Composition comprising Saccharomyces cerevisiae MGE 3400 applied distilled soju and by-product and uses thereof

Abstract

The present invention relates to a novel strain of the genus Saccharomyces, a culture thereof, and uses thereof, specifically to a novel strain of Saccharomyces cerevisiae, a culture thereof, distilled soju and by-products incorporating the Saccharomyces cerevisiae MGE 3400 strain, and uses thereof, for example, for improving skin health functions.

Description

Technology Field

[0001] The present invention relates to a novel strain of the genus Saccharomyces, a culture thereof, and uses thereof, specifically to a novel strain of Saccharomyces cerevisiae, a culture thereof, distilled soju and by-products incorporating the Saccharomyces cerevisiae MGE 3400 strain, and uses thereof, for example, for improving skin health functions. Background Technology

[0003] The distilled soju industry is expanding amidst a recent surge in consumer preference for the drink. Nevertheless, brewing yeast is often purchased for use, making it difficult to highlight product differentiation. Therefore, discovering and applying brewing yeast to products enables the development of unique items and guarantees consumer choice.

[0004] On the other hand, when producing distilled soju, by-products (sediment) are generated, and since these by-products are rich in organic matter, they must be treated as wastewater, which may result in additional costs.

[0005] In particular, by-products of distilled soju produced using rice as a raw material are rich in nutrients, and since various metabolites are produced during the fermentation process involving lactic acid bacteria and yeast, they can serve as excellent materials for health benefits.

[0006] Therefore, if yeast strains that enhance the value of distilled soju and its by-products are discovered and utilized as starter cultures, it will be possible to manufacture unique distilled soju and produce functional by-products.

[0007] Against this technical background, the inventors of the present application isolated a novel Saccharomyces cerevisiae strain and utilized it as a starter culture for the production of distilled soju, thereby developing a unique distilled soju. Furthermore, by confirming that the by-products can be utilized for the purpose of improving skin health functions, the present invention was completed. The problem to be solved

[0009] To solve the above-mentioned problems, the objective of the present invention is to provide a novel Saccharomyces cerevisiae strain or a culture thereof.

[0010] The object of the present invention is to provide distilled soju produced using a novel Saccharomyces cerevisiae strain or a culture thereof.

[0011] The object of the present invention is to provide a byproduct of distilled soju produced by fermenting grains with the above strain or a culture thereof.

[0012] The objective of the present invention is to provide a use for improving skin health functions comprising the above-mentioned distilled soju byproduct. means of solving the problem

[0014] To achieve the above objective, the present invention relates to Saccharomyces cerevisiae MGE 3400 of accession number KCTC 16217BP ( Saccharomyces cerevisiae Provides the MGE 3400) strain or a culture thereof.

[0015] The present invention provides distilled soju produced by fermenting grains with the strain or culture thereof.

[0016] The present invention provides a byproduct of distilled soju produced by fermenting grains with the above strain or culture thereof.

[0017] The present invention provides a composition for improving skin health functions comprising the above-mentioned distilled soju byproduct. Effects of the invention

[0019] According to the present invention, it is necessary to discover yeast strains with excellent alcohol-producing capabilities for the efficient production of distilled soju. Although by-products generated during the production of distilled soju are treated as wastewater, incurring additional costs, by-products produced using rice as a raw material are rich in nutrients and can serve as materials with excellent health benefits, as various metabolites are produced during the fermentation process involving lactic acid bacteria and yeast. Therefore, it is believed that if yeast strains that enhance the value of distilled soju and its by-products are discovered and utilized as starter cultures, it will be possible to manufacture unique distilled soju and produce functional by-products. Brief explanation of the drawing

[0021] Fig. 1. Process diagram for distilled soju manufacturing and byproduct production. Fig. 2. Number of lactic acid bacteria during makgeolli fermentation with Saccharomyces cerevisiae MGE 3400 strain (days 2 and 3). MGE3400, Saccharomyces cerevisiae Isolated strain; commercial strain, La Parisienne, Saccharomyces cerevisiae (DB ingredients, UK). Fig. 3. Inhibitory activity of pathogenic microbial growth in distilled soju by-products treated with Saccharomyces cerevisiae MGE 3400 strain. MGE3400, Saccharomyces cerevisiae Isolated strain; commercial strain, La Parisienne, Saccharomyces cerevisiae (DB ingredients, UK). Fig. 4. Antioxidant activity of distilled soju byproducts treated with Saccharomyces cerevisiae MGE 3400 strain. Cont, Ascorbic acid. Fig. 5. Cytotoxicity of distilled soju byproducts applied to Saccharomyces cerevisiae MGE 3400 strain on skin keratinocytes (HaCaT). Fig. 6. Effect of distilled soju byproduct applied with Saccharomyces cerevisiae MGE 3400 strain on alleviating skin damage caused by ultraviolet rays. Fig. 7. Analysis of skin moisturizing efficacy (keratinocyte HAS2 expression) of distilled soju byproducts treated with Saccharomyces cerevisiae MGE 3400 strain. MGE3400, Saccharomyces cerevisiae Isolated strain; commercial strain, La Parisienne, Saccharomyces cerevisiae (DB ingredients, UK). Specific details for implementing the invention

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled expert in the art to which this invention pertains. In general, the nomenclature used herein is well known and commonly used in the art.

[0023] In one aspect, the present invention relates to Saccharomyces cerevisiae MGE 3400 of accession number KCTC 16217BP ( Saccharomyces cerevisiae This relates to the MGE 3400) strain or a culture thereof.

[0024] The strain according to the present invention was deposited with the Korea Research Institute of Biotechnology and Bioengineering on January 2, 2025. The Saccharomyces cerevisiae strain of accession number KCTC16217BP is described as MGE 3400.

[0025] With respect to the culture of the strain above, "culture" may mean cultured in a culture medium or culture solution containing the strain. The culture contains the strain. The culture may be in a liquid or solid form, but is not limited thereto.

[0026] Various forms of the above culture may include, for example, concentrates and dried products of the culture.

[0027] The present invention also relates to distilled soju produced by fermenting grains with a strain or a culture thereof.

[0028] The above distilled soju may be characterized by being produced by, for example, mixing koji and a strain or a culture thereof for a first fermentation, then adding white rice and water for a second fermentation, and then fermenting and distilling.

[0029] The above koji is prepared by washing, soaking, and steaming the rice, and then adding Saccharomyces cerevisiae MGE 3400 of accession number KCTC 16217BP to the steamed rice ( Saccharomyces cerevisiae A single-stage immersion can be prepared by inoculating the MGE 3400) strain or a culture thereof.

[0030] After washing, soaking, and steaming white rice, steamed white rice and water are added to the first stage of fermentation to produce a second stage of fermentation, which can then be fermented and distilled. After fermentation is complete, the alcohol content of the finished fermented liquor can be, for example, about 15%.

[0031] After the above distillation, the distilled soju byproduct may include "sake lees," "distillation lees," or "freeze-dried material."

[0032] Based on this, the present invention may be a byproduct of distilled soju produced by fermenting grains with the above strain or a culture thereof.

[0033] In the present invention, "sulk" refers to the distillation residue of soju, which is the residue remaining after obtaining soju through fermentation and distillation. In the present invention, "distillation sulk" may be a distillation byproduct, for example, a distilled soju byproduct. In the present invention, "freeze-dried material" refers to a product that has been frozen and dried, meaning that the supernatant of a distilled soju byproduct is filtered through a filter after centrifugation and then freeze-dried.

[0034] The above fermentation refers to the process of denaturing or modifying raw materials using strains through physical or chemical methods, and can refer to a metabolic process that allows obtaining a desired result in a short period of time, whereas it would naturally take a long time to change.

[0035] The above fermented product refers not only to a product fermented by a strain but may also include a fermented filtrate filtered after fermentation or a fermented extract obtained by extracting the fermented product with a solvent.

[0036] The above fermented product is a biotransformed product, and the biotransformed product itself may be included in the composition according to the present invention.

[0037] The present invention relates to a composition for improving skin health functions comprising the above-mentioned distilled soju byproduct.

[0038] "Improvement" refers to any action that alleviates the symptoms of skin-related diseases or changes them for the better.

[0039] The above-mentioned improvement of skin health may include, for example, the improvement of skin diseases. Specifically, it may include the improvement of acne, an inflammatory skin disease in which comedones, papules, pustules, nodules, pseudocysts, etc. occur on the face, neck, chest, back, and shoulders. If left untreated, acne can leave permanent scars, which is a cosmetic issue and can cause psychological burden to the patient.

[0040] Through the present invention, one or more selected from the group consisting of the following are provided. Staphylococcus aureus KCTC 3881, Listeria monocytogenes KCTC 3569, Cutibacterium acnes KCTC 3314 and Candida albicans KCTC 7270 may be characterized by exhibiting antibacterial activity against skin and intestinal pathogenic microorganisms.

[0041] The present invention may also be characterized by representing one or more selected from the group consisting of the following.

[0042] (1) Skin antioxidant activity;

[0043] (2) Activity to repair skin damage caused by ultraviolet rays; and

[0044] (3) Skin moisturizing effect.

[0045] The above composition may be used as a pharmaceutical composition. The above pharmaceutical composition may be prepared in a unit dose form or contained in a multi-dose container by formulation using a pharmaceutically acceptable carrier and / or excipient, according to a method that can be easily carried out by a person skilled in the art to which the present invention belongs. In this case, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granule, tablet, capsule, or gel (e.g., hydrogel), and may additionally include a dispersant or a stabilizer.

[0046] Pharmaceutically acceptable carriers may include, but are not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, acacia, rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, which are commonly used in formulations. In addition, in addition to the above components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. may be additionally included.

[0047] The above pharmaceutical composition can be administered orally or parenterally and can be used in the form of general pharmaceutical formulations. That is, the pharmaceutical composition of the present invention can be administered in various oral and parenteral dosage forms during actual clinical administration; when formulated, it is prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with herbal extracts or fermented herbal products. In addition, lubricants such as magnesium stirate and talc are also used in addition to simple excipients. Liquid formulations for oral administration include suspensions, oral liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. Formulations 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 suspension solvents. Witepsol, Macrogol, Tween 61, cacao oil, laurin oil, glycerol, and gelatin may be used as bases for suppositories.

[0048] The concentration of the active ingredient included in the above composition may be determined by considering the purpose of treatment, the patient's condition, the duration of treatment, etc., and is not limited to a specific range of concentrations. The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, a "pharmaceutically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with a therapeutic agent for a disease caused by other contaminants, and may be administered simultaneously, separately, or sequentially with conventional therapeutic agents, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by considering all of the above factors, and this can be easily determined by a person skilled in the art. The effective dose may vary depending on the patient's age, gender, condition, body weight, absorption rate, inactivation rate, and excretion rate of the active ingredient in the body, the type of disease, and concomitant drugs.

[0049] The above composition can be used as a food composition. Based on this, the present invention relates to a health functional food comprising the above composition.

[0050] "Health functional food" means food manufactured and processed using raw materials or ingredients that have functional properties useful to the human body pursuant to Article 6727 of the Act on Health Functional Foods, and "functionality" means consuming for the purpose of obtaining useful effects for health purposes, such as regulating nutrients or physiological actions on the structure and function of the human body.

[0051] The above composition may include ordinary food additives, and unless otherwise specified, suitability as a "food additive" shall be determined in accordance with the specifications and standards for the relevant item, in accordance with the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety.

[0052] Examples of items listed in the above "Food Additives Codex" include chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid; natural additives such as persimmon dye, licorice extract, crystalline cellulose, sorghum dye, and guar gum; and mixed preparations such as L-sodium glutamate preparations, alkaline noodle additives, preservative preparations, and tar dye preparations.

[0053] The above food composition can be manufactured and processed in the form of tablets, capsules, powder, granules, liquid, pills, etc.

[0054] The above-mentioned health functional food in tablet form may be granulated by a conventional method with a mixture of excipients, binders, disintegrants, and other additives in addition to the active ingredient, and then compressed by adding a lubricant, etc., or the mixture may be compressed directly. In addition, the above-mentioned health functional food in tablet form may contain a binder, etc., as needed, and may be coated with a suitable coating agent as needed.

[0055] Among the above-mentioned capsule-type health functional foods, hard capsules can be manufactured by filling a conventional hard capsule with a mixture of active ingredients and additives such as excipients, or the granules thereof, or coated granules, and soft capsules can be manufactured by filling a capsule base such as gelatin with a mixture of active ingredients and additives such as excipients. The above-mentioned soft capsules may contain plasticizers such as glycerin or sorbitol, coloring agents, preservatives, etc., as needed.

[0056] Health functional foods in the form of pills can be prepared by molding a mixture of active ingredients, excipients, binders, disintegrants, etc., in a suitable way, and if necessary, the pills may be coated with sucrose or other suitable coating agents, or coated with starch, talc, or a suitable substance.

[0057] A granular health functional food can be manufactured into a granular form by a suitable method using a mixture of active ingredients, excipients, binders, disintegrants, etc., and may contain flavoring agents, mellowing agents, etc. as needed. When the granular health functional food is subjected to the following particle size test using No. 12 (1680 μm), No. 14 (1410 μm), and No. 45 (350 μm) sieves, the entire amount may pass through the No. 12 sieve, and the amount remaining on the No. 14 sieve may be 50% or less of the total amount, and the amount passing through the No. 45 sieve may be 15% or less of the total amount.

[0058] The above composition may be a cosmetic composition. The above composition may be used as a cosmetic.

[0059] The above cosmetic composition may further include vitamins, peptides, polysaccharides, lipids, etc., and may also include, but is not limited to, oil components, moisturizers, surfactants, pigments, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, fragrances, purified water, etc., which are commonly incorporated into cosmetics.

[0060] The above cosmetic composition may be a formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, oils, waxes, powders, sprays, soaps, cleansing products, packs, foundations, makeup bases, and hair cosmetics, but is not limited thereto. More specifically, it may be prepared in the form of softening lotions, nourishing lotions, nourishing creams, massage creams, essences, eye creams, hair tonics, shampoos, rinses, cleansing creams, cleansing foams, cleansing waters, packs, sprays, and powders.

[0061] In the case where the formulation is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0062] In the case where the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and especially in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0063] When the formulation is a solution or emulsion, a solvent, a solubilizing 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, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0064] In the case where the formulation is a suspension, liquid diluents such as water, ethanol, or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used as carrier components.

[0065] In the case where the formulation is a cleansing product containing a surfactant, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, 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.

[0067] The present invention will be described in more detail below through examples. These examples are intended solely to illustrate the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be interpreted as being limited by these examples.

[0069] 실시예

[0071] 1. Preparation of Takju for Distilled Soju

[0073] Rice grown in Chungju was purchased and used, and white koji was purchased from Joeun Goksik Co., Ltd. For the preparation of the mash, Sumizyme (Shin Nihon Chemical, Anjyo, Aichi, Japan) was used as the enzyme agent, and commercial yeast (La Parisienne, Saccharomyces cerevisiae , DB ingredients, UK) or isolated strains (derived from Nuruk Makgeolli; Sacharomyces cerevisiae MGE 3400) was used. The process flow diagram for distilled soju production and byproduct production is as shown in Fig. 1.

[0075] 2. Measurement of Takju alcohol content and lactic acid bacteria count

[0077] The alcohol content and number of lactic acid bacteria in makgeolli treated with a commercial strain of Saccharomyces cerevisiae and the MGE 3400 strain were compared and analyzed.

[0078] Strain isolated during small-scale immersion (3L) and trial production immersion (600L) S. cerevisiae When MGE 3400 was used as a starter culture, the alcohol content increased compared to commercial strains, confirming that it has excellent alcohol production ability (Table 1).

[0080] Table 1. Alcohol content of Makgeolli treated with Saccharomyces cerevisiae MGE 3400 strain

[0081]

[0082] MGE 3400, Saccharomyces cerevisiae Isolated strain; commercial strain, La Parisienne, Saccharomyces cerevisiae (DB ingredients, UK).

[0084] In addition, it was confirmed that the number of lactic acid bacteria during fermentation was also improved when the isolated strain was applied (Fig. 2).

[0086] 3. Analysis of Aroma Components in Distilled Soju

[0087] Saccharomyces cerevisiae The aroma components of distilled soju treated with the MGE 3400 strain were compared and analyzed. The analysis conditions are as follows.

[0089]

[0090] 60% of the distilled spirit was transferred to a 20ml Headspace vial, extracted, and analyzed under the conditions shown in the table below.

[0092] Table 2. Headspace Extraction Method

[0093]

[0095] Table 3. Headspace Extraction Analysis Method

[0096]

[0098] <Solvent extraction gas chromatography / mass spectrometry>

[0099] Distilled soju prepared by adjusting the alcohol content to 30% using water from distilled base liquor was used. 10 ml of dichloromethane (Sigma Aldrich Co.) was added to 50 ml of the soju sample and extracted for a total of 3 hours. The final extract was used as an analytical sample after removing excess water using 3 g of anhydrous sodium sulfate. The analytical method is as follows.

[0101] Table 4. Solvent Extraction Analysis Method

[0102]

[0104] Volatile fragrance components were identified using retention indices (RI), the Wiley / 7n mass spectral database (Agilent Co., Palo Alto, CA, USA) and fragrance characteristics.

[0106] Table 5. Saccharomyces cerevisiae Analysis of Aroma Components in Distilled Soju Prototype Using MGE 3400 Strain (HS Extraction Method)

[0107]

[0109] Table 6. Saccharomyces cerevisiae Analysis of Aroma Components in Distilled Soju Prototype Using MGE 3400 Strain (Solvent Extraction Method)

[0110]

[0112] As a result of the analysis, distilled soju using commercial yeast LP showed high levels in the order of isobutyl alcohol (36.271%), 1-propanol (34.6%), and isoamyl alcohol (29.129%), while distilled soju using another commercial yeast H1 showed high levels in the order of isobutyl alcohol (46.573%), isoamyl alcohol (30.857%), and 1-propanol (22.57%), and distilled soju using the selected MGE 3400 strain showed high levels in the order of isobutyl alcohol (47.065%), isoamyl alcohol (34.528%), and 1-propanol (18.407%) (Table 5).

[0113] Furfural, an aldehyde compound, was detected only in LP, and is known as a representative compound that imparts a burnt smell to distilled spirits (Table 6).

[0114] Ester compounds were detected at high levels in the order of LP, H1, and MGE 3400 strain-treated distilled soju. Ethyl caproate, ethyl caprylate, and isoamyl acetate, which produce a sweet fruity aroma, were detected at the highest levels in LP, followed by H1 and MGE 3400 strain-treated distilled soju. These compounds are formed by the ester bonding of higher fatty acids and higher alcohols and are known to produce strong flavors due to their low threshold values ​​(Table 6).

[0115] Distilled soju using the MGE 3400 yeast strain has a low 1-propanol content and a high ratio of isoamyl alcohol and isobutyl alcohol, so it has less spiciness and bitterness, while emphasizing banana aroma or wine flavor, which is thought to be sensorially advantageous.

[0117] 4. Antimicrobial activity

[0119] The antimicrobial activity of distilled soju byproducts against pathogenic microorganisms was compared and analyzed using a commercial strain of Saccharomyces cerevisiae and the MGE 3400 strain.

[0120] The byproduct was prepared by centrifuging (15,000 rpm, 10 min) the lees remaining after the production of distilled soju, collecting the supernatant, centrifuging it again using the same method, sterile filtering (0.45 µm membrane filter), and freeze-drying the supernatant. It was stored in a freezer (-20℃) and dissolved in 1x PBS for testing when necessary.

[0121] Final concentration 50,000 ppm ( Cutibacterium acnes As a result of comparing the antimicrobial activity against each byproduct using 25,000 ppm for KCTC 3314, all pathogenic microbial strains used ( Staphylococcus aureus KCTC 3881, Listeria monocytogenes KCTC 3569, Cutibacterium acnes KCTC 3314, Candida albicansThe antibacterial activity of the distilled soju byproduct treated with the Saccharomyces cerevisiae MGE 3400 strain was found to be excellent in KCTC 7270 (Fig. 3). Therefore, it is believed that the distilled soju byproduct treated with the MGE 3400 strain can effectively control pathogenic microorganisms affecting the intestines and skin.

[0123] 5. Antioxidant activity

[0125] The antioxidant activity of distilled soju byproducts treated with the Saccharomyces cerevisiae MGE 3400 strain was analyzed. The byproduct (final concentration 50,000 ppm) was treated with ABTS reagent, and the reaction was carried out in the dark for 20 minutes. After centrifugation, only the supernatant was transferred to a 96-well plate, and the absorbance was measured (ABTS: OD734).

[0126] Antioxidant activity was expressed as the ratio of the value of the sample (50,000 ppm) to the concentration of 50 ppm, which shows a similar value on the Sodium L-ascorbate standard curve.

[0127] Antioxidant capacity (%) = OD(byproduct) / OD(L-Ascorbate) x 100

[0128] As a result of the test, it was confirmed that it has antioxidant activity at the concentration used (Fig. 4).

[0130] 6. HaCaT Toxicity

[0132] Skin keratinocytes 1.0 x 10 5100 µl of cells / ml was dispensed into each well of a 96-well plate. The samples were incubated for 24 hours at 37°C under 5% CO2 conditions, and after confirming that the cells had adhered to the bottom, they were washed twice with 1x PBS. 100 µl of DMEM (free of FBS and antibiotics) was added, and the samples were incubated (starvated) for 24 hours under the same conditions, followed by washing twice with 1x PBS. The samples were mixed with DMEM (free of FBS and antibiotics) to adjust the final concentrations to 10,000, 7,500, 5,000, 2,500, 1,000, and 500 ppm. 100 µl was dispensed into each well and incubated for 12 and 24 hours under the same conditions. MTT reagent was prepared at a concentration of 5,000 ppm, 10 µl was dispensed into each well, and the reaction was carried out for 2 hours under the same conditions. After removing the medium, 100% DMSO 100 µl was dispensed and the absorbance (OD570) was measured after shaking incubation (150 rpm).

[0134] Cytotoxicity (%) = OD Sample / OD Control x 100

[0136] Test results confirmed that the byproduct did not show toxicity to skin keratinocytes up to 7,500 ppm at 12 hours and up to 5,000 ppm at 24 hours (Fig. 5).

[0138] 7. Effect of alleviating skin damage caused by UV rays

[0140] Skin keratinocytes 1.0 x 10 5100 µl of cells / ml were dispensed into each well of a 96-well plate. The samples were incubated for 24 hours at 37°C under 5% CO2 conditions. After confirming that the cells had adhered to the bottom, the plates were washed twice with 1x PBS. 100 µl of 1x PBS was added, followed by exposure to UV light for 30 minutes. After removing the 1x PBS, the samples were mixed with DMEM (free of FBS and antibiotics) to adjust the final concentrations to 10,000, 7,500, 5,000, 2,500, 1,000, and 500 ppm. 100 µl of each solution was dispensed into each well and incubated for 12 and 24 hours under the same conditions. MTT reagent was prepared at a concentration of 5,000 ppm, 10 µl was dispensed into each well, and the reaction was carried out for 2 hours under the same conditions. After removing the medium, 100% DMSO 100 µl was dispensed and the absorbance (OD570) was measured after shaking incubation (120 rpm, 30 minutes).

[0141] Test results showed that the byproduct had an effect of alleviating skin damage caused by ultraviolet rays up to 7,500 ppm for 12 hours and up to 5,000 ppm for 24 hours (Fig. 6).

[0143] 8. Skin moisturizing benefits

[0145] Skin keratinocytes 8.0x10 42 ml of cells / ml was dispensed into each well of a 6-well plate. After incubating for 24 hours at 37°C under 5% CO2 conditions, cells attached to the bottom were checked, and the plates were washed twice with 1x PBS. 2 ml of DMEM (free of FBS and antibiotics) was added, followed by starvation under the same conditions for 24 hours, after which the plates were washed twice with 1x PBS. The samples were mixed with DMEM (free of FBS and antibiotics) to adjust the final concentrations to 5,000, 2,500, 1,000, and 500 ppm, and 2 ml was dispensed into each well. The plates were then incubated under the same conditions for 6 hours. After washing twice with 1x PBS, 100 µl of RIPA buffer (containing 1x protease inhibitor) was added to each well, and the cells were collected using a cell scraper. After centrifugation (13,000 rpm, 4°C, 10 min), only the supernatant (protein) was collected and stored at -20°C. Protein transfer was performed via a PVDF membrane after SDS-PAGE (90V, 2 hours 20 min). Subsequently, blocking was performed (5% skim milk in TBST), followed by washing 2-3 times with TBST and overnight treatment with the primary antibody (mouse). After washing with TBST, the secondary antibody (mouse) was applied (2-3 hours). Finally, the sample was developed after EPL treatment.

[0146] As a result, the expression of HAS2 (hyaluronan synthase) protein in distilled soju byproducts treated with the MGE 3400 strain increased significantly compared to the control group, and the expression level was also higher than that of byproducts treated with commercial yeast strains (Fig. 7).

[0147] This meant that applying distilled soju byproduct material containing the MGE 3400 strain to the skin could improve moisturizing power.

[0149] Foregoing, specific parts of the content of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

[0151] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC) Trustee Number: KCTC16217BP Date of Deposit: 2025-01-02

Claims

Claim 1 Saccharomyces cerevisiae MGE 3400 of deposit number KCTC 16217BP ( Saccharomyces cerevisiae A cosmetic composition for skin antibacterial, antioxidant, alleviation of UV-induced damage, or moisturization comprising a byproduct of distilled soju produced by fermenting grains with the MGE 3400 strain. Claim 2 delete Claim 3 A composition according to claim 1, characterized in that the distilled soju is prepared by mixing koji and a strain or a culture thereof for a first fermentation, adding white rice and water for a second fermentation, and then fermenting and distilling. Claim 4 delete Claim 5 delete Claim 6 delete Claim 7 In paragraph 1, Cutibacterium acnes KCTC 3314 or Candida albicans A composition characterized by exhibiting antibacterial activity against KCTC 7270 bacteria. Claim 8 delete

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