KR102171334B1 - Composition for improving skin condition containing a fermented soybean - Google Patents

Abstract

A composition according to an aspect includes a fermented Rhynchosia nulubilis product fermented with lactic acid bacteria of Korean infants, and may exhibit skin condition improvement effects such as reducing skin wrinkles, exhibiting skin antioxidant effects, and strengthening skin barriers. The fermented Rhynchosia nulubilis product is obtained by inoculating Rhynchosia nulubilis with a Bifidobacterium animalis subsp. lactis LDTM 8102 strain (KCTC13392BP), followed by fermenting.

Description

Composition for improving skin condition containing a fermented soybean}

It relates to a cosmetic composition for skin improvement, a food composition, a feed composition, and a pharmaceutical composition containing a fermented soybean product.

Retinol is the most representative ingredient used as a functional material for skin wrinkle improvement. Retinol inhibits collagen degradation due to ultraviolet rays and at the same time promotes the formation of new collagen and has an effective wrinkle improvement effect. However, retinol is a very unstable chemical, and when its content is high, its instability increases, and when it is exposed to air, its efficacy is poor. In addition, since it is decomposed into chemical substances that are irritating to the skin by sunlight, there is a limitation that it must be used only at night. For this reason, it is known that 9 out of 10 people who used cosmetics containing retinol experienced side effects such as dry skin, redness, and keratinization. For this reason, consumers are in need of new alternative materials for wrinkle improvement, and in particular, as interest in skin beauty and health increases, not only seeking high efficacy of functional substances for skin improvement, but harmless to the human body, and side effects. We are pursuing materials that can exclude the danger of such as. Furthermore, a global consumption trend that wants natural cosmetics, organic cosmetics, and anti-animal cosmetics is forming.

On the other hand, among the materials of various natural cosmetics, soybeans contain about 18% fat, most of which are unsaturated fatty acids, of which more than half is linolenic acid (Kim YH, Hawrylewiez EJ, 1992, Klein BP, 1995, Sirtori CR, etc. 1995). In addition, it contains a large amount of functional ingredients such as isoflavones, soy saponins, and oligosaccharides (Kennedy AR 1995). Among the various types of beans, barren bean has been called Yak-bean because of its excellent pharmacological efficacy from ancient times, and it is introduced as a medicinal with excellent detoxification effect in the herbal medicine book. These weak soybeans contain 40% of essential amino acids and contain antioxidants such as vitamin E, carotenoids, saponins, and anthocyanins in an amount 4 times more than that of regular soybeans. According to recent studies, it has been reported to prevent senile dementia and improve kidney function, along with pharmacological mechanisms that protect blood circulation and liver function (Kim MH et al., 2008). Due to the efficacy of these medicinal beans, they are used in various ways as an active ingredient in health functional foods and cosmetic compositions.

Under this background, the present inventors have made efforts to maximize the efficacy of Yak Soy . As a result, when fermenting Yak Soy with lactic acid bacteria ( Bifidobacterium animalis subsp. Lactis LDTM 8102) derived from Korean infants and young children, it was confirmed that skin improvement and antioxidant efficacy were remarkably increased. The present invention has been completed.

One aspect is a cosmetic for improving skin wrinkles containing fermented soybean fermented product, which is fermented by inoculating Yak bean with Bifidobacterium animalis subsp. Lactis. LDTM 8102 (KCTC13392BP) strain as an active ingredient. It is to provide a composition.

Another aspect is to provide a food composition for improving skin wrinkles containing fermented soybeans as an active ingredient, fermented by inoculating and fermenting a strain of Vitidobacterium animalis subspecs lactis LDTM 8102 (KCTC13392BP) in Yak Kong.

Another aspect is to provide a feed composition for improving skin wrinkles containing fermented soybeans as an active ingredient, fermented by inoculating and fermenting a strain of Vitidobacterium animalis subspecis lactis LDTM 8102 (KCTC13392BP) in Yak Kong.

Another aspect is to provide a pharmaceutical composition for improving skin wrinkles containing fermented soybeans as an active ingredient, fermented by inoculating and fermenting a strain of Vitidobacterium animalis subspecis lactis LDTM 8102 (KCTC13392BP) in Yak Kong.

One aspect is a cosmetic composition for skin improvement containing fermented soybean fermented product by inoculating and fermenting the Bifidobacterium animalis subsp.Lactis. LDTM 8102 (KCTC13392BP) strain in Yak Soybean as an active ingredient Provides.

In this specification, the term “Yakkong” refers to small black beans with a black skin and smaller than ordinary black beans, and there are bark beans, Socheongja, Yakseon beans, Dawon beans, Socheong No. 2, etc., which are much smaller than ordinary black beans, but 　iso The flavone series of useful ingredients, genistein and daidzein, are contained far more than other black beans.

The yak bean may be pre-treated before fermentation. Specifically, the yak bean may be pretreated by performing roasting or grinding. In one embodiment, when the pretreatment is roasting, the roasting may be performed for 5 to 30 minutes at a temperature condition of 100 to 300°C. Specifically, the roasting is 100 to 300°C, 100 to 250°C, 100 to 200°C, 100 to 150°C, 150 to 300°C, 150 to 250°C, 150 to 200°C, 200 to 300°C, 160 to 300°C Alternatively, it may be performed under conditions of 160 to 250°C. In addition, the roasting may be performed for 5 to 30 minutes, 5 to 25 minutes, 5 to 20 minutes, 5 to 15 minutes, 5 to 10 minutes, 7 to 30 minutes, 7 to 20 minutes, or 10 to 30 minutes. have. At this time, when the roasting temperature is less than the above range, there is a problem in that the flavor of the yak bean is not maintained and the skin improvement effect is deteriorated.If the roasting temperature is exceeded, the bitter taste of the yak bean becomes strong, and consumer preference decreases. have. In addition, when the roasting time is less than the above range, there is a problem that taste stability is not maintained when applied as a food composition, and when it exceeds the above range, there is a problem that the weak beans are carbonized.

As used herein, the term " Bifidobacterium animalis subsp. Lactis.) LDTM 8102 (KCTC13392BP) strain" is isolated from feces of Korean infants and young children, and beta-glucosidase (β- glucosidase) is excellent in production capacity, so it can be effectively used for biotransformation into useful substances through microbial metabolism for natural biomass products including rice, barley, milk, soybeans, ginseng, seaweeds, woody, etc.

The term "fermented product" in the present specification means that raw materials are denatured or transformed in a physical or chemical way using lactic acid bacteria, and in nature, the desired result is obtained in just a few days or months that can be changed in tens of thousands of years. It refers to the decomposition process that can be obtained. Specifically, the fermentation product is a fermentation product produced by mixing Yak bean into a lactic acid bacteria culture medium, inoculating and fermenting lactic acid bacteria, and the lactic acid bacteria are Bifidobacterium animalis subsp. Lactis. LDTM 8102 (KCTC13392BP) may be a strain.

The fermentation product may refer not only to a product fermented by the lactic acid bacteria, but may include a fermentation filtrate or a fermentation extract obtained by extracting the fermented product from the fermented soybean as a solvent.

In the present specification, the term "fermentation extract" may refer to a useful substance itself produced by extracting the fermentation product, and more specifically, the fermentation strain and the fermentation product are removed by centrifugation from the fermentation strain culture medium coexisting A fermented product, a fermented product obtained by filtering a fermentation strain from the culture medium, a fermented product obtained by sterilizing a fermentation strain from the culture medium and filtering it, a dilution solution obtained by diluting the fermentation product, and a dried product obtained by drying the fermentation product. It may contain all of the materials. In the present specification, the term fermented extract may be used interchangeably with the fermented soybean extract.

The extraction may be obtained by extracting the yak bean fermented with the strain with water, C1 to C4 lower alcohol, or a mixture thereof. For example, the lower alcohol used in the extraction of the yak bean may be ethanol or methanol. During extraction, after slicing the yak bean, water, alcohol, or a mixture thereof may be extracted by adding 2 to 20 times the weight of the yak bean. For example, it may be extracted by adding 3 to 10 times. Extraction temperature is 30 ℃ to 100 ℃, 60 ℃ to 100 ℃. It may be 30 ℃ to 90 ℃, 30 ℃ to 80 ℃, 60 ℃ to 90 ℃, or 60 ℃ to 80 ℃. The extraction time may be 1 hour to 10 hours, 2 hours to 8 hours, 2 to 5 hours, 3 to 7 hours, or 4 to 6 hours. As for the extraction method, cold sedimentation, ultrasonic extraction, or reflux cooling extraction method can all be used. The number of extractions may be 1 to 5 times, 2 to 4 times, and 3 to 5 times of repeated extraction.

The fermented soybean extract may be extracted with 60 to 80% ethanol. In one embodiment, as a result of comparing the skin wrinkle improvement effect and antioxidant ability according to the extraction conditions of the fermented soybean fermented product, both the hot water fermented extract and the fermented extract extracted with 5% to 70% ethanol exhibited excellent effects, especially , It was confirmed that the fermented extract extracted with 70% ethanol showed the highest anti-wrinkle effect and antioxidant activity.

The cosmetic composition may exhibit a skin improvement effect. In the present specification, the term "skin improvement" may be used in a generic sense to improve skin condition, and specifically, the skin improvement may be skin wrinkle improvement, skin antioxidant activity, or skin barrier strengthening, but is not limited thereto. . The term "wrinkle" refers to fine lines caused by deterioration of the skin, and may be caused by genes, a decrease in collagen present in the skin dermis, an external environment, and the like. The term "wrinkle improvement" refers to inhibiting or inhibiting the occurrence of wrinkles on the skin or relieving wrinkles that have already been created. It includes alleviating skin wrinkles by improving or increasing skin elasticity, and may include, for example, improving skin wrinkles by reducing the pore area. The term "antioxidant activity" refers to an action of inhibiting, reducing or controlling the generation or reaction of free radicals generated in the body, hydrogen peroxide or peroxide generated from the free radicals, and hydroxy radicals generated from the hydrogen peroxide in a narrow range. And, in a broad range, it may mean an action of inhibiting, reducing or controlling the generation of oxidation reactions occurring in nature. The term "strengthening the skin barrier" may refer to any action that enhances the function of the skin barrier, which is located on the outermost side of the skin and prevents loss of moisture and nutrition. In one embodiment, it was confirmed that the fermented yak bean exhibited significant skin improvement effect and antioxidant effect compared to the non-fermented yak bean extract.

The cosmetic composition may include fermented soybeans at a concentration of 1 to 30 μg/mL based on the total weight of the composition. For example, the fermented soybean product is 1 to 30 μg/mL, 1 to 25 μg/mL, 1 to 20 μg/mL, 3 to 30 μg/mL, 5 to 25 μg/mL, 5 to 20 μg/mL Or, it may be included in a concentration of 10 to 20 ㎍ / mL. At this time, if the fermented soybean product is less than the above parts by weight, it is difficult to expect a clear skin improvement effect, and if the fermented soybean product exceeds the above parts by weight, it is difficult to increase the apparent effect due to the increase in content, and the formulation and stability of the product It may cause some problems, and there may be problems such as toxicity in the body.

The cosmetic composition may include, without limitation, ingredients that are generally allowed in addition to the active ingredient, and may include conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and a carrier. If necessary, ingredients such as surfactants, antioxidants, preservatives, pH adjusters, moisturizers, and lubricants may be further added. In addition, the cosmetic 　 composition additionally includes a substance that can supplementaryly provide essential nutrients to the skin, and preferably includes a natural scent, 　 cosmetics scent, or plant extracts, but may contain adjuvants that are not limited thereto. .

The cosmetic composition may be prepared in any formulation commonly manufactured in the art, for example, solution, emulsion, suspension, paste, cream, lotion, gel, powder, spray, surfactant-containing cleansing, oil , Soap, liquid detergent, bath agent, foundation, makeup base, essence, 　 lotion, foam, pack, softening water, sunscreen cream, sun oil, etc. 　 formulations, preferably skin ointment, softening lotion, nutrition It can be prepared as a 　 formulation of a lotion, nutrition cream, massage cream, essence, pack, emulsion or oil gel, in which case, the carrier used may be selectively used depending on the 　 formulation of the cosmetics. For example, in the case of manufacturing ointments, pastes, creams, or gels, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. are used as carrier components. Can be used alone or in combination; In the case of manufacturing a cosmetic product in the form of powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, chlorofluorohardcarbon, propane/butane, dimethyl ether, etc. are used as carrier components. Can be used either individually or in combination; When manufacturing cosmetics in the form of a solution or emulsion, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, cottonseed oil, peanuts as carrier components Oil, corn seed oil, olive oil, castor oil, sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan, etc. may be used alone or in combination; When manufacturing cosmetics in the form of a suspension, water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide as carrier components , Bentonite, agar, tracanth, and the like may be used alone or in combination; When manufacturing cosmetics in the form of cosmetic soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars, etc. are used as carrier components. They can be used alone or in combination.

Another aspect is a food composition for skin improvement containing fermented soybeans as an active ingredient, fermented by inoculating Yak bean with Bifidobacterium animalis subsp . Lactis LDTM 8102 (KCTC13392BP) strain. to provide. Details of the composition are as described above.

In the food composition for skin improvement according to an embodiment, when the fermented soybean is used as an additive to the food composition, it may be added as it is or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. have. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

The formulation of the food composition may be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

The type of food is not particularly limited, and examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes, and may include all foods in the usual sense.

In general, when preparing food or beverage, the fermented soybean may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on 100 parts by weight of raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and the present invention has no problem in terms of safety in terms of using fractions from natural products. It can also be used in the above amount.

In the food composition according to an embodiment, the beverage may contain various flavoring agents or natural carbohydrates as an additional component, like a conventional beverage. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.

In addition to the above, the food composition for improving skin according to an embodiment includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, It may contain preservatives, glycerin, alcohols, carbonates used in carbonated beverages. In addition, the composition for improving skin of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not limited, but it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the food composition.

Another aspect is a feed composition for skin improvement containing fermented soybean fermented product by inoculating and fermenting Yak bean with Bifidobacterium animalis subsp . Lactis LDTM 8102 (KCTC13392BP) strain as an active ingredient Provides. Details of the composition are as described above.

The feed 　 composition is an organic acid such as citric acid, fumaric acid, adipic acid, lactic acid in addition to the fermented soybean for administration; Phosphates such as potassium phosphate, sodium phosphate, and polymerized phosphate; One or more of natural antioxidants such as polyphenol, catechin, tocopherol, vitamin C, green tea extract, chitosan, and tannic acid may be mixed and used. In addition, a diluent, a dispersant, a surfactant, a binder, or a lubricant may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., capsules, granules or tablets.

In addition, the 　 feed 　 composition is an auxiliary ingredient, various auxiliary agents such as amino acids, inorganic salts, vitamins, antioxidants, antifungal agents, antibacterial agents, and vegetable proteins such as crushed or crushed wheat, barley, corn 　 feed, blood meal, meat meal, fish meal, etc. In addition to the main ingredients such as animal protein and feed, animal fat and vegetable fat, it can be used with nutritional supplements, growth promoters, digestive and absorption promoters, and disease prevention agents.

When the 　 feed 　 composition is used as a 　 feed 　 additive, the 　 feed 　 composition may be added as it is or used with other ingredients, and may be appropriately used according to a conventional method. The dosage form of the feed composition can be prepared as an immediate release or sustained release formulation in combination with a non-toxic pharmaceutically acceptable carrier. Such edible carriers may be corn starch, lactose, sucrose, propylene glycol. In the case of a solid carrier, it may be a dosage form such as a tablet, a powder, or a toroki agent, and in the case of a liquid carrier, a dosage form such as a syrup, a liquid suspension, an emulsion, or a solution may be used. In addition, the administration agent may contain a preservative, a lubricant, a solution accelerator, a stabilizer, and other inflammatory disease ameliorating agents and substances useful for virus prevention.

The 　 feed 　 composition can be applied to a number of animal diets, that is, 　 feed, including mammals, poultry, fish and crustaceans. It can be used for commercially important mammals such as pigs, cows, goats, zoo animals such as elephants and camels, and livestock such as dogs and cats. Commercially important poultry includes chickens, ducks and geese, and may include commercially raised fish and crustaceans such as trout and shrimp.

Another aspect is a pharmaceutical composition for skin improvement containing a fermented soybean fermented product by inoculating and fermenting the Bifidobacterium animalis subsp . Lactis LDTM 8102 (KCTC13392BP) strain in Yak Soybean as an active ingredient Provides. Details of the composition are as described above.

As used herein, the term "pharmaceutical composition" refers to a product prepared for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to respective conventional methods. For example, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, and the like, and can be formulated in the form of external preparations, suppositories, and sterile injectable solutions. Specifically, it may be formulated and used in a form suitable for eye drop administration, for example, eye drops, creams, ointments, gels or lotions.

The pharmaceutical composition may be prepared in the form of a pharmaceutical composition for preventing or treating wounds, further comprising an appropriate carrier, excipient, or diluent commonly used in the preparation of pharmaceutical compositions, wherein the carrier is a non-natural carrier. naturally occuring carrier).

Carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils.

In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient, such as starch, calcium carbonate, in the upper leaf extract and its fractions. , Sucrose (sucrose) or lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include water and liquid paraffin, which are simple diluents commonly used for suspensions, liquid solutions, emulsions, syrups, etc., and various excipients such as wetting agents, sweetening agents, fragrances, and preservatives. have. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like may be used.

The composition according to an aspect may be used to improve skin conditions. Specifically, by effectively suppressing the decomposition of collagen constituting the skin barrier due to photoaging, which is the main cause of exogenous skin aging, it can exhibit a remarkable effect on skin wrinkle improvement.

The composition according to another aspect may be used in an antioxidant composition. Specifically, the fermented yak bean may exhibit a significantly higher level of antioxidant activity compared to the non-fermented yak bean extract.

1 is a view schematically showing a process for preparing a fermented soybean product using Bifidobacterium animalis subsp. Lactis. LDTM 8102 (Accession No. KCTC13392BP).
FIG. 2 is a non-fermented yak bean extract (UFY: Unfermented Yak-kong) from skin keratinocytes exposed to UV rays, and a yak bean fermented product of Example 1 (EFY: 70% EtOH extract of fermented Yak-kong with Bifidobacterium animalis subsp.Lactis) LDTM 8102 ), and the fermented Yak-kong of Comparative Example 1 (Ref.: 70% EtOH extract of fermented Yak-kong with Bifidobacterium animalis subsp. Lactis KCTC 5854 ) is a view showing the result of comparing the reduction efficacy of collagenase .
Figure 3 is a hot water extract of fermented soybean fermented with Bifidobacterium animalis subsposis lactis LDTM 8102 in skin keratinocytes exposed to ultraviolet light (H ₂ O), 5%, 10%, 30%, 50%, And 70% alcohol extract (5%, 10%, 30%, 50%, 70% EtOH) is a view showing the result of comparing the reduction efficacy of collagenase.
4 is a graph comparing the antioxidant activity of non-fermented yak bean extract (UFY), hot water extract of yak bean fermented product (WFY), and 70% alcohol extract (EFY) of yak bean fermented product.
5 is a graph showing the results of comparing the total polyphenol content of the non-fermented yak bean extract (UFY), the hot water extract of yak bean fermentation (WFY), and the 70% alcohol extract (EFY) of yak bean fermentation product.
6 is a view showing the result of comparing the reduction effect of MMP-1 according to the treatment concentration of the 70% alcohol extract of fermented Yak bean fermented with Bifidobacterium animalis subsposis lactis LDTM 8102.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Preparation of fermented soybeans

A fermented soybean product was prepared using Bifidobacterium animalis subsp. Lactis. LDTM 8102 (accession number: KCTC13392BP) isolated from the feces of Korean infants. Specifically, Bifidobacterium animalis subsp. Lactis ( Bifidobacterium animalis subsp. Lactis.) LDTM 8102 (accession number: KCTC13392BP) preservative solution mixed with glycerol and stored frozen at -80°C was inoculated in MRSC medium to , The entire cultivation process was performed by standing for 20 to 24 hours in anaerobic conditions. Thereafter, 50 g/L of yak bean powder was inoculated with the strain culture solution having increased activity by performing the entire culturing process, and then cultured with shaking under anaerobic conditions maintained at 37°C. Samples collected in the fermentation process were lyophilized through a heat treatment process, and 0, 5, 10, 30, 50, and 70% fermented alcohol (vegetable alcohol) were added to the sample at a rate of 10 ml per 0.5 g. After bathing for 2 hours at °C, each was extracted. Thereafter, it was allowed to stand at room temperature for about 1 hour to cool, and then centrifuged at 5000 rpm for 10 minutes to separate the supernatant. The separated supernatant was filtered using a 0.2 µm syrine filter, and all alcohol was removed in a vacuum concentrator at 40°C, and then lyophilized to powder. Thereafter, the lyophilized powder was dissolved in 50% DMSO and used as fermented soybeans. Fig. 1 shows an overall process for preparing a fermented soybean product using Bifidobacterium animalis subsp. Lactis. LDTM 8102 (accession number: KCTC13392BP).

Comparative Example 1. Preparation of fermented soybeans

Except that yak beans were fermented using Bifidobacterium animalis subsp.Lactis KCTC5854 lactic acid bacteria (Ref.), a fermented soybean product was prepared in the same manner as in Example 1 .

[Experimental Example]

Experimental Example 1. Confirmation of collagenase reduction effect

In order to confirm the effect of improving skin wrinkles of the fermented soybeans of Example 1, and to compare the effects with the fermented soybeans of Comparative Example 1 prepared using other lactic acid bacteria from Example 1, skin keratinocytes (HaCaT, immortalized human keratinocyte cell line). Specifically, the secretion of the collagen-degrading enzyme MMP-1 (Matrix metalloproteinase-1) was measured using Western blot analysis. HaCaT cells were cultured in DMEM medium containing 10% (v/v) FBS and 1% (v/v) penicillin/streptomycin at 5% CO ₂ and 37°C. HaCaT cells were cultured for 24 hours, and the medium was replaced with a serum-free DMEM medium. After culturing at 37° C. for 24 hours, HaCaT cells were treated in a serum-free medium at a concentration of 80 µg/ml and not fermented Yak bean extract, Yak bean fermented product of Example 1, or Yak bean fermented product of Comparative Example 1 1 Incubated for hours. After irradiation with sUV of 25 kJ/m ² intensity, it was cultured for 48 hours. The medium was harvested on ice and then centrifuged at 18,620 g for 10 minutes. The protein concentration of the solvent was measured using a protein assay reagent kit as described by the manufacturer. Protein was separated by electrophoresis by SDS-PAGE using 10% SDS-polyacrylamide gel, and transferred to Immobilon-P PVDE (MERK Millipore, Darmstadt, Germany). After blocking treatment for 1 hour in 5% skim milk, the primary antibody was attached overnight at 4°C. Protein bands were visualized using a chemiluminescence detection kit (GE healthcare, Little Chalfont, UK) after treatment with a secondary antibody (Life techonologies, Waltham, MA, USA). The test was repeated 3 times.

FIG. 2 is a non-fermented yak bean extract (UFY: Unfermented Yak-kong) from skin keratinocytes exposed to UV rays, and a yak bean fermented product of Example 1 (EFY: 70% EtOH extract of fermented Yak-kong with Bifidobacterium animalis subsp.Lactis) LDTM 8102 ), and the fermented Yak-kong of Comparative Example 1 (Ref.: 70% EtOH extract of fermented Yak-kong with Bifidobacterium animalis subsp. Lactis KCTC 5854 ) is a view showing the result of comparing the reduction efficacy of collagenase .

As a result, as shown in FIG. 2, compared to the control group not irradiated with ultraviolet rays, when ultraviolet rays are irradiated, the expression of collagen-degrading enzyme is increased, which inhibits the biosynthesis of protocol ragen and reduces the content of collagen, thereby reducing the skin's wrinkles. It was confirmed that it promotes production and causes pigmentation to make skin tone dull. In this case, it was confirmed that the treatment of the non-fermented yak bean extract showed an effect of improving skin wrinkles by reducing the expression of collagenase. In addition, the fermented soybean fermented product (Comparative Example 1) fermented using the Bifidobacterium animalis subspasis lactis KCTC 5854 strain exhibited an effect of improving skin wrinkles at a level similar to that of the non-fermented soybean extract. The fermented soybean fermented product (Example 1) fermented using the Bifidobacterium animalis subspecis lactis LDTM 8102 strain significantly increased skin wrinkle improvement compared to the non-fermented soybean extract and the fermented soybean fermented product of Comparative Example 1. It was confirmed that it represents.

That is, it can be seen from the above results that the fermented Yak-bean according to one aspect exhibits a significantly increased skin wrinkle improvement effect compared to the fermented Yak-bean fermented product fermented using not only the non-fermented Yak bean extract, but also other types of lactic acid bacteria.

Experimental Example 2. Comparison of collagenase reduction effect of fermented soybeans according to extraction conditions

In order to compare the effect of improving skin wrinkles of fermented soybeans according to the extraction conditions, the reduction effect of MMP-1 was compared in the same manner as described in Experimental Example 1. Specifically, hot water extract of fermented soybean (H ₂ O), 5%, 10%, 30%, 50%, and 70% alcohol extract (5%, 10%, 30%, 50%, 70% EtOH) 80 After treatment with μg/ml, the effect of reducing MMP-1 was confirmed.

Figure 3 is a hot water extract of fermented soybean fermented with Bifidobacterium animalis subsposis lactis LDTM 8102 in skin keratinocytes exposed to ultraviolet light (H ₂ O), 5%, 10%, 30%, 50%, And 70% alcohol extract (5%, 10%, 30%, 50%, 70% EtOH) is a view showing the result of comparing the reduction efficacy of collagenase.

As a result, as shown in FIG. 3, hot water extract of fermented soybean fermented with Bifidobacterium animalis subspecis lactis LDTM 8102 when irradiated with ultraviolet rays, 5%, 10%, 30%, 50%, and 70% When the alcohol extract was treated, both showed excellent skin wrinkle improvement effect, and in particular, it was confirmed that the skin wrinkle improvement effect was most excellent when the 70% alcohol extract of the fermented soybean was treated.

Experimental Example 3. Comparison of antioxidant activity of fermented soybeans according to extraction conditions

In order to compare the antioxidant activity of fermented soybeans according to the extraction conditions, the antioxidant effect and total phenol content of the hot water extract and 70% alcohol extract of the fermented soybean fermented with Bifidobacterium animalis subsposis lactis LDTM 8102 were compared. Specifically, DPPH scavenging activity was measured in order to measure the antioxidant activity according to the extraction conditions (hot water extraction or 70% alcohol extraction) of the fermented soybean product. A 0.1 mM DPPH solution (7.89 mg DPPH/200 mL 80% MeOH) was prepared by diluting with 80% methanol in a bottle wrapped with silver paper, and this was mixed for 20 minutes. Vitamin C 100 mg/mL was used as the standard antioxidant solution, and diluted by 1/2 to draw a standard curve. Blank (blank, 80% methanol), DPPH solution, and samples to measure antioxidant activity (unfermented yak bean extract (UFY), hot water extract from yak bean fermentation (WFY), 70% alcohol extract from yak bean fermentation (EFY)) It was reacted 1:1. The final volume of the DPPH solution was adjusted to 200 uL using 80% methanol so that the absorbance at the 517 nm wavelength band was 1.000±0.020. Use silver paper to prevent light, and shake at 23°C for 30 minutes. At the final time point after 30 minutes, the absorbance decreased compared to the blank group in the 517 nm wavelength band was measured.

In addition, the total polyphenol content (total phenolic content) was measured in order to measure the antioxidant activity according to the extraction conditions (hot water extraction or 70% alcohol extraction) of the fermented Yak. Specifically, 80 µl tertiary distilled water, 10 µl of the sample and standard solution, and 10 µl folin-ciocalteu phenol reagent were sequentially added to each well of a 96-well multi-plate, and then for 6 minutes. Gave me a shake. After the reaction was over, 100 µl of a 7% Na ₂ CO ₃ solution was dispensed into each well and shaken for 90 minutes. As the reaction proceeded, the solution turned blue, and the absorbance was measured at a wavelength of 750 nm.

4 is a graph comparing the antioxidant activity of non-fermented yak bean extract (UFY), hot water extract of yak bean fermented product (WFY), and 70% alcohol extract (EFY) of yak bean fermented product.

5 is a graph comparing the total polyphenol content of non-fermented yak bean extract (UFY), hot water extract of yak bean fermented product (WFY), and 70% alcohol extract (EFY) of yak bean fermented product.

As shown in FIGS. 4 and 5, the fermented Yak bean showed higher antioxidant activity compared to the non-fermented Yak bean extract, and in particular, the 70% alcohol extract of the fermented Yak bean showed higher antioxidant activity compared to the hot water extract of the fermented Yak bean, It was confirmed that the total polyphenol content was also significantly increased.

Experimental Example 4. Confirmation of wrinkle improvement effect according to treatment concentration of fermented soybeans

As confirmed in Experimental Example 2, 70% alcohol extract of fermented soybean fermented with Bifidobacterium animalis subspasis lactis LDTM 8102 showing the best skin wrinkle improvement effect in a concentration-dependent manner In order to confirm whether it is indicated, the 70% alcohol extract of the fermented soybean was treated with 20 µg/ml, 40 µg/ml, or 80 µg/ml, respectively, and then MMP-1 in the same method as described in Experimental Example 1 The reduction effect of was compared.

6 is a view showing the result of comparing the reduction effect of MMP-1 according to the treatment concentration of the 70% alcohol extract of fermented Yak bean fermented with Bifidobacterium animalis subsposis lactis LDTM 8102.

As a result, as shown in FIG. 6, it was confirmed that the fermented soybean fermented with Bifidobacterium animalless subspecis lactis LDTM 8102 reduced MMP-1 in a concentration-dependent manner. From the above results, it can be seen that the fermented Yak-bean, in particular, the 70% alcohol extract of the fermented Yak-bean, exhibits a concentration-dependent effect of improving skin wrinkles.

Claims (9)

A cosmetic composition for skin improvement containing fermented soybeans as an active ingredient, fermented by inoculating and fermenting a strain of Bifidobacterium animalis subsp. Lactis. LDTM 8102 (KCTC13392BP) in Yak Bean. The cosmetic composition of claim 1, wherein the fermented product comprises a fermented filtrate or a fermented extract. The cosmetic composition of claim 2, wherein the fermentation extract is extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The cosmetic composition of claim 3, wherein the fermentation extract is extracted with 60% to 80% ethanol. The cosmetic composition of claim 1, wherein the skin improvement is at least one selected from the group consisting of skin wrinkle improvement, skin antioxidant and skin barrier strengthening. The cosmetic composition of claim 5, wherein the wrinkle improvement is achieved by promoting collagen production or inhibiting expression of matrix metalloproteinase (MMP). A food composition for skin improvement containing a fermented soybean fermented product by inoculating and fermenting the Bifidobacterium animalis subspecis lactis LDTM 8102 (KCTC13392BP) strain in Yak Bean as an active ingredient. A feed composition for skin improvement containing fermented soybeans as an active ingredient, obtained by inoculating and fermenting the Bifidobacterium animalis subspesis lactis LDTM 8102 (KCTC13392BP) strain in Yak Bean. delete

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