JP2000505642A - Novel adhesion factors of non-pathogenic microorganisms and their application for screening microorganisms for specific symbiotic properties; novel pharmaceutical compositions and food additives containing such microorganisms and adhesion factors - Google Patents

Abstract

  (57) [Summary] Disclosed are proteins having a mucosal binding promoting activity and a molecular weight of 20-40 kD, which can be obtained from non-pathogenic microorganisms. A method for screening non-pathogenic microorganisms for microorganisms capable of specifically binding to the mucosa, said method being known per se of the presence of a particular protein on or in a microorganism or in a culture of the microorganism , Wherein said specific protein is an already identified protein), such as from a non-pathogenic microorganism. Kits suitable for such screening methods are also disclosed. Expression comprising a nucleic acid encoding such a protein or peptide as a pharmacologically active component in a pharmaceutical composition for the prevention and / or treatment of diseases or conditions associated with mucosal colonizing pathogenic microorganisms A vector; a recombinant microorganism expressing such a protein or peptide or a part of said microorganism (the part expresses a mucosal binding promoting activity); a non-pathogenic microorganism capable of expressing such a protein or peptide; Use of one component selected from the group of components containing a protein or peptide identified as: a portion of a microorganism (the portion expresses a mucosal binding promoting activity). Food additives containing such components and the use of such components as compositions are described.

Description

DETAILED DESCRIPTION OF THE INVENTION Screen microorganisms for new adhesion factors and specific symbiotic properties of non-pathogenic microorganisms Application for cleaning; novel pharmacology including such microorganisms and adhesion factors Compositions and food additives Summary of the Invention   The present invention relates to a cell capable of adhering to a specific site of mucosa called a receptor. It relates to the screening of bacteria for fungi, especially non-pathogenic bacteria. More specifically The present invention relates to non-pathogenic Gram-positive bacteria, especially lactic acid bacteria (LAB) species. Specially Lactobacillus (Lactobacillus) And Bifidak Terium (Bifidobacterium) Genetic screening. Agriculture It is preferable to screen for bacteria that are endemic to field animals, pets, and humans. No.   The present invention relates to a specific group of previously unrecognized non-pathogenic bacterial adhesion factors. Screening methods. In particular, adhesion of things such as lactobacilli The factor is the goal. This new class of non-pathogenic bacterial adhesion factors is a class of diseases. Includes proteins structurally related to virulence factors of protozoa.   The present invention can further be obtained via the screening method of the present invention. Application of bacteria, especially Lactobacillus producing the adhesion factor, as an adhesion factor itself Application, application of its bacterial parts, and from a new group for various pharmaceutical applications It also concerns the application of parts of the adhesion factor. Such applications include gastrointestinal tract, respiratory tract, genitourinary tract Colonization of the mouth, mouth, or any other part of the body, especially pathogenic microorganisms May include the treatment or prevention of infections in any internal part of the body. May be.   Other suitable examples of this application include targeting specific compounds to cells of the mucosa, Elicits a specific mucosal immune response to the compound, or The purpose is to modulate the response.   For example, it can be obtained through recombinant DNA technology, Novel microorganisms expressing or overexpressing the offspring or an effective part thereof are also included in the present invention. Is done.   Nucleic acid sequences encoding these adhesion factors, and encoding mucosa-associated expression products A fragment of the sequence is also similar to a recombinant product resulting from expression of the nucleic acid sequence. Is part of the present invention.   Express or overexpress nucleic acids or their expression products, or novel types of adhesion factors Novel pharmaceutical compositions containing the emerging microorganisms are also within the scope of the invention. Background of the Invention   Pathogenic viruses and bacteria adhere to specific sites on the mucosa called receptors. And invading the cells underlying them via their receptors, It can result in cell illness or even death. Public health care and Effective and inexpensive methods for human and animal care for animal health care It is essential to be able to prevent and / or cure infectious diseases in the United States.   The mucosa is, for example, the genus Escherichia of Gram-negative bacteria.Escher ichia ), Campylobacter (Campylobacter), Hayemofi Ruth (Haemophilus), Shigella (Shigella), Vibrio (V ibrio ), Pasteurella (Pasteurella), Yersinia (Ye rsinia ), Salmonella (Sa lmonella ), Such as Mycobacterium (Mycobacterium) , Listeria (Listeria), Clostridium (Clostridu m ), Staphylococcus (Staphylococcus) Such as gram (G ram) Positive bacteria, and viruses such as rotavirus, poliovirus, and deer Ruths and many other microorganisms well known to those skilled in the field of microbial infection The entrance of the pathogenic bacteria (the port dentr6e) consisting of Form.   For example, Campylobacter (CampylobacterBacteria of the genus are taken orally May cause severe enteritis in humans and animals. C. Jee Juni (C.jejuni) Is a major source of diarrhea in humans and sometimes animals Cause. In addition to diarrhea, C.I. Jejuni (C.jejuni) Is sometimes human It can also cause appendicitis, meningitis, miscarriage, and urinary tract infections in children. Developing In upper nations, people of all ages are sick, and Campylobacter (Ca mpylobacter ) Infections are Salmonella (Salmonella), Shige La (Shigella) Or Vibrio cholerae (Vibrio cho lerae ) Are as common as infections caused by.   Mycobacterium tuberculosis (Mycobacteriumtuber culosis ) And Mycobacterium leprae (Mycobacterium lepraeMycobacteria such as tuberculosis and leprosy, respectively. Cause various diseases. These bacteria are found in poorly developed countries Cause death of many individuals. These microorganisms pass through the mucous membrane of the respiratory tract. Attacks the body.   Pathogenic microorganisms adhere to parts of the body, for example the gastrointestinal tract, Thus, the disease can be started. Pathogenic microbes on body parts of the host organism Studies of object adhesion have provided a wealth of data. These studies show that It has been shown that adhesion of protozoa can be mediated by proteins. For example Extracellular mats such as collagen, fibronectin, or proteoglycans More information on proteins from pathogenic bacteria that bind to components of Rix is available. Available. A special example is the mycobacterial fibronectin binding protein. , Fibronect of streptococcal and staphylococcal specific enterobacteria of the pilus type Tin-binding protein, collagen-binding protein, and Yersinia surface protein Aeromonas (Aeromonas) A-protein (A-protein) (For a review, see Westerlund and Korhonen, Mol . Microbiol. 9: 687-694 1993).   Adhesion of Gram-positive, non-pathogenic bacteria to the surface of the host organism Information is more limited, especially the characteristics of mucosal receptors by non-pathogenic microorganisms. Information on heterogeneity is scarce.   In the normal human gastrointestinal tract, Lactobacillus (Lactobacillus), Streptocox (Streptococcus), Enterocox (En terococcus ), Bifidobacterium (Bifidobacteri um ), Clostridium (Clostridium), Bacterioidis (BacteriodesA wide variety of non-pathogenic, including bacteria of the genus, and others Sexual microorganisms are established. These microorganisms are part of the microflora that is unique to humans. Form a minute. For this reason, considerable interest is in the mechanism of adhesion and the gastrointestinal tract. Established in It has been aimed at elucidating the role of adhesion in the process. However, with humans Very Investigated Among Nonpathogenic Bacteria Present in Animal Intestinal Microflora The mechanism of adhesion of the LAB, a group of people, generally compares with that of gastrointestinal pathogens This is more complicated (Hasty et al., Infect. Immun. . 60: 2147-2152 1992).   Adhesion of non-pathogenic bacteria is specific or non-specific. ). Hydrophobic and the electrostatic adhesion mechanism is non-specific (n involved in on-specific adhesion. In specific adhesion, the adhesion factor is specific Characterized by the so-called “key and keyhole mechanism” that binds to a specific receptor You. Specific adhesion is usually associated with the attachment of microorganisms to receptors on living tissue You. Adhesins or adhesins are generally surface-bound molecules It is. Adhesins bind tightly or loosely to bacterial surfaces. Can be. The receptor is that the active site of the adhesin will bind to bacteria, Components or structures on the surface of cells (Rutter et al., 1 984 Mechanisms of adhesin in "Microb ial adhesion and aggregation "Marsha 11, K .; C. ed. pp5-19, Springer-Verlag, Ber. lin).   Lactic acid bacteria, especially Lactobacillus (Lactobacillus) And entero Cox (Enterococcus) Is a micrograph of the gastrointestinal tract between humans and animals Nonpathogenic Gram Yang plays a major role in establishing and maintaining Laura It is an example of a bacterium. Lactobacillus (Lac tobacillus ) Species have been isolated from various regions of the human gastrointestinal tract (M olin et al. J. Appl. Bactriol. 74, 314-3 23 1993).   Determinants that may be responsible for the adhesion of some strains have been studied and It has been reported that certain structures are involved in the mechanism of adhesion. However, because of the complexity of the intestinal ecosystem, why are specific areas of the gastrointestinal tract And how most bacterial strains attach and colonize. It has not been elucidated.   In publications, adhesion of lactobacilli to the gastrointestinal mucosa is also a major mechanism. Certainly, there is a discrepancy. Fuller to chicken gizzard epithelium Lactobacillus (Lactobacillus) Describe the adhesion, and adhesion Are mediated by polysaccharides (Fuller, J. Gen. Mi.) crobiol. 87: 245-250 1975). However Conwa y and Adams (J. Gen. Microbiol. 135: 1167-117). 3 1989) found no evidence for the role of polysaccharides in the adhesion of lactobacilli. Instead, it suggested that other components could be involved. Other researchers are also Lipotaiko Lactobacillus acid (LTA)Lactobacillus) And streptocco Cass (StreptococcusThat is very important when bonding LTA causes association of fibronectin and plaque with streptococci (Hogg and Manning, J. Appl. B acterio. 65: 483-489; Vickerman and Jon es, Infect. Immun. 60: 400104008 1992). S eugara (Infect. Immun. 12: 173-179) reported that proteinaceous substances It describes mediating the adhesion of S. bacilli to rat gastric epithelium.   Current Microbiology, Vo1.28 (1994) p. 231-236, see Aleljung P. et al. Bind to collagen type I L. Reuteri (L.reuteri) Two collagens of NC1B 11951 It describes the purification of binding proteins. The sequence for the 31 kD version is XSNKPIIV GSK * XV. The sequence for the 29 kD is ASS * AVNSELV. The closest homology is E. coli (E.coli) Trigger factor (trig) ger factor). TIG is 79% relative score Is located at 27-33. L. Reuteri (L.reuteri) Of CnBP is A protein type that has been described to be involved in the adhesion to the chicken digestive tract. It is also said that it is unlikely to be an S protein. They say, "Nonpathogenic here Unique intestinal microflora binds to extracellular matrix-binding proteins It is explained as ". However, mucous membrane or mucin It does not describe toro or in vivo binding. They simply exist in the mucous membrane It merely describes binding to the components themselves known to do so. No explanation is provided for binding to such components, which take the form present in the mucosa. Not. Whether such binding occurs to collagen when present in the mucosa Is obvious due to the fact that it is unclear where its binding site is But not to mucin or collagen when present in the mucosa Such sites are available or It is not clear whether it exists. Non-pathogenic microbial adhesion to ECM or mucosa No description is provided for This editorial is rather uncertain in nature.   Recently Toba et al., Lactobacillus Crispatous (Lactobacill us crispatus) Is attached to the extracellular matrix by the S-layer (S -Layer) protein (Toba et a l. Appl. Environm. Microbiol. 61: 1995).   In EP 0 210 579 (priority date November 1984), the mouse Allegedly responsible for promoting bacterial adhesion to squamous epithelium in pigs and pigs Preparations containing the known 14 kD Mw protein have been described. 14kD protein This preparation, which contains lactobacillus fermentation, in a medium rich in sugars and amino acids Ntum (Lactobacillus fermentumCulturing) Was obtained by From EP 0 210 579, this adhesion promoter is Is specific for non-pathogenic bacteria, or is it a pathogen that does not normally adhere It is not clear whether it can promote body adhesion. In addition, European Patent No. 0 210 From No. 579, this adhesion promoter or specific site (receptor) or non-specific It is also not clear whether it promotes adhesion to specific sites. So In addition, EP 0 210 579 describes what is the origin of the 14 kD protein. I haven't even made it clear. This 14 kD protein is L. Fermentum (L .fermentum) Whether it is synthesized by itself, or This is L. Fermentum (L.fermentum) The composition of the medium Arising from the minute Is still unresolved. Therefore, identification of the 14 kD protein and Both applicability remain ambiguous in this publication.   Numerous subsequent publications also suggest that different proteinaceous components are involved Yes, but no definitive data was provided.   WO 90/09398 by Conway and Kjelleberg Exhibit anti-pathogenic activity, Crispatis (L.crispatis) 1 A fragment of greater than 30 kD from 04 is described. This break exceeds 30kD The strip retains its activity after treatment with pronase or trypsin. This This is L. in a complex medium. Crispatis (L.crispatis) Is obtained. This application also shows that the corresponding fragments of 8000 to 30,000 have anti-pathogen activity. States that she did not exhibit sex.   This application describes the exact characteristics of the causal component (s) No mention is made. The application is for the adhesion of pathogens to human and animal gastrointestinal epithelium It also suggests inhibition of They have addressed this issue, andE.coli) High molecular weight metabolite of lactobacilli isolated from pigs with adhesion of K88 strain to pig intestinal mucosa Lactobacillus from the digestive tract of mice Indicates that no event occurred. According to subsequent studies, this was It is not a compound that inhibits growth in cells, and the mechanism of inhibition of adhesion needs to be examined. It was shown to be. Lactobacillus metabolites are probably pathogenic for many strains It may inhibit the pathogen colonization of mucosal surfaces, a prerequisite for gender. Consequently, other factors should be considered in addition to growth inhibitory activity. Mucous membrane No explanation is given about the combined inhibition. Description What is being done is Lactobacillus fermentum (Lactobacil rus fermentum) KLD is E. coli (E.coli) Strain, Campylova Ktel Jejeuni (Campibacter jejeuni),monkey Monera Sofia (Salmonella sofia), And Streptococci Cass Faecium (Streptococcus faecium) Growth Was inhibited in vitro. For example, L with glucose in BHI medium . Fermentum (L.fermentum) Can be obtained during the growth The supernatant is then dialysed and cut off at 10,000 and 30,000 Mw. Can be induced by fractionation through ultrafiltration of It is described as possible.   The most recent publication on the aforementioned features is Blomberg L. et al. ; Henr iksson A. Conway P .; (Appl. Env. Microbi ol. feb. 91, p499-502). Exceeds 250,000 present in the retentate fraction of Lactobacillus fermentum by protein with different MWLactob acillus fermentum) Protein-mediated contact of strains to mouse squamous epithelium Assumptions are made about the wearing mechanism. This publication describes the nature of the protein. Has not been described, and the protein has not yet been isolated, Clearly state that its efficacy should be demonstrated by more detailed experiments are doing.   As a result: the role and properties of the protein in bacterial pathogen adhesion Although clear and well-reported, non-pathogenic The role of proteins in fungal adhesion is still at least controversial and unresolved It is a state.   Although many diseases can be treated with antibiotics or drugs, There is a general trend to limit the use of certain compounds, because pathogenic organisms It is becoming increasingly resistant to antibiotics and drugs. Cure of bowel disease A very promising surrogate for therapeutic drugs is the probiotic It is the use of non-pathogenic bacteria that combine properties.   Symbiotic substances are defined as "cells dead or fermented by humans or animals. Applied to the host by improving the properties of the inherent microflora A single or mixed culture of living organisms that have a useful effect. You.   Lactobacillus (Lactobacillus) Strains and bifidobacteria (BifidobacteriumSome of the strains have a probiotic character Reported to have. These beneficial effects include, for example, E. coli (Esch erichia coli)-Gram-negative pathogen growth This is due to a decrease in pH, which is a condition for lowering the pH. In addition, lactic acid bacteria Species are oligopeptides with anti-bacterial properties called bacteriocins To produce These compounds are, for example, Clostridium (Clostridu m ), Listeria (ListeriaGram-positive, such as) It is bacteriostatic or antibacterial for bacterial pathogens.   Some lactobacilli inhibit adhesion of pathogens in animal and in vitro models It has been suggested. These inhibitory effects are usually Described by the nonspecific steric hindrance of the receptor. In contrast, each pathogen It has a specific intestinal receptor (Falkow et al. Ann. Rev. C ell. Biol. 8: 333-363 1992).   Lactobacillus or preparations made with Lactobacillus can therefore cause intestinal and urinary tract diseases. Widely used to treat (eg, WO 9 516 461; Russian Federation No. 2000 116; WO 9418997: Europe State Patent 0 577 903; British Patent No. 2 261 372; 9 301 823; WO 0 921 475; U.S. Pat. No. 2505; Canadian Patent No. 1 298 556: European Patent No. 0 199 5 35; EP 0 210 579). Such a preparation The beneficial effects of a substance can be attributed to various factors. Properties and modes of action have not yet been published, or at best are only hypotheses. It has only been advocated. On the mechanism of symbiotic substances The answer is to discover new enhanced symbiotic substances and optimize their use. Is crucially important.   A starter showing the effects of scientifically proven symbiotic substances The economic importance of using strains to food companies and the development of mucosal vaccines GRAS (generally recognized as safe as a carrier for release (Gene rally Recognized As Safe)) The use of organisms Given the equally great benefits of pharmaceutical companies, bacteria, especially GRAS organisms, Separately, lactobacilli are screened for symbiotic and / or immunomodulatory properties. When training Great effort is paid. A major drawback of existing screening programs is that They are laborious, time consuming and therefore very expensive. That is to be great. About the symbiotic or immunomodulatory properties of the bacterial strain No concise and reliable testing system is available for all screening. Not available. Purpose of the invention   It is an object of the present invention to overcome the above difficulties. Now 29kD proteinaceous The compound was administered to the mucosal receptor site of pig and mouse at the receptor site. Fermentum (L .fermentum) Is clearly responsible for the specific adhesion To synthesize proteins with similar structures and functions Provided herein are methods for screening for other microorganisms. L . Fermentum (L.fermentum) Is the required pathogenicity Demonstrate that it is a 29 kD protein structurally related to bacterial adhesion factors The nucleotide sequence of the gene encoding the adhesion factor Thus, the present invention provides a rapid screening of microorganisms containing a gene encoding a novel type of adhesion factor. By using methods for leaning and standard protein and nucleic acid techniques Provided is a method for screening a microorganism that produces such an adhesion factor. You.   Virulence factors of pathogenic bacteria and non-pathogenic bacterial adhesion factors, ie (ic) lactobacilli The present invention provides the gastrointestinal tract, urogenital Selective for pathogen adhesion to receptors on the tract, oral, respiratory and nasal mucosa The ability to specifically interfere with and prevent the adhesion of pathogens of the type described. To screen for microorganisms To provide a way.   Many possible applications are described in more detail below. i) more rapid for bacteria with symbiotic and / or immunomodulatory properties A method for screening for and directed bacteria is now possible. The present invention Speeds up bacteria for their ability to interfere with pathogen adhesion to mucosal receptors It becomes possible to screen. In particular, the invention relates to the gastrointestinal tract of humans and animals. Bacterial receptors on the mucosa of the genitourinary tract, respiratory tract, and oral / nasal cavity (one or Specific adhesion of non-pathogenic Gram-positive bacteria to more than one), more particularly milk To screen bacteria for the presence of adhesion factors that promote bacillus adhesion To provide a way. Microorganisms to be screened in humans and animals Preferably, the microorganism is non-pathogenic. Such microorganisms are human and / or Or unique to the animal and therefore in the environment in which they will be applied It is already possible to withstand, and even against the special species from which they are obtained Preferably have no toxicity. Suitable non-pathogenic microorganisms Examples of the genus Lactobacillus (Lactobacillus), Streptococcus Su (Streptococcus), Enterococcus (Enterococc us ), Bifidobacterium (Bifidobacterium), Crost Rhidium (Clostridium), And bacterioides (Bacter ioides )including.   This screening may include, for example, polypeptide or protein probes and Using antibodies specific for the adhesion factor (s) to be detected Nucleic acid amplification and hybridization technology As well as protein or peptide detection, such as protein or peptide assays. Are proteins and / or nucleic acids using standard techniques known for nucleic acid detection. Can be done at the level of The present invention therefore further has the desired adhesive properties. For screening microorganisms for the presence of nucleic acids encoding different proteins It also provides the law. Protein and nucleic acid assays may be appropriate Easily performed by those skilled in the art once the amino acid and nucleic acid sequences have been determined Is done. With such information, for example, the nucleic acid sequence of the protein and / or Once the appropriate amino acid sequence has been determined and isolated or synthesized, the probe and It is possible to build a limer. Isolation and / or isolation of pure protein Expression of various proteins allows the production of antibodies in a manner known per se .   In accordance with the present invention, bacteria are isolated from a new class of non-pathogenic microorganisms described in detail below. Presence of a protein included in the definition of adhesion factor, or such a protein or its Can be screened for the presence of a DNA sequence encoding the active portion of Wear. Thus, the application of the present invention is particularly relevant for its ability to adhere to living tissue. Avoid the painstaking and expensive routes of screening bacteria. More specially books Application of the invention applies to animal and / or human volunteers for screening purposes Avoid using.   Screen for non-pathogenic microorganisms for those that can specifically bind to the mucosa. Culturing, on or in microorganisms, or cultures of microorganisms The presence of certain special proteins, either on or in microbial cultures, is not known per se. Wherein the specific protein is within the scope of claims 1-13. Methods that are proteins that follow Included in the protection scope. Alternatively, microorganisms that can specifically bind to mucous membranes A method for screening all non-pathogenic microorganisms, comprising: The presence of a particular gene in an organism or on or in a culture of microorganisms; Detecting the presence in a manner known per se, wherein the special gene A method for encoding a protein according to any one of ranges 1 to 13 is within the scope of the present invention. include.   The invention further relates to the detection of non-pathogenic microorganisms that can specifically bind to mucous membranes. Claims 1 to 13 also include suitable kits, such as antibodies. And a component capable of specifically binding to a protein according to any of the above. In another aspect, the present invention provides a method for identifying a non-pathogenic microorganism capable of binding specifically to a mucosa. A kit suitable for detection, said kit comprising, for example, a nucleic acid probe or a primer. A nucleic acid sequence encoding a protein according to any one of claims 1 to 13 A component capable of specifically binding to a portion of the array. ii) a protein or polypeptide capable of specifically binding to mucosa Or by applying to animals, or to such proteins or poly Microorganisms capable of expressing the peptide or cultures of such microorganisms are human Alternatively, by applying to animals, the pathogen to mucosa or mucin is used herein. It is possible to hinder the adhesion of sex microorganisms. In particular, the urogenital tract of humans and animals Prevents pathogenic microbial adhesion to the gastrointestinal tract, respiratory tract, and / or oral / nasal cavity It can be prevented or reduced. Of particular interest is the present invention Effectively and selectively interferes with the attachment of certain types of pathogens to bacterial receptors Microorganisms that can interfere with the adhesion of certain types of pathogens The purpose of the present invention is to provide a method for performing screening. In this specification Pathogens that can be excluded are Gram-positive and Gram- Includes both negative bacteria, especially those that specifically bind to mucosal receptors. Control Examples of pathogens to be found are the genus Escherichia (Escherichia), Campylovak Tell (Campylobacter), Haemophilus (Haemophilu s ), Shigella (Shigella), Vibrio (Vibrio), Pasteurella (Pasteurella), Yersinia (Yersinia), Salmonella (Salmonella), Mycobacterium (Mycobacterium ), Listeria (Listeria), Clostridium (Clostridi um ), Staphylococcus (Staphylococcus), And analogy Includes strains of the virus such as rotavirus, poliovirus, and measles.   The present invention relates to Fermentum (L.fermentum) Adhesion of 104R Feeling of pathogens adhering to mucosal receptors through adhesion factors similar to those of proteins Dyeing is a receptor rather than a more general mechanism of steric hindrance Specific interaction with L. Fermentum (L.fermentum) 1 Probiotic cells harboring an adhesion protein having a structure similar to that of the 04R adhesion protein It takes advantage of the result that it will be reduced by fungi.   According to the present invention, certain pathogens (L. fermentum (L.ferment um )) Which adheres to the adhesion protein by an adhesive factor that is structurally related Eg, such an adhesion protein or a microorganism producing such an adhesion protein. By administration into food or feed Or specifically inhibited by administration as a pharmaceutical composition Strategies can be devised. This application can be applied to pharmaceutical compositions and / or feed additives. Topical, oral, or intravenous at any of the usual applied doses be able to. The dosage form chosen depends on the type of infectious agent to be controlled. Is done. This dosage form can be solid or liquid. Purity and hygiene I.e. the prescribed standards for sterility normally applied to such compositions are adhered to It must be. Such situations are well-known to those skilled in the art.   Pharmaceutically acceptable dosage forms include pharmaceutically active components- A protein or peptide according to any one of ranges 1-21 -An expression vector according to claims 24 or 25 -A recombinant microorganism according to claims 26 or 27, or a part of said microorganism. (The part expresses mucosal binding promoting activity) Expressing a protein or peptide according to any of claims 1 to 21 A non-pathogenic microorganism, or a portion of said microorganism, wherein said portion is mucosal-bound Expresses promoting activity) And a pharmaceutically acceptable carrier selected from the group of components comprising Is included in the present invention. Take a form suitable for use as a food additive Compositions containing the aforementioned components are also considered to be within the scope of the present invention. Mucous membrane For the prevention and / or treatment of diseases or illnesses associated with Pharmaceutical compositions include, as a pharmaceutically active component, A protein or peptide according to any of claims 1 to 21 -An expression vector according to claims 24 or 25 -A recombinant microorganism according to claims 26 or 27, or a part of said microorganism. (The part expresses mucosal binding promoting activity) Expressing a protein or peptide according to any of claims 1 to 21 A non-pathogenic microorganism, or a portion of said microorganism, wherein said portion is mucosal-bound Expresses promoting activity) Use of a component selected from the group of components including .   As will become apparent from the above, claims to improve food products Products according to any of boxes 1-21 or 24-27, and / or claims May express a protein or peptide according to any of ranges 1 to 21 A non-pathogenic microorganism or a part of said microorganism (the said part is The method comprising the addition of a compound to a food product forms one aspect of the present invention. . Such a method may be in accordance with any of claims 1-21 or 24-27. It is preferred to include the addition of carbohydrate products to food products.   A product according to any of claims 1-21 or 24-27, and / or Or expressing a protein or peptide according to any of claims 1 to 21 A non-pathogenic microorganism or a portion of said microorganism, wherein said portion is a mucosal Food products containing (expressing a binding renewal activity) as an additive are clearly included in the present invention. It is. Add product according to any of claims 1-21 or 24-27 Food products comprising as a product are a particularly suitable embodiment.   One of skill in the art will appreciate that this inhibitory effect may also be specific for, for example, L. is found Fermentum (L.fermen tum )) Of the portion of the adhesion protein such as the peptide derived from the 29 kD adhesion protein. It will be appreciated that the addition may also be obtained. these Active peptides are chemically synthesized or engineered microorganisms Can be either made microbiologically. Alternatively, the protein White matter can be produced by non-recombinant or recombinant microorganisms, and And then via, for example, proteolytic digestion and optical resolution of the proteolytic fraction. To obtain the desired polypeptide. Its 29 kD adhesion factor, and Escherichia coli, a pathogenic organismEscherichia coli) And helicopter Bactel pylori (Helicobacter pyloriAdhesion factors of strains) And the common sequence KKXXXK (SEQ ID NO: 30) is assumed from the analysis of cholera toxin Wherein X represents any amino acid and K represents lysine. A 29 kD protein according to the present invention contains three such sequences. These are mature proteins White matter positions 47-52 (KKMGLK), 173-178 (KKNSTK), And 223-238 (KKLSEK), which are approximately throughout the protein molecule Evenly distributed. Numbering is performed for amino acids 54 to 59, 1 80 to 185, and 230 to 235; The quality starts at Ala at position 8. Within the protein or peptide according to the invention. At least one of the KKXXXK sequences, preferably two of these sequences Is preferably present. Most preferably, there are three such sequences No. In a particular embodiment, this consensus sequence is present in the 29 kD protein disclosed above. It may be one of the naturally occurring amino acid sequences. Against what exists in the natural environment That the corresponding array is used. Preferably, however, such a sequence is, for example, a DNA synthesis as described above. Adult machine, Merrifield synthesis, and cloning techniques Through genetic engineering means commonly known in the art. Alternatively, it can be reached through means for synthesis. Such an array is even more It is also preferred that the three-dimensional structure of the New This is confirmed using computer technology in a manner known per se can do. Such a sequence would bind to a negatively charged intestinal receptor. Get involved. For example, Aspergillus (Aspergillus), Lactobaki Ruth (Lactobacillus), And Lactococcus (Lactoco ccus Microorganisms having a GRAS state, such as) are very suitable for such purposes. I do. Those skilled in the art will recognize that other microorganisms may also be involved in the production of adhesion factors or peptides derived therefrom. It will be appreciated that it can be used. However, GRAS organisms are not A human application would be preferred for use. List of GRAS organisms is food And / or readily available to those skilled in the pharmaceutical arts, and Incorporated herein by reference. For example, the US Food and Drug Administration (US FDA) Maintains a list of such organisms.   L. Fermentum (L.fermentum) 104R or L.R. Fe Lumentum (L.fermentum) Similar to 104R adhesion promoting protein Proteins such as adhesion-promoting proteins of microorganisms that produce proteins with similar structures Will interfere with specific adhesion of pathogens carrying adhesion proteins with similar structure The result is that such an adhesion-promoting protein or an adhesion-producing protein-producing microorganism Object has a similar structure Must not interfere with the adhesion of pathogens that do not produce Nor does it suggest. Those skilled in the art will be familiar with L.A. Fermentum (L.ferment um ) Coloration of the use of a microorganism having an adhesion promoting protein, such as that of 104R -(Colori) is that adhesion of such bacteria to specific receptors Beam, L. Fermentum (L.fermentumOther than those of the) -like adhesion factor Of pathogens with multiple adhesion factors is also limited by the general mechanism of steric hindrance You will immediately recognize that there may be things that will happen. Therefore, control Capable pathogenic microorganisms are specifically bound by adhesion factors from non-pathogenic organisms It does not include only those microorganisms that bind to the mucosal receptor that receives it. iii) a group of proteins exhibiting the desired activity is now known, and Since the sequence and nucleic acid sequence have already been determined, the desired protein or polypeptide Develop and / or select microorganisms that have improved production of Options are made possible here. This allows for the usual optimization of culture conditions. Strains that express proteins with improved receptor binding properties Can be achieved through selection in the manner that is being done.   Nucleic acid sequence (s) into a better microorganism through genetic engineering Is incorporated and thereby causes the microorganism to become free of the mucosal binding promoting component. It is also possible to make it possible to carry out (redundant) expression and preferably secretion . Preferably, the microorganism is a GRAS organism such as, for example, a lactic acid bacterium. Change Contains the sequence (s) to be coded Manipulation of regulatory sequences that allow for higher expression than with linked regulatory sequences Operatively linked It is also possible to take in as follows. Numerous high expression vectors, such as lactic acid bacteria And various microorganisms within a particular GRAS microorganism such as Appropriate Novel adhesion factors from non-pathogenic microorganisms or recombinant expression vectors containing nucleic acids May also express a polypeptide or protein capable of promoting mucosal binding of the group. Alternatively, recombinant microorganisms that can be overexpressed are also included in the scope of the present invention. Heredity Microorganisms created by child engineering may already express the adhesion factor Good, but does the microorganism not express the new group of adhesion proteins in its native state? Can be selected from The microorganism can be used as a drug or as a food / feed additive Production plant for proteins or polypeptides that can be subsequently isolated and applied (Plant) may be used simply or the microorganism itself may be used. The body may be used as a drug or as a food / food additive . Preferably, the protein or polypeptide producing microorganism is non-pathogenic. No. In particular, GRAS microorganisms are active components of pharmaceutical compositions or food / food additives. For the purpose of allowing the application of its expression product and / or microorganism Good.   In any of the embodiments known per se in recombinant DNA technology, the nucleic acid Sequence can be integrated into a plasmid vector or integrated into a chromosome It may be. Most transformation and expression vectors and techniques are It is known in the state of the art and is also currently available commercially. Food grade transformation and expression vectors and methods suitable for GRAS microorganisms The method is preferred.   Preferably, the microorganism to be selected and / or transformed has the following characteristics: Good:   -Survival in environmental conditions where activity must be exhibited   -Proliferation and / or colonization at sites showing activity   -Absence of an immune response to the probiotic strain   -The probiotic strain itself, its fermentation products, or its cellular constituents after bacterial death Absence of pathogenic, toxic, allergic, mutagenic or carcinogenic reactions   -Genetic stability, absence of plasmid transfer reaction   -Easy and reproducible production   -Viability during processing and storage.   Generally, the term "recombinant microorganism" refers to a nucleic acid sequence according to claim 23, and And / or the expression vector according to claim 24 or 25, Acid sequences and / or expression vectors are absent in the corresponding non-recombinant microorganism Or its surrogate is present at a low copy number or is expressed to a lesser extent You. In yet another aspect, the invention is just as specified (according to claim 26) Recombinant microorganisms, wherein said microorganisms are non-pathogenic microorganisms, preferably human Or microorganisms that are endemic to animal microflora, more preferably human microphones. It is a microorganism unique to Loflora.   The present invention further provides a protein or peptide according to any of claims 1 to 21. A nucleic acid sequence encoding any of the nucleotides, and a nucleic acid sequence comprising such a nucleic acid sequence. An expression vector operably linked to the current regulatory sequence, wherein said expression vector is For example, the nucleic acid can be expressed in a non-pathogenic microorganism such as a GRAS microorganism. And said expression vector preferably comprises a nucleic acid derived from a GRAS microorganism. New In yet another aspect, In the expression vector according to the present invention, the nucleic acid sequence of the expression It is a vector that is not linked to the resulting gene encoding an adhesion factor. iv) The 3D structure, amino acid sequence, and nucleic acid sequence of the adhesion protein have now been confirmed. And similarities with other protein groups have been determined and The design of proteins or polypeptides that exhibit improved binding properties is also a matter of skill in the art. Included in the surgical field, and such improvements may lead to pharmaceutical applications or food / feed This results in application as an additive. Accordingly, the present invention has the amino acid sequence of FIG. Better than mucosal proteins, and were identified in the experimental part of Mutated polypeptide exhibiting better mucosal binding than any of the polypeptides IV Also includes polypeptides and proteins. The present invention further provides that it can be obtained in its natural state. And the equivalent sequence as a mutant, ie a 29 kD protein Has at least the mucosal binding activity of a protein and such a protein or polypeptide. A nucleic acid sequence encoding a protein or polypeptide Or their application in any of the claimed methods. v) The discovery of a group of proteins and peptides that can specifically bind to mucous membranes And target microorganisms that express the protein or peptide to the mucous membrane Not only that, but also to eliminate such microorganisms To target additional compounds, such as drugs, mucus modulators, or antigens Can also be used as a carrier. This microorganism already has this special property Or the microorganism is a desired drug, immunomodulatory Agent or antigen Can also be created by genetic engineering to produce later. Moreover Must be targeted to the mucosa-promoting amino acid sequence and to the mucosa. An additional desired amino acid sequence or molecule having the characteristic activity of It also allows for the development of fusion proteins or peptides containing. So the mucous membrane To develop all strains of novel pharmaceutical compounds that are specifically targeted Wear. The present invention relates to such novel microorganisms and molecules, and pharmaceutical compositions. Including their application. Thus, the present invention provides, for example, encoding antigens of a pathogenic organism. Bacteria that express a gene of interest, such as Target, and thereby elicit a specific immune response against the antigen And / or modulating the immune response. The present invention Claims 1 to 21 linked to a drug, an immunomodulator or an antigen A fusion protein or peptide also comprising a protein or peptide according to any of including.   Additional pharmacologically active components for the mucosa (the additional pharmacological components are labeled Pharmaceutical set for targeting (which is physically linked to a targeting component) As a target component in the product -A protein or peptide according to any of claims 1-21 -An expression vector according to claims 24 or 25 -A recombinant microorganism according to claims 26 or 27, or a part of said microorganism. (The part expresses mucosal binding promoting activity) Expressing a protein or peptide according to any of claims 1 to 21 A non-pathogenic microorganism or a portion of said microorganism, wherein said portion is Express the activity) Use of a component selected from the group of components comprising is included within the scope of the present invention. .   The promotion of the specific adhesion of Lactobacillus to mucosal receptors according to the invention is, for example, For example, lactic acid bacteria expressing antigens of pathogenic organisms or human proteins Opportunities to specifically target cells bearing the compound to cells in the mucosa And thereby modulates the immune response to the antigen / human protein, and This is a preferred embodiment of the present invention.   According to the present invention, the adhesive capacity of a probiotic strain is altered by altering the properties of the adhesive protein. It may be adjusted more. This property is attributed to the mucosal receptor of the adhesive protein. Requires interaction with putters or with other (auxiliary) proteins There is also. Detailed description of the invention   According to the invention, in particular, from the porcine gastrointestinal tract, which has never been described before The isolated strain L. Fermentum (L.fermentum) 104R Encoding a protein having a Mw of 29 kD and / or its adhesion protein Use of the DNA sequence is performed. This novel protein has an adhesion promoting activity. In particular, This adhesion promoting activity involves exhibiting binding to mucosa or mucin. This bond The protein is present on the surface and L. Fermentum (L.fermentum ) 104R also sheds this adhesion protein into its culture medium.   The invention more particularly relates to the special properties of the adhesion promoting protein, For example, Campylobacter Jejuni (Campylobacter jej uni ), Pasteurella haemolitica (Past eurella haemolytica), And Mycobacterium (Myc obacterium ) Toxicity of numerous pathogenic bacteria such as to adhesion factors from It takes advantage of the special property of being structurally similar to proteins. These features are: Described in the clause. According to the present invention, features similar to those of the 29 kD protein are provided. The presence of a protein having the property is determined by Western (a technique well known to those skilled in the art). This can be determined by using a Western blot technique.   L. Fermentum (L.fermentum) Adhesion promoting protein from 104R White matter is a class of proteins called class III solute transporters And its basic types are histidine transporter (HisJ), glutamine transporter (G lunH), and lysine, arginine, and ornithine transporter (LAO) is there. The three-dimensional structures of two of these proteins, HisJ and LAO, are known. Have been. L. Fermentum (L.fermentum) Adhesion promotion of 104R Depending on the amino acid sequence of the advanced protein, C.I. Jejuni (C.jejuni) Peb1 and p. Haemolitica (P.haemolyticaLap) With adhesion proteins of pathogens such as T and on the other hand LAO or His A member of a Class III solute transporter protein such as J Are shown. By modeling proteins, L. Fermentum (L .fermentum) The expected 3-D structure of the adhesion factor is also LAO and HisJ It is shown to be similar to Classes that are essential for ligand binding Amino acids in proteins in domain I of class III solute transporter Acids are conserved among all members of this class of protein. These Mino acids are also L.A. Fermentum (L.fermentum) 104R contact Adhesion promoting protein and C.I. Jejuni (C.jejuni) Toxicity within the toxic protein Was found in the position to do. In other words, L. Fermentum (L.ferm entum ) 104R are adhesion promoter proteins from C. Jejuni (C.jeju ni ) And P. Haemolitica (P.haemolytica) Like pathogens It has a 3-D structure similar to that of the adhesion factor.   Are the proteins belonging to the group of novel proteins identified according to the invention non-pathogenic microorganisms? Identified as a protein that can be obtained from And a molecular weight of 20-40 kD. The molecular weight should be between 20-30 kD Is preferred. Specific embodiments are disclosed in claims 1 to 13. In particular, the invention A protein according to comprises one or more of the following properties: a) Molecular weight between 20 kD and 40 kD b) Class III solute transporter and / or C.I. Jessie Euni (C.jejuni) Peb1, P.I. Haemolitica (P.haemol ytica ) LapT, as well as Mycobacterium tuberculosis (My cobacterium tuberculosis) Or mycobacterium Mu Leprae (Mycobacterium leprae) 85K complex protein More than 20% identical amino acids to the amino acid sequences of the toxic proteins A, B, and C Amino acid sequences exhibiting amino acids and more than 40% similar amino acids c) C.I. Jejuni (C.jejuni), P. Haemolitica (P.haem olytica ) Or Mycobacterium (Mycob acterium Characteristics of mucosal receptors also used by any of Promotes heterogeneous binding d) 3-way with two lobes (round protrusion) like LAO or HisJ Has D structure e) one or more that show 90% or more similarity to the following amino acid sequences: Contains multiple amino acid sequences   I) AASAVNSELVHK   II) ANFVPTK   III) DTAIQSSYNK   IV) ISALNK   V) IAGTGTNNA (amino acid sequences II-V are preferred).   A specific embodiment is that the protein exhibits the consensus sequence illustrated in FIGS. Formed by a group of proteins further characterized by: So claimed The protein to be isolated may be a virulence factor of a pathogenic microorganism or a class III (Class I). II) No solute transporter is involved and that type of recombinant protein is probably Recombinant toxicity that may form a report of the art on the filing date of the application May include a factor or a recombinant Class III reporter Absent.   Proteins belonging to the group of proteins suitable for application according to the invention are described in detail in the examples. Having the amino acid sequence of FIG. 3 which can be determined by a mucosal binding assay L. Fermentum (L.fermentum) 104 29 kD protein C. higher or equal to that presented by C.I. Jejuni (C.jejuni ), P. Haemolitica (P.haemolytica), Maybe Is mycobacterium (MycobacteriumA) mucosal receptor used by any of It is preferable to exhibit the activity of promoting binding.   Since the nucleotide structure of the adhesion promoting protein is known, non-pathogenic . Fermentum (L.fermentum) 104R adhesion protein Screenini for the presence of DNA sequences encoding proteins with similar structures Can also be used. It is a protein that exhibits at least the same mucosal binding activity A so-called equivalent sequence that encodes a protein or polypeptide. Especially like this The nucleic acid sequence is Fermentum (L.fermentum) 104k of 29R 2 is a nucleic acid sequence encoding the amino acid sequence of FIG. 2 corresponding to that of the D protein. Figure The nucleic acid sequence itself encoding the common amino acid sequence of 4 and 5 is also included in the scope of the present invention. I will. In particular, it encodes a 20-40 kD protein containing a consensus amino acid sequence, And a nucleic acid corresponding to a further 29 kD sequence (the only difference being that the hydrophobic properties are similar) And the high degree of conformation of the resulting protein or polypeptide Amino acid substitution by other similar amino acids such that no conformational change can be expected. Is present within the scope of the present invention. I will. Such a sequence, when compared to the sequence according to SEQ ID NO: 2, It is known that a conservative change has occurred. The present invention further relates to Stringent hybridization when performing blot assays using Also includes any nucleic acid sequence capable of hybridizing under conditions. Therefore Such a sequence comprises, in a manner known per se, an amino acid sequence according to SEQ ID NO: 2 Oligonucleotide probes, preferably screening, Consider the preferred codon usage of the microorganism to be Other non-diseases through cross-hybridization techniques using inserted probes Includes sequences encoded by nucleic acid sequences that can be obtained from protozoan microorganisms. In order to obtain such an equivalent array, for example, Molecular Cloning , A Laboratory Manual, Cold Spring Har Bor Laboratories New York Maniatis, T.W. F ritsch, E .; F. and Sambrook, J, (1982). Stringent hybridization conditions can be applied as appropriate . The cited reference cited may also appear elsewhere in this description. It also provides information on a number of other standard technologies and is incorporated herein by reference. I will. In particular, from non-pathogenic microorganisms belonging to the genus already mentioned in this description Are preferred. In a preferred embodiment, furthermore, at least one according to SEQ ID NO. There are two common arrays. Alternatively or additionally, polypeptides IV There is one of the sequences Amino acid mutation outside of the consensus sequence Having at least the mucosal binding activity of the mature protein of SEQ ID NO: 2 A protein or polypeptide having the amino acid sequence of the mature protein is also within the scope of the present invention. Contained within. Any arrangement combining any of the above definitions is also a subject of the present invention. And forms a preferred embodiment. An equivalent array is itself Although not available from certain microorganisms, for example, recombinant DNA technology, PCR It is also possible that they can be produced in alternative ways, such as. The previous aspect is And fall within the scope of the present invention. . Proteins or polypeptides according to the present invention may be used in cell extracts and other contaminating proteins. Does not include quality. Substantive The high purity is preferably, for example, a purity exceeding 80%. Its purity depends on the drug and And food additives, as well as activities required in applications according to the present invention. Is enough to achieve sex.   In accordance with the present invention, bacteria are non-living organisms, such as plastic or metal surfaces. L. can adhere to the body surface Fermentum (L.fermentum ) Being screened for the presence of a protein such as the 104R adhesion protein There may be. Such a screen was performed as described above, and was performed at 29 kD. It can be performed using an oligo probe based on the amino acid sequence of the adhesion protein. Preferably, such a probe encodes a portion of the consensus sequence of FIGS. New In addition, a suitable probe is a sequence encoding the consensus sequence of SEQ ID NO: 30. including. This consensus sequence or part thereof has at least 5 contiguous amino acids. Length, and the probes preferably comprise completely consensus sequences. New 29kD protein or its active part required for mucosal binding activity Sequences encoding proteins or polypeptides that exhibit the closest possible identity It is also possible to use such probe combinations for the purpose of obtaining. Further details of embodiments of the invention i)Lactobacillus fermentum Production and Purification of Adhesion Protein from Rentum) In a preferred embodiment of the present invention, Fermentum (L.fermentum) The adhesion-promoting protein of 104R is obtained by converting bacteria into MRS broth or LDM medium (Con way and Kjelleberg, J. et al. Gen. Microbiol. 1 35: 1175-1186 1989). It is produced by culturing for 14 to 24 hours. This 29 kD adhesion protein From the medium, ammonium sulfate precipitation, gel filtration, and affinity chromatography It is purified until it is apparently homogeneous. Adhesion promoting activity is A and dot blot assays detected in the fractions and PAGE, SD using mucus or mucin labeled with viperoxidase Visualized by S-PAGE and Western blot. Purified protein Under non-denaturing conditions and under reducing and denaturing conditions (using standard proteins) SDS-PAGE for isocratic denaturation using the obtained calibration curve, and its standard song Gel Filtration Chromatography Correlating to Line) Expected Mw of 29 kD And is sensitive to pronase, and is therefore No. 210 579 and WO 90/09398 and / or Suggested adhesion proteins, as well as Conway and Kjellenberg (J.G. en. Microbiol. 135, 1175-1186), Blomberg (Appl. Environm. Microbiol. 59, 34-391). 993), and Aleljung et al. (Current Microbiolo). gy vol 28 (1994) p. 231-236) Is also different. Proteins specifically disclosed as such in the cited references Quality does not fall within the protection scope of the claims for proteins or peptides. Quoted The compositions specifically described as such in the cited references are claimed in the composition. It is not included in the scope of protection. In particular, such as pathogen inhibition Application of the composition of WO 90/09398, specifically described as Not included. When specifically stated in this clause, this may be an example or its In the following description, the materials and methods of the cited references are referred to. like this The general disclosure scope of the genes in the various references covers some aspects of the invention. The present invention nevertheless forms the invention of choice against the cited document. I do.   Adhesion promoting proteins are described in L. et al. Fermentum (L.fermentum) Cell table Surface, extraction of the bacteria with 1 M LiCl and low concentrations of lysozyme I was able to get out. Both small intestinal mucus and gastric mucin from pigs or mice An adhesion promoter having an affinity for is grown on the culture after growing for 24 hours. Released into the fresh liquid. ii)L. In the presence of L. fermentum-like adhesion protein Microbial screening   In another preferred embodiment of the invention, the lactobacilli are L. lactis. Fermentum (L.fe rmentum )), Which have characteristics similar to those of the adhesion-promoting protein from For the presence of proteins, SDS-PAGE, and L. et al. Fermentum (L.fermentum ) Created in rabbit for 104R purified adhesion protein Western blot using the isolated polyclonal antibody The overnight culture is screened by separation of the protein from the culture medium. iii)L. Coating L. fermentum-like adhesion protein Screening microorganisms for the presence of transfecting genes   In another preferred embodiment of the present invention, the DNA is subjected to screening, and From microorganisms subjected to PCR analysis, Fermentum (L.ferment um ) The gene encoding the 104R adhesion protein It is isolated using a pair of primers based on the leotide sequence. The product formed is a theory Standards described in the written statement (eg, cited elsewhere in this statement) It is analyzed by molecular biology techniques or commercially available kits. iv)L. Organisms other than L. fermentum 104R Of adhesion-promoting proteins in yeast   In another specific aspect of the present invention, Fermentum (L.fermentu m ) Isolated from 104R or from other selected strains by the methods described above. The gene coding for the adhesion protein is, for example, Aspergillus niger (As pergillus niger), Lactobacillus (Lactobacill us ), Etc., in a GRAS-producing organism. Cloned after the promoter and secretion signals. This culture medium is And / or as a food / feed additive or pharmaceutical composition Or the adhesion-promoting protein is first purified (by standard techniques) and then It is either added to the feed preparation or the pharmaceutical composition. The nucleic acid sequence , Whose sequence is the 29 kD L. Fermentum (L.fermentum ) May be regulated to encode the same amino acid sequence of the 104R adhesion protein May be present, but codons are adjusted to the preferred codon usage of the host where the incorporation takes place Have been. Details of preferred codon usage are known to those skilled in the art of nucleic acid expression. Available from commercial sources. v)Production of peptides with adhesion promoting properties   In another preferred embodiment of the present invention, L. var. Fermen Tum (L.fermentum) Peptide derived from 104R adhesion protein Chemically synthesized and used as food / feed additives. Alternatively, the DNA sequence encoding such a peptide may be, for example, A. Nigel (A .niger) Or Lactobacillus (Lactobacillus)Such After a strong and preferably inducible promoter in a GRAS producing organism such as Cloned. Sequence encoding the peptide encodes a secretory signal Cloned after the sequence that is to be secreted, and the peptide is secreted into the medium. If so, the medium can be used as a food / feed additive. The peptide Is not secreted into the medium, the whole organism, or from such an organism The resulting extract can be used as a food / feed additive. Otherwise desired Is a protein or polypeptide to be isolated, for example. For example, to isolate proteins or polypeptides in combination with antibodies specific for Isolation in a manner known per se, for example by using chromatography May be. Epitope or protein according to any of claims 1 to 20 Alternatively, antibodies or antibody fractions capable of binding to peptides are within the scope of the present invention. Contained within. Such antibodies are polyclonal antibodies (see Examples). Alternatively, it can be a monoclonal antibody. Among the cited references cited earlier The antibodies specifically disclosed in any of the claims Out of range of enclosure protection. vi)Targeting antigens or human proteins to the mucosa   In another aspect of the invention, an antigen of a pathogen is immunized to mucosal immunity to the antigen. Mucosal set to target to mucosa to enhance response The ability of the adhesion protein to specifically adhere to the weave is used. Glued for this purpose A microorganism capable of synthesizing proteins and the antigen of interest is constructed. Otherwise To regulate the immune response to human proteins in order to suppress the autoimmune response Are microorganisms that carry genes encoding human proteins, and those microorganisms Fermentum (L.fermentum) Properties similar to those of adhesive proteins It is created by genetic engineering in such a way as to synthesize an adhesion protein having Example i)Lactobacillus fermentation binds small intestinal mucus from pigs and gastric mucin Of surface proteins from Lactobacillus fermentum Purification and characterization   Centrifuge the spent culture from the 14-24 hour culture at 6000 g for 20 minutes. Collected by separation and dialyzed against ultrapure water at 4 ° C. Retention (ret was concentrated by ultrafiltration through a 14 kDa molecular weight cut-off membrane . The high molecular weight fraction was lyophilized and stored at 4 ° C. Add more spent culture Concentrate 10 times by hollow fiber ultrafiltration apparatus and remove ammonium sulfate Dissolved in condensate (40, 60, and 100% saturation at 4 ° C). These deposits Is collected by centrifugation (18000 × g / 30 min), dissolved in ultrapure water, and Dialysis was performed against 0.01 M ammonium bicarbonate. This solution is lyophilized and And stored at 4 ° C.   Lyophilized preparation from 24 hour spent culture concentrated by ultrafiltration Dissolve the material in HEPES-Hanks and filter (0.2 2 μm) to remove insoluble particles. this A 4 ml aliquot of the solution (2.1 mg of protein) was subjected to gel filtration chromatography For XK-26 column (Pharmacia-LKB, Uppsala) Sweden) in Sephadex G200. Hepe The column was equilibrated with HEPES-Hanks buffer and To elute the sample. Dot blot of fractions with each absorption peak at 280-nm Assay and crude in microtiter plate adhesion inhibition assays Upon inhibition of lactobacilli binding to the purified mucus, HPR-mucin and HPR- The ability to bind to the fluid was assayed. Activity with each absorption peak at 280 nm Fractions were pooled, dialyzed and subjected to SDS-PAGE and Western blot Lyophilized for analysis. Alternative purification method.   Activate mucin against CH-Sepharose 4B by its manufacturer (Pharm conjugate according to the instructions of Acia-LKB, Biotechnology) For coupling. Column C10 / 40 (30 ml separation bed volume) And equilibrated with HEPES-Hanks It was made. L. Fermentum (L.fermentum) Used culture broth Fractions from cell extracts, cell extracts, or gel filtration chromatography Over The column was equilibrated at a flow rate of 6 mL / hr to equilibrate a double bed volume. Wash with buffer, then add a different solution (0.1 M glycine pH 3.0, 0.1 M Wash continuously with Tris pH 8 and 0-2M sodium chloride gradient) Was.   This adhesion-promoting activity is determined by the adhesion inhibition assay and dot blot assay. A) in their fractions, and PAGE using mucus or mucin labeled with ze, SDS-PAGE, And visualized by Western blot. This adhesion promoting protein is Treatment of the bacteria with 1 M LiCl and low concentrations of lysozyme caused Ferment Pum (L.fermentum) Could be extracted from the cell surface. Small intestine sticky An adhesion-promoting protein with affinity for both fluid and gastric mucin grows for 24 hours After being released, it was released into the culture supernatant. This active fraction is separated into periodic acid Sc Hiff staining method (SIGMA) and DIG glycan detection kit (Boehrin) ger Manheim (Germany) in the presence of carbohydrates To characterize it, and heat the active region of the adhesion-promoting protein. Stability was determined. This adhesion promoting activity lacks carbohydrates, and Fe Lumentum (L.fermentum) At 100 ° C. For 5 minutes and tested live by the dot blot adhesion assay. Physically it remained completely active.   The purified protein is used under non-denaturing conditions and under reducing and denaturing conditions (standard protein). SDS-PAGE using a calibration curve obtained using white matter and its standard curve Gel filtration chromatography correlating with M; w.   The N-terminal amino acid sequence of this adhesion promoting protein was further determined. And the following sequence:             AXXAVNXELV (V) (K) It has been shown.   This adhesion promoting protein is digested with modified porcine trypsin and formed. The purified peptide was purified by reversed-phase HPLC, and many peptides were found to be mucus and Specific adherence to mucin is confirmed by dot blot assay and mucin adhesion It was found when measured by ISSEY. The amino acid sequence of this peptide is: ANFVPTK, II: DTAQSSYNK, III: ISALFNK, IV : IIAG (T) G (T) NNA. In these sequences, X represents serine (S) Most often it represents. ii)Cloning and sequencing of proteins encoding adhesion factors   The adhesion-promoting protein gene was expressed as L. Fermentum (L.fermentum) 1 It was cloned from the 04R genome bank. Probe to identify adhesion gene sequence The amino acid sequence of the peptide for which the sequence of the adhesion-promoting protein has been determined Oligonucleotide primers were synthesized based on the data. These oligonucleotides Reotide was used in various combinations in the PCR reaction. N-terminal peptide Oligonucleotide 42 corresponding to the peptide and peptide I (sense: 5'-CTI.GC I.GTI.AAC / T.TCI.GAG / A.TTG / A.GT-3 ′) and 105 (antisense: 5′-GCC. GGGA.TCC.TTT.G / A / T / CGT.G / TGG.G / TAC.G / AAA.G / ATT.G / A / TGC-3 ')EcoRI andBamThis produces a 183 bp (7) PCR product flanked by HI sites. Is 3.5 kbSstI-PstI chromosome L. Fermentum (L.f ermentum ) Southern blots on fragments and hybrida I did it. E. coli (E.coli) Cloned into pGEM3 I let you. The location of the gene encoding the adhesion factor is determined by restriction enzyme analysis, and The appropriate of the 3.5 kb fragment The nucleotide sequence of the site was determined (FIG. 2). The expected amino acid of this adhesion protein The noic acid sequence is shown in FIG. iii)L. L. fermentum 104R adhesion protein Analysis of amino acid sequence   L. Fermentum (L.fermentum) 104R adhesion protein Computer-assisted analysis of the acid sequence was performed. FIG. 4 shows that the protein is C. Jessie Euni (C.jejuni) And Peb1 from P.). Haemolitica (P.ha emorytica ) Shows significant similarity to the toxic protein LapT from And FIG. Fermentum (L.fermentum) Adhesion protein The quality should also show similarity to Class III solute transporters. And FIG. 6 shows that this adhesion protein is mycobacterium leprae (Mycob acterium leprae) And Mycobacterium tuberculosis (Mycobacterium tuberculosis) 85K complex toxicity Shows similarity to proteins. By protein modeling studies, L. Fe Lumentum (L.fermentum) Putative 3 of 104R adhesion protein The -D structure is shown to be similar to that of LAO and HisJ. These laboratories Furthermore, Peb1 has a 3-D structure similar to that of LAO and HisJ. Is also shown. iv)Adhesion protein-like protein in Lactobacillus strain White matter determination   About 20 Lactobacillus (Lactobacillus) Strain in LDM medium Culturing, recovering the culture medium, and performing SDS-PAGE. The protein was separated. The presence of adhesion protein-like proteins follows standard molecular biology techniques. Determined by Western blot. Results shown in Table 1 Some Lactobacillus (Lactobacillus) Strain is adhesion protein While others produce the same protein, others do not. BRIEF DESCRIPTION OF THE FIGURES Figure 1 Adhesion promoting protein using mucus labeled with HRP for blotting White matter (APP) SDS-PAGE and Western block G. A) Molecular weight marker (lane 1); APP after affinity chromatography ( Lanes 2, 6, and 7); APP from native PAGE (lanes 3, 4, and 7). And 5). B) Molecular weight marker (lane 1); from gel filtration chromatography APP (lanes 2 and 3). Arrow in lane 3 indicates location of APP; C) gel Western (W) of APP from SDS-PAGE after filtration chromatography ester) blot (lane 1); after affinity chromatography (lane 2) L.) after growing for 14 hours. Fermentum (L.fermentum) 1M LiCl extraction (lane 3); from PAGE (lane 4). FIG. Fermentum (L.fermentum) 104R adhesion promoting protein White matter nucleotide sequence. The open reading frame starts at nucleotide 1 and Ends with Reotide 734. FIG. Fermentum (L.fermentum) 104R adhesion promoting protein White matter amino acid sequence. FIG. Fermentum (L.fermentum) 104R Adhesion promoting protein, C.I. Jejuni (C.jejuniPeb1), and P. Haemolitica (P.haemolyticaAmino acids of LapT from Sequence comparison. The consensus sequence is shown in the sequence below. Bold type indicates the same amino acid or Means existing substitutions. FIG. Fermentum (L.fermentum) 104R adhesion promoting protein White matter and the class III (Class IIII) solute transporter protein (Atuno p, agrobacterium tumefakiens (Agrobacter tumefa ciens ) Nopaline; Atuoct, Agrobacterium tumefakiens (Agrobacter tumefaciens) Octopine; GlnH, large Enterobacteria (E.coliGlutamine binding protein; HisJ, histidine binding Protein, LAO, Salmonella typhimurium (Salmonella th yphimurium ) Of lysine, arginine, ornithine binding protein Comparison of noic acid sequences. The consensus sequence is shown in the sequence below. Adhesion promotion also occurs in other proteins The amino acids of the hex protein appear in bold capital letters; the colons represent conserved substitutions, and The asterisk indicates a less conservative substitution. FIG. Fermentum (L.fermentum) 104R adhesion promoting protein White matter and mycobacterium (Mycobacterium)) Of the 85K complex Comparison of white matter amino acid sequences. The consensus sequence is shown in the sequence below. Adehesin and one One or more mycobacteria (Mycobacterium) With protein Identical amino acids in bold uppercase letters. The colon represents a saved substitution , And the asterisk represent less conservative substitutions. Table 1 L. as a probe Fermentum (L.ferment um ) Lactobacillus using antibodies raised against 104R adhesion promoting protein (LactobavacillusWestern culture medium of the strain n) Blot     signal L. Gasselli (L.gasseri) NCK 89 + L. Reuteri (L.reuteri) ML1 ++ L. Murinus (L.murinus) + L. Fermentum (L.fermentum) 2399 +/- L. Plantarum (L.plamtarum) ++ L. Fermentum (L.fermentum) KLD − L. Animalis (L.animalis) 364T + L. Animalis (L.animalis) 364 +/- L. Kasei (L.casei) ATCC 393 +/- L. Achidophilus (L.acidophilus) NCK 65- L. Animalis (L.animalis) 362- L. Plantarum (L.plantarum) 8014 +/- L. Plantarum (L.plantarum) LP80 + L. Brevis (L.brevis) R3 + L. Brevis (L.brevis) ML12 + E. coli (E.coli) − L. Fermentum (L.fermentum) 104R ++ L. Plantarum (L.plantarum) 256 ++

[Procedure for Amendment] Article 184-8, Paragraph 1 of the Patent Act [Date of Submission] October 17, 1997 (1997.10.17) [Details of Amendment] Claims 1. A protein obtainable from a non-pathogenic microorganism and having a mucosal binding activity and a molecular weight of 20-40 kD, preferably 20-30 Kd, or a polypeptide equivalent thereto, comprising the following amino acid sequence: I) AASAVNSELVHK II) ANFPVPTK III Iv) a protein or polypeptide comprising one or more amino acid sequences identical to IAQGTNNNA by more than 80%. 2. Mucosal binding activity, pathogenic microorganisms, preferably the pathogen Kanpiro tapir ether (Campylobacter), Mycobacterium (Mycobacteriu m), or Pasuteurera (Pasteurella), for example C. C. jejuni , P.J. 2. The protein or polypeptide of claim 1, comprising binding to a receptor recognized on mucosa by any of P. haemolytica . 3. The mucosa-binding activity is determined by the L. lactis having the amino acid sequence of SEQ ID NO: 2. A protein or polypeptide according to any of the preceding claims, which is equal to or greater than that of the L. fermentum 104R 29 kD adhesion factor. 4. A protein or polypeptide according to any of the preceding claims, comprising the amino acid consensus sequence set forth in detail in SEQ ID NO: 6. 5. A protein or polypeptide according to any of the preceding claims, comprising the amino acid consensus sequence set forth in detail in SEQ ID NO: 13. 6. The protein or polypeptide according to any of the preceding claims, comprising the amino acid consensus sequence specifically set forth in SEQ ID NO: 6 and SEQ ID NO: 13. 7. A protein or polypeptide according to any of the preceding claims, wherein the amino acid sequence present is more than 90% identical to any of IV. 8. A protein or polyprotein according to any of the preceding claims, wherein the amino acid sequence present is more than 80%, preferably more than 90%, identical to one or more of the amino acid sequences II-V. peptide. 9. A protein or polypeptide according to any of the preceding claims, comprising one or more of the following amino acid sequences: I) AASAVNSELVHK II) ANFVPTK III) DTAIQSSYNK IV) ISALNK V) IAGTGTNNA. 10. A protein or polypeptide according to any of the preceding claims, comprising one or more preferably the amino acid sequences II-V. 11. A protein or polypeptide according to any of the preceding claims having a two lobe structure. 12. A protein or polyprotein according to any of the preceding claims having an amino acid sequence that is more than 20% identical to any of the following proteins: noc, occ, GlnH, HisJ, LAO. peptide. 13. The protein or polyprotein according to any one of the preceding claims, which has an amino acid sequence that is more than 40% identical to any one of the following proteins: noc, occ, GlnH, HisJ, and LAO. peptide. 14． The 29 kD L. of SEQ ID NO: 2. Ferme Ntumu (L. Fermentu m) 104R a higher proportion of 40% to the amino acid sequence of the adhesion factor, more preferably a higher proportion 6 0%, according to the foregoing and more preferably include the same amino acid sequence at a rate in excess of 80% A protein or polypeptide according to any of the ranges. 15. The 29 kD L. of SEQ ID NO: 2. Ferme Ntumu (L. Fermentu m) 104R protein or polypeptide according to any one of the claims above comprising the identical amino acid sequence to the amino acid sequence of the adhesion factor. 16. The 29 kD L. of SEQ ID NO: 2. Ferme Ntumu (L. Fermentu m) 104R with the amino acid sequence of adhesion factors, the scope of the claims above, which can be obtained by binding to either antibodies of the created polyclonal or monoclonal against the protein or polypeptide A protein or polypeptide according to any one of the above. 17． A polypeptide fragment which is a protein or polypeptide fragment according to any of the preceding claims, consisting of at least 5 and preferably 7 amino acids having mucosal binding activity, wherein at least 5 and preferably 7 Wherein said polypeptide fragment comprising 7 amino acids is present as a continuous sequence in any of the proteins or polypeptides of the preceding claims, wherein said fragment has a length of less than 60 amino acids. Peptide fragments. 18. 18. A polypeptide fragment according to claim 17 having a length of less than 40 amino acids. 19. Peptide I) AASAVNSELVHK II) ANFVPTK III) DTAIQSSYNK IV) ISALNK V) IAGTGTNNA VI) common sequence of SEQ ID NO: 6 and VII) common sequence of SEQ ID NO: 13, preferably an amino acid sequence selected from peptides II to VII. 19. Polypeptide fragment according to claims 17 or 18, comprising an amino acid sequence consisting of at least a series of 5 amino acids which is at least 80% identical, preferably at least 90% identical. 20. A peptide comprising: I) AASAVNSELVHK II) ANFVPTK III) DTAIQSSYNK IV) ISALNK V) IAGTGTNNA VI) consensus sequence of SEQ ID NO: 6; and VII) consensus sequence of SEQ ID NO: 13; A polypeptide fragment according to any one of claims 17 to 19. 21. A recombinant protein, polypeptide or polypeptide fragment comprising an amino acid sequence of any of the proteins, polypeptides or polypeptide fragments according to any of the preceding claims. 22. A fusion protein or polypeptide comprising a protein, polypeptide, or polypeptide fragment having mucosal binding activity according to any of the preceding claims, linked to a better drug, immunomodulator, or antigen. . 23. Binds to an epitope on a protein, polypeptide or polypeptide fragment as defined in any one of claims 1 to 21 and preferably in any one of claims 1 to 16 Antibodies or antibody fragments, preferably monoclonal antibodies. 24. A nucleic acid sequence encoding any of the proteins, polypeptides, or polypeptide fragments according to any one of claims 1 to 22. 25. An expression vector comprising the nucleic acid of claim 24 operably linked to an expression control sequence, wherein said nucleic acid is a non-pathogenic microorganism such as a GRAS microorganism. An expression vector that can be expressed and preferably contains a nucleic acid from a GRAS microorganism. 26. 26. The expression vector according to claim 25, wherein the expression vector regulatory sequence is not naturally linked to a gene encoding an adhesion factor from which the nucleic acid sequence has been obtained. 27. A recombinant microorganism comprising a nucleic acid sequence according to claim 24 and / or an expression vector according to claim 25 or 26, wherein said nucleic acid sequence and / or expression vector is absent or low copy. A recombinant microorganism that is present in numbers or is expressed to a low degree in non-recombinant microorganisms. 28. 28. The recombinant microorganism according to claim 27, which is a non-pathogenic microorganism, preferably a microorganism specific to human or animal microflora, more preferably a microorganism specific to human microflora. 29. As a pharmaceutically active composition in a pharmaceutically acceptable dosage form-as in any of claims 1 to 22, preferably in any of claims 1 to 16 29. The protein or polypeptide or polypeptide fragment according to claim 27, wherein said protein, polypeptide or polypeptide fragment has mucosal binding activity. 29. The expression vector according to claim 25 or 26. 29. The expression vector according to claim 27 or 28 A recombinant microorganism, or a part of said microorganism, said part expressing a mucosal binding activity-a non-pathogenic agent capable of expressing a protein or polypeptide or a peptide according to any of claims 1-22. Or a portion of the microorganism, wherein the portion expresses mucosal binding activity. One of the constituents, as well as compositions comprising a pharmaceutically One acceptable carrier. 30. As a pharmaceutically active component in a pharmaceutical composition for the prevention and / or treatment of diseases or illnesses associated with mucosal colonizing pathogenic microorganisms, having a mucosal binding activity. A protein or polypeptide or polypeptide fragment according to any of the above; an expression vector according to claim 25 or 26; a recombinant microorganism according to claim 27 or 28; A portion expressing mucosal binding activity)-a non-pathogenic microorganism capable of expressing a protein or polypeptide or peptide according to any of claims 1-22, or a portion of said microorganism (where said portion is Which exhibits mucosal binding activity), the use of one component selected from the group consisting of: 31. As a targeting component in a pharmaceutical composition for targeting one additional pharmacologically active component to the mucosa (the additional pharmacological component being physically The protein or polypeptide or polypeptide fragment according to any one of claims 1 to 22, which has a mucosal binding activity; the expression vector according to claim 25 or 26; A recombinant microorganism according to claims 27 or 28, or a part of said microorganism, said part expressing a mucosal binding activity-a protein or polypeptide or peptide according to any of claims 1-22. Selected from the group of components comprising a non-pathogenic microorganism capable of expressing, or a portion of said microorganism, said portion expressing mucosal binding activity. Use of one of the constituents that. 32. Claims 1. A product according to any of claims 1 to 22 or 25 to 28 and / or a protein or polypeptide or polypeptide fragment according to any of claims 1 to 22 to a food product. A method for improving a food product comprising the addition of a non-pathogenic microorganism capable of being expressed or a part of said microorganism, said part expressing mucosal binding activity. 33. A method for improving a food product comprising adding to the food product a product according to any of claims 1-22 or 25-28. 34. A product according to any of claims 1-22 or 25-28 and / or a protein or polypeptide or polypeptide fragment according to any of claims 1-22 can be expressed. A food product comprising a non-pathogenic microorganism or a part of said microorganism, said part expressing a mucosal binding activity. 35. A food product comprising, as an additive, the product according to any one of claims 1 to 22 or 25 to 28. 36. A method of screening a non-pathogenic microorganism for a microorganism capable of specifically binding to a mucosa, comprising detecting the presence of a particular protein or polypeptide on or in the microorganism, or in a culture of the microorganism. 23. A method comprising detecting by a method known per se, wherein said specific protein or polypeptide is a protein or polypeptide according to any of claims 1-22. 37. A method of screening a non-pathogenic microorganism for a microorganism capable of specifically binding to a mucosa, comprising detecting the presence of a particular gene in a microorganism in a manner known per se, A method wherein said special gene encodes a protein or polypeptide according to any of claims 1-22. 38. A kit suitable for the detection of a non-pathogenic microorganism capable of binding specifically to a mucosa, such as an antibody, specific for a protein or polypeptide according to any of claims 1 to 22. A kit comprising one component capable of binding to a. 39. A kit suitable for detecting a non-pathogenic microorganism capable of specifically binding to a mucosa, such as a nucleic acid probe or a primer, such as a protein or polypeptide according to any one of claims 1 to 22. A kit comprising one component capable of specifically binding to a portion of a nucleic acid sequence encoding

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C07K 14/205 C07K 14/285 14/285 14/335 14/335 14/35 14/35 16/12 16 / 12 19/00 19/00 C12N 1/21 C12N 1/21 C12P 21/02 C C12P 21/02 C12Q 1/68 A C12Q 1/68 G01N 33/53 D G01N 33/53 33/569 Z 33/569 A61K 35/66 // A61K 35/66 C12P 21/08 C12P 21/08 A61K 37/02 (81) Designated country EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE , IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (KE, LS, MW, SD, SZ, U ), UA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU , CZ, DE, DK, EE, ES, FI, GB, GE, HU, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, TJ, TM, TR, TT, UA, UG, US, uz , VN (72) Inventor Conway, Patricia Lynn Australia New South Wales 2036 Ruperous Gouraire Avenue 22

Claims (1)

[Claims] 1. A protein obtainable from a non-pathogenic microorganism, which has a mucosal binding promoting activity and a molecular weight of 20 to 40 kD, preferably 20 to 30 kD, or a polypeptide equivalent thereto. 2. Mucosal binding promoting activity, pathogenic microorganisms, preferably Kanpiro tapir ether (C ampylobacter), mycobacterium (Mycobacterium), or Pasuteurera (Pasteurella), for example C. C. jejuni , P.J. 2. The protein or polypeptide of claim 1, comprising promoting binding to a receptor recognized on mucosa by any of P. haemolytica . 3. Mucosal binding-promoting protein activity is determined by L. Ferme Ntumu (L. Fermen tum) 104R 29kD protein or polypeptide according to any of equal to or scope of the claims above over it to that of the adhesive agent. 4. A protein or polypeptide according to any of the preceding claims, comprising the amino acid consensus sequence specifically described in FIG. 4 and / or FIG. 5. The protein or polypeptide according to any of the preceding claims, comprising the amino acid consensus sequence specifically set forth in SEQ ID NO: 30. 6. The following amino acid sequences I) AASAVNSELVHK II) ANFVPTK III) DTAIQSSYNK IV) ISALFNK V) IAGTGTNNA more than 80%, preferably more than 90%, more preferably one or more of the amino acid sequences II-V A protein or polypeptide according to any of the preceding claims, comprising one or more identical amino acid sequences in a plurality of more than 80%, preferably more than 90%. 7. The following claim comprising one or more of the following amino acid sequences: I) AASAVNSELVHK II) ANFVPTK III) DTAIQSYNK IV) ISALNK V) IAGTGTNNA, more preferably one or more of the amino acid sequences II-V A protein or polypeptide according to any of the ranges. 8. A protein or polypeptide according to any of the preceding claims having a two lobe structure. 9. The protein according to any one of the preceding claims, which has an amino acid sequence that is more than 20% identical to the amino acid sequence of any of the following proteins: Atuoct, Atunop, GlnH, HisJ, LAO or Polypeptide. 10. The protein according to any one of the preceding claims, which comprises an amino acid sequence that is more than 40% identical to any one of the following proteins: Atuoct, Atunop, GlnH, HisJ, LAO, or Polypeptide. 11. The L.D. of 29 kD in FIG. Claims as described above comprising an amino acid sequence of more than 40%, more preferably more than 60%, more preferably more than 80% identical to the amino acid sequence of the L. fermentum 104R adhesion factor. A protein or polypeptide according to any one of the above. 12. The L.D. of 29 kD in FIG. The protein or polypeptide according to any of the preceding claims, comprising an amino acid sequence identical to the amino acid sequence of L. fermentum 104R adhesion factor. 13. The L.D. of 29 kD in FIG. Within the scope of the preceding claims obtained by binding the amino acid sequence of L. fermentum 104R adhesion factor to either a polyclonal or monoclonal antibody generated against the protein or polypeptide. The protein or polypeptide according to any one of the above. 14． A polypeptide fragment which is a fragment of a protein or polypeptide according to any of the preceding claims, which comprises at least 5 and preferably 7 amino acids which do not necessarily have to have mucosal binding promoting activity, A protein or polypeptide wherein at least 5 and preferably 7 amino acids of the peptide fragment are amino acid sequences present as a contiguous sequence in any of the proteins or polypeptides according to the preceding claims. 15. 15. The polypeptide fragment of claim 14, which is less than 60 amino acids in length. 16. 16. A polypeptide fragment according to claim 15 having a length of less than 40 amino acids. 17． The polypeptide sequence according to one of claims 14 to 16, which comprises, as a continuous sequence, an amino acid sequence of at least one sequence consisting of at least 5 amino acids present in the consensus sequence of SEQ ID NO: 30. 18. Peptides, I) AASAVNSELVHK II) ANFVPTK III) DTAIQSYNK IV) ISALNK V) IAGTGTNNA VI) common sequence of FIG. 4 and VII) common sequence of FIG. 5, preferably at least 80 amino acids selected from peptides II-VII A polypeptide fragment according to any of claims 14 to 18, which is selected from peptides comprising an amino acid sequence which is% identical, preferably at least 90% identical. 19. A peptide comprising: I) AASAVNSELVHK II) ANFVPTK III) DTAIQSYNK IV) ISALNK V) IAGTGTNNA VI) the consensus sequence of FIG. 4, and VII) the consensus sequence of FIG. The polypeptide fragment according to any one of the ranges 14 to 18. 20. A polypeptide fragment which is a combination of a plurality of polypeptide fragments according to any one of claims 14 to 18. 21. A recombinant protein, polypeptide or polypeptide fragment comprising an amino acid sequence of any of the proteins, polypeptides or polypeptide fragments according to any of the preceding claims. 22. A fusion protein comprising a protein, polypeptide, or polypeptide fragment having mucosal binding promoting activity according to any of the preceding claims, which is linked to a better drug, an immunomodulator, or an antigen, or Polypeptide. 23. An antibody capable of binding to an epitope on a protein, polypeptide or polypeptide fragment according to any of claims 1 to 21, and preferably to any of claims 1 to 13. Alternatively, an antibody fragment, preferably a monoclonal antibody. 24. A nucleic acid sequence encoded by any of the proteins, polypeptides or polypeptide fragments according to any one of claims 1 to 22. 25. An expression vector comprising the nucleic acid of claim 24 operably linked to an expression control sequence, for example, expressing the nucleic acid of claim 21 in a non-pathogenic microorganism such as a GRAS microorganism. An expression vector that can, and preferably contains, nucleic acid from a GRAS microorganism. 26. 26. The expression vector according to claim 25, wherein the expression control sequence is not naturally linked to a gene encoding an adhesion factor from which the nucleic acid sequence has been obtained. 27. A recombinant microorganism comprising a nucleic acid sequence according to claim 24 and / or an expression vector according to claim 25 or 26, wherein said nucleic acid sequence and / or expression vector is absent or low copy. A recombinant microorganism that is present in numbers or is expressed to a lesser extent in non-recombinant microorganisms. 28. 28. The recombinant microorganism according to claim 27, which is a non-pathogenic microorganism, preferably a microorganism specific to human or animal microflora, more preferably a microorganism specific to human microflora. 29. As a pharmaceutically active composition in a pharmaceutically acceptable dosage form-as defined in any of claims 1 to 21, preferably in any of claims 1 to 13 29. The protein or polypeptide or polypeptide fragment according to the above (the protein or the polypeptide or polypeptide fragment has a mucosal binding promoting activity);-the expression vector according to claim 25 or 26; Or a part of said microorganism (the part expresses a mucosa-binding promoting activity)-a non-protein capable of expressing the protein or polypeptide or peptide according to any one of claims 1 to 21. A group of components comprising a pathogenic microorganism, or a portion of said microorganism, wherein said portion expresses a mucosal binding promoting activity. Is selected - one component, as well as pharmaceutically acceptable - one composition comprising a carrier. 30. As a pharmaceutically active component in a pharmaceutical composition for preventing and / or treating a disease or illness related to mucosal colonizing pathogenic microorganisms, having a mucosal binding promoting activity. 22. A protein or polypeptide or polypeptide fragment according to any of 22; an expression vector according to claim 25 or 26; a recombinant microorganism according to claim 27 or 28, or a part of said microorganism ( The portion expresses a mucosal binding promoting activity)-a non-pathogenic microorganism capable of expressing the protein or polypeptide or peptide according to any of claims 1 to 21, or a part of the microorganism (the Wherein the portion expresses a mucosal binding promoting activity), the use of one component selected from the group of components comprising: 30. As a targeting component in a pharmaceutical composition for targeting one additional pharmaceutically active component to the mucosa (the additional pharmaceutically active component is The protein or polypeptide or polypeptide fragment according to any one of claims 1 to 22, which has a mucosal binding promoting activity; the expression vector according to claim 25 or 26; -A recombinant microorganism according to claims 27 or 28, or a part of said microorganism (the part expresses a mucosal binding promoting activity)-a protein or polypeptide according to any of claims 1-22 Or a non-pathogenic microorganism capable of expressing a peptide, or a portion of said microorganism, wherein said portion expresses a mucosal binding promoting activity. Use of one of the constituents to be more selective. 32. Claims 1. A product according to any of claims 1 to 22 or 25 to 28 and / or a protein or polypeptide or polypeptide fragment according to any of claims 1 to 22 to a food product. A method for improving a food product comprising the addition of a non-pathogenic microorganism that can be expressed or a part of said microorganism, said part expressing a mucosal binding promoting activity. 33. A method for improving a food product comprising adding to the food product a product according to any of claims 1-22 or 25-28. 34. The product according to any of claims 1 to 22 or 25 to 28 and / or the protein or polypeptide or polypeptide fragment according to any of claims 1 to 22 can be expressed. A food product comprising a non-pathogenic microorganism or a part of said microorganism, said part expressing a mucosal binding promoting activity. 35. A food product comprising, as an additive, the product according to any one of claims 1 to 22 or 25 to 28. 36. A method of screening a non-pathogenic microorganism for a microorganism capable of specifically binding to a mucosa, comprising detecting the presence of a particular protein or polypeptide on or in the microorganism, or in a culture of the microorganism. 23. A method comprising detecting by a method known per se, wherein said specific protein or polypeptide is a protein or polypeptide according to any of claims 1-22. 37. A method of screening a non-pathogenic microorganism for a microorganism capable of specifically binding to a mucosa, comprising detecting the presence of a particular gene in a microorganism in a manner known per se, A method wherein the special gene encodes a protein or polypeptide according to any of claims 1 to 22. 38. A kit suitable for the detection of a non-pathogenic microorganism capable of binding specifically to a mucosa, such as an antibody, specific for a protein or polypeptide according to any of claims 1 to 22. A kit comprising one component capable of binding to a. 39. A kit suitable for detecting a non-pathogenic microorganism capable of specifically binding to a mucosa, such as a nucleic acid probe or a primer, such as a protein or polypeptide according to any one of claims 1 to 22. A kit comprising one component capable of specifically binding to a portion of a nucleic acid sequence encoding