Antimicrobial composition containing fermented Melissa officinalis or its extract as effective component

Abstract

The present invention relates to an antibacterial composition containing a lemon balm fermented product or an extract thereof as an active ingredient. Compared to a non-fermented lemon balm extract, the lemon balm fermented product or the extract thereof of the present invention showed excellent antibacterial effects against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, Salmonella enterica, Pseudomonas aeruginosa, and Cutibacterium acnes.

Description

Technology Field

[0001] The present invention relates to an antibacterial composition containing a lemon balm ferment or an extract thereof as an active ingredient. Background Technology

[0002] Cosmetics contain various nutrients, including water, making them susceptible to contamination by bacteria or fungi. The negative consequences of microbial contamination in cosmetics can be summarized into two points: first, the risk to consumer health, and second, the decline in product value. Since most cosmetics are applied to the skin, applying a product contaminated with microorganisms can lead to inflammation or skin problems caused by microorganisms entering through wounds. In particular, contamination by pathogenic microorganisms can cause serious health issues. Furthermore, products overgrown with mold or bacteria result in a decrease in product value by causing changes in odor, breakdown of emulsification, and changes in product properties due to pH fluctuations. Therefore, various types of antimicrobial agents are used to suppress this, but it is important to use substances that are relatively harmless to the human body, as these can cause skin allergies. While most conventional antimicrobial agents used in the past were manufactured through chemical synthesis, the emergence of bacteria resistant to such agents is becoming increasingly frequent. Moreover, due to the high toxicity inherent in most of these chemically synthesized agents, there is a trend toward avoiding their use. Therefore, efforts to develop natural materials with antibacterial activity have been ongoing recently.

[0003] Meanwhile, lemon balm ( Melissa officinalisLemon balm is a perennial dicotyledonous plant belonging to the Lamiaceae family. Native to the Mediterranean coast, it resembles mint in appearance and is also called "bee balm" because bees are attracted to the strong lemon scent emanating from its leaves. Lemon balm has been reported to have various benefits related to bodily functions; it alleviates depression, stress, fever, and indigestion, and it is known that applying fresh, kneaded leaves to wounds can exert a detoxifying effect, thereby preventing infection.

[0004] As prior art related to lemon balm, Korean Published Patent No. 2024-0099588 discloses a cosmetic composition containing lemon balm extract as an active ingredient, and Korean Registered Patent No. 1282335 discloses a composition for improving skin whitening containing lemon balm extract as an active ingredient; however, no antibacterial composition containing the lemon balm fermentation product or extract thereof of the present invention has been disclosed to date. The problem to be solved

[0005] The present invention was derived from the above-mentioned requirements. The present invention provides an antibacterial composition containing a lemon balm ferment or an extract thereof as an active ingredient, and completes the present invention by confirming the antibacterial effect of the lemon balm ferment or its extract against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Enterobacter cloaca, Salmonella enterica, Pseudomonas eruginosa, and Cutibacterium acnes. means of solving the problem

[0006] To solve the above problem, the present invention provides an antibacterial composition containing a lemon balm ferment or an extract thereof as an active ingredient.

[0007] In addition, the present invention provides a cosmetic composition for preventing or improving acne containing a lemon balm ferment or an extract thereof as an active ingredient.

[0008] In addition, the present invention provides an antibacterial health functional food composition containing lemon balm ferment or an extract thereof as an active ingredient.

[0009] In addition, the present invention provides an antibacterial pharmaceutical composition containing a lemon balm ferment or an extract thereof as an active ingredient.

[0010] In addition, the present invention provides an antibacterial quasi-drug containing lemon balm ferment or an extract thereof as an active ingredient.

[0011] In addition, the present invention provides an antibacterial feed additive containing lemon balm ferment or an extract thereof as an active ingredient. Effects of the invention

[0012] The present invention relates to an antibacterial composition containing a lemon balm fermented product or an extract thereof as an active ingredient. Compared to a non-fermented lemon balm extract, the lemon balm fermented product or the extract thereof of the present invention showed excellent antibacterial effects against Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Escherichia coli, Enterobacter cloaca, Salmonella enterica, Pseudomonas eruginosa, and Cutibacterium acnes. Specific details for implementing the invention

[0013] To achieve the objective of the present invention, the present invention provides an antibacterial composition containing a lemon balm ferment or an extract thereof as an active ingredient.

[0014] The above lemon balm fermented product is Bacillus subtilis ( Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plants Fermentation is performed using any one strain selected from among the strains, but is not limited thereto.

[0015] In one embodiment of the present invention, the lemon balm fermented product

[0016] 1) A step of sterilizing water in which glucose and peptone are dissolved;

[0017] 2) Bacillus subtilis (in the sterilized water of step 1) above) Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plants A step of obtaining a culture solution by inoculating with one strain selected from among the strains, adding lemon balm, and culturing at 28–40°C for 12–48 hours; and

[0018] 3) After step 2) above, the obtained culture medium is sterilized and then filtered to obtain a filtrate; may be manufactured including this step, and

[0019] Preferably

[0020] 1) A step of dissolving 1~10g of glucose and 0.5~2.5g of peptone in 250mL of water, and then pressurizing and high-temperature sterilizing the water in which the glucose and peptone are dissolved;

[0021] 2) Activated Bacillus subtilis in the sterilized water of step 1) above ( Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plants A step of obtaining a culture solution by inoculating 1.2 to 6.5 mL of any one strain selected from among the strains, adding 6 to 20 g of lemon balm, and culturing at 30 to 40°C for 18 to 36 hours; and

[0022] 3) After step 2) above, the obtained culture medium may be manufactured by including the step of pressurizing and high-temperature sterilizing the culture medium and then filtering it to obtain a filtrate; and

[0023] More preferably

[0024] 1) A step of dissolving 3~8g of glucose and 0.7~1.7g of peptone in 250mL of water, and then pressurizing the water containing the dissolved glucose and peptone at high temperature;

[0025] 2) Activated Bacillus subtilis in the sterilized water of step 1) above ( Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plants A step of obtaining a culture solution by inoculating 1.7 to 4.5 mL of any one strain selected from among the strains, adding 10 to 15 g of lemon balm, and culturing at 30 to 38°C for 22 to 30 hours; and

[0026] 3) After step 2) above, the obtained culture medium may be manufactured by including the step of pressurizing and high-temperature sterilizing the culture medium and then filtering it to obtain a filtrate; and

[0027] Most preferably

[0028] 1) A step of dissolving 5g of glucose and 1.25g of peptone in 250mL of water, and then pressurizing the water containing the dissolved glucose and peptone at 121℃ for 15 minutes;

[0029] 2) Activated Bacillus subtilis in the sterilized water of step 1) above ( Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plants A step of obtaining a culture solution by inoculating 2.5 mL of any one strain selected from the strains, adding 12.5 g of lemon balm, and then culturing at 30°C or 37°C for 24 hours; and

[0030] 3) After step 2) above, the obtained culture medium may be manufactured by including the step of pressurizing the culture medium at 121°C for 15 minutes and then filtering it to obtain the filtrate; but is not limited thereto.

[0031] The extraction solvent of the above extract may be water, C1 to C4 lower alcohols, or a mixture thereof, preferably ethanol, but is not limited thereto.

[0032] The above extraction method may utilize any conventional method known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. It is preferable to add the extraction solvent in an amount of 1 to 20 times the weight of the lemon balm fermented product, and more preferably 5 to 15 times the weight. The extraction temperature is preferably 40 to 80°C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, and more preferably 0.5 to 5 hours, but is not limited thereto.

[0033] In one embodiment of the present invention, the antibacterial agent is Bacillus subtilis ( Bacillus subtilis ), Bacillus cereus( Bacillus cereus ), Staphilococcus aureus( Staphylococcus aureus ), E. coli( Escherichia coli ), Enterobacter cloa ( Enterobacter cloacae ), Salmonella enterica( Salmonella enterica ), Pseudomonas eruginosa( Pseudomonas aeruginosa ) and Cutibacterium acnes( Cutibacterium acnes It inhibits the growth of one or more bacteria selected from among, but is not limited to.

[0034] In one embodiment of the present invention, a lemon balm fermented using a Bacillus subtilis strain exhibits an antibacterial effect against Enterobacter cloaca, a lemon balm fermented using a Reviractobacillus brevis or Lactiplantibacillus plantarum strain exhibits an antibacterial effect against Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Escherichia coli, Enterobacter cloaca, Salmonella enterica, and Pseudomonas eruginosa, an extract of a lemon balm fermented using a Bacillus subtilis strain exhibits an antibacterial effect against Bacillus subtilis, Staphylococcus aureus, and Cutibacterium acnes, and an extract of a lemon balm fermented using any one strain selected from Saccharomyces cerevisiae, Reviractobacillus brevis, Lacticaseibacillus casei, and Lactiplantibacillus plantarum strain It is desirable to exhibit an antibacterial effect against Cutibacterium acnes, but is not limited thereto.

[0035] An antibacterial composition according to one embodiment of the present invention is preferably Reviractobacillus brevis ( Levilactobacillus brevis ) or Lactibactibacillus plantarum ( Lactiplantibacillus plants Bacillus subtilis (containing lemon balm fermented using the ) strain as an active ingredient Bacillus subtilis ), Bacillus cereus( Bacillus cereus ), Staphilococcus aureus( Staphylococcus aureus ), E. coli( Escherichia coli ), Enterobacter cloa ( Enterobacter cloacae ), Salmonella enterica( Salmonella enterica ) and Pseudomonas eruginosa ( Pseudomonas aeruginosa It may exhibit an antibacterial effect against one or more bacteria selected from ), but is not limited thereto.

[0036] An antimicrobial composition according to one embodiment of the present invention is preferably Bacillus subtilis ( Bacillus subtilis ), Reviractobacillus brevis( Levilactobacillus brevis), Lacticasey Bacillus casei ( Lactobacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plantarum Bacillus subtilis (containing as an active ingredient an extract of lemon balm fermented using any one strain selected from ) strains Bacillus subtilis ) and Cutibacterium acnes( Cutibacterium acnes It may exhibit an antibacterial effect against one or more bacteria selected from ), but is not limited thereto.

[0037] In addition, the present invention provides a cosmetic composition for preventing or improving acne containing a lemon balm ferment or an extract thereof as an active ingredient.

[0038] The above acne may be caused by acne-causing bacteria, and the acne-causing bacteria are preferably Staphylococcus aureus ( Staphylococcus aureus ), Pseudomonas eruginosa( Pseudomonas aeruginosa ) and Cutibacterium acnes( Cutibacterium acnes It is one or more bacteria selected from among ), but is not limited to this.

[0039] In a cosmetic composition according to one embodiment of the present invention, the cosmetic composition may have any one formulation selected from skin, skin softener, skin toner, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, eye cream, moisture cream, hand cream, essence, nourishing essence, pack, cleansing foam, cleansing water, cleansing cream, body lotion, body cleanser, soap, and powder, but is not limited thereto. A cosmetic composition made of each of these formulations may contain various bases and additives that are necessary and appropriate for the formulation of the formulation, and the types and amounts of these ingredients can be easily selected by a person skilled in the art.

[0040] In the case where the formulation of the cosmetic composition of the present invention is a paste, cream, or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0041] When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane-butane, or dimethyl ether.

[0042] When the formulation of the cosmetic composition of the present invention is a solution or an emulsion, a solvent, a solvating agent, or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0043] In the case where the formulation of the cosmetic composition of the present invention is a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracanth may be used as carrier components.

[0044] In the case where the formulation of the cosmetic composition of the present invention is a cleansing product containing a surfactant, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0045] In addition, the present invention provides an antibacterial health functional food composition containing lemon balm ferment or an extract thereof as an active ingredient.

[0046] The health functional food composition of the present invention may be prepared in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0047] When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The active ingredient may be used appropriately according to its intended use (prevention or improvement). Generally, when manufacturing food or beverages, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, relative to the raw material. However, in the case of long-term consumption for health purposes, the above amount may be less than the above range, and since there are no issues regarding safety, the active ingredient may be used in an amount greater than the above range.

[0048] There are no special restrictions on the types of the above-mentioned health functional foods. Examples of foods to which the above-mentioned health functional food composition may be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

[0049] In addition, the health functional food composition of the present invention may be manufactured into a food, particularly a functional food. The functional food of the present invention may include ingredients that are typically added. For example, it includes proteins, carbohydrates, fats, nutrients, and flavoring agents. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredients. The natural carbohydrate is preferably a monosaccharide (e.g., glucose, fructose, etc.), a disaccharide (e.g., maltose, sucrose, etc.), an oligosaccharide, a polysaccharide (e.g., dextrin, cyclodextrin, etc.), or a sugar alcohol (e.g., xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., taumatin, stevia extract, etc.) or a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

[0050] In addition to the above health functional food composition, it may further contain various nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Although the proportion of these added ingredients is not of great importance, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health functional food composition of the present invention.

[0051] In addition, the present invention provides an antibacterial pharmaceutical composition containing a lemon balm ferment or an extract thereof as an active ingredient.

[0052] The above composition is Bacillus subtilis ( Bacillus subtilis ), Bacillus cereus( Bacillus cereus ), Staphilococcus aureus( Staphylococcus aureus ), E. coli( Escherichia coli ), Enterobacter cloa ( Enterobacter cloacae ), Salmonella enterica( Salmonella enterica ), Pseudomonas eruginosa( Pseudomonas aeruginosa ) and Cutibacterium acnes( Cutibacterium acnes It has the effect of preventing or treating diseases caused by infection with one or more bacteria selected from ).

[0053] The pharmaceutical composition according to the present invention can be formulated and used in the form of oral formulations such as capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols, external formulations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

[0054] The pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent.

[0055] The carrier, excipient, and diluent that may be included in the pharmaceutical composition of the present invention are preferably selected from lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto.

[0056] When formulating, the formulation is prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, humectants, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules; these solid dosage forms are prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin, with the above pharmaceutical composition. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid dosage forms for oral administration include suspensions, liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. Witepsol, Macrogol, Tween 61, cocoa paste, laurin paste, glycerogelatin, etc. may be used as bases for suppositories.

[0057] Suitable dosages of the pharmaceutical composition of the present invention can be prescribed in various ways depending on factors such as the formulation method, mode of administration, patient's age, body weight, sex, pathological condition, food, time of administration, route of administration, excretion rate, and response sensitivity.

[0058] The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by topical application to the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

[0059] In addition, the present invention provides an antibacterial quasi-drug containing lemon balm ferment or an extract thereof as an active ingredient.

[0060] The above quasi-drugs may include external skin preparations and personal hygiene products. For example, they may be, but are not limited to, disinfectants, shower foams, mouthwash, wet wipes, detergent soaps, hand washes, or ointments.

[0061] When the above-described quasi-drug according to the present invention is used as an additive to a quasi-drug, the composition may be added as is or used together with other quasi-drugs or quasi-drug ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient can be appropriately determined according to the purpose of use.

[0062] The quasi-drug of the present invention may be manufactured in the form of a general emulsion formulation and a solubilized formulation, for example. For example, it may have a formulation such as a lotion, cream, ointment, spray, oil gel, gel, oil, or aerosol, but it is not limited to any other form as long as it exhibits the antibacterial effect of the present invention.

[0063] In addition, the above-mentioned quasi-drug may be used by appropriately incorporating oils, water, surfactants, moisturizers, lower alcohols having 1 to 4 carbon atoms, thickeners, chelating agents, colorants, preservatives, or fragrances, which are generally incorporated into quasi-drugs, into each formulation as needed.

[0064] In addition, the present invention provides an antibacterial feed additive containing lemon balm ferment or an extract thereof as an active ingredient.

[0065] The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act. In the present invention, the term "feed" may refer to any natural or artificial prescribed food, single meal, etc., or the components of said single meal, intended for or suitable for animals to eat, consume, and digest. The types of said feed are not particularly limited, and feeds commonly used in the relevant technical field may be used. Non-limiting examples of said feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, single-cell proteins, zooplankton, or food waste. These may be used individually or in a mixture of two or more types.

[0066] The present invention will be described in more detail below using manufacturing examples and embodiments. These manufacturing examples and embodiments are intended solely to explain the present invention more specifically, and it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

[0067] Preparation Example 1. Preparation of lemon balm fermented product

[0068] The lemon balm used in the present invention was purchased from Herb Market Co., Ltd. in August 2023.

[0069] The strain used for fermentation is Bacillus subtilis KACC 14549(BS) and Saccharomyces cerevisiae KACC 48234(SC), Levilactobacillus brevis KACC 18270(LB), Lacticaseibacillus casei KACC 12413(LC) and Lactiplantibacillus plantarum KACC 18510(LP) was obtained from the Agricultural Genetic Resources Center and used, and was grown and cultured under the conditions of Table 1 below.

[0070] BS was cultured in nutrient broth (NB), SC in YM broth, and LP, LC, and LB in Lactobacilli MRS broth. After subculturing a total of three times at 30 or 37°C for 24 hours, the absorbance was measured at 600 nm, and the absorbance values ​​were set to be within the range of 0.4 to 0.6 to be used as fermentation strains.

[0071] strain badge Temperature (°C) Bacillus subtilis KACC 14549(BS) NB 30 Saccharomyces cerevisiae KACC 48234(SC) YM 30 Levilactobacillus brevis KACC 18270(LB) MRS 37 Lacticaseibacillus casei KCTC 12413(LC) MRS 37 Lactiplantibacillus plantarum KCTC 18510(LP) MRS 37

[0072] The manufacturing process of the lemon balm fermentation product is specifically as follows.

[0073] 5g of glucose and 1.25g of peptone were dissolved in 250mL of water, and then the water containing the dissolved glucose and peptone was pressurized at 121℃ for 15 minutes. After that, 2.5mL of the activated fermentation strain was inoculated into the sterilized water, 12.5g of lemon balm was added, and then cultured at 30 or 37℃ for 24 hours to obtain a culture solution. Afterwards, the culture medium was pressurized at high temperature at 121°C for 15 minutes, filtered using filter paper (Whatman No. 2, Whatman International Ltd., Maidstone, UK), stored at -50°C for 12 hours, and then freeze-dried in a freeze-dryer (FD8508, Ilshin Lab Co., Ltd., Yangju-si, Gyeonggi-do, South Korea) to obtain a powder (lemon balm fermentation product), which was used as the sample for the experiment.

[0074] The preparation process of the lemon balm water extract used as a non-fermented control is as follows.

[0075] 12.5g of lemon balm powder was mixed with 250mL of distilled water and extracted at room temperature for 24 hours. The resulting liquid was filtered using filter paper (Whatman No. 2), cooled under the same conditions as the fermentation group, and freeze-dried to obtain a powder (lemon balm water extract), which was used as the control sample for the experiment.

[0076] Each powder was dissolved in distilled water for the experiment.

[0077] Preparation Example 2. Extract of fermented lemon balm

[0078] Ten times the amount of 70% (v / v) ethanol was added relative to the lemon balm fermentation powder prepared in Preparation Example 1, followed by ultrasonic extraction at 60°C for 1 hour, and centrifugation (4,000 rpm, 5 min) was performed three times. Subsequently, the supernatant obtained from centrifugation was filtered (Whatman No. 4, GE Healthcare Life Sciences), concentrated under reduced pressure, and then freeze-dried to obtain a powder (lemon balm fermentation extract), which was used as the sample for the experiment.

[0079] Example 1. Analysis of culture characteristics of lemon balm fermented product according to fermentation strain

[0080] 1) Yield measurement

[0081] The lemon balm fermentation product cultured for 24 hours was filtered (Whatman No. 2) and then dried using a freeze-dryer (FD8518, Ilshin Lab Co. Ltd), and the yield was calculated by substituting it into Equation 1 below.

[0082] [Equation 1]

[0083] Yield of lemon balm ferment = (Weight of freeze-dried fermented powder (g) / Weight of raw material before fermentation (g)) × 100

[0084] The change in yield due to fermentation is as disclosed in Table 2 below.

[0085] zymogen transference number(%) Control (non-fermented lemon balm extract) 13.19 BS 53.53 SC 25.27 LC 45.86 LB 43.74 LP 46.46

[0086] 2) pH measurement

[0087] Changes in pH of the culture medium due to the growth of the strain were measured by filtering the culture medium through filter paper (Whatman No. 2) after 24 hours of culture and then using a pH meter (Neomet pH-200L, Istek Co., Seoul, Korea).

[0088] As a result, as disclosed in Table 3, it was confirmed that fermented lemon balm extract underwent overall fermentation compared to non-fermented lemon balm extract, and that the pH was lowered and the growth of harmful bacteria could be inhibited.

[0089] zymogen pH Control (non-fermented lemon balm extract) 5.76±0.01 a BS 5.44±0.01 b SC 5.32±0.01 c LC 5.18±0.00 d LB 3.59±0.01 f LP 3.63±0.00 e

[0090] Different characters a~f within the columns of the table indicate that there are statistically significant differences between them, p It is <0.05.

[0091] 3) Turbidity measurement

[0092] To determine the degree of microbial fermentation, turbidity was measured by measuring the absorbance at 600 nm of the filtered fermented liquid after 24 hours of culture (UV-1800, Shimadzu, Kyoto, Japan).

[0093] As a result, it was confirmed that the turbidity varied depending on the type of fermentation bacteria, as disclosed in Table 4.

[0094] zymogen Turbidity Control (non-fermented lemon balm extract) 0.29±0.00 f BS 0.84±0.02 b SC 0.56±0.00 e LC 0.61±0.00 d LB 0.76±0.001 c LP 1.05±0.01 a

[0095] Different characters a~f within the columns of the table indicate that there are statistically significant differences between them, p It is <0.05.

[0096] 4) Measurement of viable cell count

[0097] Changes in viable cell count: After diluting the lemon balm fermented product cultured for 24 hours in physiological saline and mixing thoroughly, BS ( B. subtilis KACC 14549), LB( L. brevis KACC 18270), LC( L. casei KACC 12413), LP( L. plantarum KACC 18510) is used for PCA (plate count agar), SC ( S. cerevisiae KACC) was confirmed by measuring the viable cell count after dispensing onto PDA (potato dextrose agar) and incubating at 37°C and 30°C for 24 hours, respectively.

[0098] As a result, it was confirmed that bacterial growth occurred in all strain fermented products, as disclosed in Table 5.

[0099] zymogen Viable cell count (log CFU / mL) Control (non-fermented lemon balm extract) - BS 6.57±0.02 d SC 7.85±0.05 b LC 7.55±0.04 c LB 7.99±0.008 a LP 7.77±0.07 b

[0100] Different characters a~f within the columns of the table indicate that there are statistically significant differences between them, p It is <0.05.

[0101] Example 2. Antimicrobial activity

[0102] The antimicrobial activity of lemon balm ferment and its extracts was measured against each strain using the disk diffusion assay.

[0103] The strain used to measure antimicrobial activity was a Gram-positive bacterium, Bacillus cereus ( Bacillus cereus, B. cereus ) KCTC 1012, Bacillus subtilis( Bacillus subtilis, B. subtilis ) KACC 14549, Staphylococcus aureus( Staphylococcus aureus, S. aureus ) KCTC 3881 and Cutibacterium acnes( Cutibacterium acnes , C. acnes ) KACC 19984, as a Gram-negative bacterium, Escherichia coli ( Escherichia coli , E. coli ) KCTC 2441, Enterobacter cloa( Enterobacter cloacae, E. cloacae ) KCTC 1685, Salmonella enterica( Salmonella enterica subsp. enterica ) KCTC 2515 and Pseudomonas eruginosa( Pseudomonas aeruginosa, P. aeruginosa ) KCTC 1636 was used.

[0104] Dormant strains were inoculated into NA liquid medium (nutrient broth, Difco Laboratories) and Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Enterobacter cloaca were subcultured three times for 24 hours each at 30°C, and Salmonella enterica, Pseudomonas eruginosa, and Cutibacterium acnes were subcultured at 37°C. After confirming that the growth of the strains had occurred by ensuring that the absorbance value at 600 nm fell within the range of 0.4 to 0.6, they were used.

[0105] 0.1 mL of each activated strain was dispensed onto NA solid medium and spread using a sterile spreader to prepare a plate medium for antimicrobial testing. The sample was slowly absorbed onto paper discs (8 mm) to obtain 10 mg per disc, and after drying to evaporate the solvent, the discs were placed in close contact with the plate medium and incubated at 30°C and 37°C for 24 hours. Subsequently, the diameter (mm) of the clear zone formed around the disc was measured to compare the antimicrobial activity.

[0106] As a result of measuring the antibacterial effect of the lemon balm fermentation product of the present invention against seven types of microorganisms including Bacillus cereus, as disclosed in Table 6, the non-fermented lemon balm extract (Control) did not show an antibacterial effect, but the Reviractobacillus brevis (LB; L. brevis ) or Lactibacillus plantarum (LP; L. plantarum Lemon balm fermented product prepared by fermentation with the ) strain exhibited antibacterial effects against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Enterobacter cloaca, Salmonella enterica, and Pseudomonas eruginosa, and Bacillus subtilis (BS; B. subtilis Lemon balm fermented product prepared by fermenting with a strain showed an antibacterial effect against Enterobacter cloaca.

[0107] In addition, the antibacterial effect of the extract of the lemon balm fermentation product against eight types of microorganisms, including Cutibacterium acnes, was measured. As disclosed in Table 7, the extract of the fermentation product fermented with Bacillus subtilis, Lacticasei, Bacillus casei, Reviractobacillus brevis, or Lactiplantibacillus plantarum showed an antibacterial effect against Bacillus subtilis, and in particular, the extract of the lemon balm fermentation product fermented with Bacillus subtilis showed an antibacterial effect against Bacillus subtilis, which had not shown an antibacterial effect in the lemon balm fermentation product fermented with Bacillus subtilis.

[0108] In addition, the extract of the fermented product prepared by fermentation with Bacillus subtilis or Lactibacillus plantarum showed an antibacterial effect against Staphylococcus aureus, and it was confirmed that this was an effect opposite to that of the lemon balm fermented with Bacillus subtilis, which did not show an antibacterial effect against Staphylococcus aureus.

[0109] Meanwhile, the extract of the fermented product fermented with Lactibacillus plantarum showed an antibacterial effect against Salmonella enterica, but the effect was not superior compared to the fermented product fermented with Lactibacillus plantarum.

[0110] In terms of antibacterial effects against Cutibacterium acnes, the extracts of lemon balm fermented by fermentation with Bacillus subtilis, Lacticasei, Bacillus casei, Reviractobacillus brevis, Lactiplantibacillus plantarum, or Saccharomyces cerevisiae showed excellent antibacterial effects, and in particular, the extracts of lemon balm fermented by fermentation with Bacillus subtilis or Lactiplantibacillus plantarum strains showed significant antibacterial effects (Table 7).

[0111] microorganism Sample Size of growth inhibition zone (mm) 5mg / disc 10mg / disc Bacillus cereus Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 1.88±0.29 * 5.03±0.16 * LP 2.34±0.28 * 6.22±0.20 * Bacillus subtilis Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 3.03±0.22 * 8.80±0.21 * LP 2.58±0.11 * 7.57±0.15 * Staphilococcus aureus Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 1.57±0.17 * 3.97±0.10 * LP 1.68±0.12 * 3.84±0.08 * E. coli Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 2.66±0.07 * 5.75±0.18 * LP 2.42±0.06 * 4.12±0.09 * Enterobacter claw bird Control (non-fermented lemon balm extract) - - BS 1.37±0.07 * 2.17±0.06 * SC - - LC - - LB 2.60±0.18 * 6.65±0.16 * LP 1.38±0.10 * 2.24±0.13 * Salmonella enterica Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 2.15±0.06 * 4.92±0.10 * LP 2.24±0.07 * 5.41±0.10 * Pseudomonas eruzinosa Control (non-fermented lemon balm extract) - - BS - - SC - - LC - - LB 4.20±0.07 * 6.03±0.07 * LP 4.93±0.36 * 6.94±0.12 *

[0112] * This means that there is a statistically significant difference compared to non-fermented lemon balm extract, p It is <0.05.

[0113] microorganism Sample Size of growth inhibition zone (mm) 10mg / disc Bacillus cereus BS - SC - LC - LB - LP - Bacillus subtilis BS 1.73±0.43 SC - LC 1.95±0.19 LB 1.68±0.10 LP 2.18±0.39 Staphilococcus aureus BS 3.93±0.05 SC - LC - LB - LP 2.59±0.14 E. coli BS - SC - LC - LB - LP - Enterobacter claw bird BS - SC - LC - LB - LP - Salmonella enterica BS - SC - LC - LB - LP 2.18±0.39 Pseudomonas eruzinosa BS - SC - LC - LB - LP - Cutibacterium acnes BS 20.40±0.77 SC 13.65±0.78 LC 11.25±0.29 LB 13.53±0.35 LP 36.27±1.36

Claims

Claim 1 Lemon balm Melissa officinalis An antibacterial composition containing a fermented product or an extract thereof as an active ingredient. Claim 2 In paragraph 1, the lemon balm fermented product is Bacillus subtilis ( Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lacticaseibacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plantarum An antimicrobial composition characterized by being fermented using any one strain selected from among ) strains. Claim 3 In claim 1, the lemon balm fermented product comprises: 1) a step of sterilizing water in which glucose and peptone are dissolved; 2) Bacillus subtilis (in the sterilized water of step 1). Bacillus subtilis ), Saccharomyces cerevisiae( Saccharomyces cerevisiae ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lacticaseibacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plantarum An antibacterial composition characterized by comprising: a step of inoculating one strain selected from among strains, adding lemon balm, and then culturing at 30 to 40°C for 12 to 48 hours to obtain a culture solution; and a step of, after step 2), sterilizing the obtained culture solution and then filtering it to obtain a filtrate. Claim 4 An antibacterial composition according to claim 1, wherein the extraction solvent of the extract is water, a C1-C4 lower alcohol, or a mixture thereof. Claim 5 In paragraph 1, the antibacterial agent is Bacillus subtilis ( Bacillus subtilis ), Bacillus cereus( Bacillus cereus ), Staphilococcus aureus( Staphylococcus aureus ), E. coli( Escherichia coli ), Enterobacter cloa ( Enterobacter cloacae ), Salmonella enterica( Salmonella enterica ), Pseudomonas eruginosa( Pseudomonas aeruginosa ) and Cutibacterium acnes( Cutibacterium acnes An antimicrobial composition characterized by being antimicrobial against one or more bacteria selected from ). Claim 6 In paragraph 1, Reviractobacillus brevis ( Levilactobacillus brevis ) or Lactibactibacillus plantarum ( Lactiplantibacillus plantarum Bacillus subtilis (containing lemon balm fermented using the ) strain as an active ingredient Bacillus subtilis ), Bacillus cereus( Bacillus cereus ), Staphilococcus aureus( Staphylococcus aureus ), E. coli( Escherichia coli ), Enterobacter cloa ( Enterobacter cloacae ), Salmonella enterica( Salmonella enterica ) and Pseudomonas eruginosa ( Pseudomonas aeruginosa An antimicrobial composition characterized by exhibiting an antimicrobial effect against one or more bacteria selected from ). Claim 7 In paragraph 1, Bacillus subtilis ( Bacillus subtilis ), Reviractobacillus brevis( Levilactobacillus brevis ), Lacticasey Bacillus casei ( Lacticaseibacillus casei ) and Lactibacillus plantarum ( Lactiplantibacillus plantarum Bacillus subtilis (containing as an active ingredient an extract of lemon balm fermented using any one strain selected from ) strains Bacillus subtilis ) and Cutibacterium acnes( Cutibacterium acnes An antimicrobial composition characterized by exhibiting an antimicrobial effect against one or more bacteria selected from ). Claim 8 A cosmetic composition for preventing or improving acne containing lemon balm ferment or an extract thereof as an active ingredient. Claim 9 Antibacterial health functional food composition containing lemon balm ferment or its extract as an active ingredient. Claim 10 Antimicrobial pharmaceutical composition containing lemon balm ferment or its extract as an active ingredient. Claim 11 An antibacterial quasi-drug containing lemon balm ferment or an extract thereof as an active ingredient. Claim 12 Antimicrobial feed additive containing lemon balm ferment or its extract as an active ingredient.