KR101785487B1 - Endolysin lysbps13, and uses thereof - Google Patents

Abstract

The present invention relates to a novel endorhynchine having an inhibitory activity against a specific microorganism, wherein an endolysin LysCPAS15 consisting of the amino acid sequence of SEQ ID NO: 1 is provided.

Description

Endolysin LysCPAS15, and its use {ENDOLYSIN LYSBPS13, AND USES THEREOF}

The present invention relates to a novel endorhynic LysCPAS15 having an inhibitory activity against a specific microorganism.

Antibiotics are the primary means of treating infectious diseases caused by bacterial infections. However, since the 1980s, excessive use of antibiotics has caused more antibiotic resistant strains. In 1986, a number of Staphylococcus aureus strains resistant to Vancomycin, which is the last antibiotic, and many resistant strains were found. .

Vancomycin Resistant Enterococci (VRE), which is resistant to vancomycin, was first reported in France in 1986 and has been isolated since 1988 in the United States and is the causative agent of nosocomial infections that are difficult to treat, .

On the other hand, Endolysin has recently attracted attention as a promising antibiotic. Endolizine is an enzyme that breaks down the peptidoglucan in the bacterial cell wall and is encoded in the genetic material of the bacteriophage. The endolysin can remove specific microorganisms in a short time without inducing tolerance to microorganisms.

It has been reported that endolizine-treated beverages are not contaminated by Staphylococcus aureus or some gram-negative bacteria, but only specifically for Clostridium perprtensis and Clostridium difficile No enzyme has been identified.

Recently, as government regulations for the use of antibiotics have been strengthened around the world, interest in means of replacing antibiotics has been increasing, and the present inventors have found that clostridium perturbis and clostridium difficile And to discover new endorhysins with silico-specific inhibitory activity.

It is an object of the present invention to provide a novel enzyme having microbial-specific lytic activity.

It is another object of the present invention to provide an industrial application method of the enzyme.

According to one aspect of the invention, an endolysin LysCPAS15 consisting of the amino acid sequence of SEQ ID NO: 1 is provided.

In one embodiment, the endolacin LysCPAS15 may have an inhibitory activity against Clostridium perfringens or Clostridium difficile .

In one embodiment, the endolacin LysCPAS15 can be derived from bacteriophage CPAS-15 (ATCC PTA 8480).

According to another aspect of the present invention, there is provided a food composition comprising the endolizine.

According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the endolizine.

According to another aspect of the present invention, there is provided a feed composition comprising the enorlizine.

According to another aspect of the present invention, there is provided a detergent composition comprising the endolizine.

According to the present invention, the endolacin LysCPAS15 has a specific lytic activity against Clostridium perfringens and Clostridium difficile, and because of its excellent heat resistance and acid resistance, it can not only be used in medical applications but also in a variety of industries such as foods and feed additives It is available.

It should be understood that the effects of the present invention are not limited to the effects described above, but include all effects that can be deduced from the description of the invention or the composition of the invention set forth in the claims.

FIG. 1 illustrates the structure of an endolysin according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

When an element is referred to as "comprising ", it means that it can include other elements, not excluding other elements unless specifically stated otherwise.

Unless otherwise defined, can be performed by molecular biology, microbiology, protein purification, protein engineering, and DNA sequencing and routine techniques commonly used in the art of recombinant DNA within the skill of those skilled in the art. These techniques are known to those skilled in the art and are described in many standardized textbooks and references. Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

Various scientific dictionaries, including the terms contained herein, are well known and available in the art. Although any methods and materials similar or equivalent to those described herein are found to be used in the practice or testing of the present application, some methods and materials have been described. It is not intended that the invention be limited to the particular methodology, protocols, and reagents, as they may be used in various ways in accordance with the context in which those skilled in the art use them.

As used herein, the singular forms include plural objects unless the context clearly dictates otherwise. Also, unless otherwise indicated, nucleic acids are written from left to right, 5 'to 3', amino acid sequences from left to right, amino to carboxyl.

Hereinafter, the present invention will be described in more detail.

According to one aspect of the invention, an endolysin LysCPAS15 consisting of the amino acid sequence of SEQ ID NO: 1 is provided.

The endolysin is an enzyme that hydrolyzes the cell wall of a bacterium and can exhibit bacterial activity against bacteria. The endolysin may pass through the cell membrane through holin in the phage infectious process and may migrate to the peptidoglycan substrate of the cell wall to hydrolyze the peptidoglycan. Holes may be formed in the cell wall by the hydrolysis, and the cell membrane may be decomposed by the action of osmotic pressure, so that the lytic action may occur.

Therefore, the endolysin can effectively control the growth of pathogenic microorganisms. In particular, the endolysin LysCPAS15 may have an inhibitory activity against Clostridium perfringens or Clostridium difficile .

The Clostridium is a genus of spore forming Gram-positive bacteria that grows under anaerobic conditions containing more than 100 species, and there are four major species involved in disease in humans and other warm-blooded animals.

The Clostridium perfringens are commonly found in soil, dust, raw materials such as spices used in food processing, and in animal fields. The enterotoxins produced by Clostridium perfringens are usually hemolytic and necrotic, and there are four major toxins (α, β, ε, i). And can be classified into toxigenic types of six types (A to F type) depending on the presence or absence of the toxin. The Clostridium Perfectionis can induce perforin type A food poisoning, enterotoxemia, necrotizing enteritis, and gas gangrene by infection, and the A strain is a representative food poisoning cause and the C strain is the causative agent of necrotizing enteritis .

Infection with Clostridium perfringens, especially in poultry farming, can cause problems with the digestive tract health, with a negative economic impact on the broiler flock.

Recently, the poultry industry has been increasingly affected by infection with Clostridium perfringens and by farmers. The Clostridium perfringens can be epidemic in a large scale and can be latent without manifesting symptoms for a long time, so that the damage can be further extended.

The Clostridium difficile is a gram-positive, anaerobic, spore-forming microorganism. The clostridium difficile has been reported to be intestinal microorganisms present in the colon of about 2 to 5% of normal adults.

The clostridium difficile is a major cause of human hospital-acquired gastrointestinal disease and can cause symptoms ranging from mild to severe diarrhea and colitis. Treatment of a wide range of antibiotics such as ampicillin, amoxicillin, cephalosporin, and clindamycin may result in the collapse of normal intestinal flora and may permit colonization of clostridium difficile in the intestine (Kelly and Lamont , (1998), Ann Rev. Med., 49: 375-90).

Certain of the Clostridium difficile strains are capable of producing toxins which can lead to severe diarrhea, toxic megacolon, perforation, sepsis, and pseudomembranous colitis. The toxins produced by the pathogenic Clostridium difficile are known as intestinal toxins (toxin A), cytotoxins (toxin B), and binary toxins, which can cause diarrhea and inflammatory reactions. In particular, the toxin A induces secretory and hemorrhagic diarrhea, and the toxin B is a cytotoxic agent and may exhibit a destructive cytopathic effect in tissue-cultured cells.

Diffissible bacterial diarrhea (CDAD) by Clostridium difficile may cause medical costs leading to prolonged hospitalization as a cause of major prevalence and mortality. In addition, the clostridium difficile has been associated with an increased recurrence rate following treatment with metronidazole and new strains can produce higher levels of antibiotic resistant toxins.

On the other hand, the endolianin LysCPAS15 can be derived from bacteriophage CPAS-15 (ATCC PTA 8480).

The present inventors have sought to discover a novel enzyme capable of effectively inhibiting the clostridium spp. Strain. The present inventors have analyzed the gene sequence of the bacteriophage CPAS-15 obtained from the American Type Culture Collection (ATCC), and have used the BlastP (Altscfhul et al, 1997) and Interproscan program (E. Quevilillon, 2005) Found an open reading frame with a high similarity to endolase and named it LysCPAS15.

The endorhynic LysCPAS15 may comprise the amino acid sequence of SEQ ID NO: 1 and the endolysin may comprise a functional equivalent of a protein comprising the amino acid sequence of SEQ ID NO:

The above-mentioned " functional equivalent "means an amino acid sequence having at least 40% or more, preferably 80% or more, more preferably 90% or more, more preferably 95 Or more homologous to the amino acid sequence of SEQ ID NO: 1, and having substantially the same physiological activity as the amino acid sequence of SEQ ID NO: 1.

 The "substantially homogenous physiological activity" means a strain-specific inhibitory activity due to the structural and functional homology of the endolysin. The "sequence homology " is determined by comparing two optimally aligned sequences with the comparison region, and some nucleic acid sequences in the comparison region may be added or deleted.

The endolysin can be used for various applications such as food compositions, pharmaceutical compositions, feed compositions, additives, detergents and detergents due to the specific inhibitory activity against Clostridium perprtensis and Clostridium difficile.

According to another aspect of the present invention, there is provided a food composition comprising the endolizine.

The food composition may comprise any form of food composition, such as functional food, nutritional supplement, health food and food additives. Food compositions of this type may be prepared according to conventional methods known in the art. Since the food composition contains the endolysin, the propagation of the clostridium perprication and clostridium dipicyll can be effectively inhibited and the preservability of the food can be remarkably improved.

The above food composition may contain food additives and, unless otherwise specified, conform to the standards and standards of the relevant items in accordance with General Regulations and General Test Methods approved by the Food and Drug Administration. Can be determined.

The food additives described in the above-mentioned Food Additives Code include chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as persimmon extract, licorice extract, crystalline cellulose, high- A preservative formulation, a tar coloring agent, and the like.

In addition, the food composition may be manufactured and processed into any one form selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings.

According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the endolizine. Since the pharmaceutical composition contains the endolysin, it has excellent inhibitory activity against Clostridium perfringens and Clostridium difficile.

The pharmaceutical compositions may further comprise carriers, excipients and diluents used in the preparation of conventional pharmaceutical compositions.

Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, But are not limited to, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

In addition, the composition may be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like, external preparation, suppository and sterilized injection solution.

The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and these solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, Lactose, or gelatin. In addition to the above excipients, lubricants such as magnesium stearate and talc may also be used.

Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, syrups, and the like. In addition to water and liquid paraffin which are simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included.

The preparation for parenteral administration may be a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, or a suppository. Examples of the non-aqueous solvent and suspensions may include injectable esters such as propylene glycol, polyethylene glycol, vegetable oil such as olive oil and ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, and glycerogelatin.

A pharmaceutically effective amount of the pharmaceutical composition comprising the endolysin may be administered to a subject. The " pharmaceutically effective amount " means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, activity of the drug, Sensitivity, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts.

The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is desirable to administer an amount that allows for all of the above factors to be maximally effected in a minimal amount without side effects, which can be readily determined by one skilled in the art.

According to another aspect of the present invention, there is provided a feed composition comprising the enorlizine. Since the feed composition contains the endolysin, the propagation of the clostridium perprication and clostridium dipicyll can be effectively inhibited and the preservability can be remarkably improved.

The feed is a substance which supplies organic or inorganic nutrients necessary for the growth of livestock. The animal is an organism which consumes organic matter as a nutrient and has digestion, embryogenesis and differentiation of the respiratory organ. Specifically, it may be an echinoderm, crustacean, mollusk, fish, amphibian, reptile, algae, mammal, It is not.

The feed composition may further comprise cereal, vegetable protein feed, animal protein feed, sugar or dairy products. Specifically, the cereal may be crushed or crushed wheat, oats, barley, corn and rice, and the vegetable protein feed may be composed mainly of rapeseed, soybean and sunflower, and the animal protein feed includes blood, meat, And the sugar or dairy product may be a dry component consisting of various powdered milk and whey powders.

The feed composition may further comprise ingredients such as nutritional supplements, digestion and absorption enhancers, constipation accelerators, or disease prophylactic agents.

In addition, the feed composition can be made into a viscous granular or granular material depending on the degree of grinding of the components, and can be fed into a mesh or formed into a desired discrete shape for further processing and packaging. In addition, the composition may be subjected to a pelletizing, expanding or extruding process for storage, and may be dried and distributed for ease of storage.

On the other hand, the endolysin may be used for food and feed additives. The additive imparts preservability to foods or feeds, and the endorhynes are cheap and harmless to the human body, so they can be safely applied to food or feed.

The additive may further contain other additives depending on the purpose and use of the additive, and may be mixed with other active ingredients, such as commonly used additives, and additives such as pigments, surfactants and preservatives. The additive may be prepared by pulverizing, granulating, tableting or liquefying according to the purpose and use of the additive, and the additive may be packaged and distributed according to a conventional method.

According to another aspect of the present invention, there is provided a detergent comprising the endolizine.

For example, the detergent may be a detergent for clothes, a dishwasher, a food cleaner, or a household appliance cleaner. The endolysin suppresses the growth of a specific strain and is harmless to the human body. Therefore, it can be used for the purpose of washing various household articles or imparting antibacterial properties.

The detergent may further contain an adversely selected additive depending on the purpose and use of the detergent. The detergent may be mixed with other active ingredients, such as detergents, and additives such as pigments, surfactants and preservatives. The detergent may be prepared by pulverization, granulation, tableting or liquefying according to the purpose and use of the detergent, and the detergent may be packaged and dispensed according to a conventional method.

According to another aspect of the present invention, there is provided a cleansing agent comprising the endolizine.

The cleaner may be used for a hand, mouth, or a ladies' cleanser to prevent infection of a pathogenic microorganism that is easily infected in daily life. The endorhindo is harmless to the human body and thus can be safely used as a hand, mouth, and lady cleanser.

The cleansing agent may further include other additives optionally selected depending on the purpose and use of the cleansing agent. The cleansing agent may be mixed with other active ingredients, such as commonly used additives, and additives such as coloring matters, surfactants and preservatives . The cleansing agent may be prepared by pulverizing, granulating, tableting or liquefying according to the purpose and use of the cleansing agent.

The present invention will be further described with reference to the following examples, but it should be apparent that the present invention is not limited by the following examples.

Example 1: Isolation and purification of endolysin LysCPAS15

The gene sequence of the bacteriophage CPAS-15 (ATCC PTA 8480) obtained from the American Type Culture Collection (ATCC) was analyzed and then analyzed using BlastP (Altscfhul et al, 1997) and Interproscan program (E. Quevilillon, 2005) And an open reading frame of 1236 base pairs with a high similarity to the open reading frame, named LysCPAS15.

LysCPAS15 was analyzed to contain two amino acid sequences, and N-acetylmuramoyl-L-alanine amidase and CBD (cell-wall), known as enzymatically active domains The amino acid sequence of SH3_3, known as the.

Example 2: Biochemical characterization of endolysin LysCPAS15

To evaluate the heat resistance of LysCPAS15, the enzyme activity was measured after storage at a temperature between -20 and 70 캜. Each enzyme stored at that temperature was diluted in an enzyme storage buffer (Tris-HCl buffer) and its activity was measured.

The enzymatic activity of LysCPAS15 was not decreased at -20 to 40 ℃, but the enzymatic activity was slightly decreased at 50 ℃ and the enzymatic activity was not observed at 60 ℃.

In order to evaluate the acid resistance of LysCPAS15, the enzyme activity was measured after storage at pH 2 to pH 12. Each enzyme stored at the corresponding pH was diluted in an enzyme storage buffer (Tris-HCl buffer) and the activity was measured.

The enzyme activity of LysCPAS15 was not decreased at pH 4 to pH 8, but little activity was observed under conditions of pH 2 or pH 10 or strong acidity.

Example 3: Antimicrobial activity analysis of endolysin LysCPAS15

After purification of LysCPAS15, the antimicrobial activity against various microorganisms was measured. The results are shown in Table 1 below.

division Enzyme activity Gram-positive bacteria (Gram-positive bacteria) C. perfringens ATCC 3624 +++ C. perfringens KTCC 3269 +++ C. perfringens NCTC 8798 +++ C. perfringens NCTC 8239 +++ C. perfringens FD-1 +++ C. difficile ATCC 9689 +++ C. indolis ATCC 25771 - C. saccharoperbutylacetonicum - C. beijerinckii - Bacillus cereus ATCC 27348 - Bacillus cereus ATCC 14579 - Bacillus cereus ATCC 13061 - Bacillus cereus ATCC 21772 - Bacillus cereus ATCC 10876 - Bacillus cereus ATCC 21768 - Bacillus licheniformis JCM 2505 - Bacillus pumilus JCM 2508 - Bacillus circulans JCM 2504 - Bacillus megaterium JCM 2506 - Bacillus mycoides ATCC 6452 - Staphylococcus aureus ATCC 12600 - Staphylococcus aureus ATCC 29213 - Staphylococcus aureus RN 4220 - Staphylococcus epdermidis ATCC 35983 - Enterococcus faecalis ATCC 29212 - Enterococcus faecium ATCC 27276 - Listeria monocytogenes ATCC 15313 - Listeria innocua ATCC 33090 - Gram-negative bacteria Salmonella Typhimurium SL 1344 - Salmonella Typhimurium DT 104 - Salmonella Typhimurium LT2c - Salmonella Typhimurium ATCC 14028 - Salmonella Enteritidis ATCC 13076 - Escherichia coli O157: H7 ATCC 43890 - Escherichia coli O157: H7 ATCC 43894 - Escherichia coli O157: H7 ATCC 43895 - Escherichia coli O157: H7 ATCC 35150 -

As a result of the activity measurement, the lytic activity against the Gram-positive bacteria Clostridium perprtensis and Clostridium difficile was confirmed, but no activity against the other microorganisms was observed.

In other words, LysCPAS15 has no inhibitory activity against intestinal flora in human body due to its specific action activity on Clostridium perfringens and Clostridium difficile, so that side effects due to antibiotic treatment are not caused.

Example 4: Preparation of feed additive and feed

Feed additives containing purified LysCPAS15 were prepared. The purified LysCPAS15 solution was obtained by adding maltodextran and trihalose followed by lyophilization to give a powder. 5% by weight of LysCPAS15 was mixed with the nutrient feed to complete the feed.

Example 5: Confirmation of effect of specification in breeding of chicken

Whether the improvement in the results of the feeding of the feed of Example 4 was investigated. This survey was conducted in terms of mortality. A total of 60 two-day-old chicks were divided into two groups (a group fed the feed of Example 4 and a feed group fed general feed), each of which was divided into 30 groups of 30 birds. As a result of feeding for 4 weeks, the mortality rate of the group fed with the feed of Example 4 was 0%, but the mortality rate of the control group (general feed group) reached 27%.

That is, the above results suggest that animal feed containing endol-lysine LysCPAS15 is effective not only in reducing mortality but also in improving the animal specification results.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

<110> Kyung Hee University <120> ENDOLYSIN LYSBPS13, AND USES THEREOF <130> 16PP10933 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 412 <212> PRT <213> Artificial Sequence <220> <223> LysCPAS15 <400> 1 Met Lys Ile Ala Val Arg Gly Gly His Asn Phe Gln Ala Lys Gly Ala   1 5 10 15 Tyr Gly Leu Ile Asp Glu Thr Thr Glu Asp Arg Lys Val Lys Asp Ser              20 25 30 Val Ile Leu Ala Leu Arg Asn Glu Gly His Ser Val Leu Asp Val Thr          35 40 45 Pro Gly Asn Cys Asp Val Asn Thr Asp Leu Ala Tyr Gly Val Asn Lys      50 55 60 Ala Glu Ala Trp Gly Ala Asp Leu Phe Ile Ser Ile His Phe Asp Lys  65 70 75 80 Ala Tyr Asp Met Tyr Glu Gly Ala Leu Gly Thr Gly Thr Trp Ile Tyr                  85 90 95 Gly Ile Gly Gly Glu Ala Glu Val Tyr Ala Thr Arg Ile Val Asn Ala             100 105 110 Val Ala Lys Gly Thr Gly Leu Lys Asn Arg Gly Val Lys Cys Asn Ala         115 120 125 Lys Leu Tyr Glu Leu Arg Lys Thr Thr Met Pro Ala Val Ile Val Glu     130 135 140 Val Cys Phe Cys Glu Ala Thr Thr Asp Val Asn Ile Tyr Lys Ala Lys 145 150 155 160 Gly Pro Lys Leu Ile Gly Glu Leu Ile Ala Gly Gly Ile Cys Asn Lys                 165 170 175 Glu Ile Ser Gly Gly Ser Thr Pro Ser Ile Pro Ala Asp Asn Asn Pro             180 185 190 Lys Gly Phe Tyr Glu Ser Asn Glu Thr Arg Thr Asn Ala Thr Leu Val         195 200 205 Gly Glu Gly Ser Ile Glu Val Leu Asp Glu Asp Cys Lys Pro Val Pro     210 215 220 Gly Arg Phe Ile Asp Ser Leu Asp Asn Leu Phe Val Leu Gly Ile Tyr 225 230 235 240 Pro Ser Arg Asn Phe Ile Glu Val Val Tyr Pro Gly Lys Asn Lys Lys                 245 250 255 Tyr His Ala Tyr Ile Asp Ile Lys His Tyr Ser Arg Leu Ser Phe Asp             260 265 270 Tyr His Met Lys Tyr Gln Asn Asp Asn Gly Val Thr Tyr Val Trp Trp         275 280 285 Asn Pro Glu Asp Val Asn Ile Lys Asp His Asn Glu Glu Leu Gln Pro     290 295 300 Gly Gln Lys Ala Ser Pro Met Tyr Arg Thr Asn Gly Trp Leu Arg Ile 305 310 315 320 Thr Phe Tyr Arg Lys Asp Gly Thr Pro Ser Asp Gly Tyr Val Arg Tyr                 325 330 335 Glu Gly Glu Gln Ser Gln Lys Phe Tyr Glu Asp Val Lys Gln Gly Ile             340 345 350 Val Lys Val Asn Thr Ser Leu Asn Val Arg Asp Asp Val Asn Gly Asn         355 360 365 Ile Ile Gly Ser Val Phe Ser Asn Glu Arg Val Thr Ile Leu Gly Ser     370 375 380 Lys Asn Gly Trp Tyr His Ile Glu Tyr Asn Thr Ser His Gly Lys Lys 385 390 395 400 Gln Gly Tyr Val Ser Ser Lys Tyr Val Glu Ile Val                 405 410

Claims (7)

An endolysin LysCPAS15 consisting of the amino acid sequence of SEQ ID NO: 1. The method according to claim 1,
Endolysin LysCPAS15, which has an inhibitory activity against Clostridium perfringens or Clostridium difficile . The method according to claim 1,
Endolysin LysCPAS15 derived from bacteriophage CPAS-15 (ATCC PTA 8480). A food composition comprising an endorhindamine according to any one of claims 1 to 3. A pharmaceutical composition for the prevention and treatment of Clostridium perfringens or clostridium difficile infectious diseases comprising endorhizine according to any one of claims 1 to 3. A feed composition comprising the enorhloisin of any one of claims 1 to 3. A detergent composition comprising an enolyisin of any one of claims 1 to 3.

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