

Val Asp Tyr Met Ile Ile Gly Glu Val Ala Lys Asp Tyr Val Pro Ser

610 615 620

Glu Val Val Ala Glu Thr Glu Gly Ser Thr Val Ile Gly Phe Thr Ile

625 630 635 640

Lys Thr Pro Glu Trp Val Thr Leu Asp Glu Asn Phe Asp Thr Glu Lys

645 650 655

Ile Lys Ile Tyr Asn Ser Asn Thr Gln Glu Glu Val Val Gly Gly Lys

660 665 670

Ser Val Ser Tyr Ala Ala Ala Phe Glu Asp Phe Ser Ile Ala Leu Glu

675 680 685

Asn Pro Leu Thr Glu Lys Gly Thr Tyr Thr Leu Phe Ile Pro Ala Glu

690 695 700

Leu Phe Gly Asn Gly Ser Trp Ile Pro Gly Ser Thr Glu Gly Ser Cys

705 710 715 720

Asn Pro Glu Leu Thr Tyr Thr Ile Val Ile Asp Glu Asn Gly Val Thr

725 730 735

Val Gly Val Asn Ser Val Leu Ala Glu Ile Gly Thr Lys Val Thr Val

740 745 750

Tyr Asn Leu Asn Gly Val Arg Val Leu Tyr Asn Ala Asp Arg Asp Ala

755 760 765

Leu Lys Ala Leu Arg Lys Gly Ile Tyr Val Val Asn Gly Lys Lys Val

770 775 780

Val Ile Lys

785

<210> 22

<211> 2364

<212> DNA

<213> Paraprevotella xylaniphila

<400> 22

atgaggaaga tgtttacttt ggaaatgaaa gactatctga tgaggatggt tctttttgtg 60

gcagtgtgta tggtgggtgt tggaggattg aaagctgccc ccggattgtc agctgatgat 120

cctttgatct ttgaaggtgg agaagaatat gatgtttcgg gaagttattt taaagatttg 180

tatgctacgt ttacggctcc ttcggacgga gtattgactt tgacatttga tagtaccgat 240

cctttggatt tatatacgga taatacatat caaacgatgg tggagccacg tcttgtttac 300

aataatgggg tttgcgaatt ggaagtaaaa gccgggatta cttattattt tcaccgttct 360

tttatgttga gcgcacgaac catttctgtg gaatttggaa aggaagctgt tgctttgaag 420

ttacaacgag tttctccaac agagggtgag acattgtttg tttccaatgc tacagtaagt 480

ttcactttca atagggatgt taaaatggat ggggctactc ttacaattgg aggaaaagaa 540

aagtctattg atgcggtttc gacatcgtct tctgtacaat ttgacctatc tgccattatg 600

atggaggcat acaatgatgg aatcctaaag gaaggagatg atatgctctt aactttgaag 660

ggagtacgga atgcgaataa agaaagtgat atttatggag aagacgggac ttgctccttg 720

gctttaaaag cagcagcaaa acctgtggtt ctaaacagtt ctgtaaatac gcctggaaat 780

ggaatggata tatttcattc ttatattatg tctggggacg agaatgttgt acagttgaat 840

tttgatggtg ctttggatac agagaagaaa ccggttgcaa cattaatata tggggatata 900

gaagcggaga atggttttta tcaagaaacc ttagatgcgc aattcttagg tgaaaatact 960

gtaactgtag atttgagcgg aaaatcgcgt cggcataaag atatgctgcc gaactattcc 1020

ggcgctgcct ttggaactat ttctttgaaa attgcagggg tgtatgcagc tgacggacaa 1080

cctgtttatt ctggtgctca atcctcagta ggttcttgta cgtttcaata tgcctatgag 1140

gaagtaaagg tggatatagc tgtggctttt gatgacttgg cagaaagtaa ttctattgac 1200

ggattagact cattaacaat ttgggttcgt ggtgatgaat acttgcatta cactggagtg 1260

cagtttgatt ttatggtaaa tggtcaagtt aattctattg tagagaaaga tattaagaaa 1320

aaagcggatt ctgaagaaga aggtgcctgt atattaagag tatgtgtacc gaaaagtgat 1380

gccgatgcca attcggaggt aaaggtcagc tttgttggcc ttgaagcagc tgacggcggg 1440

gattactccg cagatttttc tgctgttttc acaacagtag gaaacttggt aaatgtaccg 1500

gatttgatac tcactccagt gagcggttct acagtggaaa atattaaaga ggtattagtt 1560

ggttgttctg agggtattcg cgtgaatgca gacaacaagg agaaaataca gatatgggat 1620

agaatgcgga acgtagtagc tacagcggta agtatggagc ctgttattcc tgaagatgag 1680

aaggaaaatc cggattacat accgacagaa ataaaggtgg tttttgataa tgaagtgaag 1740

ctccccggag cttatatggt gaatcttccg gcagaggcat tcgttttagg aaatcagaca 1800

tctattttaa gtaaggcaac ccttgtggat tatatgatta taggggaagt tgcgaaagat 1860

tatgttccga gcgaagttgt agccgagaca gaaggaagta cagtgatagg ttttacgata 1920

aagacaccag aatgggtgac tttggatgag aattttgaca cagagaaaat caagatttat 1980

aattcgaata ctcaggaaga agttgtgggt ggaaaatcgg taagttatgc agcagctttt 2040

gaagatttta gtatcgcatt ggagaatcct ctcacagaga aaggtactta tacgttgttt 2100

attccggcag aattgtttgg aaatggttct tggattcccg gttctacaga agggtcttgt 2160

aatccggaat taacttatac gatagttata gacgagaatg gggtgacagt cggagtaaat 2220

tcagtattgg ctgaaattgg gacaaaagta acggtgtaca atcttaatgg tgtacgtgtg 2280

ctttataatg cagaccgtga tgcattgaag gctttaagaa aaggaattta tgtcgtgaat 2340

ggaaagaagg tggtgattaa gtaa 2364

<210> 23

<211> 787

<212> PRT

<213> Paraprevotella xylaniphila

<400> 23

Met Arg Lys Met Phe Thr Leu Glu Met Lys Asp Tyr Leu Met Arg Met

1 5 10 15

Val Leu Phe Val Ala Val Cys Met Val Gly Val Gly Gly Leu Lys Ala

20 25 30

Ala Pro Gly Leu Ser Ala Asp Asp Pro Leu Ile Phe Glu Gly Gly Glu

35 40 45

Glu Tyr Asp Val Ser Gly Ser Tyr Phe Lys Asp Leu Tyr Ala Thr Phe

50 55 60

Thr Ala Pro Ser Asp Gly Val Leu Thr Leu Thr Phe Asp Ser Thr Asp

65 70 75 80

Pro Leu Asp Leu Tyr Thr Asp Asn Thr Tyr Gln Thr Met Val Glu Pro

85 90 95

Arg Leu Val Tyr Asn Asn Gly Val Cys Glu Leu Glu Val Lys Ala Gly

100 105 110

Ile Thr Tyr Tyr Phe His Arg Ser Phe Met Leu Ser Ala Arg Thr Ile

115 120 125

Ser Val Glu Phe Gly Lys Glu Ala Val Ala Leu Lys Leu Gln Arg Val

130 135 140

Ser Pro Thr Glu Gly Glu Thr Leu Phe Val Ser Asn Ala Thr Val Ser

145 150 155 160

Phe Thr Phe Asn Arg Asp Val Lys Met Asp Gly Ala Thr Leu Thr Ile

165 170 175

Gly Gly Lys Glu Lys Ser Ile Asp Ala Val Ser Thr Ser Ser Ser Val

180 185 190

Gln Phe Asp Leu Ser Ala Ile Met Met Glu Ala Tyr Asn Asp Gly Ile

195 200 205

Leu Lys Glu Gly Asp Asp Met Leu Leu Thr Leu Lys Gly Val Arg Asn

210 215 220

Ala Asn Lys Glu Ser Asp Ile Tyr Gly Glu Asp Gly Thr Cys Ser Leu

225 230 235 240

Thr Leu Lys Ala Ala Ala Lys Pro Val Val Leu Asn Ser Ser Val Asn

245 250 255

Thr Pro Gly Asn Gly Met Asp Ile Phe His Ser Tyr Val Met Ser Gly

260 265 270

Asp Glu Asn Val Val Gln Leu Asn Phe Asp Gly Ala Leu Asp Thr Glu

275 280 285

Lys Lys Pro Val Ala Thr Leu Ile Tyr Gly Asp Ile Glu Ala Glu Asn

290 295 300

Gly Phe Tyr Gln Glu Thr Leu Asp Ala Gln Phe Leu Gly Glu Asn Thr

305 310 315 320

Val Thr Val Asp Leu Ser Gly Lys Ser Arg Arg His Lys Asp Met Leu

325 330 335

Pro Asn Tyr Ser Gly Ala Ala Phe Gly Thr Ile Ser Leu Lys Ile Ala

340 345 350

Gly Val Tyr Ala Ala Asp Gly Gln Pro Val Tyr Ser Gly Ala Gln Ser

355 360 365

Ser Val Gly Ser Cys Thr Phe Gln Tyr Ala Tyr Glu Glu Val Lys Val

370 375 380

Asp Ile Ala Val Ala Phe Asp Asp Leu Ala Gly Ser Asn Ser Ile Asp

385 390 395 400

Gly Leu Asp Ser Leu Thr Ile Trp Val Arg Gly Asp Glu Tyr Leu His

405 410 415

Tyr Thr Gly Val Gln Phe Asp Phe Met Val Asn Gly Gln Val Asn Ser

420 425 430

Ile Val Glu Lys Asp Ile Lys Lys Lys Ala Asp Ser Glu Glu Glu Gly

435 440 445

Ala Cys Ile Leu Arg Val Cys Val Pro Lys Ser Asp Ala Asp Ala Asn

450 455 460

Ser Glu Val Lys Val Ser Phe Val Gly Leu Glu Ala Ala Asp Gly Gly

465 470 475 480

Asp Tyr Ser Ala Asp Phe Ser Ala Val Phe Thr Thr Val Gly Asn Leu

485 490 495

Val Asn Val Pro Asp Leu Ile Leu Thr Pro Val Ser Gly Ser Thr Val

500 505 510

Glu Asn Ile Lys Glu Val Leu Val Gly Cys Ser Glu Gly Ile Arg Val

515 520 525

Asn Ala Asp Asn Lys Glu Lys Ile Gln Ile Trp Asp Arg Met Arg Asn

530 535 540

Val Val Ala Thr Ala Val Ser Met Glu Pro Val Ile Pro Glu Asp Glu

545 550 555 560

Lys Glu Asn Pro Asp Tyr Ile Pro Thr Glu Ile Lys Val Val Phe Asp

565 570 575

Asn Glu Val Lys Leu Pro Gly Ala Tyr Met Val Asn Leu Pro Ala Glu

580 585 590

Ala Phe Val Leu Gly Asn Gln Thr Ser Ile Leu Ser Lys Ala Thr Leu

595 600 605

Val Asp Tyr Met Ile Ile Gly Glu Val Ala Lys Asp Tyr Val Pro Ser

610 615 620

Glu Val Val Ala Glu Thr Glu Gly Ser Thr Val Ile Gly Phe Thr Ile

625 630 635 640

Lys Thr Pro Glu Trp Val Thr Leu Asp Glu Asn Phe Asp Thr Glu Lys

645 650 655

Ile Lys Ile Tyr Asn Ser Asn Thr Gln Glu Glu Val Val Gly Gly Lys

660 665 670

Ser Val Ser Tyr Ala Ala Ala Phe Glu Asp Phe Ser Ile Ala Leu Glu

675 680 685

Asn Pro Leu Thr Glu Lys Gly Thr Tyr Thr Leu Phe Ile Pro Ala Glu

690 695 700

Leu Phe Gly Asn Gly Ser Trp Ile Pro Gly Ser Thr Glu Gly Ala Cys

705 710 715 720

Asn Pro Glu Leu Thr Tyr Thr Ile Val Ile Asp Glu Asn Gly Val Thr

725 730 735

Val Gly Val Asn Ser Val Leu Ala Glu Ile Gly Thr Lys Val Thr Val

740 745 750

Tyr Asn Leu Asn Gly Val Arg Val Leu Tyr Asn Ala Asp Arg Asp Ala

755 760 765

Leu Lys Ala Leu Arg Lys Gly Ile Tyr Val Val Asn Gly Lys Lys Val

770 775 780

Val Ile Lys

785

<210> 24

<211> 2364

<212> DNA

<213> Paraprevotella xylaniphila

<400> 24

atgaggaaga tgtttacttt ggaaatgaaa gactatctga tgaggatggt tctttttgtg 60

gcagtgtgta tggtgggtgt tggaggattg aaagctgccc ccggattgtc agctgatgat 120

cctttgatct ttgaaggtgg agaagaatat gatgtttcgg gaagttattt taaagatttg 180

tatgctacgt ttacggctcc ttcggacgga gtattgactt tgacatttga tagtaccgat 240

cctttggatt tatatacgga taatacatat caaacgatgg tggagccacg tcttgtttac 300

aataatgggg tttgcgaatt ggaagtaaaa gccgggatta cttattattt tcaccgttct 360

tttatgttga gcgcacgaac catttctgtg gaatttggaa aggaagctgt tgctttgaag 420

ttacaacgag tttctccaac agagggtgag acattgtttg tttccaatgc tacagtaagt 480

ttcactttca atagggatgt taaaatggat ggggctactc ttacaattgg aggaaaagaa 540

aagtctattg atgcggtttc gacatcgtct tctgtacaat ttgacctatc tgccattatg 600

atggaggcat acaatgatgg aatcctaaag gaaggagatg atatgctctt aactttgaag 660

ggagtacgga atgcgaataa agaaagtgat atttatggag aagacgggac ttgctccttg 720

actttaaaag cagcagcaaa acctgtggtt ctaaacagtt ctgtaaatac gcctggaaat 780

ggaatggata tatttcattc ttatgttatg tctggggacg agaatgttgt acagttgaat 840

tttgatggtg ctttggatac agagaagaaa ccggttgcaa cattaatata tggggatata 900

gaagcggaga atggttttta tcaagaaacc ttagatgcgc aattcttagg tgaaaatact 960

gtaactgtag atttgagcgg aaaatcgcgt cggcataaag atatgctgcc gaactattcc 1020

ggcgctgcct ttggaactat ttctttgaaa attgcagggg tgtatgcagc tgacggacaa 1080

cctgtttatt ctggtgctca atcctcagta ggttcttgta cgtttcaata tgcctatgag 1140

gaagtaaagg tggatatagc tgtggctttt gatgacttgg caggaagtaa ttctattgac 1200

ggattagact cattaacaat ttgggttcgt ggtgatgaat acttgcatta cactggagtg 1260

cagtttgatt ttatggtaaa tggtcaagtt aattctattg tagagaaaga tattaagaaa 1320

aaagcggatt ctgaagaaga aggtgcctgt atattaagag tatgtgtacc gaaaagtgat 1380

gccgatgcca attcggaggt aaaggtcagc tttgttggcc ttgaagcagc tgacggcggg 1440

gattactccg cagatttttc tgctgttttc acaacagtag gaaacttggt aaatgtaccg 1500

gatttgatac tcactccagt gagcggttct acagtggaaa atattaaaga ggtattagtt 1560

ggttgttctg agggtattcg cgtgaatgca gacaacaagg agaaaataca gatatgggat 1620

agaatgcgga acgtagtagc tacagcggta agtatggagc ctgttattcc ggaagatgag 1680

aaggaaaatc cggattacat accgacagaa ataaaggtgg tttttgataa tgaagtgaag 1740

ctccccggag cttatatggt gaatcttccg gcagaggcat tcgttttagg aaatcagaca 1800

tctattttaa gtaaggcaac ccttgtggat tatatgatta taggggaagt tgcgaaagat 1860

tatgttccga gcgaagttgt agccgagaca gaaggaagta cagtgatagg ttttacgata 1920

aagacaccag aatgggtgac tttggatgag aattttgaca cagagaaaat caagatttat 1980

aattcgaata ctcaggaaga agttgtgggt ggaaaatcgg taagttatgc agcagctttt 2040

gaagatttta gtatcgcatt ggagaatcct ctcacagaga aaggtactta tacgttgttt 2100

attccggcag aattgtttgg aaatggttct tggattcccg gttctacaga aggggcttgt 2160

aatccggaat taacttatac gatagttata gacgagaatg gggtgacagt cggagtaaat 2220

tcagtattgg ctgaaattgg gacaaaagta acggtgtaca atcttaatgg tgtacgtgtg 2280

ctttataatg cagaccgtga tgcattgaag gctttaagaa aaggaattta tgtcgtgaat 2340

ggaaagaagg tggtgattaa gtaa 2364

<210> 25

<211> 542

<212> PRT

<213> Prevotella rara

<400> 25

Met Lys Arg Leu Ile Phe Thr Leu Ile Gly Ile Val Leu Gly Leu Thr

1 5 10 15

Gly Leu Lys Ala Val Thr Phe Thr Pro Ile Glu Ile Asn Asp Pro Ser

20 25 30

Glu Ser Val Thr Ile Lys Leu Gln Pro Gly Ala Asn Tyr Leu Gln Leu

35 40 45

Lys Ala Ile Glu Ser Asn Thr Ile His Phe Asn Val Gly Tyr Phe Gly

50 55 60

Ile Met Met Phe Glu Cys Asn Ala Met Gly Glu Glu Gly Asn Asn Leu

65 70 75 80

Ser Ile Ser Tyr Asp Ala Glu Gly His Lys Ile Phe Ser Ser Glu Val

85 90 95

Glu Glu Gly Lys Thr Tyr Tyr Phe Ser Thr Ser Met Ile Thr Glu Pro

100 105 110

Ser Ile Asp Val Thr Ile Tyr Tyr Gly Asn Gly Glu Gly Met Pro Ile

115 120 125

Thr Leu Thr Ser Asn Phe Ser Asp Gly Asp Thr Tyr Thr Val Ser Gly

130 135 140

Ser Asn Leu Glu Leu Ala Phe Asp Arg Thr Val Asp Ile Ala His Asn

145 150 155 160

Trp Ile Glu Tyr Gly Glu Glu Ala Asp Gly Val Phe Lys Thr Lys Glu

165 170 175

Glu Ile Pro Ala Ala Tyr Ile Asn Gly Thr Tyr Thr Thr Gln Tyr Phe

180 185 190

Tyr Ser Ile Glu Leu Ser Lys Phe Ile Arg Glu Met Ala Glu Asp Gly

195 200 205

Lys Leu Glu Val Gly Asp Lys Phe Lys Ile Thr Leu Glu Gly Ile Cys

210 215 220

Asp Ala Asn Asp Glu Ser Val Ile Tyr Gly Glu Asp Gly Asn Tyr Ser

225 230 235 240

Val Thr Leu Ile Met Gly Glu Met Pro Gly Glu Leu Val Ser Ile Asp

245 250 255

Ser Ala Gln Gly Thr Thr Leu Tyr Thr Tyr Tyr Pro Glu Thr Gly Glu

260 265 270

Glu Gly Leu Leu Thr Phe Thr Phe Thr Asp Glu Leu Asn Thr Asp Lys

275 280 285

Ser Lys Val Thr Val Asp Leu Ser Tyr Gly Asp Ser Glu Ala Gly Ser

290 295 300

Met Gly Ser Phe Asn Pro Asp Phe Thr Ile Glu Gly Lys Thr Val Val

305 310 315 320

Val Asp Ile Arg Gly Tyr Arg Phe Pro Glu Thr Val Glu Thr Ser Arg

325 330 335

Asn Gly Glu Val Gln Thr Val Ile Thr Met Asn Ile Lys Gly Leu Thr

340 345 350

Thr Ala Asp Gly Arg Ala Ile Leu Thr Asn Asn Ala Ser Ala Gly Thr

355 360 365

Ser Gly Ile Val Val Thr Tyr Pro Leu Lys Lys Gln Glu Ile Ser Phe

370 375 380

Tyr Tyr Asp Phe Leu Pro Thr Glu Gly Ser Ser Leu Ala Asp Cys Ser

385 390 395 400

Glu Ile Ile Ile Trp Leu Pro Glu Glu Val Gly Val Leu Thr Phe Asp

405 410 415

Lys Val Leu Leu Gln Trp Leu Asn Asn Arg Gly Thr Leu Gln Thr Lys

420 425 430

Glu Phe Lys Ala Glu Asp Val Pro Phe Ala Tyr Asp Lys Ser Tyr Ala

435 440 445

Gly Tyr Val Ser His Ile Pro Leu Ala Gly Val Ser Lys Asp Arg Glu

450 455 460

Val Thr Leu Thr Val Glu Gly Gly Met Leu Ser Asn Gly Asp Ser Val

465 470 475 480

Glu Ile Thr Gly Lys Phe Asn Thr Asp Pro Thr Gly Ile Asp Ser Val

485 490 495

Leu Gly Asp Asp Ala Asn Ala Val Val Lys Leu Tyr Thr Ile Asp Gly

500 505 510

Ile Leu Val Lys Glu Ala Pro Ala Ala Thr Val Leu Thr Gly Val Lys

515 520 525

Lys Gly Val Tyr Ile Met Asn Gly Lys Lys Val Val Val Lys

530 535 540

<210> 26

<211> 1629

<212> DNA

<213> Prevotella rara

<400> 26

atgaaacgac ttatttttac tttaatcggt attgtattgg ggttgacagg tttaaaggct 60

gttactttta ctccgattga gattaatgac ccgtctgagt ctgtaactat taaattgcag 120

ccaggtgcaa actatttaca gctgaaggct attgagagca acaccatcca ctttaatgtc 180

ggatattttg gcattatgat gtttgaatgt aatgccatgg gtgaagaggg caacaacctt 240

tctataagct atgacgctga aggtcacaaa atcttcagta gtgaggttga ggaaggaaaa 300

acttattatt tctcaaccag tatgattact gagccttcaa tagatgttac aatttattat 360

ggtaacggag aaggcatgcc tatcacattg acatctaact tttcggatgg tgatacctat 420

acagtttctg gctcaaacct tgaattggct tttgaccgta cggttgatat agcccacaac 480

tggattgagt atggtgagga ggctgacggc gtatttaaaa caaaggaaga gattcccgca 540

gcatatatca atggtaccta tactactcag tatttctata gcatcgagct gagcaagttt 600

atccgtgaga tggctgaaga cggtaaactt gaggttggtg ataagtttaa aatcactctt 660

gaaggtattt gcgatgctaa cgatgagagc gtaatctatg gtgaggacgg aaactattct 720

gttacactga ttatgggcga gatgcccggt gagctggttt ctatagattc ggcacaaggt 780

acaactcttt atacatatta tcctgaaacc ggtgaagaag gtttgctgac atttactttc 840

acagatgaac ttaatactga taagagtaag gtaactgtag acctgtctta tggcgattca 900

gaggccggtt ctatgggttc atttaatcct gattttacta tcgaaggcaa gactgttgtt 960

gttgatatcc gcggttaccg cttcccagag acagtagaga cttcaagaaa cggtgaagtg 1020

cagactgtga taacaatgaa catcaagggc cttacaactg cagacggacg tgctattctc 1080

acaaacaacg cttctgccgg aacaagtggc atcgttgtta cttatcctct gaagaagcag 1140

gaaatatctt tttattatga cttcttgcct actgaaggca gctctctggc tgattgcagc 1200

gagatcataa tctggttgcc ggaggaagtt ggtgttctta ctttcgacaa ggttctgttg 1260

cagtggctca acaaccgcgg cacactccag acaaaggaat tcaaggcaga ggatgttccg 1320

tttgcttatg ataaatctta tgcgggttat gtaagccata ttccacttgc cggcgtaagc 1380

aaggaccgtg aggttacatt gactgtagaa ggtggaatgc tcagcaatgg cgacagcgtt 1440

gagattacag gtaaattcaa cactgatcct acaggtatag attctgttct tggtgacgat 1500

gccaatgccg ttgttaagct ttacacaatc gacggtatcc tcgttaagga ggctcctgct 1560

gcaactgttc ttacaggtgt taagaagggc gtttatatca tgaacggcaa gaaggttgtt 1620

gtaaaataa 1629

<210> 27

<211> 1027

<212> PRT

<213> Prevotella rodentium

<400> 27

Met Phe Ser Phe Met Gly Ile Gly Gln Ala Ile Ala Asp Val Val Gly

1 5 10 15

Ser Gly Ser Lys Glu Asp Pro Tyr Val Leu Glu Asn Gly Thr Thr Leu

20 25 30

Thr Leu Lys Ala Tyr Gln Ser Phe Tyr Ala Lys Phe Thr Ala Pro Ala

35 40 45

Asp Gly Ile Phe Ser Leu Tyr Thr Lys Asp Ser Tyr Ala Leu Tyr Thr

50 55 60

Asp Glu Ser Phe Ser Gln Ile Asp Glu Ser Ala Asn Ile Thr Phe Asn

65 70 75 80

Gly Ser Tyr Ser Glu Thr Ala Tyr Tyr Phe Asn Cys Lys Ala Gly Val

85 90 95

Thr Tyr Tyr Ile Gly Asn Ser Phe Val Met Val Gly Gly Thr Val Thr

100 105 110

Ala Lys Phe Asn Thr Glu Ala Glu Pro Leu Val Leu Arg Glu Ile Ser

115 120 125

Pro Glu Ala Gly Ser Val Phe Asn Ala Gly Lys Gly Ser Val Glu Leu

130 135 140

Thr Phe Asn Gln Lys Val Gln Ile Ala Ser Val Thr Val Asn Ala Gly

145 150 155 160

Thr Ser Ser Lys Thr Met Ser Ala Asn Val Asn Gly Val Tyr Val Ser

165 170 175

Val Asp Val Lys Asp Trp Leu Asn Gln Met Tyr Asp Glu Gly Lys Val

180 185 190

Lys Glu Gly Asn Glu Ile Ser Phe Lys Phe Lys Gly Val Ala Pro Thr

195 200 205

Ile Ser Pro Ser Lys Leu Tyr Asn Gly Asn Gly Glu Leu Asp Ile Thr

210 215 220

Tyr Thr Ala Gly Lys Lys Pro Leu Gln Leu Val Ser Ser Thr Asn Thr

225 230 235 240

Pro Met Ser Thr Pro Ala Val Thr Thr Phe Lys Ser Phe Tyr Thr Ser

245 250 255

Asp Asp Asn Gln Gly Ile Val Thr Leu Glu Phe Ser Asp Glu Val Asn

260 265 270

Phe Ser Glu Gly Asn Lys Pro Thr Ala Thr Leu Thr Tyr Gly Asn Met

275 280 285

Asp Gln Asp Ala Asp Gly Glu Tyr Tyr Thr Glu Ser Leu Pro Ile Leu

290 295 300

Pro Leu Gly Ser Asn Ile Leu Met Val Asn Leu Lys Asp Lys Leu Arg

305 310 315 320

Arg Ala Gln Asp Met Val Thr Ser Gly Thr Val Tyr Asp Gly Ile Thr

325 330 335

Leu Ser Ile Ser His Val Lys Asp Met Asp Gly Asn Tyr Ala Tyr Gly

340 345 350

Ser Gly Ser Gly Val Leu Gly Ser Phe Ala Phe Asn Tyr Lys Leu Ser

355 360 365

Glu Ile Asn Tyr Thr Val Asp Thr Asp Trp Ala Leu Leu Asp Ala Thr

370 375 380

Gly Asn Pro Thr Asn Lys Asp Val Ile Asp Ser Asp Thr Lys Ser Ile

385 390 395 400

Glu Leu Trp Met Ser Glu Asn Asp Gly Gln Ile Thr Phe Asn Gly Val

405 410 415

Glu Phe Lys Tyr Thr Glu Ser Gly Thr Glu Lys Val Lys Thr Met Thr

420 425 430

Leu Ser Glu Ile Lys Val Asp Asn Asn Gly Asn Glu Thr Thr Ile Thr

435 440 445

Ile Pro Val Pro Asn Ile Ala Ala Asp Ala Gly Ser Asp Ile Ala Ile

450 455 460

Ser Leu Lys Asp Val Glu Arg Pro Asp Gly Ile Thr Thr Ser Ile Asp

465 470 475 480

Pro Ala Ala Leu Ser Tyr Phe Thr Lys Thr Phe Thr Thr Thr Gly Ile

485 490 495

Thr Glu Ser Lys Phe Asp Ile Thr Ser Ala Ile Trp Met Tyr Glu Glu

500 505 510

Asn Glu Gly Asn Ile Val Glu Val Asn Met Ile Asn Gly Asn Ile Gly

515 520 525

Val Leu Thr Arg Gly Ser Lys Ser Val Ile Lys Thr Asn Lys Asp Asn

530 535 540

Glu Ile Gly Tyr Val Glu Trp Glu Ile Arg Gly Val Asp Asn Pro Asp

545 550 555 560

Met Glu Tyr Ile Arg Ala Gly Tyr Val Asp Thr Thr Met Thr Gly Lys

565 570 575

Leu Val Asp Gly Phe Thr Ile Glu Trp Arg Gly Glu Gly Leu Thr Ala

580 585 590

Gly Lys Asp Tyr Thr Phe Thr Leu Gln Ala Trp Lys Asn Glu Ala Asp

595 600 605

Lys Asn Ser Gly Ala Glu Pro Asn Val Gly Glu Ala Met Phe Ile Ile

610 615 620

His Gly Thr Lys Gln Ala Tyr Ile Tyr Ser Asp Val Val Met Lys Thr

625 630 635 640

Asp Ile Ser Gln Pro Ile Arg Leu Ala Ser Ala Asn Asp Asn Tyr Arg

645 650 655

Thr Ile Glu Phe Ser Ala Pro Val Thr Leu Asn Ala Val Val Asn Leu

660 665 670

Gly Met Gly Thr Ser Ala Asp Cys Thr Val Glu Pro Ser Ala Asp Arg

675 680 685

Thr Ala Trp Thr Val Thr Ile Pro Glu Tyr Val Met Ser Gln Tyr Gly

690 695 700

Glu Phe Ser Val Asn Val Phe Ala Lys Asp Asp Glu Gly Arg Ala Val

705 710 715 720

Asn Lys Thr Glu Asn Gly Leu Gly Ile Ile Met Gly Thr Glu Asp Asn

725 730 735

Thr Trp Phe Gln Ile Asp Phe Val Ser Glu Ser Leu Ala Pro Asp Phe

740 745 750

Thr Val Thr Pro Ala Asn Glu Ser Val Leu Glu Ser Leu Ser Thr Val

755 760 765

Thr Phe Gly Tyr Glu Gly Ser Ile Ser Ile Asn Trp Asn Asn Ser Glu

770 775 780

Lys Ile Thr Ile Tyr Asn Arg Thr Thr Arg Glu Lys Ile Ala Glu Phe

785 790 795 800

Ser Gly Asp Asp Val Val Leu Asp Glu Asp Pro Asp Asp Tyr Trp Ala

805 810 815

Pro Ile Leu Ser Cys His Ile Thr Leu Pro Glu Pro Val Thr Ala Ile

820 825 830

Gly Val Tyr Asp Val Asn Val Pro Ala Gly Phe Phe Val Leu Gly Glu

835 840 845

Gln Phe Asp Ser Asn Leu Ser Lys Ala Thr Ser Ile Val Tyr Glu Ile

850 855 860

Lys Glu Pro Val Glu Pro Phe Gly Ile Glu Ile Ser Pro Thr Ala Gly

865 870 875 880

Leu Val Ser Glu Ile Pro Ser Lys Leu Ile Val Thr Val Thr Asp Arg

885 890 895

Ser Ser Cys Asn Phe Thr Ala Asn Pro Thr Leu Thr Asp Asn Thr Gly

900 905 910

Asn Ser Tyr Pro Val His Asn Asp Phe Asp Trp Gly Ile Pro Glu Met

915 920 925

Asn Lys Phe Val Ile Ile Leu Asp Asn Gly Ala Ile Thr Ala Asp Gly

930 935 940

Ile Tyr Thr Leu Thr Ile Pro Ala Gly Ser Ile Ile Gly Asp Asp Glu

945 950 955 960

Thr Asp Leu Asn Lys Glu Asp Phe Val Phe Ile Tyr Thr Ile Asn Leu

965 970 975

Ala Gly Ile Thr Glu Leu Val Asn Asn Glu Gly Gly Lys Val Asp Val

980 985 990

Tyr Thr Leu Lys Gly Thr Leu Leu Met Lys Asp Ala Asp Ala Ser Ala

995 1000 1005

Val Asn Lys Leu Ala Lys Gly Met Tyr Ile Ile Asn Gly Lys Lys

1010 1015 1020

Val Phe Ile Arg

1025

<210> 28

<211> 3084

<212> DNA

<213> Prevotella rodentium

<400> 28

atgttttcat ttatgggcat cggacaggcc atagccgatg ttgtgggttc gggatcaaaa 60

gaagatccgt acgtgcttga aaacggcaca acactgacat tgaaagcata ccagtctttc 120

tatgccaaat tcacggcccc ggctgacggg atcttctcat tatatacaaa agacagctat 180

gccctatata ccgacgagag tttcagtcag attgacgagt cggcaaacat tacattcaac 240

ggaagctaca gtgagacggc atactacttt aactgtaaag caggtgttac ctattatata 300

ggaaacagtt ttgtaatggt cggaggtacg gttacagcca aattcaacac agaggctgaa 360

ccattggttc tgcgtgaaat atcacctgag gccggttctg ttttcaatgc aggcaaagga 420

agcgtagaac tgacattcaa ccaaaaagtc caaatagctt ctgttactgt caatgctggc 480

acatcctcaa agacaatgtc agccaatgta aatggagtat atgtatctgt agacgtaaaa 540

gactggctga accaaatgta tgatgaaggt aaggtcaaag aaggaaatga gatcagcttc 600

aagttcaaag gcgtggcccc aaccatttct ccaagcaaat tatacaacgg caatggagag 660

ttggacatta catacacagc aggcaagaaa cccctgcaac ttgtaagcag taccaacacc 720

ccaatgagca ctcctgccgt aaccacattc aagtcattct atacatccga tgacaaccaa 780

ggcatcgtaa cactggaatt cagtgacgaa gtgaactttt cggaaggcaa caaaccaacc 840

gcaaccctta catatggaaa tatggaccaa gatgccgacg gcgaatacta cacagaatca 900

ctccccatac ttccacttgg aagcaacatt ttaatggtta atctcaagga caagctgcgc 960

cgtgcacagg acatggtgac ttccggcact gtttacgacg gaataacact atccatatca 1020

cacgtaaagg atatggacgg aaactatgcc tatggctcgg gctccggtgt actcggatcg 1080

ttcgcattca actacaaact atctgaaata aactataccg ttgatacaga ctgggcacta 1140

cttgacgcaa ccggcaatcc gacaaacaaa gacgtcatag attcagacac caaaagcata 1200

gaactatgga tgagcgaaaa cgacggtcag ataacattca acggagtaga attcaaatat 1260

accgaaagcg gtactgaaaa ggtcaagaca atgaccctca gcgagattaa agttgacaac 1320

aatggaaacg aaacaacaat aacaatcccc gttcccaaca tcgcagccga tgccggtagc 1380

gacatcgcca tatctctcaa agacgtggaa cgccccgacg gtattaccac cagtatcgat 1440

ccggccgcac taagctattt cacaaaaaca ttcacaacaa caggcataac tgaaagcaaa 1500

ttcgacatta caagtgccat atggatgtat gaagagaatg aaggcaatat tgtcgaagtc 1560

aatatgataa acggtaatat cggtgtactg acaagaggca gcaaatctgt aatcaaaacc 1620

aacaaagaca atgagattgg ttatgtagaa tgggaaatca gaggcgttga caatcctgat 1680

atggaatata tcagagctgg atatgtagat acaaccatga caggcaagtt ggtagacggc 1740

tttaccatag aatggagagg agaaggcttg acagccggca aagactacac tttcacgctt 1800

caggcatgga agaatgaagc cgacaaaaac agtggtgccg agcctaatgt aggtgaagca 1860

atgttcatca tacacggtac aaaacaagcc tacatttaca gcgatgttgt gatgaagact 1920

gatataagcc agcctatcag acttgcttct gccaatgaca attaccgtac catcgagttc 1980

agcgctcctg taacgttaaa tgccgtggta aaccttggca tggggacatc ggcagattgc 2040

accgttgaac cctcggccga ccgtactgca tggacagtca caatacctga atatgtaatg 2100

tcacaatatg gagagttcag cgtgaatgtc tttgcaaaag atgacgaagg gcgtgcagta 2160

aacaagactg agaacggact gggtataata atggggacag aagacaatac gtggttccag 2220

attgattttg tcagtgagag ccttgccccg gactttacag taacaccggc aaacgaatca 2280

gtgctcgaga gtctcagcac cgtcactttc ggttatgaag gcagtatcag catcaactgg 2340

aacaacagtg agaaaatcac catatataac agaacaacac gtgagaagat agcagagttc 2400

agcggtgatg acgtcgtgtt ggatgaagat ccggatgact attgggctcc aatactctca 2460

tgccacatca cacttcccga gcctgtcact gctattggag tctacgatgt aaatgttcct 2520

gccggtttct tcgttcttgg tgagcagttc gacagtaact taagtaaggc aacaagcata 2580

gtatacgaaa taaaggaacc ggttgagccg ttcggaatag aaataagccc tactgccgga 2640

ttggtaagcg agattccgtc aaaactcata gtaacggtaa cggacagaag cagttgtaac 2700

tttactgcca accccacatt gaccgacaat accggtaaca gctatcccgt acacaacgat 2760

ttcgactggg gtatcccgga aatgaacaag ttcgtcatca ttcttgacaa cggagccatc 2820

acggctgacg gaatatacac actcactatt cctgccggct cgattatagg tgacgacgag 2880

acagacctca acaaggaaga ctttgtattc atatatacaa tcaatctggc cggaatcaca 2940

gaactcgtca acaatgaggg cggaaaggtt gacgtataca ctctcaaagg aacattgttg 3000

atgaaagacg ccgatgcatc agcagtaaac aagctggcaa aaggcatgta cataatcaat 3060

ggtaaaaaag tgttcataag ataa 3084

<210> 29

<211> 1528

<212> DNA

<213> Paraprevotella clara

<400> 29

atgaagagtt tgatcctggc tcaggatgaa cgctagctac aggcttaaca catgcaagtc 60

gaggggcagc atggactcag ctttgctgag tttgatggcg accggcgcac gggtgagtaa 120

cgcgtatcca acctgccctt tactccggga tagtctcctg aaagggagtt taataccgga 180

tgtgtttgtt tttccgcatg ggagcgacaa ataaagatta attggtaaag gatggggatg 240

cgtcccatta gcttgttggc ggggtaacgg cccaccaagg cgacgatggg taggggttct 300

gagaggaagg tcccccacat tggaactgag acacggtcca aactcctacg ggaggcagca 360

gtgaggaata ttggtcaatg ggcgggagcc tgaaccagcc aagtagcgtg aaggacgacg 420

gccctacggg ttgtaaactt cttttataag ggaataaagt tcgccacgcg tggtgttttg 480

tatgtacctt atgaataagc atcggctaat tccgtgccag cagccgcggt aatacggaag 540

atgcgagcgt tatccggatt tattgggttt aaagggagcg taggcgggct tttaagtcag 600

cggtcaaatg ccacggctca accgtggcca gccgttgaaa ctgtaagcct tgagtctgca 660

cagggcacat ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 720

tcgcgaaggc attgtgccgg ggcagcactg acgctgaggc tcgaaagtgc gggtatcaaa 780

caggattaga taccctggta gtccgcacgg taaacgatga atgctcgcta tgggcgatat 840

attgtccgtg gccaagcgaa agcgttaagc attccacctg gggagtacgc cggcaacggt 900

gaaactcaaa ggaattgacg ggggcccgca caagcggagg aacatgtggt ttaattcgat 960

gatacgcgag gaaccttacc cgggcttgaa ttgcaggtgc atgagtcaga gacggctctt 1020

tccttcggga ctcctgtgaa ggtgctgcat ggttgtcgtc agctcgtgcc gtgaggtgtc 1080

ggcttaagtg ccataacgag cgcaaccctt ctccccagtt gccatcgggt aatgccgggc 1140

cctctgggga cactgccatc gtaagatgcg aggaaggtgg ggatgacgtc aaatcagcac 1200

ggcccttacg tccggggcta cacacgtgtt acaatggggg gtacagaggg ccgctgtccg 1260

gtgacggtcg gccaatccct aaaacccctc tcagttcgga ctggagtctg caacccgact 1320

ccacgaagct ggattcgcta gtaatcgcgc atcagccatg gcgcggtgaa tacgttcccg 1380

ggccttgtac acaccgcccg tcaagccatg aaagccgggg gtgcctgaag tccgtgaccg 1440

cgagggtcgg cctagggtaa aactggtgat tggggctaag tcgtaacaag gtagccgtac 1500

cggaaggtgc ggctggaaca cctccttt 1528

<210> 30

<211> 1481

<212> DNA

<213> Paraprevotella xylaniphila

<400> 30

gatgaacgct agctacaggc ttaacacatg caagtcgagg ggcagcatga acttagcttg 60

ctaagtttga tggcgaccgg cgcacgggtg agtaacgcgt atccaacctg ccctttacgc 120

ggggatagcc ttctgaaagg aagtttaata cccgatgaat tcgtttagtc gcatggcttg 180

atgaataaag atttatcagt aaaggatggg gatgcgtccc attagcttgt tggcggggta 240

acggcccacc aaggcgacga tgggtagggg ttctgagagg aaggtccccc acattggaac 300

tgagacacgg tccaaactcc tacgggaggc agcagtgagg aatattggtc aatgggcgcg 360

agcctgaacc agccaagtag cgtggaggac gacggcccta cgggttgtaa actcctttta 420

taaggggata aagttggcca tgtatggcca tttgcaggta ccttatgaat aagcatcggc 480

taattccgtg ccagcagccg cggtaatacg gaagatgcga gcgttatccg gatttattgg 540

gtttaaaggg agcgtaggcg ggcagtcaag tcagcggtca aatggcgcgg ctcaaccgcg 600

ttccgccgtt gaaactggca gccttgagta tgcacagggt acatggaatt cgtggtgtag 660

cggtgaaatg cttagatatc acgaggaact ccgatcgcgc aggcattgta ccggggcatt 720

actgacgctg aggctcgaag gtgcgggtat caaacaggat tagataccct ggtagtccgc 780

acagtaaacg atgaatgccc gctgtcggcg acatagtgtc ggcggccaag cgaaagcgtt 840

aagcattcca cctggggagt acgccggcaa cggtgaaact caaaggaatt gacgggggcc 900

cgcacaagcg gaggaacatg tggtttaatt cgatgatacg cgaggaacct tacccgggct 960

tgaatcgcag gtgcatgggc cggagacggc cctttccttc gggactcctg cgaaggtgct 1020

gcatggttgt cgtcagctcg tgccgtgagg tgtcggctta agtgccataa cgagcgcaac 1080

ccccctcccc agttgccatc gggtaatgcc gggcactttg gggacactgc caccgcaagg 1140

tgcgaggaag gtggggatga cgtcaaatca gcacggccct tacgtccggg gcgacacacg 1200

tgttacaatg gggggtacag agggccgctg cccggtgacg gttggccaat ccctaaagcc 1260

cctctcagtt cggactggag tctgcaaccc gactccacga agctggattc gctagtaatc 1320

gcgcatcagc catggcgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcaagc 1380

catgaaagcc gggggtgcct gaagtccgtg accgcgaggg tcggcctagg gtaaaaccgg 1440

tgattggggc taagtcgtaa caaggtagcc gtaccggaag g 1481

<210> 31

<211> 1535

<212> DNA

<213> Prevotella rara

<400> 31

ttttacagtg gagagtttga tcctggctca ggatgaacgc tagctacagg cttaacacat 60

gcaagtcgcg gggcagcatg aaggatgctt gcattctttg atggcgaccg gcgcacgggt 120

gagtaacgcg tatccaacct gcctgttacc acggcacagc ccgtcgaaag gcggattaat 180

gccgtatgtg gtcttgaaag ggcatctgat catgactaaa ggttcagcgg taacggatgg 240

ggatgcgtcc gattagctag acggcggggt aacggcccac cgtggcgacg atcggtaggg 300

gttctgagag gaaggtcccc cacactggaa ctgagacacg gtccagactc ctacgggagg 360

cagcagtgag gaatattggt caatgggcgt aagcctgaac cagccaagta gcgtgaggga 420

agactgccct atgggttgta aacctctttt gtgcggggat aaagtgaggg acgtgtccct 480

tattgcaggt accgcacgaa taaggaccgg ctaattccgt gccagcagcc gcggtaatac 540

ggaaggtccg ggcgttatcc ggatttattg ggtttaaagg gagcgcaggc cgtcctttaa 600

gcgtgctgtg aaatgccgcg gctcaaccgt ggcactgcag cgcgaactgg aggacttgag 660

tacgcacgag gtaggcggaa ttcgtggtgt agcggtgaaa tgcttagata tcacgaagaa 720

ctccgattgc gaaggcagct taccggagcg caactgacgc tgaggctcga aagcgcgggt 780

atcgaacagg attagatacc ctggtagtcc gcgcggtaaa cgatggatgc ccgccgttgg 840

gatattgatt tcagcggcca agcgaaagcg ttaagcatcc cacctgggga gtacgccggc 900

aacggtgaaa ctcaaaggaa ttgacggggg cccgcacaag cggaggaaca tgtggtttaa 960

ttcgatgata cgcgaggaac cttacccggg cttgaattgc aggagaacga tccagagatg 1020

gtgaggccct tcggggctcc tgtgaaggtg ctgcatggtt gtcgtcagct cgtgccgtga 1080

ggtgtcggct caagtgccat aacgagcgca acccctgtcc atagttgcca tcaggtttgc 1140

tgggcactct gtggagactg ccgccgtaag gtgtgaggaa ggtggggatg acgtcaaatc 1200

agcacggccc ttacgtccgg ggctacacac gtgttacaat ggggcataca gcgagttgga 1260

tgtgcgcaag tacgtccgga tcaataaagt gcctctcagt tcggactggg gtctgcaacc 1320

cgaccccacg aagctggatt cgctagtaat cgcgcatcag ccatggcgcg gtgaatacgt 1380

tcccgggcct tgtacacacc gcccgtcaag ccatgaaagc cgggggcgcc tgaagtccgt 1440

gaccgcgagg gtcggcctag ggtgaaaccg gtgattgggg ctaagtcgta acaaggtagc 1500

cgtaccggaa ggtgcggctg gaacacctcc tttct 1535

<210> 32

<211> 1438

<212> DNA

<213> Prevotella rodentium

<400> 32

tgatggcgac cggcgcacgg gtgagtaacg cgtatccaac ctgcccttta ctatgggaca 60

gcccgtcgaa aggcggatta ataccgtatg ttgtcatgtt gatgcatatt ttcatgacca 120

aaggcttcgg ccggtaaagg atggggatgc gtctgattag cttgccggcg gggtaacggc 180

ccaccggggc gacgatcagt aggggttctg agaggaaggt cccccacatt ggaactgaga 240

cacggtccaa actcctacgg gaggcagcag tgaggaatat tggtcaatgg gcgcgagcct 300

gaaccagcca agtagcgtgc aggacgacgg ccctatgggt tgtaaactgc ttttatacgg 360

ggataaagta tgccacgtgt ggtttattgc aggtaccgta tgaataagga ccggctaatt 420

ccgtgccagc agccgcggta atacggaagg tccgggcgtt atccggattt attgggttta 480

aagggagcgt aggccgggtt ttaagcgtgc cgtgaaatgt cggggctcaa ccttgacact 540

gcggcgcgaa ctggagtcct tgagtgcgcg gaacgtatgc ggaattcgtg gtgtagcggt 600

gaaatgctta gatatcacga agaaccccga ttgcgaaggc agcatacggc agcgctactg 660

acgctgaagc tcgaaggcgc gggtatcgaa caggattaga taccctggta gtccgcgcgg 720

taaacgatgg atgcccgctg ttggcgatat attgtcagcg gccaagcgaa agcgttaagc 780

atcccacctg gggagtacgc cggcaacggt gaaactcaaa ggaattgacg ggggcccgca 840

caagcggagg aacatgtggt ttaattcgat gatacgcgag gaaccttacc cgggcttgaa 900

ctgccggcga acgatccaga gatggtgagg cccttcgggg cgccggcgga ggtgctgcat 960

ggttgtcgtc agctcgtgcc gtgaggtgtc ggcttaagtg ccataacgag cgcaacccct 1020

gttctcagtt gccatcgggt aatgccgggc actctgtgaa gactgcctcc gtaaggagtg 1080

aggagggtgg ggatgacgtc aaatcagcac ggcccttacg tccggggcta cacacgtgtt 1140

acaatgggtg gtacagacgg tcggcgtccg gcaacgtacg tccaatccgt aaagccgccc 1200

tcagttcgga ctggggtctg caacccgacc ccacgaagct ggattcgcta gtaatcgcgc 1260

atcagccatg gcgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcaagccatg 1320

aaagccgggg gcgcctgaag tccgtgaccg cgagggtcgg cctagggtga aaccggtgat 1380

tggggctaag tcgtaacaag gtagccgtac cggaaggtgc ggctggaaca cctccttt 1438

<210> 33

<211> 1531

<212> DNA

<213> Prevotella muris

<400> 33

acaatgtaga gtttgatcct ggctcaggat gaacgctggc tacaggctta acacatgcaa 60

gtcgaggggc agcatgacat gttttcggac gtgtcgatgg cgaccggcgc acgggtgagt 120

aacgcgtatc cgacctgccc cgtaccaggg cacagcccgt cgaaagacgg attaatgccc 180

tatgttctcc ggaagatcca tgttttccgg agcaaaggtt attccggtac gggatgggga 240

tgcgtccgat tagcttgctg gcggggtaac ggcccaccag ggcaccgatc ggtaggggtt 300

ctgagaggaa ggtcccccac actggaactg agacacggtc cagactccta cgggaggcag 360

cagtgaggaa tattggtcaa tggggggaac cctgaaccag ccaagtagcg tgcaggatga 420

cggccctatg ggttgtaaac tgcttttgtg cgtggataaa gtaggccact tgtggccttt 480

tgcaggtacc gcacgaataa ggaccggcta attccgtgcc agcagccgcg gtaatacgga 540

aggtccgggc gttatccgga tttattgggt ttaaagggag cgtaggccgc ctgtcaagcg 600

tgctgtgaaa cgccgtggct caaccacggt cctgcagcgc gaactggcgg gcttgagtgt 660

gcggaaggca cgcggaattc gtggtgtagc ggtgaaatgc ttagatatca cgaagaactc 720

cgattgcgaa ggcagcgtgc cgcagcatta ctgacgctga tgctcgaaag cgcgggtatc 780

gaacaggatt agataccctg gtagtccgcg cggtaaacga tggatgcccg ctgtcggcga 840

tatacagtcg gcggccaagc gaaagcgtta agcatcccac ctggggagta cgccggcaac 900

ggtgaaactc aaaggaattg acgggggccc gcacaagcgg aggaacatgt ggtttaattc 960

gatgatacgc gaggaacctt acccgggctt gaattgcaga tgaatgattc agagatgatg 1020

aagtccttcg ggacatctgt gaaggtgctg catggttgtc gtcagctcgt gccgtgaggt 1080

gtcggcttaa gtgccataac gagcgcaacc ccttccctca gttgccatcg ggtcatgccg 1140

ggcactctgt gggtactgcc tccgcaagga gcgaggaagg cggggatgac gtcaaatcag 1200

cacggccctt acgtccgggg ctacacacgt gttacaatgg ggcatacagc gagcaggtgc 1260

cgtgcaaacg gtgtcgaatc ttgaaagtgc ccctcagttc ggactggggt ctgcaacccg 1320

accccacgaa gctggattcg ctagtaatcg cgcatcagcc atggcgcggt gaatacgttc 1380

ccgggccttg tacacaccgc ccgtcaagcc atgaaagccg ggggcgcctg aagtccgtga 1440

ccgtgagggt cggcctaggg tgaaaccggt gattggggct aagtcgtaac aaggtagccg 1500

taccggaagg tgcggctgga acacctcctt t 1531

<210> 34

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 34

Gly Gly Gly Gly Ser

1 5

<210> 35

<211> 24

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 35

gtcgtggagt ctactggtgt cttc 24

<210> 36

<211> 24

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 36

gtcatatttc tcgtggttca cacc 24

<210> 37

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 37

tgtgaccctc aatgccagag 20

<210> 38

<211> 19

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 38

agcactgggg catcaacac 19

<210> 39

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 39

agrgtttgat ymtggctcag 20

<210> 40

<211> 22

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 40

tacggytacc ttgttacgac tt 22

<210> 41

<211> 90

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<220>

<221> misc_feature

<222> (30)..(37)

<223> n is a, c, g, or t

<400> 41

aatgatacgg cgaccaccga gatctacacn nnnnnnnaca ctctttccct acacgacgct 60

cttccgatct agrgtttgat ymtggctcag 90

<210> 42

<211> 85

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<220>

<221> misc_feature

<222> (25)..(32)

<223> n is a, c, g, or t

<400> 42

caagcagaag acggcatacg agatnnnnnn nngtgactgg agttcagacg tgtgctcttc 60

cgatcttgct gcctcccgta ggagt 85

<210> 43

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 43

cgtaggagtt tggaccgtgt 20

<210> 44

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 44

gcatgggagc gacaaataaa 20

<210> 45

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 45

aagagtgatt ggcgtccgta c 21

<210> 46

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 46

atggacacgt cactggcaga g 21

<210> 47

<211> 38

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 47

gtggatcccc cgggctgcag catccctcct ccatctgt 38

<210> 48

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 48

ctggaagata ggcaattagt ttgggacgta ggggcaatc 39

<210> 49

<211> 27

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 49

tcccgaccga ctgatccttt ttaaccc 27

<210> 50

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 50

ctgtgcccag taattttgtg 20

<210> 51

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 51

gtggatcccc cgggctgcac cccttgtcca tgttagacg 39

<210> 52

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 52

ctggaagata ggcaattagt gaagtgacag gggatgcta 39

<210> 53

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 53

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 54

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 54

ctggaagata ggcaattagt gaagtgacag gggatgcta 39

<210> 55

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 55

gtggatcccc cgggctgcaa cactgatgtc gttgccttt 39

<210> 56

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 56

ctggaagata ggcaattagg caaccttacc acagggaaa 39

<210> 57

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 57

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 58

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 58

ctggaagata ggcaattagg caaccttacc acagggaaa 39

<210> 59

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 59

gtggatcccc cgggctgcac aagtagagtt ccggttcgg 39

<210> 60

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 60

ctggaagata ggcaattaga aatcacgacc ctcgatgtg 39

<210> 61

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 61

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 62

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 62

ctggaagata ggcaattaga aatcacgacc ctcgatgtg 39

<210> 63

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 63

gtggatcccc cgggctgcaa gttcgttctt ctcgatggc 39

<210> 64

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 64

ctggaagata ggcaattagc aaccgcgggt ataacaaga 39

<210> 65

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 65

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 66

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 66

ctggaagata ggcaattagc aaccgcgggt ataacaaga 39

<210> 67

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 67

gtggatcccc cgggctgcat ggggcgtttg tatttgtca 39

<210> 68

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 68

ctggaagata ggcaattaga aaccgctgac ggtctattc 39

<210> 69

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 69

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 70

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 70

ctggaagata ggcaattaga aaccgctgac ggtctattc 39

<210> 71

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 71

gtggatcccc cgggctgcat ccagtcgtca gactttcct 39

<210> 72

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 72

ctggaagata ggcaattagg aagggactaa aatcgccgt 39

<210> 73

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 73

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 74

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 74

ctggaagata ggcaattagg aagggactaa aatcgccgt 39

<210> 75

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 75

gtggatcccc cgggctgcag ggtcgcttac attttgcac 39

<210> 76

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 76

ctggaagata ggcaattagt cggtgattat gctttcgca 39

<210> 77

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 77

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 78

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 78

ctggaagata ggcaattagt cggtgattat gctttcgca 39

<210> 79

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 79

gtggatcccc cgggctgcac ttgcttcaaa tcacgctgg 39

<210> 80

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 80

ctggaagata ggcaattagg ttgcgtaatg tgaacggtg 39

<210> 81

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 81

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 82

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 82

ctggaagata ggcaattagg ttgcgtaatg tgaacggtg 39

<210> 83

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 83

gtggatcccc cgggctgcat gtcgaaaggc atcatctgg 39

<210> 84

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 84

ctggaagata ggcaattaga ccattacggc cattaccac 39

<210> 85

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 85

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 86

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 86

ctggaagata ggcaattaga ccattacggc cattaccac 39

<210> 87

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 87

gtggatcccc cgggctgcag tgccagtttc aaccaatcg 39

<210> 88

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 88

ctggaagata ggcaattagc ttgtacccca gcagatgtc 39

<210> 89

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 89

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 90

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 90

ctggaagata ggcaattagg taagctggcg cttgaaatc 39

<210> 91

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 91

gtggatcccc cgggctgcat tgttttggtt gtaactgcc 39

<210> 92

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 92

ctggaagata ggcaattagg tcaatacgaa ctgcaactg 39

<210> 93

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 93

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 94

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 94

ctggaagata ggcaattagg tcaatacgaa ctgcaactg 39

<210> 95

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 95

gtggatcccc cgggctgcag accaattaag cacgtcaag 39

<210> 96

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 96

ctggaagata ggcaattagt ggtacaattc gctcaaaga 39

<210> 97

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 97

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 98

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 98

ctggaagata ggcaattagt ggtacaattc gctcaaaga 39

<210> 99

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 99

gtggatcccc cgggctgcat actcccgtgt ttttagtgg 39

<210> 100

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 100

ctggaagata ggcaattagg gatgccagtt acctgatag 39

<210> 101

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 101

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 102

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 102

ctggaagata ggcaattagg gatgccagtt acctgatag 39

<210> 103

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 103

gtggatcccc cgggctgcat caaaccacac acagagaaa 39

<210> 104

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 104

ctggaagata ggcaattagg aattcggcca aaccttatg 39

<210> 105

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 105

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 106

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 106

ctggaagata ggcaattagg aattcggcca aaccttatg 39

<210> 107

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 107

gtggatcccc cgggctgcaa gacaatcgag gcatcatac 39

<210> 108

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 108

ctggaagata ggcaattaga tagcgtgaat ggaaaacct 39

<210> 109

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 109

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 110

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 110

ctggaagata ggcaattaga tagcgtgaat ggaaaacct 39

<210> 111

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 111

gtggatcccc cgggctgcaa tctatctcaa cctcctgct 39

<210> 112

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 112

ctggaagata ggcaattagg aaggaatcag cgtagatgt 39

<210> 113

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 113

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 114

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 114

ctggaagata ggcaattagg aaggaatcag cgtagatgt 39

<210> 115

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 115

gtggatcccc cgggctgcat gtgccgataa cctgaatgg 39

<210> 116

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 116

ctggaagata ggcaattagc agagaaaagg gctggtgtt 39

<210> 117

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 117

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 118

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 118

ctggaagata ggcaattagc agagaaaagg gctggtgtt 39

<210> 119

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 119

tcccgaccga ggtctttgtc cccttctccg 30

<210> 120

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 120

tcttacaccg gcgggttgct ttttatgcct 30

<210> 121

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 121

gcctgatgca tggaaaaacc 20

<210> 122

<211> 19

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 122

ctggaagata ggcaattag 19

<210> 123

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 123

gtggatcccc cgggctgcac ggataacgga attacccat 39

<210> 124

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 124

ctggaagata ggcaattagt ctgacaaagg ttggttgaa 39

<210> 125

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 125

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 126

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 126

ctggaagata ggcaattagt ctgacaaagg ttggttgaa 39

<210> 127

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 127

gtggatcccc cgggctgcat gtataacccc cagaaggaa 39

<210> 128

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 128

ctggaagata ggcaattaga acaataccgt caacctctc 39

<210> 129

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 129

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 130

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 130

ctggaagata ggcaattaga acaataccgt caacctctc 39

<210> 131

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 131

gtggatcccc cgggctgcat ttattagccg tggtatcgg 39

<210> 132

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 132

ctggaagata ggcaattaga ctccaacagc atcttcaat 39

<210> 133

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 133

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 134

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 134

ctggaagata ggcaattaga ctccaacagc atcttcaat 39

<210> 135

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 135

gtggatcccc cgggctgcag tagtgtatcg ctttgtcct 39

<210> 136

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 136

ctggaagata ggcaattaga ttcattactg gtcggaacc 39

<210> 137

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 137

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 138

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 138

ctggaagata ggcaattaga ttcattactg gtcggaacc 39

<210> 139

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 139

gtggatcccc cgggctgcag ggatactgaa tgtctgtcc 39

<210> 140

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 140

ctggaagata ggcaattaga cagtgtaatc atcaacgct 39

<210> 141

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 141

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 142

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 142

ctggaagata ggcaattaga cagtgtaatc atcaacgct 39

<210> 143

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 143

gtggatcccc cgggctgcaa ttttcaatcc agtccctcc 39

<210> 144

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 144

ctggaagata ggcaattagt acagtttgcc gataccaaa 39

<210> 145

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 145

aaacaaataa atgaagaaaa ttttatcttt gct 33

<210> 146

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 146

ctggaagata ggcaattagt acagtttgcc gataccaaa 39

<210> 147

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 147

gtggatcccc cgggctgcat atgaactaca aagtcatcg 39

<210> 148

<211> 27

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 148

ttcaagaagc accaccgtta tgatata 27

<210> 149

<211> 27

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 149

taacggtggt gcttcttgaa aattaag 27

<210> 150

<211> 35

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 150

ctggaagata ggcaattagg acgatttcgt ggaca 35

<210> 151

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 151

gtggatcccc cgggctgcat atgaactaca aagtcatcg 39

<210> 152

<211> 35

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 152

ctggaagata ggcaattagg acgatttcgt ggaca 35

<210> 153

<211> 19

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 153

tcaacgcacc attactttc 19

<210> 154

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 154

atgaaaaaca gtttacttgc 20

<210> 155

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 155

gtggatcccc cgggctgcat acaatccgtt ggaactgtc 39

<210> 156

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 156

attagtattt cagcattcat ttgctggtga 30

<210> 157

<211> 40

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 157

atgaatgctg aaatactaat tttagataga ctataatttg 40

<210> 158

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 158

ctggaagata ggcaattagg aactggtcgt aaaaatcac 39

<210> 159

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 159

gtggatcccc cgggctgcat acaatccgtt ggaactgtc 39

<210> 160

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 160

ctggaagata ggcaattagg aactggtcgt aaaaatcac 39

<210> 161

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 161

gtggatcccc cgggctgcat ccagtcgtca gactttcct 39

<210> 162

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 162

ctggaagata ggcaattagg aagggactaa aatcgccgt 39

<210> 163

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 163

gtggatcccc cgggctgcaa tccttccact tttcctctc 39

<210> 164

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 164

aattgatcgt tctaaagccg acgatttaag 30

<210> 165

<211> 35

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 165

cggctttaga acgatcaatt ttaaaatgaa tatat 35

<210> 166

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 166

ctggaagata ggcaattagt tcaacgaaaa cagaacatc 39

<210> 167

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 167

gtggatcccc cgggctgcaa tccttccact tttcctctc 39

<210> 168

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 168

ctggaagata ggcaattagt tcaacgaaaa cagaacatc 39

<210> 169

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 169

gtggatcccc cgggctgcaa gacaatcgag gcatcatac 39

<210> 170

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 170

ctggaagata ggcaattaga tagcgtgaat ggaaaacct 39

<210> 171

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 171

attcgtaaaa caccaccacc accaccactg 30

<210> 172

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 172

tcacgccgtc catggtatat ctccttctta 30

<210> 173

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 173

atataccatg gacggcgtga gccaacctac 30

<210> 174

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 174

ggtggtggtg ttttacgaat actttcttca 30

<210> 175

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 175

gatggtaaaa caccaccacc accaccactg 30

<210> 176

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 176

cttcaccaac catggtatat ctccttctta 30

<210> 177

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 177

atataccatg gttggtgaag atttacattt 30

<210> 178

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Synthesized oligonucleotide

<400> 178

ggtggtggtg ttttaccatc atcttcttac 30

Claims

1. A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria:

a bacterium having 00502 protein or a bacterium having a protein which has 30% or more of sequence identity with the amino acid sequence of 00502 protein and has trypsin-binding ability; or alternatively

2. A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria:

a bacterium having 00502 protein or a bacterium having a protein which has 30% or more of sequence identity with the amino acid sequence of 00502 protein and has trypsin-binding ability; and

3. A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria:

a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 1 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 1 and encodes a protein having trypsin binding ability; or alternatively

Bacteria having a gene comprising a base sequence shown as SEQ ID NO. 2 or bacteria having a gene which has 30% or more sequence identity with a base sequence shown as SEQ ID NO. 2 and encodes a protein having trypsin binding ability.

4. A composition for degrading trypsin or TMPRSS2, the composition comprising as active ingredients the following bacteria:

a bacterium having a gene comprising a base sequence shown in SEQ ID NO. 1 or a bacterium having a gene which has 30% or more sequence identity with a base sequence shown in SEQ ID NO. 1 and encodes a protein having trypsin binding ability; and

5. The composition for degrading trypsin or TMPRSS2 of any one of claims 1-4, wherein the bacteria has a type IX secretion system (T9 SS).

6. The composition for degrading trypsin or TMPRSS2 of claim 5, wherein the T9SS comprises PorV protein, porU protein, porN protein, porM protein, porL protein, porK protein, or PorP protein.

7. The composition for degrading trypsin or TMPRSS2 according to any of claims 1-6, wherein the bacterium is a bacterium of the genus Paraprevatella, prevoltella, prevoltelamasiia or Bactoidetes.

8. The composition for degrading trypsin or TMPRSS2 according to claim 7, wherein the bacterium of the genus Paraprevatella is Paraprevotella clara and the bacterium of the genus Prevoltella is at least one bacterium selected from the group consisting of Prevoltella rara, prevotella rodentium and Prevoltella muris.

9. The composition for degrading trypsin or TMPRSS2 according to any of claims 1-8, wherein the bacterium is a bacterium having a 16S rRNA gene comprising a base sequence as shown in SEQ ID NO. 3 or SEQ ID NO. 4 or a bacterium having a 16S rRNA gene comprising a base sequence having 97% or more sequence identity with the base sequence as shown in SEQ ID NO. 3 or SEQ ID NO. 4.

10. The composition for degrading trypsin or TMPRSS2 according to any one of claims 1-6, wherein the bacteria is at least one bacteria selected from the group consisting of paraprevatella sp.msp 0303, paraprevatella sp.msp 0335, prevotellamassilia timonensis, bacteriodes sp.msp 0288, bacteriodes sp.msp 0410, bacteriodes sp.msp 0435 and Porphyromonas gingivalis.

11. The composition for degrading trypsin or TMPRSS2 of any one of claims 1-10, wherein the bacteria is a viable bacteria.

12. The composition for degrading trypsin or TMPRSS2 of any one of claims 1-10, wherein the bacteria is dead bacteria.

13. A composition for degrading trypsin or TMPRSS2, comprising as active ingredients the following proteins:

14. A composition for degrading trypsin or TMPRSS2, comprising as active ingredients the following proteins:

15. A composition for degrading trypsin or TMPRSS2 according to any one of claims 1-14 for use in treating a disease caused by trypsin or TMPRSS 2.

16. The composition for degrading trypsin or TMPRSS2 of any one of claims 1-15, wherein the disease caused by trypsin or TMPRSS2 is inflammatory bowel disease (ulcerative colitis, crohn's disease), irritable bowel disease, infection, acute pancreatitis, or chronic pancreatitis.

17. The composition for degrading trypsin or TMPRSS2 of claim 16, wherein the infection is a viral infection or a bacterial infection.

18. The composition for degrading trypsin or TMPRSS2 of claim 16 or 17, wherein the inflammatory bowel disease, irritable bowel disease or infection is a disease involving TMPRSS2 or IgA.

19. A diagnostic agent for a disease caused by trypsin or TMPRSS2, the diagnostic agent comprising a specific binding substance for detecting:

00502 protein or a protein having 30% or more sequence identity to the amino acid sequence of 00502 protein and having trypsin binding capacity; or alternatively

20. A diagnostic agent for a disease caused by trypsin or TMPRSS2, the diagnostic agent comprising a primer set or probe for detecting:

21. A quasi-drug for diseases caused by trypsin or TMPRSS2, which contains bacteria having the following proteins as active ingredients:

22. A quasi drug for diseases caused by trypsin or TMPRSS2, which contains as active ingredients the following proteins: