Bacillus amyloliquefaciens strain MU2 and use thereof

Abstract

The present invention relates to a Bacillus amyloliquefaciens strain MU2 and use thereof. The Bacillus amyloliquefaciens strain MU2 of the present invention can be effectively utilized in various bioindustries such as in the production of fermentation starters and heat resistant enzymes, and preparation of microorganism for extracting nutrients from raw food sources.

Description

Bacillus amyloliquefaciens MU2 strain and its use

[0001] The present invention relates to Bacillus amyloliquefaciens MU2 strain and its use. * The present invention is an invention that received support from the Busan Technopark's Innovative Growth Field-oriented Technology Development Project for Processed Fishery Foods, Development of Future High Value-Added Fishery Foods Based on Blue Food Tech (Project No. 202303370001, Research Period: 2023.04. 01 - 2024.03. 31) and the National Research Foundation of Korea's Key Research Institute Support Project, Marine Bionics Convergence Technology Center (Project No. 2021R1A6A1A03039211, Research Period: 2023.03. 01 - 2024.02. 29).

[0002] Traditional fermented foods are highly valued for their nutritional and health benefits, and can be considered a genetic resource pool containing a diverse array of microorganisms, including lactic acid bacteria and Bacillus species. Microorganisms derived from these traditional fermented foods have been reported to possess beneficial health functionalities, including anticancer, cholesterol-lowering, blood pressure-lowering, thrombolytic, and diarrhea-preventing effects. Therefore, the discovery and utilization of starter strains capable of achieving stable quality and enhanced functionality from traditional fermented foods with superior health functionalities is essential.

[0003] Fermentation technology using Bacillus amyloliquefaciens is being utilized in diverse fields, including agriculture, food, and life sciences. Bacillus amyloliquefaciens is known to improve the properties of raw materials through metabolic activity, extracting and producing desired substances, and thereby enhancing product quality. Furthermore, fermentation technology is recognized as an environmentally friendly production method with a minimal impact on the environment, unlike other food processing techniques.

[0004] Accordingly, the inventors of the present invention have completed the present invention by confirming that a novel Bacillus amyloliquefaciens MU2 strain (KCTC15499BP) isolated from salted seafood, a safe traditional food, has excellent alkali tolerance, heat resistance, extracellular enzyme secretion ability, antibiotic susceptibility, and antibacterial activity against harmful microorganisms, and does not produce biogenic amines or have hemolysis.

[0005] The purpose of the present invention is to provide a novel Bacillus amyloliquefaciens MU2 strain.

[0006] Another object of the present invention is to provide a fermentation starter, a composition for inhibiting the growth of pathogenic microorganisms, a probiotic preparation, a health food, and a feed additive, which contain the strain and its culture solution as effective ingredients.

[0007] To achieve the above purpose, the present invention provides a Bacillus amyloliquefaciens MU2 strain (KCTC15499BP) having excellent protein and cell wall decomposing enzyme activity.

[0008] In addition, the present invention provides a fermentation starter, a composition for inhibiting the growth of pathogenic microorganisms, a probiotic preparation, a health food, and a feed additive, which contain the strain and its culture solution as effective ingredients.

[0009] The present invention relates to a Bacillus amyloliquefaciens MU2 strain and its use. The Bacillus amyloliquefaciens MU2 strain of the present invention can be usefully utilized in various bio-related industries, such as a fermentation starter, a microbial preparation for producing heat-resistant enzymes, and for extracting nutrients from food raw materials.

[0010] The present invention is described in detail below.

[0011] The present invention provides a Bacillus amyloliquefaciens MU2 strain (KCTC15499BP) having excellent protein and cell wall decomposing enzyme activity.

[0012] The Bacillus amyloliquefaciens MU2 strain of the present invention is a novel strain isolated from a traditional scorpion, and has excellent alkali tolerance, heat tolerance, extracellular enzyme secretion ability, antibiotic susceptibility, and antibacterial activity against harmful microorganisms, and has been confirmed to have mycological characteristics of not producing hemolysin and biogenic amines. It was deposited with the Korean Collection for Type Cultures (KCTC) of the Korea Research Institute of Bioscience and Biotechnology on July 11, 2023, and was assigned the accession number KCTC15499BP.

[0013] In a strain according to one embodiment of the present invention, the strain is characterized in that it does not produce a biogenic amine.

[0014] The above biogenic amines are formed from amino acids by the action of amino acid decarboxylase of microorganisms, and when biogenic amines exceeding the human body's decomposition limit are consumed in food, symptoms such as rash, local skin inflammation, allergy, vomiting, nausea, and diarrhea are induced.

[0015] The above biogenic amine may be, but is not limited to, histamine, tyramine, putrescine or cadaverine.

[0016] The Bacillus amyloliquefaciens MU2 strain of the present invention has a 16S rRNA base sequence of sequence number 1.

[0017] In addition, the present invention provides a fermentation starter comprising the strain and its culture solution as an effective ingredient.

[0018] In the present invention, a fermentation starter refers to a preparation or composition containing microorganisms involved in fermentation. When a fermentation starter is added during the production of a fermented food, the quality of the fermented food can be consistently controlled by the Bacillus amyloliquefaciens MU2 strain or its culture solution included in the fermentation starter, and the speed of fermentation can be controlled by the amount of starter added or the initial number of bacteria.

[0019] The above fermentation starter may further contain additives commonly used.

[0020] The method for culturing the strain of the present invention can be performed according to a method commonly used in the art, and is not limited to a specific method.

[0021] The culture solution used in the present invention may include not only the culture itself in which the strain of the present invention is cultured, but also a culture filtrate obtained by physically filtering the supernatant containing the organic acid produced by the strain of the present invention during culture, after removing the strain (cells) of the present invention to some extent by centrifugation or the like.

[0022] The term "comprising as an active ingredient" as used herein means that the strain of the present invention or a culture medium thereof is included in an amount sufficient to achieve its efficacy or activity. The active ingredients used in the present invention are strains isolated from salted seafood or a culture medium thereof, and since there are no adverse effects on the subject even when administered in excess, the quantitative upper limit included in the composition of the present invention can be selected and implemented within an appropriate range by those skilled in the art.

[0023] In addition, the present invention provides a composition for inhibiting the growth of pathogenic microorganisms, which comprises the strain and its culture solution as an effective ingredient.

[0024] In the present invention, the pathogenic microorganism may be, but is not limited to, Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and brio parahaemolyticus.

[0025] In the present invention, the pathogenic microorganism may be an antibiotic-resistant pathogenic microorganism.

[0026] In the present invention, the antibiotic may be, but is not limited to, gentamicin, kanamycin, streptomycin, tetracycline, erythromycin, chloramphenicol, and vancomycin.

[0027] The composition of the present invention can be administered in various oral and parenteral dosage forms during actual clinical administration. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing Bacillus amyloliquefaciens MU2 strain and its culture medium with at least one excipient, such as starch, calcium carbonate, and gelatin. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.

[0028] The dosage range varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The daily dosage is about 0.001 to 100 mg / kg of Bacillus amyloliquefaciens MU2 strain or its culture solution, preferably 5 to 50 mg / kg, and it is more preferably administered once or in several divided doses per day.

[0029] Since the composition of the present invention comprises the Bacillus amyloliquefaciens MU2 strain of the present invention or a culture solution thereof as described above, description of duplicate contents is omitted to avoid excessive complexity of the present specification.

[0030] In addition, the present invention provides a probiotic preparation comprising the Bacillus amyloliquefaciens MU2 strain and its culture solution as an active ingredient.

[0031] The above probiotic preparation can be prepared and administered in various formulations and methods according to methods known in the art. For example, the Bacillus amyloliquefaciens MU2 strain of the present invention or a culture solution thereof can be prepared and administered in the form of powders, liquids and solutions, tablets, capsules, syrups, suspensions, or granules by mixing with a carrier commonly used in the pharmaceutical field. The carrier may be, for example, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a colorant, a flavoring agent, etc., but is not limited thereto. In addition, the administration dosage can be appropriately selected depending on the absorption rate of the active ingredient in the body, the inactivation rate, the excretion rate, the age, sex, livestock species, condition of the recipient, the severity of the disease, etc.

[0032] Since the probiotic preparation of the present invention comprises the Bacillus amyloliquefaciens MU2 strain of the present invention or a culture solution thereof as described above, description of duplicated contents is omitted to avoid excessive complexity of the present specification.

[0033] In addition, the present invention provides a health food for inhibiting the growth of pathogenic microorganisms, which comprises the Bacillus amyloliquefaciens MU2 strain and its culture solution as an effective ingredient.

[0034] Health foods containing the Bacillus amyloliquefaciens MU2 strain of the present invention or its culture solution include folk remedies for inhibiting the growth of pathogenic microorganisms, such as tea, jelly, juice, extract, and beverage, containing the strain or its culture solution as an active ingredient. The health foods for inhibiting the growth of antibiotic-resistant pathogenic microorganisms or harmful intestinal microorganisms of the present invention, processed in various forms as described above, are excellent in inhibiting the growth of pathogenic microorganisms without causing side effects on the human body, and are also easy to take and can be stored for a long period of time.

[0035] The Bacillus amyloliquefaciens MU2 strain of the present invention or its culture solution can be added to health foods for the purpose of inhibiting the growth of pathogenic microorganisms. When the Bacillus amyloliquefaciens MU2 strain of the present invention or its culture solution is used as a food additive, each of the above can be used as is or in combination with other foods or food ingredients, and can be used appropriately according to a conventional method. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverage, the Bacillus amyloliquefaciens MU2 strain of the present invention or its culture solution is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, per 100 parts by weight of the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can also be used in an amount above the above range.

[0036] Since the health food of the present invention includes the Bacillus amyloliquefaciens MU2 strain of the present invention or a culture solution thereof as described above, description of duplicated contents is omitted to avoid excessive complexity of the present specification.

[0037] In addition, the present invention provides a feed additive for inhibiting the growth of pathogenic microorganisms, which comprises the Bacillus amyloliquefaciens MU2 strain and its culture solution as an effective ingredient.

[0038] The Bacillus amyloliquefaciens MU2 strain of the present invention and its culture solution can be used for animals including livestock and can be added to animal feed.

[0039] The feed additive of the present invention may be prepared by using the strain of the present invention or its culture solution as is, or additionally adding known carriers, stabilizers, etc., such as grains and their by-products, which are allowed in feed additives, and, if necessary, organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polyphosphate), natural antioxidants such as polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid, antibiotics, antimicrobial agents, and other additives, and the feed additive may be prepared in an appropriate form such as powder, granules, pellets, and suspension. When the feed additive is supplied, it may be supplied alone or mixed with feed.

[0040] Since the feed additive of the present invention includes the Bacillus amyloliquefaciens MU2 strain of the present invention or a culture solution thereof described above, description of duplicated contents is omitted to avoid excessive complexity of the present specification.

[0041] Hereinafter, the present invention will be described in detail by examples.

[0042] However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0043] <Example 1> Isolation and cultivation of strains

[0044] Samples of three types of traditionally manufactured salted seafood (flounder sikhae, anchovy jeotgal, and squid jeotgal) were collected from a traditional market in Jeju Island, and strains were isolated using these samples.

[0045] Specifically, 25 g of each sample was taken, mixed with 225 mL of sterile phosphate-buffered saline (PBS), and homogenized using a bag mixer. Next, the homogenized sample was diluted stepwise using 9 mL of PBS, inoculated onto all culture agar (AC agar) using the spread method, and cultured at 37°C for 24 hours. After that, the morphological characteristics of the fungi were confirmed on the AC agar plate after the culture was completed, and the strains were isolated and cultured. After that, the morphological characteristics of the fungi were confirmed on the AC agar plate after the culture was completed, and the strains were isolated and cultured.

[0046] <Example 2> Confirmation of protein and cell wall degrading enzyme activity and selection of strains

[0047] An experiment was conducted to confirm the protein and cell wall decomposing enzyme activity of the strain identified in Example 1 above.

[0048] Specifically, it was confirmed using skim milk agar and carboxymethyl-cellulose agar (CMC agar). The pure isolated strain was inoculated into AC broth and cultured at 37℃ for 24 hours, and then inoculated onto each plate using the spot inoculation method using a sterilized cotton swab, and the size of the clear zone was confirmed after culture for 24 hours at 37℃. The proteolytic enzyme activity and cell wall degrading enzyme activity were expressed as the ratio of the clear zone on the plate after culture to the colony size of each strain. As a result, as shown in Table 1, the F1-2-1 strain showed the highest proteolytic enzyme activity of 3.0 and cell wall degrading enzyme activity of 4.0, and thus the F1-2-1 strain was selected and used in the subsequent experiments.

[0049]

[0050] The selected strains were extracted and purified using a DNA extraction kit, amplified using a 16s rRNA PCR kit, sequenced, and identified using NCBI's Blast program.

[0051] The DNA base sequence encoding 16S rRNA of the above strain (16S rDNA; sequence number 1) was analyzed for phylogenetic relationships by examining the homology with the 16S rRNA base sequences of strains registered in the GeneBank database using the Basic local alignment search tool (Blast, NCBI). As a result, the selected isolated strain of the present invention was confirmed to have 99% homology with Bacillus amyloliquefaeciens, confirming that it is a strain belonging to Bacillus amyloliquefaeciens.

[0052] Accordingly, the inventors of the present invention named the isolated Bacillus amyloliquefaeciens MU2 and deposited it with the Korea Research Institute of Bioscience and Biotechnology (KCTC) Biological Resource Center on July 11, 2023, and were assigned the accession number KCTC15499BP.

[0053] Therefore, the strain of the present invention, Bacillus amyloliquefaciens MU2, is a novel strain that has been found to have differences in base sequence when compared to the published strain, and since the strain exhibits excellent mycological characteristics such as protein and cell wall decomposing enzyme activity, it can achieve effects not only as the strain itself but also as its culture metabolites.

[0054] <Example 3> Confirmation of heat resistance of Bacillus amyloliquefaciens MU2 strain

[0055] An experiment was conducted to confirm the heat resistance of the selected strain Bacillus amyloliquefaciens MU2.

[0056] Specifically, the strains were inoculated into sterilized AC broth and cultured at temperatures of 30, 37, 40, 50, and 60°C for 24 hours, and the cultured strains were diluted stepwise using 9 mL of PBS, and then distributed onto all culture agar (AC agar) using the spread method and cultured again at 37°C for 24 hours to measure the number of viable cells.

[0057] As a result, the strain of the present invention showed the highest growth rate of 7.74±0.06 Log CFU / mL at a culture temperature of 30℃, as shown in Table 2, and also showed a high growth rate of 6.98±0.03 Log CFU / mL at 50℃.

[0058]

[0059] General fermentation takes place in a medium temperature range of 20-40℃, which is in line with the growth temperature of most harmful microorganisms such as food poisoning bacteria, and may lower the quality or safety of the product. On the other hand, it was confirmed that the strain of the present invention is capable of fermentation even at 50℃ or higher.

[0060] <Example 4> Confirmation of biogenic amine production by Bacillus amyloliquefaciens MU2 strain

[0061] Experiments were conducted to determine whether the selected strain Bacillus amyloliquefaciens MU2 produces biogenic amines harmful to the human body.

[0062] Specifically, decarboxylase broth was prepared by adding 1% arginine, 2% lysine, and 1% orinithine, respectively. Next, the prepared medium was inoculated with the strain and Escherichia coli KCTC 2571 (E. coli) as a positive control, and sterilized mineral oil was applied and sealed on the medium. It was checked every 24 hours at 37℃. If the color of the medium changed from purple to yellow and then back to purple, it was considered positive, and if the color of the positive control remained yellow until it turned purple again, it was considered negative.

[0063] As a result, it was confirmed that the strain of the present invention does not produce biogenic amines for cadaverine and putricin, as shown in Table 3.

[0064]

[0065] Biogenic amines are a representative problematic substance in fermented foods. Cadaverine and putrescine are known to react with nitrite added to foods to produce carcinogenic nitrosamines. The strain of the present invention was confirmed not to produce these biogenic amines, indicating a positive safety impact.

[0066] <Example 5> Confirmation of alkalinity tolerance of Bacillus amyloliquefaciens MU2 strain

[0067] An experiment was conducted to confirm the salt tolerance of the selected strain Bacillus amyloliquefaciens MU2.

[0068] Specifically, the pH of AC broth was adjusted to 7, 8, 9, and 10, respectively, and sterilized. The strains were inoculated into each medium and cultured at 37°C for 24 hours. The cultured strains were diluted stepwise using 9 mL of PBS, and then inoculated into all culture agar (AC agar) using the spread method. The strains were then cultured again at 37°C for 24 hours, and the number of viable cells was measured.

[0069] As a result, the strain of the present invention had the highest viable cell count at pH 7 of 7.79±0.01 Log CFU / mL, as shown in Table 4, and the lowest viable cell count at pH 10 of 3.30±0.19 Log CFU / mL.

[0070]

[0071] In general, most bacteria exhibit optimal growth at a pH of 6.5 to 7.5. This is a pH commonly found in most environments and also promotes the growth of pathogens harmful to the human body. The strain of the present invention exhibits alkalinity, allowing it to grow well even at pH 9, confirming its ability to perform fermentation even in environments where pathogens are not well-suited for growth.

[0072] <Example 6> Confirmation of antibiotic susceptibility of Bacillus amyloliquefaciens MU2 strain

[0073] To confirm the antibiotic susceptibility of the selected strain Bacillus amyloliquefaciens MU2, the MIC (Minimum Inhibitory Concentration) was analyzed.

[0074] Specifically, MIC was performed using the dilution method of CLSI M07-A9, and antibiotic susceptibility was confirmed using the cut-off value designated by the EU. Seven antibiotics (Gentamicin, Kanamycin, Streptomycin, Tetracycline, Erythromycin, Chloramphenicol, Vancomycin) used to confirm antibiotic susceptibility of Bacillus spp. were selected and diluted to match the cut-off value.

[0075]

[0076] As a result, the strain of the present invention was found to have no antibiotic resistance as it did not exceed the cut-off value for 7 types of antibiotics as shown in Table 6.

[0077]

[0078] Antibiotic-resistant strains pose a risk of spreading antibiotic resistance through horizontal gene transfer. The strain of the present invention was confirmed to not exhibit antibiotic resistance, confirming its safety from the aforementioned threats during fermentation.

[0079] <Example 7> Confirmation of hemolytic activity of Bacillus amyloliquefaciens MU2 strain

[0080] An experiment was conducted to determine whether the selected strain Bacillus amyloliquefaciens MU2 has hemolytic activity.

[0081] Specifically, sheep blood agar was used, and the pure isolated strain and Bacillus cereus ATCC 10876 were inoculated into AC broth and cultured at 37℃ for 24 hours. Then, a sterilized cotton swab was used to inoculate sheep blood agar using the spot inoculation method, and after culture at 37℃ for 24 hours, the size of the clear zone was confirmed.

[0082] As a result, the strain of the present invention was confirmed to be γ-hemolytic since no clear zone was confirmed as shown in Table 7.

[0083]

[0084] Hemolysis is a phenomenon in which red blood cells in the blood are destroyed for some reason, and is broadly divided into α-hemolysis, β-hemolysis, and γ-hemolysis. α-hemolysis is incomplete hemolysis, which partially dissolves hemoglobin, turning the inoculated medium greenish-gray. β-hemolysis is complete hemolysis, which completely dissolves hemoglobin, and a transparent clear zone can be confirmed in the inoculated medium. γ-hemolysis is non-hemolytic, which means it does not cause hemolysis, so it can be confirmed that there is no change in the medium. Hemolysis destroys red blood cells in the human body, which can have a negative effect on organs and the circulatory system, causing diseases such as anemia, jaundice, and organomegaly, and therefore the hemolysis of a strain serves as an important factor in confirming safety. The strain of the present invention was confirmed to not exhibit hemolysis by γ-hemolysis, confirming its safety from the above threats.

[0085] <Example 8> Confirmation of antibacterial activity of Bacillus amyloliquefaciens MU2 strain against pathogenic microorganisms

[0086] An experiment was conducted to confirm whether the selected strain Bacillus amyloliquefaciens MU2 has antibacterial activity against pathogenic microorganisms.

[0087] Specifically, four pathogenic microorganisms (Eschericha coli KCTC 2571, Bacillus cereus ATCC 10876, Staphylococcus aureus ATCC 6538, Listeria Monocytogenes KCTC 3569) were selected and the disk diffusion method was performed. Each pathogenic microorganism and the selected strain were inoculated into tryptic soy broth and cultured at 37°C for 24 h. Then, the pathogenic microorganisms were plated on sterilized mueller hinton agar using 9 mL of PBS and 10 5 The culture medium was diluted to log CFU / mL and inoculated with bacteria using the spread method. After that, a sterilized paper disk was placed on the plate inoculated with pathogenic microorganisms, and the selected strains were inoculated with 10 7 Diluted with 9 mL of PBS to log CFU / mL, 20 μL was dispensed onto a paper disk and cultured at 37°C for 24 h. After incubation, the size of the clear zone was measured based on the center of the paper disk to confirm antibacterial activity.

[0088] As a result, it was confirmed that the strain of the present invention exhibited antibacterial activity of 17 mm against E. coli KCTC 2571, 15 mm against S. aureus ATCC 6538, and 23 mm against L. monocytogenes KCTC 3569, as shown in Table 8, and no antibacterial activity against B. cereus ATCC 10876 was observed.

[0089]

[0090] In summary, the Bacillus amyloliquefaciens MU2 strain of the present invention has excellent protein and cell wall decomposing enzyme activity, alkalinity, heat resistance, antibiotic sensitivity, and antibacterial activity against harmful microorganisms, and does not produce biogenic amines or cause hemolysis, so it is expected to be utilized in various bio-related industries such as fermentation starters, production of heat-resistant enzymes, and microbial preparations for nutrient extraction from food raw materials.

[0091] Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0092] [Accession number]

[0093] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0094] Accession number: KCTC15499BP

[0095] Date of acceptance: 20230711

[0096] [Correction pursuant to Rule 91, May 3, 2024]

Claims

1. Bacillus amyloliquefaeciens MU2 strain (KCTC15499BP) with excellent protein and cell wall decomposition enzyme activity.

2. In paragraph 1, The strain is characterized by having a 16S rRNA base sequence of sequence number 1.

3. In paragraph 1, The strain is characterized by excellent heat resistance, alkali resistance, antibiotic sensitivity, and antibacterial activity against harmful microorganisms, and does not produce biogenic amines or cause hemolysis.

4. In paragraph 3, A composition characterized in that the biogenic amine is histamine, tyramine, putrescine or cadaverine.

5. A starter for fermentation comprising the Bacillus amyloliquefaciens MU2 strain of paragraph 1 or a culture solution thereof as an active ingredient.

6. A composition for inhibiting the growth of pathogenic microorganisms, comprising the Bacillus amyloliquefaciens MU2 strain of paragraph 1 or a culture solution thereof as an active ingredient.

7. In paragraph 6, A composition characterized in that the pathogenic microorganisms are Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria Monocytogenes, and Brio parahaemolyticus.

8. In paragraph 6, A composition characterized in that the pathogenic microorganism is an antibiotic-resistant pathogenic microorganism.

9. In paragraph 8, A composition characterized in that the antibiotic is Gentamicin, Kanamycin, Streptomycin, Tetracycline, Erythromycin, Chloramphenicol, Vancomycin.

10. A probiotic preparation containing the strain of paragraph 1 or a culture solution thereof as an active ingredient.

11. A health food containing the strain of paragraph 1 or its culture solution as an active ingredient.

12. A feed additive containing the Bacillus amyloliquefaciens MU2 strain of paragraph 1 or a culture solution thereof as an active ingredient.

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