WO2023106829A2 - Fermentate, produced by fermenting ginseng sprout trituration with novel strain, for antioxidant or anti-inflammatory cosmetic composition, preparation method therefor, and antioxidant or anti-inflammatory cosmetic composition comprising same fermentate as active ingredient - Google Patents

Abstract

The present invention relates to an antioxidant or anti-inflammatory cosmetic composition that comprises as an active ingredient a fermentate produced by fermenting a mixture of a ginseng sprout trituration and a ginseng sprout extract with the (Saccharomyces cerevisiae strain, and as such, exhibits an excellent antioxidant and anti-inflammatory effect.

Description

A fermented product obtained by fermenting softened leaves of ginseng sprouts with a new strain for antioxidant or anti-inflammatory cosmetic composition and a manufacturing method thereof, and an antioxidant or anti-inflammatory cosmetic composition containing the fermented product as an active ingredient

The present invention relates to a fermented product obtained by fermenting softened ginseng sprout leaves with a new strain for an antioxidant or anti-inflammatory cosmetic composition, a method for producing the same, and an antioxidant or anti-inflammatory cosmetic composition containing the fermented product as an active ingredient. .

As modern people become more interested in health, various studies on natural and eco-friendly cosmetics are being actively conducted. In functional cosmetics, interest in cosmetics using materials derived from natural products is increasing, rather than in the case of chemical ingredient-based whitening, antioxidants, UV protection, preservatives, and antibacterial agents in the past.

Among various natural materials, animal-derived materials are actively developing functional cosmetics using plant-based ingredients as consumers' resistance to animal-derived materials increases due to animal welfare, mad cow disease, swine cholera, and bird flu. Consumer demand for complex functional cosmetics such as wrinkle improvement and sun protection is increasing.

Functional cosmetics refers to cosmetics at various stages of use, such as essence, serum, cream, powder, and base, to which specific functions such as whitening, wrinkle improvement, and UV protection are added. Multifunctional cosmetics with one function have also been developed to increase the ease of use, and specialized products for various purposes such as pore care, nutrition supply, elasticity enhancement, acne treatment, whitening and UV protection are also being released.

Inflammation is an immune response of the human body in response to wounds or diseases, and oxidative stress such as ultraviolet rays, active oxygen, and free radicals activates inflammatory factors to cause various diseases and aging of the skin. Vasoactive polypeptides, such as kinin, plasmin, or complement, act on vasodilation, contraction, and chemotaxis, and other lymphocytes such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized into prostaglandin or leukotrienes, which are inflammatory mediators, through two pathways, cyclooxygenase and lipooxygenase, to mediate various inflammatory responses.

An anti-inflammatory agent that acts to remove the inflammatory source and reduce the physiological response and symptoms in order to disappear the inflammation. Substances that have been used for anti-inflammatory purposes so far include flufenamic acid, ibuprofen, benzydamine, or indomethacin as nonsteroidal agents, and prednisolone as steroidal agents. or dexamethasone. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have a problem in that the amount of use is limited due to safety issues on the skin, or the effect is insignificant, so that the anti-inflammatory effect cannot be expected. there is.

Therefore, there is a demand for the development of excellent ingredients that are safe to the body, stable active ingredients, and have anti-inflammatory or antioxidant effects.

[Prior art literature]

[Patent Literature]

Republic of Korea Patent No. 1484884

Republic of Korea Patent Publication No. 2020-0079212

An object of the present invention is to provide a cosmetic for antioxidant or anti-inflammatory use containing, as an active ingredient, a fermented product prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with Saccharomyces cerevisi ae strain. to provide a composition.

In addition, another object of the present invention is to provide a fermented ginseng sprout prepared by fermenting Saccharomyces cerevisi ae strain, soft ginseng sprout leaf, and ginseng sprout leaf extract.

In addition, another object of the present invention is to inoculate and ferment a Saccharomyces cerevisi ae strain in a mixture of ginseng sprout leaf softening and ginseng sprout leaf extract; fermenting sprout ginseng including It is to provide a method for producing water.

Antioxidant or anti-inflammatory cosmetic composition of the present invention for achieving the above object is fermentation prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with Saccharomyces cerevisi ae strain It may contain water as an active ingredient.

The above Saccharomyces cerevisiae (Saccharomyces cerevisi ae) The strain was Saccharomyces cerevisiae purchased from ATCC (American Type Culture Collection). (Saccharomyces cerevisi ae, ATCC ^® 204508) strain.

The above Saccharomyces cerevisiae (Saccharomyces cerevisi ae) The strain is Saccharomyces cerevisiae deposited under the accession number KFCC11825P (Saccharomyces cerevisi ae) 28-7 strain.

The Saccharomyces cerevisiae strain may be Saccharomyces cerevisi ae 28-7 strain having the nucleotide sequence represented by SEQ ID NO: 1 [Accession Number: KFCC11825P] .

The mixture of the ginseng sprout leaf softener and the ginseng sprout leaf extract may be inoculated with the Saccharomyces cerevisi ae strain at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU/ml and fermented.

The number of viable cells after inoculation and fermentation of the Saccharomyces cerevisi ae strain at a concentration of 1X10 ⁶ CFU/ml may be 1X10 ⁸ to 9X10 ⁸ CFU/ml.

The ginseng sprout leaf softening step of extracting the ginseng sprout leaf; Impregnating the filtered solid after the extraction with an aqueous solution containing an acid and purified water; and softening by warming the aqueous solution.

The extraction may be performed under water, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The acid is an organic acid including at least one selected from the group consisting of citric acid, tartaric acid, lactic acid, glycolic acid, formic acid, malic acid, oxalic acid, salicylic acid, ascorbic acid, citric acid, and gluconodeltalactone; Or an inorganic acid containing at least one member selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid; may be included.

The softening may be performed at 40 to 95 °C for 30 to 240 minutes.

The ginseng sprout leaf softening may have a pH of 2.5 to 6.5.

The ginseng sprout leaf softener and the ginseng sprout leaf extract may be mixed in a weight ratio of 1: 1-3.

The composition can inhibit IL-6 and IL-8 production.

In addition, the fermented ginseng sprouts of the present invention for achieving the above other object may be prepared by fermenting Saccharomyces cerevisi ae strain, soft ginseng sprout leaves, and ginseng sprout leaf extract. .

In addition, the method for preparing the fermented ginseng sprout of the present invention for achieving the above other object is a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract mixed with Saccharomyces cerevisiae (Saccharomyces cerevisi ae) strain It may include; inoculating and fermenting.

The cosmetic composition of the present invention contains as an active ingredient a fermented product prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with a novel Saccharomyces cerevisi ae 28-7 strain. , It is non-toxic, and has excellent antioxidant and anti-inflammatory effects on inflammation.

1 is a photograph of fermented products of strain 28-7 (yeast A), strain 25-6 (yeast B), and strain 57-3 (yeast C) in softened ginseng sprout leaves.

Figure 2 is a molecular phylogenetic diagram showing the analysis of the ITS gene sequence of Saccharomyces cerevisiae 28-7 strain of the present invention.

3 is a photograph of samples 1-6 of fermented ginseng sprouts prepared according to Example 1 of the present invention and ginseng sprout leaves prepared according to Comparative Example 1, softened ginseng sprout leaves, and fermented ginseng sprouts after softening.

4 is a graph showing the DPPH radical scavenging ability of fermented ginseng sprouts prepared according to Example 1 of the present invention and ginseng sprout leaves prepared according to Comparative Example 1, softened ginseng sprout leaves, and fermented ginseng sprouts after softening.

5 is a graph measuring HaCaT cell viability of fermented ginseng sprouts prepared according to Example 1 of the present invention and softened ginseng sprout leaves prepared according to Comparative Example 1.

6 is a graph measuring the level of IL-6, an inflammatory cytokine, when treated with the fermented ginseng sprouts prepared in Example 1 of the present invention and the softened ginseng sprout leaves prepared in Comparative Example 1.

7 is a graph measuring the level of IL-8, an inflammatory cytokine, when treated with the fermented ginseng sprouts prepared according to Example 1 of the present invention and the softened ginseng sprout leaves prepared according to Comparative Example 1.

The present invention relates to an antioxidant or anti-inflammatory cosmetic composition containing , as an active ingredient, a fermented product prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with Saccharomyces cerevisi ae strain. it's about

Hereinafter, the present invention will be described in detail.

The cosmetic composition of the present invention contains, as an active ingredient, a fermented product prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with Saccharomyces cerevisi ae strain.

Saccharomyces cerevisiae S288c strain

The Saccharomyces cerevisiae strain is a Saccharomyces cerevisiae standard strain (ATCC ^® 204508) purchased from ATCC (American Type Culture Collection).

Saccharomyces cerevisiae strain 28-7

The Saccharomyces cerevisiae strain is a Saccharomyces cerevisiae 28-7 strain, preferably isolated from yeast, but is not limited thereto.

As a result of performing ITS sequencing for the identification and classification of microorganisms, strain 28-7, a novel strain derived from Nuruk through the following preparation example, was found to have the nucleic acid sequence of SEQ ID NO: 1. As a result of analyzing the nucleic acid sequence of the strain, it was investigated that it was a strain belonging to Saccharomyces cerevisi ae, and as a result of confirming homology to the nucleic acid sequence, Saccharomyces cerevisiae It was confirmed that it was 98% identical to the standard strain of Saccharomyces cerevisi ae.

Ginseng sprout leaf softener and ginseng sprout leaf extract

The softened ginseng sprout leaf is prepared by (a) extracting the ginseng sprout leaf; (b) impregnating the extracted and filtered solids into an aqueous solution containing an acid and purified water; and (c) warming and softening the aqueous solution.

Ginseng sprout leaves maintain their shape in cosmetics to visually increase the degree of preference, and softening materials are used to allow the ingredients of ginseng sprout leaves to permeate into the skin by melting on the epidermis only when the user uses it.

First, in step (a), sprout ginseng leaves are boiled at 80 to 120 ° C. for 1 to 5 hours, preferably 2 to 3 hours, using water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water. extract over time

When the ginseng sprout leaves are used as they are without extraction, fermentation does not proceed smoothly, so it is preferable to use the solids and the extract separately after extracting the ginseng sprout leaves.

If the temperature and time during the extraction are less than the lower limit, the softening may not be performed well when softening the solid, and if it exceeds the upper limit, the shape of the ginseng sprout leaf is not maintained and the desired effect is not obtained due to the decomposition of physiologically active substances may not work at all.

After the extraction, it is separated into solids and extracts through filtration, and then the solids are softened through the following steps (b) and (c), and the extract is a ginseng sprout leaf extract. Used with cargo.

Next, in the step (b), the solid is impregnated so as to be immersed in an aqueous solution containing an acid and purified water. The solid material may be dried to a moisture content of 5 to 15% before being impregnated in an aqueous solution in order to facilitate softening and maintain the original shape.

The acid can soften the solid by partially cutting the bonds between the celluloses present in the solid by an acidic functional group such as a carboxyl group or sulfonic acid included in the acid. As a result, the solid material is softened while maintaining its shape, and when the user uses cosmetics, it is possible to disintegrate the cellulose only with the force of rubbing with a finger. As a result, when the user uses it while maintaining its shape in a softened state, the solid is completely disintegrated with only a small amount of force, allowing most of the active ingredients of the solid to permeate into the user's skin.

The acid used in the present invention includes at least one selected from the group consisting of citric acid, tartaric acid, lactic acid, glycolic acid, formic acid, malic acid, oxalic acid, salicylic acid, ascorbic acid, citric acid, and gluconodeltalactone. organic acids; or an inorganic acid containing at least one selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid;

The concentration of the acid is 0.1 to 4% by weight, preferably 0.5 to 2% by weight, based on the total weight of the aqueous solution. If the acid concentration is less than the lower limit above, the solid may not be softened, and if it exceeds the upper limit above, toxicity may occur.

Next, in step (c), the aqueous solution is softened by warming to 40 to 95 °C, preferably 45 to 55 °C for 30 to 240 minutes.

In the present specification, the term "softening" is used, and the softening may be interpreted in the sense of softening, softening, properly dissolving tissues, and the like, and for convenience, the term softening is used implicitly in the present specification.

The softening is performed by immersing and impregnating a solid in an aqueous solution prepared in advance to satisfy the temperature range and then softening for the above time, or after immersing and impregnating a solid in an aqueous solution and raising the temperature to satisfy the above temperature range, and then softening for the above time. can

If the temperature and time during softening are less than the lower limit, the degree of softening may decrease and may not be completely decomposed during use by the user. .

In addition, after the softening step, it can be confirmed whether the solid is softened as desired, and the degree of softening is determined by hardness, adhesiveness, springiness, cohesiveness, and gumminess of the solid. , It can be measured by selecting any one of Chewiness and Resilience.

The hardness may be 10 to 100 g, preferably 15 to 70 g in unit of gravity. Here, 1g of the unit of gravity means 9.8 m/s ² . In addition, the adhesiveness may be -0.2 to -1.2 g.sec, preferably -0.3 to -1.0 g.sec; The springiness may be 0.03 to 0.22, preferably 0.04 to 0.11; The cohesiveness may be 0.2 to 0.5, preferably 0.35 to 0.45; The gumminess may be 1 to 100, preferably 5 to 40; The chewiness (Gumminess) may be 0.1 to 4, preferably 0.5 to 2.5; The resilience may be 0.1 to 1.8, preferably 0.3 to 1. The Hardness, Adhesiveness, Springiness, Cohesiveness, Gumminess, Chewiness and Resilience are measured using a Texture Analyzer (Model TAXT Express: Lamyrheology, France) equipment , and probe NO. This is done by using a P/2 θ (diameter) 2 mm rod.

The softened solid material, that is, the softened sprout ginseng leaf, is washed with purified water to remove acid and at the same time bring the pH to 2.5 to 6.5. If the pH is less than the lower limit, there is a possibility of disrupting the network of the cosmetic formulation.

The soft ginseng sprout leaf extract prepared in step (c) and the ginseng sprout leaf extract separated from the solid material in step (a) are mixed at a weight ratio of 1: 0.4-1, preferably 1: 0.8-1. . In the case of fermenting only the softened ginseng buds with a new strain, fermentation is not easily performed due to the lack of nutrients contained in the softened material, so it is fermented together with the ginseng sprout leaf extract containing free sugars and nutrients.

If the content of ginseng sprout leaf extract is less than the lower limit based on softened ginseng sprout leaf, fermentation may not be performed, and if it exceeds the upper limit, the anti-inflammatory effect may be reduced.

fermentation

Saccharomyces cerevisiae (Saccharomyces cerevisi ae) 28-7 strain is added to the mixture of the ginseng sprout leaf softener and the ginseng sprout leaf extract in an amount of 1X10 ⁶ to 9X10 ⁷ CFU/ml, preferably 1X10 ⁶ to 1X10 ⁷ CFU / ml, fermentation is carried out for 60 to 80 hours, preferably 70 to 75 hours at 20 to 40 ° C, preferably 25 to 30 ° C and 80 to 200 rpm, preferably 110 to 150 rpm .

When the temperature, rate, and time during fermentation are less than the lower limit value, the number of viable cells may be small after fermentation, and when the number of viable cells exceeds the upper limit value, there may be almost no viable cell number.

For example, the number of viable cells after fermentation is 1X10 ⁸ to 9X10 ⁸ CFU / ml, based on fermentation by inoculation of the Saccharomyces cerevisi ae 28-7 strain at a concentration of 1X10 ⁶ CFU / ml, It can be seen that the strain has increased compared to the first inoculation.

The cosmetic composition containing the fermented product prepared in this way has excellent DPPH radical scavenging ability and thus excellent antioxidant activity, and inhibits the production of IL-6 and IL-8, so it is excellent in anti-inflammatory properties.

In addition, the cosmetic composition of the present invention not only has high marketability with excellent visual effects because the fermented ginseng sprout leaf softener maintains its shape, but also contains the fermented ginseng sprout leaf softener whose shape remains. If the user puts the cosmetic composition on his/her skin and applies only a little force (rubbing), it completely disintegrates and disappears, so most of the active ingredients remaining in the fermented ginseng sprout leaf softener can be absorbed into the user's skin, increasing its efficacy. very excellent

That is, the fermented ginseng sprout leaf softening material in the cosmetic composition prepared according to the present invention maintains its shape while allowing almost all ingredients to melt or remain in or on the user's skin during use, so that the fermented ginseng sprout leaf softening material It maximizes the efficiency of the cargo itself.

In this specification, the term 'extract' used while referring to ginseng sprout leaves includes not only a crude extract obtained by treating an extraction solvent, but also a processed product of ginseng sprout leaf extract. For example, ginseng sprout leaf extract may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

On the other hand, the term 'contained as an active ingredient' in this specification means that it contains a sufficient amount to achieve the efficacy or activity of the fermented product. For example, the fermented product is used at a concentration of 10 to 1500 μg/ml, preferably 100 to 1000 μg/ml. Since the fermented product is a natural product and does not have side effects on the human body even when used in excess, the upper limit of the amount of the fermented product included in the composition of the present invention can be selected and implemented by those skilled in the art within an appropriate range.

In the cosmetic composition of the present invention, in addition to the above cosmetic composition, if necessary, other ingredients commonly used in cosmetics may be mixed. Examples of these ingredients include oils and fats, humectants, emollients, surfactants, organic and inorganic pigments, Organic powders, UV absorbers, preservatives, bactericides, antioxidants, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, water-soluble vitamins, fat-soluble vitamins, high-molecular peptides, polysaccharides, sphingolipids, and seaweed An extract etc. are mentioned.

The cosmetic composition of the present invention may be prepared in the form of an emulsified formulation and a solubilized formulation commonly used in the art.

In addition, the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the above ingredients as active ingredients, for example, conventional adjuvants such as stabilizers, pigments and natural flavors, and A carrier may be further included.

Products to which the composition of the present invention can be added include, for example, mist, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, and sunscreen cream. , Moisture Cream, Hand Cream, Foundation, Essence, Nutritional Essence, Mask Pack, Press Powder, Loose Powder, Eye Shadow, etc. Cosmetics and Soap, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion and Body Cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or 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, additional chlorofluorohydrocarbon, propane , butane or a propellant such as dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or 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, fatty acid esters of glycerol, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

Hereinafter, preferred embodiments are presented to aid understanding of the present invention, but the following examples are merely illustrative of the present invention, and various changes and modifications are possible within the scope and spirit of the present invention. It is obvious to those skilled in the art, It goes without saying that these variations and modifications fall within the scope of the appended claims.

Preparation Example 1. Isolation and selection of microorganisms from Nuruk

In order to select a strain of Saccharomyces boulardi among yeast derived from traditional yeast, in order to secure a strain that cannot use alactose, has acid resistance, and is sequence-identified as Saccharomyces cerevisiae, the following The same experiment was performed. First, about 200 yeast colonies of about 200 candidates were isolated from 60 types of traditional Nuruk prepared and collected according to the manufacturing method of the old literature, and galactose utilization analysis was performed for the primary screening of the isolated 200 colonies. Among them, a total of 12 yeasts of the genus Saccharomyces were selected. Thereafter, in order to select strains capable of surviving in a medium environment of 37 ° C. and pH 2.0, three species of Saccharomyces boulardii (strain 25-6, strain 25-6, 28-7 strains, 57-3 strains) were selected (Fig. 1).

The three strains were selected as acid-resistant yeast capable of growing at pH 2-3 because acid treatment is essential during the softening process.

Production Example 2. Identification of isolated strains

In order to confirm that the three types of yeast (strain 25-6, strain 28-7, and strain 57-3) can survive and grow on their own without adding additional sugar in the softened material, the above three types of yeast were tested on the leaves of ginseng sprouts. After inoculating the cargo at the level of 1X10 ⁶ CFU/ml, culturing at 30 °C in a shaking incubator (120 rpm) for 72 hours, then diluting to the level of 1X10 ⁶ CFU/ml to check viable cells, spreading 1 ml on an agar plate and culturing It was confirmed whether the yeast survived.

As shown in Figure 1, the viability of yeast was confirmed in the soft material itself without the addition of additional sugar. In addition, as a result of measuring the alcohol concentration in order to confirm whether the yeast fermented alcohol, it was confirmed that alcohol almost did not exist at the level of 0.023, 0.016, and 0.029%, respectively.

In addition, looking at the number of colonies in the plate, it was confirmed that they grew at relatively high concentrations on the 28-7 strain plate and the 57-3 strain plate, and when looking at the color and shape of the colonies, several colonies were mixed in the 57-3 strain plate. It was confirmed that a single colony appeared on the 28-7 strain plate. Through ITS resion 4 and 5 identification of each colony, it was confirmed that most of the white spherical colonies were inoculated Saccharomyces cerevisi ae (Table 1).

As a result of additionally identifying colonies of different shapes, the pink colonies that appeared on the 57-3 strain plate were Sporidiobolus pararoseus , which was confirmed to exist in the soft material itself, which is a fungus, a kind of yeast widely present in the surrounding environment. Turned out.

Therefore, it was confirmed that the growth of strain 25-6 was not higher than the other two strains, and strain 57-3 was superior in competition with environmental microorganisms, but it was not at the level of inhibiting their growth itself. did

Therefore, in future experiments , the Saccharomyces cerevisi ae 28-7 strain, which has excellent growth and high dominance efficiency under the conditions, was used in the fermentation process.

The identification was performed after extracting the chromosomal DNA of each strain, sequencing the base sequence using ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') primers, and BLAST ( http://www. ncbi.nlm.nih.gov ) program was used to identify the corresponding microorganisms as shown in Table 1 below.

inoculation strain	colony color	Identification result
S288c	white	Saccharomyces cerevisiae
28-7	white	Sporidiobolus cerevisiae

Preparation Example 3. Identification of Saccharomyces boulardii 28-7 strain

ITS sequencing was performed using the chromosomal DNA of the 28-7 strain, and as a result of the sequencing, the 28-7 strain of the present invention was found to be a strain belonging to Saccharomyces cerevisi ae. , It was found to have the nucleotide sequence of SEQ ID NO: 1 (see FIG. 2). In addition, as a result of analyzing the base sequence through BLAST Search, it showed about 98% homology with Saccharomyces cerevisi ae strains previously uploaded to the database, and the sequence It was confirmed that a strain having 100% homology with No. 1 did not previously exist.

Therefore, the present inventors deposited the 28-7 strain of Saccharomyces cerevisi ae with the Korea Microorganism Conservation Center on April 1, 2019 and received accession number KFCC11825P.

Example 1. Fermentation

Ginseng sprout leaves and water were mixed at a weight ratio of 1:9, extracted at 90 °C for 3 hours, and filtered to separate solids and extracts. The solid was impregnated with an aqueous solution (98% by weight of purified water, 2% by weight of citric acid), heated to a temperature of 50 ° C. and softened for 60 minutes, and then washed with water to obtain softened ginseng sprout leaves having a pH of 3.0.

As a sprout ginseng leaf extract, the extract separated from the solid after the extraction was used.

The ginseng sprout leaf softener and the ginseng sprout leaf extract were mixed at a weight ratio of 1: 1, and then the Saccharomyces cerevisiae 28-7 strain was added to the mixture at a concentration of 1X10 ⁶ CFU/ml. After inoculation, the fermented product was obtained by culturing at 30 °C in a shaking incubator (120 rpm) for 72 hours.

Comparative Example 1. Sprouts ginseng leaf softening

Ginseng sprout leaves and water were mixed at a weight ratio of 1:9, extracted at 90 °C for 3 hours, and filtered to separate solids and extracts. The solid was impregnated with an aqueous solution (98% by weight of purified water, 2% by weight of citric acid), heated to a temperature of 50 ° C. and softened for 60 minutes, and then washed with water to obtain softened ginseng sprout leaves having a pH of 4.5.

<Test Example>

Test Example 1. Measurement of viable cell count after fermentation

As in Example 1, after mixing the ginseng sprout leaf softener and ginseng sprout leaf extract at a weight ratio of 1: 1, strain 28-7 was inoculated into the mixture at a concentration of 1X10 ⁶ CFU/ml, and then cultured. As a result, all plates were incubated. Only white spherical colonies that appeared to be the same strain were searched for on the image, and the number of viable cells after fermentation was 4.5x10 ⁸ CFU/ml, confirming that the strain had grown more than at the time of initial inoculation.

Test Example 2. Measurement of DPPH radical scavenging activity

DPPH scavenging ability (%) was measured by modifying Blois' method, and DPPH (2,2-Diphenyl-1-picrylhydrazyl, Sigma-Aldrich) using the principle of color fading from dark purple to yellow in response to substances with antioxidant activity The reducing power of the sample was measured through the free radical scavenging activity. After adding 1.25 ml of DPPH solution to 0.25 ml of the sample and reacting in the dark for 20 minutes, the absorbance was measured at 517 nm. indicated by

[Equation 1]

DPPH radical scavenging activity (%)={1-(Abs(test)-Abs(color))/Abs(control)}X100

3 is a S288c strain (yeast A, ATCC purchase standard) in the fermented ginseng sprouts prepared according to Example 1 of the present invention, ginseng sprout leaves, softened ginseng sprout leaves, and extracts after softening ginseng sprouts prepared according to Comparative Example 1 strain) and 28-7 strain (yeast B) were inoculated and cultured.

Sample 1 is an extract of ginseng sprouts as a dry raw material, sample 2 is an extract after hydration of ginseng sprouts leaves as a dried raw material, and sample 3 is a bioconversion to yeast B after hydration, which is fermented with strain 28-7 after hydration of ginseng sprouts leaves. Sample 4 is an extract after softening the leaves of ginseng sprouts as an extract after softening the dried raw material, and sample 5 is an extract after softening the leaves of ginseng sprouts after softening the leaves, and sample 5 is an extract after softening the leaves of ginseng sprouts and fermenting with yeast A (S288c). 6 shows the bioconversion to yeast B after softening, and the extract after softening sprout ginseng leaves and fermenting with yeast B (28-7).

4 is a graph showing the DPPH radical scavenging ability of fermented ginseng sprouts prepared according to Example 1 of the present invention and ginseng sprout leaves prepared according to Comparative Example 1, softened ginseng sprout leaves, and fermented ginseng sprouts after softening.

Description of DPPH drawing: Among the six samples, it was confirmed that sample No. 6, which was bioconverted to yeast B after softening, had the most excellent antioxidant activity.

Sample 3 applied with softening technology was superior to sample 3 in which ginseng sprout leaves were simply bioconverted, and sample 6 bioconverted after softening was superior to sample 4 in which only softening technology was applied to ginseng sprout leaves. In addition, it was confirmed that sample No. 6 fermented with yeast B (28-7) had the best antioxidant activity than the sample fermented with yeast A (S288c), a standard yeast strain.

Test Example 3. Measurement of skin anti-inflammatory efficacy

Figure 5 is a graph measuring the HaCaT cell viability of fermented ginseng sprouts prepared according to Example 1 of the present invention and softened ginseng sprout leaves prepared according to Comparative Example 1; 6 is a graph measuring the level of IL-6, an inflammatory cytokine, when treated with fermented ginseng sprouts prepared in Example 1 of the present invention and softened ginseng sprout leaves prepared in Comparative Example 1; 7 is a graph measuring the level of IL-8, an inflammatory cytokine, when treated with the fermented ginseng sprouts prepared according to Example 1 of the present invention and the softened ginseng sprout leaves prepared according to Comparative Example 1.

HaCaT cell line, which is a skin cell, to confirm whether the fermented ginseng sprouts prepared according to Example 1 of the present invention and the softened ginseng sprout leaves prepared according to Comparative Example 1 have an improvement effect on skin inflammation induced by fine dust, etc. was utilized. HaCaT skin cells were placed in a culture container at a concentration of 4x10 ⁵ cells/mL and cultured in an animal cell incubator at 37 °C and 5% CO ₂ for 24 hours, and then the Ca ²⁺ concentration of the medium was changed to a high concentration. After 24 hours, replace the medium with fetal bovine serum-free medium, incubate in an incubator for 18 hours, process the sample, and after 1 hour, treat fine dust (PM; particulate matter, 10 μg/ml), and then incubate for 24 hours. did The cell medium was collected and centrifuged at 13000 rpm for 2 minutes to obtain a supernatant, and then MTS reagent was added to the culture vessel and reacted for 1 hour, and then cell viability was measured by measuring absorbance with a plate reader.

In the collected supernatant, ELISA kits were used to detect IL-6 and IL-8, which are representative skin inflammatory cytokines in the experimental group. Inflammatory cytokines such as IL-6 and IL-8 are produced when exposed to a stressful environment to protect the body from external stimuli such as fine dust or ultraviolet rays, which are involved in immune/inflammatory responses. However, when cytokines are chronically produced due to continuous skin damage, it causes skin dryness and causes skin diseases such as atopy. Therefore, IL-6, IL-8, TNF-a, and the like are being used as target materials for searching for materials that have the effect of suppressing the inflammatory response by reducing the secretion of inflammatory cytokines after exposure to a stressful environment.

First, in order to set concentrations that do not affect the survival of HaCaT cells when the fermented ginseng sprouts prepared according to Example 1 of the present invention and the soft ginseng sprouts prepared according to Comparative Example 1 were treated, 10 and 20 μg/ Cell survival was confirmed by treating the MTS reagent under the same conditions as those inducing skin inflammation due to fine dust at a concentration of ml.

As shown in FIG. 5, as a result of confirming cytotoxicity through the MTT assay, it was confirmed that both the fermented ginseng sprouts of Example 1 and the softened ginseng sprouts leaf softening material of Comparative Example 1 had excellent cell viability up to 20 μg/ml.

In addition, as shown in FIGS. 6 and 7 , it was confirmed that the fermented ginseng sprouts of Example 1 showed significant inhibition of L-6 and IL-8 production, which was not observed in the softened ginseng sprout leaf of Comparative Example 1.

This means that the softened sprout ginseng leaf has an effect of improving skin inflammation induced by fine dust after fermentation.

That is, the fermented ginseng sprouts of Example 1 did not undergo alcohol fermentation during fermentation, but it was confirmed that yeast growth appeared normally, and the antioxidant ability and the effect of reducing the secretion of inflammatory cytokines derived from fine dust were confirmed.

Preparation examples of cosmetic compositions suitable for application of the present invention will be presented.

Table 2 below shows preparation examples of lotion among cosmetics containing the fermented product of Example 1.

ingredient	Content (% by weight)
Fermented product of Example 1	5.0
glycerin	6.0
1,3-butylene glycol	3.0
PEG 1500	1.0
allantoin	0.1
DL-panthenol	0.3
EDTA-2Na	0.02
Benzophenone-9	0.04
sodium hyaluronate	5.0
ethanol	10.0
Polysorbate 20	0.2
preservatives, fragrances, colors	a very small amount
Distilled water	balance
Sum	100

Preparation Example 1: Lotion

Examples of preparation of a lotion among cosmetics containing the fermented product of Example 1 are shown in Table 3 below.

ingredient	Content (% by weight)
Fermented product of Example 1	3.0
propylene glycol	6.0
glycerin	4.0
triethanolamine	1.2
Tocopheryl Acetate	3.0
liquid paraffin	5.0
squalane	3.0
macadamia nut oil	2.0
polysorbate 60	1.5
Sorbitan sesquioleate	1.0
carboxyvinyl polymer	1.0
preservatives, fragrances, colors	a very small amount
Distilled water	balance
Sum	100

Preparation Example 2: Lotion

Examples of preparation of nutritional cream among cosmetics containing the fermented product of Example 1 are shown in Table 4 below.

ingredient	Content (% by weight)
Fermented product of Example 1	1.0
Pro-type glycerin monostearate	1.5
cetearyl alcohol	1.5
stearic acid	1.0
polysorbate 60	1.5
Sorbitan Stearate	0.6
isostearyl isostearate	5.0
squalane	5.0
mineral oil	35.0
dimethicone	0.5
Hydroxyethyl Cellulose	0.12
glycerin	6.0
triethanolamine	0.7
preservatives, fragrances, colors	a very small amount
Distilled water	balance
Sum	100

Preparation Example 3: Nutrition Cream

Examples of preparation of essence among cosmetics containing the fermented product of Example 1 are shown in Table 5 below.

ingredient	Content (% by weight)
Fermented product of Example 1	1.5
glycerin	10.0
betaine	5.0
PEG 1500	2.0
allantoin	0.1
DL-panthenol	0.3
EDTA-2Na	0.02
Benzophenone-9	0.04
Hydroxyethyl Cellulose	0.1
sodium hyaluronate	8.0
carboxyvinyl polymer	0.2
triethanolamine	0.18
Octyldodecanol	0.3
Octyldodeces-16	0.4
ethanol	6.0
preservatives, fragrances, colors	a very small amount
Distilled water	balance
Sum	100

Preparation Example 4: Essence

Examples of preparation of emulsion for mask pack among cosmetics containing the fermented product of Example 1 are shown in Table 6 below.

ingredient	Content (% by weight)
Fermented product of Example 1	1.0
polyvinyl alcohol	15.0
cellulose gum	0.15
glycerin	3.0
PEG 1500	2.0
cyclodextrin	0.15
DL-panthenol	0.4
allantoin	0.1
Monoammonium glycyrrhizinate	0.3
nicotinamide	0.5
ethanol	6.0
PEG 40 hydrogenated castor oil	0.3
preservatives, fragrances, colors	a very small amount
Distilled water	balance
Sum	100

Preparation Example 5: Emulsion for mask pack

[Accession number]

Name of Depository Organization: Korea Microbial Conservation Center

Accession number: KFCC11825P

Entrusted date: 20190401

[Copy of Deposit]

Claims (20)

1. Antioxidant or anti-inflammatory cosmetic composition comprising a fermented product prepared by fermenting a mixture of ginseng sprout leaf softener and ginseng sprout leaf extract with Saccharomyces cerevisi ae strain as an active ingredient .
2. According to claim 1, wherein the Saccharomyces cerevisiae (Saccharomyces cerevisi ae) strain is purchased from ATCC (American Type Culture Collection) Saccharomyces cerevisiae (Saccharomyces cerevisi ae, ATCC ^® 204508) strain Characterized in antioxidant or anti-inflammatory cosmetic composition.
3. According to claim 1, wherein the Saccharomyces cerevisiae (Saccharomyces cerevisi ae) strain is Saccharomyces cerevisiae having a nucleotide sequence represented by SEQ ID NO: 1 (Saccharomyces cerevisi ae) 28-7 strain [accession number : KFCC11825P] Antioxidant or anti-inflammatory cosmetic composition, characterized in that.
4. The method of claim 1, wherein the Saccharomyces cerevisi ae strain is inoculated into a mixture of the ginseng sprout leaf softener and the ginseng sprout leaf extract at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU / ml, Antioxidant or anti-inflammatory cosmetic composition, characterized in that the viable cell count after fermentation is 1X10 ⁸ to 9X10 ⁸ CFU / ml.
5. The cosmetic composition for antioxidant or anti-inflammatory according to claim 1, wherein the fermented product is used at a concentration of 10 to 1500 μg/ml.
6. The method of claim 1, wherein the softened ginseng sprout leaves are extracted from ginseng sprout leaves; Impregnating the filtered solid after the extraction with an aqueous solution containing an acid and purified water; Antioxidant or anti-inflammatory cosmetic composition comprising the steps of softening and warming the aqueous solution.
7. [Claim 7] The cosmetic composition for antioxidant or anti-inflammatory according to claim 6, wherein the extraction is performed under water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
8. The method of claim 6, wherein the acid comprises at least one selected from the group consisting of citric acid, tartaric acid, lactic acid, glycolic acid, formic acid, malic acid, oxalic acid, salicylic acid, ascorbic acid, citric acid and gluconodeltalactone. organic acids; Or an inorganic acid comprising at least one selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid; antioxidant or anti-inflammatory cosmetic composition comprising a.
9. [Claim 7] The cosmetic composition for antioxidant or anti-inflammatory according to claim 6, wherein the softening is performed at 40 to 95 °C for 30 to 240 minutes.
10. The cosmetic composition for antioxidant or anti-inflammatory according to claim 6, wherein the soft ginseng sprout has a pH of 2.5 to 4.5.
11. The antioxidant or anti-inflammatory cosmetic composition according to claim 1, wherein the softened ginseng sprout leaf and the ginseng sprout leaf extract are mixed in a weight ratio of 1:0.4-1.
12. The cosmetic composition for antioxidant or anti-inflammatory according to claim 1, wherein the composition inhibits the production of IL-6 and IL-8.
13. A fermented ginseng sprout obtained by mixing sprout ginseng leaf softening material and sprout ginseng leaf extract, inoculating the mixture with a Saccharomyces cerevisi ae strain, and culturing the mixture.
14. The method of claim 13, wherein ginseng sprout leaves are mixed with a solvent (water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof) in a weight ratio of 1: 9, extracted at 80 to 95 ° C for 2 to 4 hours, and then filtered. Separated into solids and extracts, the solids were impregnated with an aqueous solution (95 to 98% by weight of purified water, 5 to 2% by weight of citric acid), warmed to a temperature of 45 to 55 ° C., softened for 50 to 70 minutes, and washed with water A fermented ginseng sprout product characterized by obtaining a softened ginseng sprout leaf having a pH of 2.5 to 3.5, and using the extract separated from the solid after the extraction as a ginseng sprout leaf extract.
15. The method of claim 13, wherein the Saccharomyces cerevisiae (Saccharomyces cerevisi ae) strain is Saccharomyces cerevisiae having a nucleotide sequence represented by SEQ ID NO: 1 (Saccharomyces cerevisi ae) 28-7 strain [accession number : KFCC11825P],

    Or a fermented sprout ginseng product characterized in that it is a Saccharomyces cerevisi ae ( ATCC ^® 204508) strain purchased from ATCC (American Type Culture Collection).
16. The method of claim 13, wherein the Saccharomyces cerevisiae (Saccharomyces cerevisi ae) 28-7 strain or standard strain in a mixture of the ginseng sprout leaf softener and the ginseng sprout leaf extract at a weight ratio of 1: 0.4 to 1 S288c is inoculated at a concentration of 1X10 ⁶ to 1X10 ⁷ CFU / ml and fermentation is performed at 20 to 40 ° C. and 80 to 200 rpm for 60 to 80 hours.
17. The method of claim 13, wherein the fermented ginseng sprout leaf is contained as an active ingredient in a cosmetic composition for antioxidant or anti-inflammatory use and is used in the manufacture of any one of cosmetic lotions, lotions, nourishing creams, essences, and emulsions for mask packs. Sprout ginseng fermented product.
18. A method for producing fermented ginseng sprouts, comprising the steps of inoculating and fermenting Saccharomyces cerevisi ae in a mixture of ginseng sprout leaf softening material and ginseng sprout leaf extract.
19. The method of claim 18, wherein the soft ginseng sprout leaves,

    (a) extracting ginseng sprout leaves; (b) impregnating the extracted and filtered solids into an aqueous solution containing an acid and purified water; and (c) performing the step of warming and softening the aqueous solution to soften the leaves of ginseng sprouts.
20. The method of claim 18, wherein the fermenting step,

    The mixture of the ginseng sprout leaf softener and the ginseng sprout leaf extract was inoculated with the Saccharomyces cerevisi ae 28-7 strain at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU/ml at 20 to 40 °C and A method for producing fermented ginseng sprouts, characterized in that fermentation is performed for 60 to 80 hours at 80 to 200 rpm.

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