Selective antibacterial use of micrococcus luteus

Abstract

The present invention relates to a selective antibacterial use of Micrococcus luteus against harmful Staphylococcus sp. This composition comprising Micrococcus luteus, or a lysate, culture medium, or extract thereof exhibits selective antibacterial activity against harmful Staphylococcus sp., particularly Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis, and thus can be variously used as an antibacterial composition in cosmetics, quasi-drugs, external use skin preparations, medicines or the like.

Description

[0001] Description Title of the invention: Selective antibacterial use of Micrococcus luteus Technical field The present invention relates to the selective antibacterial use of Micrococcus luteus against harmful Staphylococcus sp. More specifically, the present invention relates to the selective antibacterial use of a composition comprising Micrococcus luteus, a lysate, a culture medium or an extract thereof against harmful Staphylococcus aureus. Background art Since the surface of human skin contains a lot of waste materials such as keratin, sweat and sebum, various microorganisms inhabit it and form the skin microbiome, which is the skin microbiome. Beneficial bacteria and harmful bacteria coexist in the skin microbiome, and the healthier the skin, the greater the distribution of beneficial bacteria, while the greater the distribution of harmful bacteria in atopic or acne-prone skin. Therefore, it is known that maintaining the balance of resident bacteria present on the skin helps maintain a healthy skin condition. Normally, skin flora that reside on the skin forms a skin barrier that prevents the penetration of other more harmful microorganisms (pathogens) and helps the skin immune function. An imbalance between beneficial and harmful skin bacteria can cause a breakdown of the skin barrier function and a decrease in the skin's immune response, and can act as a cause of various skin diseases such as atopic dermatitis, seborrheic dermatitis, acne, and psoriasis. Therefore, if the activity of only harmful bacteria could be selectively inhibited without affecting beneficial skin bacteria, it would be helpful in treating various skin diseases. In the present invention, as a result of studying skin flora, it was confirmed that Micrococcus luteus exhibits antibacterial activity selectively against harmful Staphylococcus {St aphy 1 ococcus sp.), and thus the present invention was completed. Detailed Description of the Invention Technical Problem Accordingly, the present invention provides a composition comprising Micrococcus luteus, or a lysate, culture solution, or extract thereof, and having an antibacterial activity against harmful Staphylococcus {St aphy 1 ococcus sp.) and exhibits antibacterial activity selectively against harmful Staphylococcus aureus. Specifically, the present invention provides an antibacterial cosmetic composition comprising Micrococcus luteus, or a lysate, culture or extract thereof, and exhibits antibacterial activity selectively against harmful Staphylococcus aureus. The present invention provides an antibacterial pharmaceutical composition comprising Micrococcus luteus, or a lysate, culture or extract thereof, and exhibits antibacterial activity selectively against harmful Staphylococcus aureus. The present invention provides an antibacterial skin external application composition comprising Micrococcus luteus, or a lysate, culture or extract thereof, and exhibits antibacterial activity selectively against harmful Staphylococcus aureus. The present invention provides an antibacterial quasi-drug composition comprising Micrococcus luteus, or a lysate, culture or extract thereof, and exhibits antibacterial activity selectively against harmful Staphylococcus aureus. The present invention provides an antibacterial food or health functional food composition comprising Micrococcus luteus, or a lysate, culture medium or extract thereof, which selectively exhibits antibacterial activity against harmful Staphylococcus aureus. In addition, the present invention provides a Micrococcus luteus strain deposited under accession number KACC 81251BP. In addition, the present invention provides a lysate, culture medium or extract of the Micrococcus luteus strain. Technical solution One aspect of the present invention provides an antibacterial composition comprising Micrococcus luteus, or a lysate, culture medium or extract thereof, as an active ingredient, which selectively exhibits antibacterial activity against harmful Staphylococcus aureus. In the present invention, Micrococcus luteus may be a viable bacterium (live bacterium) or a killed bacterium (dead bacterium). More specifically, the killed bacterium may be a bacterium killed by heat treatment. In one specific example, the Micrococcus luteus may be a Micrococcus luteus strain deposited under accession number KACC 81251BP.The term “lysate” as used herein means a solution or suspension in an aqueous medium of broken cells of a microorganism, such as Micrococcus luteus. A cell lysate includes macromolecules, such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc., and / or micromolecules, such as amino acids, sugars, fatty acids, etc., or fractions thereof. The lysate also includes cell debris, which may be smooth or granular in structure. Methods for achieving cell lysis of the microorganism may use various known methods, and any method capable of achieving cell lysis of the microorganism may be used. For example, cell opening / disruption may be performed enzymatically, chemically, or physically. Non-limiting examples of enzymes and enzyme mixtures are proteases, such as proteinase O, lipases, or glycosidases; non-limiting examples of chemicals are ionophores, detergents, such as sodium dodecyl sulfate, acids, or bases; Non-limiting examples of physical means are high pressure, such as French pressing, temperature, such as osmotic pressure, heat, or cold. In addition, a method using an appropriate combination of enzymes other than proteolytic enzymes, acids, bases, etc. may also be used. The term “culture medium” as used herein may be used interchangeably with “culture supernatant”, “conditioned medium”, or “conditioned medium”, and may mean the entire medium including the microorganism, its metabolites, extra nutrients, etc. obtained by culturing Micrococcus luteus for a certain period of time in a medium capable of supplying nutrients so that Micrococcus luteus can grow and survive in vitro. In addition, the culture medium may mean a culture medium obtained by removing the microorganisms from a cell culture medium obtained by culturing the microorganisms.Meanwhile, the liquid from which the bacterial cells have been removed from the culture medium is also called the “supernatant”, and can be obtained by leaving the culture medium still for a certain period of time and taking only the liquid from the upper layer excluding the portion that has settled to the lower layer, removing the bacterial cells through filtration, or centrifuging the culture medium to remove the sediment at the lower layer and taking only the liquid at the upper layer. The “bacterial cells” refers to the microorganisms of the present invention themselves, and includes the microorganisms themselves that have been separated and selected from a skin sample or the like, or the microorganisms that have been separated from the culture medium by culturing the microorganisms. The bacterial cells can be obtained by centrifuging the culture medium and taking the portion that has settled to the lower layer, or by allowing the bacterial cells to settle to the lower layer of the culture medium by gravity for a certain period of time and then removing the upper liquid. According to one aspect, the culture of Micrococcus luteus of the present invention can use a medium easily selected by a person skilled in the art according to the purpose among the media used for culturing microorganisms, and specifically, a medium used for culturing Micrococcus, such as MH (Mueller Hinton broth) medium or TSB (Tryptic soy broth), can be used, but is not limited thereto. According to one specific example, the culture of Micrococcus luteus of the present invention can be prepared by inoculating Micrococcus luteus into the microbial culture medium and according to a microbial culturing method known in the art (for example, static culture, etc.). The culture solution may include the culture solution itself obtained by culturing microorganisms, a concentrate thereof, or a lyophilized product, or a culture supernatant obtained by removing microorganisms from the culture solution, a concentrate thereof, or a lyophilized product thereof. The above culture solution may be obtained by culturing Micrococcus luteus in an appropriate medium (e.g., MH medium) at a temperature of 10 °C to 40 °C for a certain period of time, e.g., 4 to 50 hours.In one specific example, the microbial culture supernatant can be obtained by a step of removing microorganisms by centrifuging or filtering the microbial culture. In another specific example, the concentrate can be obtained by a step of concentrating the microbial culture itself, or the supernatant obtained after centrifuging or filtering the culture. The culture medium and culture conditions for culturing the Micrococcus luteus can be appropriately selected or modified and used by those skilled in the art. The term “extract” as used herein means something extracted from the lysate, culture solution, or concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a adjusted or purified product thereof, and a fraction obtained by fractionating the extract. The composition comprises 0.00001 wt% to 80 wt%, for example, 0.00001 wt% to 60 wt%, 0.00001 wt% to 40 wt%, 0.00001 wt% to 30 wt%, 0.00001 wt% to 20 wt%, 0.00001 wt% to 10 wt%, 0.00001 wt% to 5 wt%, 0.05 wt% to 60 wt%, 0.05 wt% to 40 wt%, 0.05 wt% to 30 wt%, 0.05 wt% to 20 wt%, 0.05 wt% to 10 wt%, 0.05 wt% to 5 wt%, 0.1 wt% to 60 wt%, 0.1 wt% to 40 wt%, 0.1 It may comprise from 0.1 wt% to 30 wt%, from 0.1 wt% to 20 wt%, from 0.1 wt% to 10 wt%, or from 0.1 wt% to 5 wt% of a microorganism, or a lysate, culture, or extract thereof.As used herein, the term "including as an active ingredient" means that the microorganism, or a lysate, culture medium or extract thereof, is added to an extent that the above-mentioned effect can be exhibited, and includes formulations in various forms by adding various components as auxiliary components for drug delivery and stabilization, etc. The term "antibacterial" as used herein means having an activity of (i) inhibiting, reducing or preventing the growth of bacteria; (ii) inhibiting or reducing the ability of bacteria to cause infection in a subject; or (iii) inhibiting or reducing the ability of bacteria to proliferate or maintain infectivity in the environment. In the present invention, Micrococcus luteus, or a lysate, culture medium or extract thereof, can selectively exhibit antibacterial activity against harmful Staphylococcus aureus. Specifically, Micrococcus luteus exhibits antibacterial activity against harmful Staphylococcus aureus, but has weak or no effect on harmless skin beneficial bacteria. The above harmful staphylococcus refers to staphylococcus that invades the skin surface or tissue of the host and causes troubles such as skin rashes, infections (abscesses, pus, redness, etc.). The above Micrococcus luteus, or its lysate, culture solution or extract, is a harmful staphylococcus among bacteria of the genus Staphylococcus (staphylococcus), for example, Staphylococcus aureus, a bacterium that causes skin diseases. aureus'), methicillin-resistant Staphylococcus aureus (Methici 11 in resistance Staphylococcus aureus'- MRSA), and Staphylococcus It can exhibit antibacterial activity against, but is not limited to, Micrococcus luteus, or its lysate, culture or extract. However, the above Micrococcus luteus, or its lysate, culture or extract, can exhibit antibacterial activity against, but is not limited to, epidermidisY^, and it was confirmed that there was almost no antibacterial activity against Staphylococcus hominis (Staphylococcus hominis'), a skin beneficial bacteria. In addition, the above Micrococcus luteus, or its lysate, culture solution or extract, was effective against Micrococcus luteus, Candida albicans SC5314, and Corynebacterium prophinguum, which are not Staphylococci.

[0002] ( Corynebacterium propingmini), Pseudomonas aeruginosa ATCC10145, and Escherichia coli (Escherichia col P) DH5 a were confirmed to exhibit little antibacterial activity. In one specific example, the Micrococcus luteus, or a lysate, culture or extract thereof, can selectively exhibit antibacterial activity against one or more of Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis. In one specific example, the Micrococcus luteus, or a lysate, culture or extract thereof, can selectively exhibit antibacterial activity against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis. The antibacterial composition of the present invention can be used for various purposes and uses requiring antibacterial activity, and specifically, can be used as a cosmetic, a pharmaceutical (e.g., a topical skin preparation), a quasi-drug, a food, a health functional food, a detergent, a cleaning agent, a preservative, an antiseptic, a food additive, or a feed additive, but is not limited thereto. In one specific example, the antibacterial composition comprising Micrococcus luteus, or a lysate, culture solution, or extract thereof, and selectively exhibiting antibacterial activity against harmful Staphylococcus aureus, can be a cosmetic composition. In the present invention, the cosmetic composition can be prepared in a formulation including a toner (skin lotion), skin, skin softener, skin toner, astringent jet, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, hand sanitizer, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, suspension, gel, powder, paste, mask pack, and sheet. The composition in such a formulation can be prepared according to a method conventional in the art.The mixing amount of additional ingredients such as the above moisturizer can be easily selected by a person skilled in the art within a range that does not impair the purpose and effect of the present invention. In addition to the effective ingredients disclosed in the present specification, the cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions, and may further include purified water, thickeners, preservatives, stabilizers, solubilizers, surfactants, carriers, fragrances, or combinations thereof that are commonly used. The functional additives may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular peptides, high molecular polysaccharides, sphingolipids, and seaweed extracts. Examples of the carriers include alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosity modifiers, emulsifiers, stabilizers, ultraviolet scatterers, ultraviolet absorbers, colorants, fragrances, and the like. Compounds / compositions that can be used as the above alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosity modifiers, emulsifiers, stabilizers, ultraviolet scatterers, ultraviolet absorbers, colorants, fragrances, etc. are already known in the art, so a person skilled in the art can select and use the appropriate corresponding substances / compositions. In addition, the above cosmetic composition may contain, as necessary, a sunscreen, an antioxidant (butylated hydroxyanisole, propyl gallate, ellisorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), a preservative (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinyl urea, chlorphenesin, etc.), a colorant, a pH adjuster (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, sodium malate, formaldehyde, sodium formaldehyde, succinic acid, sodium succinate, sodium hydroxide, sodium hydrogenphosphate, etc.), a moisturizer (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycereth-26, methylgluceth-20) Additional ingredients such as lubricants, etc. can be added.In addition, for each type of cosmetic composition, appropriate ingredients can be selected and combined according to the formulation or intended use of the cosmetic. Since the combining ingredients and method can follow conventional techniques, a detailed description thereof is omitted herein. In another specific embodiment of the present invention, an antibacterial composition comprising Micrococcus luteus, or a lysate, culture medium, or extract thereof, and selectively exhibiting antibacterial activity against harmful Staphylococcus aureus can be an antibacterial pharmaceutical composition. The term “pharmaceutical composition” as used herein can refer to a molecule or compound that imparts several beneficial effects upon administration to a subject. Beneficial effects can include enabling diagnostic determination; improving a disease, symptom, disorder, or condition; reducing or preventing the onset of a disease, symptom, disorder, or condition; and generally responding to a disease, symptom, disorder, or condition. As used herein, the term “prevention” refers to a method of partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its incidental symptoms, preventing the acquisition or reacquisition of a disease or disorder, or reducing the risk of acquiring a disease or disorder. For example, the prevention refers to any act of inhibiting or delaying the onset of skin damage or symptoms by administering a composition according to the present invention. The term “treatment” as used herein includes inhibition, alleviation, or elimination of the development of a disease. The term “improvement” as used herein may refer to any act of at least reducing a parameter related to the alleviation or treatment of a condition, for example, the severity of a symptom. In the pharmaceutical composition according to the present invention, Micrococcus luteus, or a lysate, culture, or extract thereof, may be included as an active ingredient of a pharmaceutical composition having antibacterial activity, or may be used as an adjuvant of an active ingredient having a medicinal use. The pharmaceutical composition may be prepared according to a method well known to those skilled in the art.The pharmaceutical composition described above can be administered orally or parenterally depending on the intended method and can be used in the form of a general pharmaceutical preparation. Parenteral administration can mean administration via a route other than oral administration, such as rectal, intravenous, peritoneal, intramuscular, intraarterial, transdermal, nasal, inhalation, ocular, and subcutaneous. When the pharmaceutical composition of the present invention is used as a pharmaceutical, it can additionally contain one or more active ingredients having the same or similar function. The pharmaceutical composition can further be prepared by including one or more pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers include saline solution, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets. Furthermore, it can be preferably formulated according to each disease or ingredient by an appropriate method in the art. When formulating the above pharmaceutical composition, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases that can be used include Witepsol, Macrogol, Tween 61, cocoa butter, liurin butter, and glycerogelatin.The pharmaceutical composition may contain carbohydrates such as glucose, sucrose or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers (Needle 3 Cutting Heart 26 Factory 8) to increase stability or absorbability. In another embodiment of the present invention, the pharmaceutical composition may be an antibacterial skin topical composition. The skin topical composition may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a lotion, or a combination thereof. The above skin external preparation may be appropriately mixed with ingredients commonly used in skin external preparations such as cosmetics or medicines, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof, as needed. The above skin external preparation may also appropriately contain metal sequestrants such as sodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, bellapamil, licorice extract, glabridin, hot water extract of the fruit of Kalin, various herbal medicines, tocopheryl acetate, glycyrrhizic acid, tranexamic acid and derivatives thereof or salts thereof; and sugars such as vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, and trehalose. The skin external preparation composition according to the present invention may contain 0.00001 wt% to 80 wt% of microorganisms, lysates, cultures, or extracts thereof based on the total weight of the composition. The above skin includes all skin areas of the body, including the face, hands, arms, legs, feet, chest, stomach, back, buttocks, and scalp.Another aspect of the present invention provides a method for preventing, improving, or treating a condition of a subject, comprising administering or applying to the subject a composition of Micrococcus luteus, or a lysate, culture, or extract thereof. The condition of the subject may be a condition requiring antimicrobial treatment, and specifically, antimicrobial treatment against harmful Staphylococcus aureus. The terms “administering,” “applying,” “introducing,” and “implanting” are used interchangeably herein and may refer to the placement of a composition of an embodiment into a subject by a method or route that results in at least partial localization of the composition to a desired site. Administration may be by any method known in the art. Administration may be directly administered to the subject by any means, such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous administration. The above administration may be systemic or local. The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an subject in need of antibacterial treatment. The administration may be 0.00001 mg to 1,000 mg of the composition according to one embodiment per subject per day, for example, 0.00001 mg to 500 mg, 0.00001 mg to 100 mg, 0.00001 mg to 50 mg, 0.It may be administered in an amount of 00001 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, the dosage may be prescribed in various ways depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity, and a person of ordinary skill in the art can appropriately adjust the dosage by considering these factors. The frequency of administration may be once a day or twice or more within the range of clinically acceptable side effects, and the administration may be done at one site or more than two sites, and the total number of administration days may be from 1 to 30 days per treatment, daily or at intervals of 2 to 5 days. If necessary, the same treatment may be repeated after an appropriate period. For animals other than humans, the same dosage as for humans per kg may be used, or the above dosage may be converted into an amount calculated by taking into account the volume ratio (e.g., average value) of the organs (such as the heart) of the target animal and the human, for example. The present invention also provides a composition comprising Micrococcus luteus, or a lysate, culture, or extract thereof, for use in antibacterial treatment against harmful Staphylococci. Another aspect of the present invention provides a selective antibacterial method against harmful Staphylococci, comprising treating the bacteria with Micrococcus luteus, or a lysate, culture, or extract thereof.At this time, the description of Micrococcus luteus, or its lysate, culture medium and extract, and the selective antibacterial activity against harmful Staphylococcus aureus is as described above. The method for treating bacteria with Micrococcus luteus, or its lysate, culture medium or extract can be appropriately used according to a conventional method well known to those skilled in the art in various fields requiring antibacterial activity. The amount of Micrococcus luteus, or its lysate, culture medium or extract to be treated can be appropriately determined depending on the intended use. In another specific embodiment of the present invention, the antibacterial composition comprising Micrococcus luteus, or its lysate, culture medium or extract, and exhibiting selective antibacterial activity against harmful Staphylococcus aureus may be a quasi-drug composition. In this specification, the term “quasi-drug” refers to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals, and have a milder effect than drugs. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products other than those used for the purpose of drugs, and include fibers and rubber products used for the treatment or prevention of diseases of humans and animals, things that have a mild or no direct effect on the human body, are not instruments or machines and similar thereto, and sterilizing and insecticides for preventing infectious diseases. The type or formulation of the quasi-drug composition of the present invention is not particularly limited, but may be a bandage, gauze, cotton wool, adhesive plaster, disinfectant cleanser, shower foam, gargle, wet tissue, detergent soap, hand wash, humidifier filler, mask, or filter filler. When the composition of the present invention is included in a quasi-drug for selective antibacterial purposes, the composition may be used as is or together with other quasi-drug ingredients, and may be used appropriately according to a conventional method. The amount of active ingredients mixed can be appropriately determined depending on the intended use, and the pharmaceutical composition according to the present invention contains 0.01 wt% to 20 wt% of a microorganism, a lysate, a culture medium or an extract thereof. In another specific embodiment of the present invention, the antimicrobial composition comprising Micrococcus luteus, or a lysate, a culture medium or an extract thereof, and selectively exhibiting antimicrobial activity against harmful Staphylococcus aureus may be an antimicrobial food composition or an antimicrobial health functional food composition. The food or health functional food composition may use Micrococcus luteus, or a lysate, a culture medium or an extract thereof, alone or in combination with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredients may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). Generally, when manufacturing a food or beverage, the composition of the present specification may be added in an amount of 15 parts by weight or less to the raw material. There is no particular limitation on the type of the food or health functional food. Examples of foods to which the above substances can be added include formulations selected from the group consisting of powders, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, infused teas, and health drinks, and include all health foods in the conventional sense. Among the types of health functional foods, beverage compositions can contain various flavorings or natural carbohydrates as additional ingredients, just like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used.The health food composition may also contain nutrients, vitamins, electrolytes, flavorings, colorings, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, or combinations thereof. The health functional food composition may also contain fruit pulp for the production of natural fruit juice, fruit juice drinks, vegetable drinks, or combinations thereof. In addition, when the antibacterial composition of the present invention is used as a detergent material, there is no particular limitation on the articles to which the antibacterial composition can be applied, and specific examples thereof include hand wash, body wash, shampoo, rinse, cleansing foam, cleansing gel, soap, gargle, toothpaste, bathroom cleaner, kitchen cleaner, etc. In addition, when the antibacterial composition of the present invention is used as a preservative or antiseptic material, the articles to which the antibacterial composition can be applied are not particularly limited, and specific examples thereof include cosmetics, foods, pharmaceuticals, etc. In addition, when the antibacterial composition of the present invention is used as an additive material, the articles to which the antibacterial composition can be applied are not particularly limited, and specific examples thereof include foods and animal feeds, etc. Another aspect of the present invention provides a Micrococcus luteus strain deposited under the accession number KACC 81251BP. The Micrococcus luteus strain may be a strain comprising 16S rRNA of SEQ ID NO: 3. The strain may selectively exhibit antibacterial activity against harmful Staphylococcus aureus. Specifically, the Micrococcus luteus strain exhibits antibacterial activity against harmful Staphylococcus aureus, but has weak antibacterial activity or has no effect on harmless skin beneficial bacteria.The above Micrococcus luteus strain can exhibit antibacterial activity against harmful Staphylococci among bacteria of the genus Staphylococcus (Staphylococci), such as, but not limited to, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis, which are skin disease-causing bacteria. However, the Micrococcus luteus strain was confirmed to have weak antibacterial activity against Staphylococcus epidermidis, which is a beneficial skin bacterium, and no antibacterial activity against Staphylococcus hominis, which is a beneficial skin bacterium. In addition, the Micrococcus luteus strain was confirmed to have no antibacterial activity against non-staphylococci, such as Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophingum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5a. In one embodiment, the Micrococcus luteus strain can exhibit antibacterial activity selectively against one or more of Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis. In one embodiment, the Micrococcus luteus strain can exhibit antibacterial activity selectively against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis. Another aspect of the present invention provides a lysate, culture medium, or extract of the Micrococcus luteus strain. The description of the Micrococcus luteus strain, the lysate, and the culture medium thereof is as described above. The lysate, culture medium, or extract of the Micrococcus luteus strain can exhibit antibacterial activity selectively against harmful Staphylococci. Herein, exhibiting antibacterial activity selectively against harmful Staphylococci is as described above with respect to the Micrococcus luteus strain.Effect of the invention A composition comprising Micrococcus luteus, or a lysate, culture or extract thereof according to one aspect of the present invention, selectively exhibits antibacterial activity against harmful staphylococci, particularly Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis, and thus can be usefully used as an antibacterial composition in cosmetics, quasi-drugs, external skin preparations or medicines. Brief description of the drawings Figure 1 shows the results of measuring the growth inhibition ability against Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus (MRSA) USA 300, Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophingum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5a when treated with a strain homogenate according to one specific example of the present invention (N-CNT: negative control; cell lysate: strain homogenate). Figure 2 is a graph showing the results of measuring the degree of biofilm formation of Staphylococcus aureus using WST-8 analysis in the case of treatment with a strain homogenate according to one specific example of the present invention (control group: PBS, 0% cell lysate; experimental group: lysate, 20% cell lysate). Figure 3 is a graph showing the results of measuring the cell viability of human keratinocytes in the case of treatment with a strain homogenate according to one specific example of the present invention (control group: 10% PBS; experimental group: 10% cell lysate).FIG. 4 is an experimental result for determining the properties of the active ingredient of the strain homogenate by measuring the antibacterial activity by treating the strain homogenate with heat or protease K according to one specific example of the present invention and then treating it with Staphylococcus aureus ATCC 25923 (heat treatment: 95 °C, 10 minutes, protease K; Img / mL) FIG. 5 is an experimental result for determining the properties of the active ingredient of the strain homogenate by measuring the antibacterial activity by treating the strain homogenate with Micrococcus luteus CBN009 according to one specific example of the present invention and its standard strain Micrococcus luteus ATCC 4698, when treated with Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus USA 300 (MRSA), Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus, Micrococcus luteus, Candida albicans SC5314, This is the result of comparing the growth inhibition ability against Corynebacterium prophinguum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5a. Mode for carrying out the invention Hereinafter, the present invention will be described in more detail through examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples. Example 1. Isolation and identification of strains A specimen was collected from the skin of a healthy Korean, mixed in a phosphate buffered saline (PBS) solution, and inoculated into an LB (Luria-Berani) medium through serial dilution. After culturing for 63 hours in a 34 °C incubator, bacterial colonies formed on a plate were selected and purified using a streaking method in the same medium and conditions. 16S rRNA gene sequence analysis was performed to confirm the molecular phylogenetic characteristics of the strain after cultivation. PCR (Polymerase chain reaction) amplification was performed 32 times at 95 °C for 30 seconds, 55 °C for 30 seconds, and 72 °C for 1 minute and 45 seconds, and finally at 72 °C for 5 minutes.At this time, primers (SEQ ID NOs: 1 and 2) designed to specifically react with bacterial common genes were used. The PCR amplification product was purified to confirm the base sequence (Macrogen, Republic of Korea), and the sequence was analyzed using the BLAST program of the National Center for Biotechnology Information (NCBI) of the United States. When compared with other previously registered strains, species with more than 99% base sequence homology were determined to be closely related species. The strain isolated in this example had the 16S rRNA sequence of SEQ ID NO: 3 (complementary DNA) and was identified as Micrococcus luteus. The present inventors named the strain Micrococcus luteus CBN009 and deposited it with the National Institute of Agricultural Genetic Resources on January 3, 2023, and were assigned the accession number KACC 81251BP. Example 2. Culture and homogenization of strain The strain isolated in Example 1 was cultured in MH (Mueller Hinton broth) medium at 30 °C incubator, and the cells were collected at the final point of the growth phase (exponenti al phase) on the growth curve. The cells were centrifuged at 8,000 × g for 30 minutes and washed once with PBS (Phosphate-Buffered Saline). The cells were dissolved in PBS and homogenized through sonication. The homogenized strain homogenate was quantified to 2 mg / ml through a 0.2 μm filter to remove the cells, and used for future evaluation and activity analysis. The evaluation and activity analysis were performed no longer than 1 day after preparation. Experimental Example 1. Antibacterial activity It was analyzed whether the strain homogenate obtained in Example 2 exhibited antibacterial activity against harmful strains.Specifically, antibacterial activity was tested against Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus USA 300 (MRSA), Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus, Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophinguum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5 a. The strains were first cultured in TSB or MH medium for use in the experiment, and bacteria were grown in an aerobic chamber at 34 °C, and yeasts were grown in an aerobic chamber at 30 °C. Subcultures were performed to l / 100 (seed / medi a) at the end of the exponenti al phase of the growth curve. The antibacterial activity was evaluated by preparing a diluted solution in which the cell line that reached the beginning of the growth phase (exponent i al phase) on the growth curve through subculture was diluted to ODxo 0.01. The antibacterial activity of the strain homogenate was measured under the following conditions. First, the diluted solutions of the 10 strains (5. aureus, MRSA, S. epidermidis, S. hominis, S. capitis, M. luteus, C. albicans, C. propinquum, P. aeruginosa, and E. co / 7) to be evaluated for antibacterial activity were spread twice using a sterile cotton swab on an MH agar plate, and then circular agar holes with a diameter of 0.8 cm were made. After inoculating 150 uL of the strain homogenate prepared in Example 2 into an agar pit, the culture was grown in an incubator at 30 °C for 24 and 48 hours, and the resulting clear zone was observed, and the results are shown in Fig. 1.Figure 1 shows the results of measuring the growth inhibition ability against Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus USA 300 (MRSA), Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus, Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophingum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5a when treated with the strain homogenate of Example 2 (N-CNT: negative control; Cell lysate: strain homogenate). As shown in Figure 1, clear zones were formed in Staphylococcus aureus ATCC 25923, MRSA, Staphylococcus epidermidis, and Staphylococcus capitus treated with the strain homogenate of Example 2. Specifically, since a clear zone is formed in Staphylococcus aureus ATCC 25923, MRSA, and Staphylococcus capitis, which are harmful skin bacteria, antibacterial activity is observed. In Staphylococcus epidermidis, which is a beneficial skin bacteria, the clear zone is reduced and appears fainter compared to Staphylococcus aureus ATCC 25923, MRSA, and Staphylococcus capitis, which are harmful skin bacteria, so the antibacterial activity is relatively weak. In addition, in Staphylococcus hominis, which is a beneficial skin bacteria, the clear zone is hardly formed, so it was confirmed that there is almost no antibacterial activity. In addition, it can be seen that there is almost no antibacterial activity in Micrococcus luteus, Candida albicans SC5314, Corynebacterium propingum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5a, which are not Staphylococci, since the clear zone is hardly formed even when the strain homogenate of Example 2 is treated. These results show that the strain homogenate of Example 2 exhibits selective antibacterial activity against harmful staphylococci, particularly Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus capitis. Experimental Example 2. Inhibitory activity against biofilm formation This experiment was conducted to determine whether the strain homogenate obtained in Example 2 inhibits the biofilm formation ability of Staphylococcus aureus ATCC 25923.Specifically, a frozen stock of Staphylococcus aureus ATCC 25923 strain was first inoculated into MH medium and cultured at 34°C for 17 hours. The strain was then inoculated into fresh MH medium and subcultured at 30°C until it reached the beginning of the exponential phase of the growth curve. Prior to the experiment, 160 iiL of MH medium containing 1% glucose was added to all available wells of a 96-well plate. Half of the available wells were then filled with 40 ng of PBS. UL was treated, and the remaining half of the available wells were treated with 40 μl of cell homogenate at a protein concentration of 2 mg / ml each. Then, 2 Il L of 5. aureus subculture was added to all wells and grown at 30 °C, 200 μg / ml for 24 hours. At this time, the wells treated with PBS were used as a negative control. After that, the wells were washed twice with distilled water, dried to fix the biofilm, and then stained with a 0.1% crystal violet solution. After that, the wells were washed three times with distilled water, and the biofilm was dissolved with a 30% acetic acid solution. The absorbance at 560 μg / ml was measured using a microplate reader, and the results are shown in Fig. 2. Fig. 2 is a graph showing the results of measuring the absorbance through WST-8 analysis for the degree of biofilm formation of Staphylococcus aureus when treated with the strain homogenate of Example 2 (control group: PBS, 0% cell lysate; experimental group: lysate, 20% cell lysate). As shown in Fig. 2, the amount of biofilm formation of Staphylococcus aureus in 5. aureus treated with the strain homogenate was reduced to 27.4% compared to the experimental group treated with PBS. From this, it can be confirmed that the strain homogenate can significantly inhibit biofilm formation of Staphylococcus aureus. Experimental Example 3. Cytotoxicity Evaluation (Relative cell viability) Cell viability was tested to determine whether the strain isolated in Example 1 was toxic to cells. Specifically, first, human epidermal keratinocyte cell line (Human epidermal keratinocyte cell line, HaCaT) was cultured at 2.5 x 10 in DMEM medium (Dubelcco's modified eagle medium) containing 10% fetal bovine serum (FBS). 4Cells were seeded into 24-well plates at a density of 10 cells / well and cultured overnight, and then cultured for 24 hours in DMEM medium without FBS. Cytotoxicity of the strain homogenate was evaluated using the WST assay. The cultured cell lines were treated with the strain homogenate with the protein concentration of 2 mg / ml at a concentration of 10% of the culture medium or with 10% PBS and cultured for 24 hours under the same conditions. The 10% PBS treatment group was used as a negative control, and the total volume per well was 500 uL. After removing the culture medium, the cells were washed once with IX PBS, and DMEM medium containing WST-8 reagent was added and cultured at 37 °C for 0.5 to 4 hours. Afterwards, the absorbance at 450 / 625 was measured using a microplate reader, and the results are shown in Figure 3. Fig. 3 is a graph showing the results of WST analysis evaluating the viability of human keratinocytes according to treatment with a homogenate of the strain isolated in Example 1 (control group: 10% PBS; experimental group: 10% cell lysate). As shown in Fig. 3, when treated with a 10% (v / v) strain homogenate, there was no significant difference in the viability of human keratinocytes compared to the negative control group treated with PBS, indicating that the strain homogenate was not cytotoxic. This result indicates that the strain homogenate is safe when applied to the skin. Experimental Example 4. Screening of antibiotics When the strain homogenate obtained in Example 2 was heated or treated with a protein-decomposing enzyme, Protase K, it was observed whether there was a change in the antibacterial activity against Staphylococcus aureus ATCC 25923. Specifically, as examined in Experimental Example 1 above, the strain homogenate of Example 2 exhibits antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, and Staphylococcus capitis. This experiment was conducted to estimate the substance from which this activity originated.The same experimental method as Experimental Example 1 was used, and Staphylococcus aureus ATCC 25923 was used as the antibacterial target strain. The strain homogenate was prepared according to Example 2, and then divided into 1.5 mL portions and placed in three E-tubes, and each tube underwent the following processing. One tube was stored in an ice bucket as is, one tube was treated at 95 °C for 10 minutes, and one tube was treated with protease at a concentration of 1 mg / mL. 150 μL of the strain homogenate that completed each processing step was inoculated into an agar hole and incubated in a 30 °C incubator for 24 hours.

[0003] After culturing for 48 hours, the clear zone that was created was observed, and the results are shown in Fig. 4. Fig. 4 shows the results of an experiment to determine what properties the effective substance of the strain homogenate has by measuring the antibacterial activity by treating the strain homogenate with heat or protease and then treating it with Staphylococcus aureus ATCC 25923 (heat treatment: 95 °C, 10 minutes, protease K; Img / mL). As shown in Fig. 4, when the strain homogenate was heat-treated or treated with protease, the clear zone became faint or disappeared, indicating that the antibacterial activity was inhibited, and in particular, when treated with protease, it was confirmed that the clear zone completely disappeared. In light of this, it is presumed that the antibacterial activity confirmed in the strain homogenate is due to any protein. Experimental Example 5. Comparison of antibacterial activity with standard strains In this experiment, the antibacterial activity of the homogenate of Micrococcus luteus CBN009 strain obtained in Example 2 was compared with that of its standard strain, Micrococcus luteus ATCC 4698. Specifically, the antibacterial activity was tested against Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus USA 300 (MRSA), Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus, Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophingum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5 a. First, the strains were cultured in TSB or MH medium for use in the experiment, and bacteria were grown in an aerobic chamber at 34 °C, and yeasts were grown in an aerobic chamber at 30 °C. Subculture was performed to reach l / 100 (seed / medi a) at the final point of the growth phase (exponent i al phase) on the growth curve. Cell lines that reached the beginning of the growth phase (exponent i al phase) on the growth curve through subculture were selected as ODxo 0.The antibacterial activity was evaluated by preparing a diluted solution diluted to 01. The antibacterial activity of the strain homogenate was measured under the following conditions. First, the diluted solution of the 10 strains (5. aureus, MRSA, 5. epidermidis, S. hominis, S. capitis, M. luteus, C. albicans, C. propinquum, P. aeruginosa, and E. co / 7) to be evaluated for antibacterial activity was spread twice on an MH agar plate using a sterilized cotton swab, and then a circular agar hole with a diameter of 0.8 cm was made. After inoculating 150 uL of the strain homogenate prepared in Example 2 into the agar hole, the plate was grown in an incubator at 30 °C for 24 and 48 hours, and the resulting clear zone was observed, and the results are shown in Fig. 5. FIG. 5 shows the results of measuring the growth inhibition ability against Staphylococcus aureus ATCC 25923, methicillin-resistant Staphylococcus aureus USA 300 (MRSA), Staphylococcus epidermidis ATCC 12228, Staphylococcus hominis, Staphylococcus capitus, Micrococcus luteus, Candida albicans SC5314, Corynebacterium prophinguum, Pseudomonas aeruginosa ATCC10145, and Escherichia coli DH5 a when treated with the homogenate of Micrococcus luteus CBN009 strain obtained in Example 2 and its standard strain, Micrococcus luteus ATCC 4698. As shown in Fig. 5, clear zones were formed in Staphylococcus aureus ATCC 25923, MRSA, Staphylococcus epidermidis, and Staphylococcus capitis treated with the strain homogenate of Micrococcus luteus CBN009, and the same results were also observed in the homogenate of its standard strain, Micrococcus luteus ATCC 4698.In light of these experimental results, the selective antibacterial activity of Micrococcus luteus against harmful Staphylococci can be considered a unique characteristic of the Micrococcus luteus species, rather than a specific characteristic of the Micrococcus luteus CBN009 strain.

[0004] [Name of the depositor: National Institute of Agricultural Genetic Resources (overseas)]Deposit number: KACC 81251BP Deposit date: January 3, 2023

[0005] Where Rule 64(d) applies, such date is the dale on which the status of iriternadonaJ depositary authority was acquired

[0006] Form BP / ^Sole page} Treaty on the International Authorization of the Deposit of Microorganisms for the Purpose of the Creation of Species of Animals

[0007] "To. QTsby Co., Ltd. This waste paper: Hi-dan is indicated in the following national deposit

[0008] 8th floor, Apgujing Building, 842 Nonhyeon-ryo, issued by the trustee under Rule 7.1, Kang Nan-gu. Seoul, Korea. Original. When Rule 6.4(d) applies, the date is the day on which the national tax base is established: Yangshi HP / M (single page)

Claims

Scope of the claim

1. An antibacterial composition comprising Micrococcus luteus, or a lysate, culture medium or extract thereof, and exhibiting antibacterial activity selectively against harmful Staphylococcus sp. [Jeongguhang 2] An antibacterial composition according to Jeongguhang 1, wherein Micrococcus luteus, or a lysate, culture solution or extract thereof, exhibits antibacterial activity against harmful Staphylococcus aureus, but has weak or no antibacterial activity against harmless skin beneficial bacteria. ［ Claim 3] In claim 1 or 2, Micrococcus luteus, or a lysate, culture solution or extract thereof, is Staphylococcus aureus. aureus), methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus An antimicrobial composition that selectively exhibits antimicrobial activity against one or more of the following: ［

4. An antibacterial composition according to any one of claims 1 to 3, wherein Micrococcus luteus is a Micrococcus luteus strain deposited under accession number KACC 81251BP. ［

5. An antibacterial composition according to any one of claims 1 to 4, wherein the antibacterial composition is used as a cosmetic, a pharmaceutical, a skin external preparation, a quasi-drug, a food, a health functional food, or a detergent. ［ Claim 6] A method for antibacterial treatment against harmful Staphylococcus aureus, comprising administering to a subject Micrococcus luteus, or a lysate, culture solution or extract thereof. ［ Claim 7] A composition comprising Micrococcus luteus, or a lysate, culture or extract thereof, for use in antibacterial treatment against harmful Staphylococcus aureus.

8. A strain of Micrococcus luteus deposited under accession number KACC 81251BP. ［ Claim 9] A lysate of the strain according to claim 8. ［

10. A culture solution of the strain according to claim 8. ［

11. An extract of the strain according to claim 8.

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