KR101101604B1 - A bacteriophage killing pathogenic enteric bacteria - Google Patents

Abstract

The present invention relates to a novel bacteriophage, specifically to a bacteriophage SS3e having a killing ability against pathogenic enterobacteria and having a putative helicase gene sequence encoding an amino acid sequence consisting of SEQ ID NO: 1. It is about.

According to the present invention, the bacteriophage can effectively kill or reduce pathogenic enterobacteriaceae as a causative agent of causing foodborne diseases. Bacteriophage, in particular, is a bacterium-based virus that is known to not work at all in humans and animals, so it can be applied to the pharmaceutical industry, food industry, and biotechnology, as well as to target harmful enterobacteriaceae without problems of antibiotic resistance. Alternatively, it can be effectively killed at the target substance.

Bacteriophage, Enterobacteriaceae, Salmonella, Pathogenic E. Coli, Heterologous bacteria, Enterobacter

Description

Bacteriophage killing pathogenic enteric bacteria

The present invention relates to a bacteriophage that kills pathogenic enterobacteria, and more particularly, to have a putative helicase gene sequence encoding an amino acid sequence consisting of SEQ ID NO: 1 having an ability to kill pathogenic enterobacteriaceae. A bacteriophage SS3e of Siphoviridae characterized by.

Bacteriophages are a type of virus that infects bacteria, often called abbreviations. Bacteriophages are simple organisms in which the protein envelope is wrapped around the core of a genetic material consisting of nucleic acids. Nucleic acids are composed of DNA or RNA, either single or double chains. Bacteriophages are known to require a host for survival, and all bacteria are known to have a particular phage. The bacteriophage penetrates the host, completes the replication process, and then expresses a group of enzymes necessary to break down the cell wall of the host bacterium. These enzymes attack the murine or peptidoglycan layers of the cell wall, responsible for the rigidity and mechanical strength of the cell wall, destroying the cell wall.

The bacteriophage was discovered in 1915 by a British bacteriologist Twort, in a study of the transparent melting of micrococcus colonies by something. Also, in 1917, French bacteriologist d'Herelle discovered that some filtrates of foreign patients had a function of dissolving Shigella disentriae, and through this study, they independently discovered bacteriophages and consumed them. In the sense, they named it bacteriophage. Since then, bacteriophages have been found for several pathogens, such as dysentery, typhoid, and cholera, and research has been focused on the bacteriophages of E. coli by Delbruck of the California Institute of Technology and European scientists who moved to the United States during World War II. However, since penicillin was discovered by Flemming in 1950, research on bacteriophages has continued only in some Eastern European countries due to the widespread use of antibiotics. Since 2000, however, the continued use of antibiotics has led to the emergence of multi-drug resistant pathogenic bacteria, which has led to the development of bacteriophages in developed countries.

On the other hand, typhoid fever, salmonella, bacterial dysentery, Escherichia coli, etc. are typical bacterial pathogens that cause foodborne diseases (food poisoning). Effective killing or reduction of bacterial pathogens that cause foodborne diseases is very important for disease prevention. In order to treat such infectious diseases or to kill causative bacteria, antimicrobial agents are administered to humans or livestock, and the causative bacteria may be reduced by food and environment by physicochemical therapy. However, the current epidemic of multidrug-resistant bacteria caused by the abuse of antimicrobial agents is a major hazard to public health, and physicochemical therapies have disadvantages of high environmental pollution and economic cost.

Accordingly, the present inventors have made intensive studies to overcome the problems of the prior art, and as a result, it was confirmed that the bacteriophage SS3e isolated from the stream shows specific killing ability against various enterobacteriaceae, and completed the present invention.

Therefore, the main object of the present invention is to provide a bacteriophage SS3e having a killing ability against a variety of pathogenic enterobacteria.

Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of pathogenic enterobacterial-induced diseases containing the bacteriophage SS3e as an active ingredient.

It is another object of the present invention to provide an antibiotic and a disinfectant comprising the bacteriophage SS3e.

According to an aspect of the present invention, the present invention is sipoviridae characterized by having a killing ability against pathogenic enterobacteria and having a putative helicase gene sequence encoding an amino acid sequence consisting of SEQ ID NO: 1 Provides Bacteriophage SS3e from Siphoviridae.

In the present invention, the bacteriophage SS3e was isolated and identified for the first time in the present invention, the sequence of the genome was also sequenced for the first time by the present inventors. Among them, a putative helicase gene encoding an amino acid sequence consisting of SEQ ID NO: 1 was identified as a gene having high homology with other species. Preferably putative helicase (Putative helicase) gene is characterized in that having a nucleotide sequence of SEQ ID NO: 2.

In the present invention, the genome of the bacteriophage SS3e was restriction enzyme mapped according to Example 3, and the results are shown in FIG. 3B. Preferably, the genome of the bacteriophage is characterized in that it has a restriction enzyme map (map) disclosed in Figure 3b.

In the present invention, the bacteriophage is characterized in that the accession number KCTC11492BP. The present inventors screened a novel bacteriophage having killing ability against various pathogenic enterobacteriaceae from river water in the metropolitan area, and separated the bacteriophage into the gene bank of the Korea Biotechnology Research Institute on March 27, 2009.

As used herein, the term “nucleic acid molecule” is meant to encompass DNA (gDNA and cDNA) and RNA molecules inclusively, and the nucleotides, which are the basic building blocks of nucleic acid molecules, are not only natural nucleotides, but also sugar or base sites. Modified analogues (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews, 90: 543-584 (1990)).

In the present invention, the pathogenic enterobacteriaceae is characterized in that the enterobacteriaceae selected from the group consisting of Salmonella, pathogenic E. coli, dysentery and Enterobacter.

As used herein, the term "enterally intestinal bacteria" refers to bacteria present in the intestine, and these intestinal bacteria include Escherichia coli, enterococci, lactic acid bacteria, staphylococci, and fungi, and others such as Salmonella and Red bacteria, modified bacteria, and the like.

Salmonella and E. coli and some Escherichia coli (pathogenic Escherichia coli) among the intestinal bacteria show pathogenicity, and these 'pathogenic enterobacteriaceae' have only enterococci in the upper part of the small intestine, and the number and types of bacteria increase as they go down. Most often in the subcapitalia. Pathogenic enterobacteriaceae are usually drinking water and foodborne and are spread through the mouth, causing disease in the intestine and sometimes spreading to systemic infections. Representatives include typhoid fever caused by Salmonella Typhi , paratyphoid fostered by Salmonella Paratyphi A, B and C, salmonellosis caused by Nontyphoidal Salmonella, bacterial dysentery caused by Shigella , and intestinal hemorrhagic colon symptoms caused by Escherichia coli . Enterobacteriaceae are gram-negative bacilli that do not form apo. It also has the ability to break down glucose and reduce nitrates to nitrites.

Among the pathogenic enterobacteriaceae, Salmonella, Escherichia coli, Heterozygos and Enterobacter are the most representative causative agents causing food-borne diseases, and bacteiophages of the present invention can effectively kill or reduce these pathogenic enterobacteriaceae.

The bacteriophage known so far is known to kill only certain bacteria, but the bacteriophage SS3e of the present invention kills various foodborne diseases causing bacteria, and is highly valuable for use as a food and environment-friendly alternative antibacterial agent as well as the health industry. The bacteriophage killing technique using the bacteriophage SS3e of the present inventors will be of great help in treating foodborne diseases and infections or improving health hygiene.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of pathogenic enterobacteriaceae-induced diseases comprising the bacteriophage as an active ingredient.

The term "treatment" as used herein refers to the prevention of diseases caused by pathogenic enterobacteriaceae; Suppression of diseases caused by pathogenic enterobacteriaceae; And alleviation of diseases caused by pathogenic enterobacteriaceae.

Diseases that may be caused by the pathogenic enterobacteriaceae include typhoid such as typhoid fever and paratyphoid caused by bacteria of the genus Salmonella; Food poisoning caused by pathogenic E. coli, dysentery, enterobacter and the like; And infections caused by Citrobacter, Serratia and the like, wherein the pharmaceutical compositions of the present invention can be effectively used for the treatment of such diseases.

Pharmaceutically acceptable carriers included in the compositions of the present invention are those commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate , Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. no.

In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The pharmaceutical composition of the present invention can be used as a method of spraying or spraying on a diseased site, and can also be administered by oral or parenteral administration. In the case of parenteral administration, intravenous administration, intraperitoneal administration, muscle Administration may be by intra-, subcutaneous or topical administration.

Suitable application, spraying, and dosage of the pharmaceutical compositions of the present invention may include factors such as formulation method, mode of administration, age, weight, sex, degree of disease symptom, food, time of administration, route of administration, rate of excretion, and reaction sensitivity of the patient. The physician, who is usually skilled in the art, can readily determine and prescribe a dosage effective for the desired treatment. In general, the pharmaceutical compositions of the present invention comprise 1 × 10 ³ to 1 × 10 ¹⁰ pfu / ml of bacteriophage.

The pharmaceutical compositions of the present invention are prepared in unit dose form by being formulated with pharmaceutically acceptable carriers and / or excipients according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

According to another aspect of the present invention, the present invention provides a cosmetic antibacterial agent and a pharmaceutical antibiotic comprising the bacteriophage as an active ingredient.

Cosmetics contain oil and water as the main ingredients, and glycerin or sorbitol, which is a carbon source of microorganisms, and amino acid derivatives and proteins, which are nitrogen sources, are easily blended, so it is easy to penetrate microorganisms such as bacteria. In addition, the use period is much longer than the food, it can be said that the risk of contamination by microorganisms is much greater. Addition of antimicrobial agent is essential for long-term protection of cosmetics from discoloration or odor caused by microbial contamination.

Bacteriophage of the present invention has the advantage of killing a large number of pathogenic enterobacterial compared to conventional antibiotics. This is very valuable because it can kill multiple pathogenic enterobacteria at once with a single prescription, rather than using different antibiotics for each pathogenic enterobacteriaceae. In addition, when using the bacteriophage of the present invention as an antibiotic has a significant advantage that does not induce resistance or resistance of pathogens unlike conventional antibiotics, the life cycle of the product compared to conventional antibiotics (long life cycle) It can be provided as a novel antibiotic. While most antibiotics face increasing resistance, the range of use is inevitably decreased. On the other hand, antibiotics containing the bacteriophage of the present invention as an active ingredient can fundamentally solve the problem of antibiotic resistance. It is expected.

Therefore, antibiotics including the bacteriophage of the present invention that specifically kill pathogenic enterobacteriaceae as an active ingredient may be usefully used as antibiotics having excellent antibacterial, bactericidal and antiseptic effects. As used herein, the term "antibiotic" refers to preservatives, fungicides and antimicrobials.

In addition, according to another aspect of the present invention, the present invention provides a disinfectant comprising the bacteriophage as an active ingredient.

Disinfectants containing the bacteriophage of the present invention having an effective killing ability against the pathogenic enterobacterial as an active ingredient can be usefully used as a disinfectant for hospitals and health care to prevent the infection of the hospital, and also a general household disinfectant, food and cooking place And it can be usefully used as a disinfectant of equipment, a livestock disinfectant of the livestock industry.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1 Isolation of Bacteriophage

Samples were collected from living streams in Seoul and Gyeonggi-do. First, the samples were filtered using a 0.45 μm filter, mixed with Salmonella Enteritidis culture grown in logarithmic order, and mixed with 10 ml of top LB agar containing 0.5% agar, and then poured into the LB agar plate to firmly. It was. After incubation in 37 ℃ incubator was observed for the presence of plaque (plaque). The phages were separated from the formed plaques, and then the same phages were repeated to amplify the phages, and the ten phages were separated and purified purely (see FIG. 1). Stock phage was dissolved in LB liquid medium and stored at 4 ° C.

Example 2 Soluble Host Range of Phages

10 kinds of phages obtained in Example 1, various bacteria such as Salmonella cultured in a nutrient medium (strains shown in Table 1) in a culture broth (Nutrient broth) after 1 hour stirring culture in nutrient agar plate (nutrient agar plate) After smearing each drop of phage solution and incubated in 37 ℃ incubator for 18 hours to determine the degree of lysis (lysis).

As a result of testing the lysing effect against a variety of enterobacteriaceae using 10 phages, 9 phages showed no lytic effect in bacteria other than Salmonella enteritidis, while one phage was shown in Table 1 below. As such, it was confirmed that the lysis effect in a variety of host range and was selected as a bacteriophage having a killing effect against a variety of intestinal bacteria (see Fig. 2 and Table 1). The present inventors deposited the selected phage into the Gene Bank (KCTC) of the Korea Biotechnology Research Institute (Accession Number: KCTC11492BP) and named it as 'SS3e' phage.

Table 1-1

TABLE 1-2

As a result of testing the host range of the phage, as shown in Table 1, it can be seen that Enterobacter cloacae , Citrobacter freundii , Seratia marcescenes are included in the host range of the phage as well as most Salmonella bacteria, many pathogenic E. coli, and many heterogeneous bacteria. there was. Therefore, the phage of the present invention was confirmed to have a specific ability to kill against a variety of pathogenic bacteria.

Example 3 Isolation of Phage Chromosome and Mapping Using Restriction Enzymes

Proteinase K and SDS (Sodium Dodecyl Sulphate) were added to 50 μg / ml and 0.5%, respectively, in the phage solution prepared in Example 1, incubated at 56 ° C. for 1 hour, and protein was removed using phenol. Centrifugation at 13,000 rpm for 10 minutes and the supernatant was carefully transferred to a new tube. 100% ethanol was added twice and left at -20 ° C. for 15 minutes, then centrifuged at 1,300 rpm for 15 minutes, the supernatant was discarded, and 100 μl of 70% ethanol was added and centrifuged at 1,300 rpm for 10 minutes. The supernatant was discarded and dried and stored in an appropriate amount of distilled water. The prepared phage DNA was treated with Sma I, Pst I, Sac I, Not I, Xho I, EcoR I, BamH I, Xba I, Hind III, and Sal I restriction enzymes to map the restriction enzyme and genome size. ) (See FIG. 3A).

As a result of restriction enzyme mapping, the genome of the SS3e phage was a double-stranded DNA with the restriction enzyme map of FIG. 3b, and the entire genome was about 41K bp in size.

Example 4 Electron Microscopy Observation of Phage

The enriched phage solution was centrifuged for 2 hours at 110,000 g of RCF using ultra centrifuge. Carefully discard the supernatant, add 1 ml of SM buffer used for phage storage, and incubate overnight to dissolve the precipitate. The purified phage was fixed with 5% formalin and then stained with 2% uranyl acetate (pH 4.0) to observe the shape of the phage using a Transmission Electron Microscope (TEM, 75 Kv). (FIG. 4).

As a result, it was found that the phage belong to Siphoviridae of Cadovirales. At first, Salmonella enteritidis was isolated as a hemolytic phage, but experiments have shown that it kills most Salmonella serotypes and Escherichia coli, dysentery and enterobacter.

As described above, according to the present invention, Salmonella, D. coli, Escherichia coli, and the like cause food-borne diseases, and effectively killing or reducing the causative pathogens is very important in terms of disease prevention. In order to treat such infectious diseases or to kill causative bacteria, antimicrobial agents may be administered to humans or livestock, and the causative bacteria may be reduced by physicochemical treatment to food materials or the environment. However, the current epidemic of multidrug-resistant bacteria caused by the abuse of antimicrobial agents is a major hazard to public health, and physicochemical therapies have disadvantages of high environmental pollution and economic cost. SS3e is a unique bacteriophage with a range of enterobacteriaceae hosts that kills most Salmonella bacteria, including both typhoid and paratyphoid bacteria, alleles, and Escherichia coli. The bacteriophage known so far is known to kill only certain bacteria, but SS3e kills bacteria causing various food-borne diseases, and is highly valued as a food and eco-friendly alternative antimicrobial agent. The bacteriophage killing method using the bacteriophage SS3e of the present invention will greatly help in treating foodborne diseases and infections or improving health hygiene.

1 is a plaque photograph of bacteriophage SS3e produced by Salmonella enteritidis.

Figure 2 shows the lysis results of bacteriophage SS3e against various enterobacteriaceae. 1: E. coli DH5α, 2: S. Paratyphi A , 3: S. Typhimurium , 4: S. Typhi , 5: S. Enteritidis , 6: Shigella sonnei .

Figure 3a is a restriction map of the SS3e genomic DNA. Lane 1: complete phage genome, lane 2: 1 kb DNA size marker (Invitrogen 1 kb plus ladder), lane 3: Sma I, lane 4: Pst I, lane 5: Sac I, lane 6: Not I, lane 7: Xho I, lane 8: EcoR I, lane 9: BamH I, lane 10: Xba I, lane 11: Hind III, lane 12: Sal I

3B shows a restriction map of SS3e genomic DNA.

4 is an electron microscope (TEM) image of SS3e phage. Purified phage particles were stained with 2% uranyl acetate. The size bar is 100 nm.

<110> Kyungpook National University Industry-Academic Cooperation Foundation <120> A bacteriophage killing pathogenic enteric bacteria <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 474 <212> PRT <213> Bacteriophage SS3e <400> 1 Met Ser Lys Phe Arg Arg Arg Glu Tyr Gln Lys Ile Met Thr Ser Phe   1 5 10 15 Met Leu Gln His Pro Arg Cys Asn Ile Trp Cys Gly Met Gly Gly Gly              20 25 30 Lys Thr Ser Ser Thr Met Trp Val Leu Ser Arg Leu Phe Arg Asn Gly          35 40 45 Gln Leu Asn Asp Asp Asp Arg Val Leu Ile Leu Ala Pro Leu Arg Val      50 55 60 Ala Ser Gly Thr Trp Pro Ala Glu Gln Glu Lys Trp Asn Phe Pro Cys  65 70 75 80 Leu Ser Val Val Asp Ala Thr Gly Ser Glu Lys Arg Arg Ile Ala Ala                  85 90 95 Leu Glu Ser Asp Ala Asn Val Val Cys Thr Asn Tyr Glu Val Ile Glu             100 105 110 Trp Leu Ile Asp Tyr Tyr Gly Lys Asp Asp Trp Pro Phe Thr Val Ile         115 120 125 Val Ala Asp Glu Ser Thr Lys Leu Lys Ser Phe Arg Ser Arg Ser Gly     130 135 140 Gly Ser Lys Arg Ala Lys Ala Leu Ser Lys Val Ala Phe Gly Lys Val 145 150 155 160 Lys Arg Phe Ile Asn Leu Thr Gly Thr Pro Ser Pro Asn Gly Leu Lys                 165 170 175 Asp Leu Trp Gly Gln Asn Trp Phe Ile Asp Ala Gly Glu Arg Leu Gly             180 185 190 Ser Ser Tyr Thr Ala Phe Thr Asp Arg Trp Phe Asn Ser Val Gln Lys         195 200 205 Gly Lys Ser Ala Met Ala Arg Glu Tyr His Ser Arg Pro Gly Ala Asp     210 215 220 Asn Glu Ile His Gln Lys Met Lys Asp Ile Ser Leu Thr Ile Asp Ala 225 230 235 240 Ala Glu Trp Phe Gly Cys Glu Ala Pro Val Ile Val Pro Val Glu Ile                 245 250 255 Asp Leu Pro Lys Lys Ala Arg Gln Ala Tyr Ile Asp Met Glu Glu Lys             260 265 270 Leu Phe Ala Glu Leu Glu Ser Gly Glu Val Glu Ala Ala Asn Ala Ala         275 280 285 Ala Lys Thr Ser Lys Cys Leu Gln Ile Ala Ser Gly Ala Val Tyr Val     290 295 300 Ser Gly Pro Asp Gly Glu Ala Thr Lys Glu Trp Glu Lys Val His Asp 305 310 315 320 Thr Lys Leu Asp Ala Leu Glu Ser Ile Val Glu Glu Leu Gln Gly Ala                 325 330 335 Pro Leu Leu Val Ala Tyr Gln Phe Lys His Glu Leu Glu Arg Ile Leu             340 345 350 Lys Arg Phe Pro Gln Ala Gln Ala Phe Ser Lys Gly Ala Lys Gly Asn         355 360 365 Lys Gln Met Glu Ser Trp Asn Arg Gly Glu Ile Glu Ile Leu Cys Val     370 375 380 His Pro Ala Ser Ala Gly His Gly Leu Asn Leu Gln Asp Gly Gly His 385 390 395 400 His Leu Ala Phe Ile Ser Gln Gly Trp Asn Leu Glu His Tyr Leu Gln                 405 410 415 Val Val Glu Arg Ile Gly Pro Val Arg Gln Lys Gln Ala Gly His Glu             420 425 430 Arg Pro Val Phe Leu Tyr His Ile Val Ala Lys Asp Thr Leu Asp Glu         435 440 445 Val Val Ala Ala Arg Thr Asp Glu Lys Lys Ser Val Gln Glu Glu Leu     450 455 460 Leu Asn Tyr Met Lys Arg Arg Gly Lys Arg 465 470 <210> 2 <211> 1425 <212> DNA <213> Bacteriophage SS3e <400> 2 atgagtaagt ttaggcgcag ggaataccag aaaataatga cgtcgtttat gctacagcac 60 ccacgttgca atatatggtg cggtatgggt ggcggcaaga cctcgtcgac aatgtgggtg 120 cttagccgcc tgttccgtaa tgggcaactt aatgacgacg accgagtgtt aattctggcc 180 cctttacgtg ttgcgtcagg tacgtggcca gcagaacaag agaaatggaa cttcccgtgt 240 ctgagtgtag tagatgcaac tggttctgag aagcgacgca tcgcggcgct ggagtcagac 300 gctaacgtgg tttgcacaaa ttacgaagtt atagaatggc ttattgacta ctacggcaaa 360 gacgactggc cttttaccgt tatcgttgcc gatgagagca cgaagctgaa atcgttccgt 420 agccgttctg gcgggagcaa gagggcaaag gcgcttagta aggtggcgtt cggtaaagtt 480 aagcgtttca ttaacctgac cggtacacca tcaccaaacg gcctcaaaga cttgtggggt 540 cagaactggt tcatcgacgc cggcgagcgc ctcgggtctt cgtacacagc attcacagac 600 cgatggttta actcggtaca gaaagggaag tcagctatgg cacgggaata ccactctcgc 660 cctggcgccg ataacgagat tcaccagaaa atgaaggata ttagcctgac cattgatgct 720 gccgagtggt tcggttgtga agcgcccgtt attgtgccgg ttgaaattga cctgccgaag 780 aaagcacgtc aggcgtacat cgatatggag gagaagttat tcgcagaact ggagagtggg 840 gaagttgaag cggctaacgc cgcggcgaag acgtcgaagt gtttgcagat tgcatcaggc 900 gccgtgtatg tatcggggcc agacggagaa gctaccaaag aatgggagaa agtgcacgat 960 acgaaactgg atgcgctcga gtccattgta gaggagttac agggtgcgcc gttgttggtg 1020 gcctatcagt tcaaacacga actggagcgc atccttaagc gattcccgca ggctcaggcc 1080 ttttccaaag gtgctaaagg taataagcag atggaatctt ggaaccgcgg ggaaatcgag 1140 attttgtgcg tgcaccctgc atcggcgggc catggtttga atttacagga cggcgggcat 1200 catctggcgt ttatttcgca aggctggaac ctggagcact atttgcaggt tgtcgagcgt 1260 ataggtcctg tacgccagaa acaggctggc cacgagcgtc cagtgttcct gtatcacata 1320 gtcgctaaag acacgctgga tgaggtcgtt gccgcgcgta cggacgagaa aaaatctgtc 1380 caggaagagt tgcttaatta tatgaagaga cgaggtaaga gatga 1425  

Claims (8)

delete delete delete delete delete Escherichia coli comprising bacteriophage SS3e of Siphoviridae deposited with the putative helicase gene sequence coding for amino acid sequence consisting of SEQ ID NO: 1 and deposited with accession number KCTC11492BP A pharmaceutical composition for preventing or treating a disease selected from the group consisting of food poisoning caused by, food poisoning caused by dysentery bacteria, food poisoning caused by enterobacter, infectious disease caused by citrobacter and infection caused by seratia. Escherichia coli, heterologous bacteria, including Bacteriophage SS3e of Siphoviridae deposited with the putative number KCTC11492BP with a putative helicase gene sequence encoding the amino acid sequence consisting of SEQ ID NO: 1 An antibiotic for killing pathogenic enterobacteriaceae selected from the group consisting of Enterobacter, Citrobacter and Serratia. Escherichia coli, heterologous bacteria, including Bacteriophage SS3e of Siphoviridae deposited with the putative number KCTC11492BP with a putative helicase gene sequence encoding the amino acid sequence consisting of SEQ ID NO: 1 A disinfectant for killing pathogenic enterobacteriaceae selected from the group consisting of Enterobacter, Citrobacter and Serratia.

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