KR101778538B1

KR101778538B1 - Cosmetic composition comprising ginseng fruit Fermentation Product by Botrytis sp.

Info

Publication number: KR101778538B1
Application number: KR1020150150214A
Authority: KR
Inventors: 이동걸; 박지은; 윤석균; 여혜림; 노윤화; 강승현; 김연준
Original assignee: 코스맥스 주식회사
Priority date: 2015-10-28
Filing date: 2015-10-28
Publication date: 2017-09-15
Also published as: KR20170049756A

Abstract

The present invention relates to a cosmetic composition containing a ginseng fermented product obtained by fermenting ginseng fruit with Botrytis sp. And a process for producing the same.
According to the present invention, in the case of a cosmetic composition containing a fermented product of ginseng berry cultivar obtained by fermenting ginseng fruit with Botrytis sp., Or a fermented product of ginseng fruit obtained by fermenting ginseng with fungus bacteria and lactic acid bacteria, , It promotes cell proliferation, restores damaged cells, inhibits MMPs, and promotes collagen synthesis, thereby inhibiting skin aging and improving wrinkles. In addition, skin whitening effect can be exhibited by inhibiting melanin production.

Description

Technical Field [0001] The present invention relates to a cosmetic composition containing ginseng fermented product of Botrytis sp.

The present invention relates to a cosmetic composition containing a fermented product of ginseng fruit, and more particularly to a cosmetic composition containing a fermented product of ginseng fruit bran obtained by fermenting ginseng fruit with Botrytis sp.

Ginseng (Panax ginseng C.A. Meyer) is a herb that belongs to the genus Ogawa and Ginseng. It has been used in Korea, China and Japan for over 2,000 years and has been used empirically to prevent disease and prolong the life span. The efficacy and effect of ginseng which has been known so far can be determined by the action of the central nervous system, the anticarcinogenic action, the anticancer activity, the immune function regulating action, the anti-diabetic action, the hyperfunction of liver function, the improvement of cardiovascular disorder, (Korean Ginseng & Tobacco Research Institute, 56-112, 1996). In addition, it has been reported that the antioxidant activity and antioxidant activity of osteoporosis are improved. Ginsenoside, which is a representative physiologically active ingredient of ginseng, is distributed evenly on the ground and underground of ginseng. Especially, ginsenoside (root), ginseng leaf and ginseng berries have not only ginsenoside content but also composition . (Attele AS et al, Biochem Pharmacol, 58: 1685-1693, 1999). In particular, the fruit of ginseng has been reported to exhibit superior antidiabetic efficacy to ginseng root based on the contents of ginseng roots and other ginsenosides (Dey L. et al., Phytomedicine, 10, 600 -605, 2003).

Recently, consumers are interested in natural cosmetics. At the same time, many cosmetics using herbal materials have been launched, and ginseng has been studied as a plant material having important skin effect of cosmetics. However, most of them are ginseng root extract or ginseng root ginsenoside Side component and ginseng polysaccharide are utilized only for the inhibition of skin aging and the wrinkle improving effect due to the ingredient of ginseng fruit. The ginseng fruit is considered to be more valuable than ginseng since ancient times and has been harvested for the purpose of obtaining seeds. Ginseng fruit is grown only once in 4 years of ginseng cultivation period. When seeds are cultivated in 3 year olds, it is difficult to produce good seedling ginseng because seeds are too small. If seeds are grown for 5 years or more, seeds are faithful and good, The quality of red ginseng can be greatly reduced when red ginseng is produced because it is not only inhibited development but also is inadequate in the tissue. When the number of seedlings is increased to more than 2 times during the growing period, the yield and the quality of red ginseng deteriorate significantly (Korean Ginseng and Tobacco Research Institute, 130-131, 1996).

US Patent No. 6524626 discloses that ginseng fruit juice is used in a cosmetic composition by freeze-drying, and that by using a combination of ginseng fruit juice and other natural substances, they exhibit moisturizing and skin-softening effects. However, It is clearly distinguished from the cosmetic use as an anti-aging effect and an anti-wrinkle effect of the extract.

The human skin undergoes changes due to various internal and external factors as it gets older. In other words, internally, secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction As the content of ultraviolet rays reaching the surface of the sunlight increases and environmental pollution is further intensified, free radicals and active noxious oxygen are increased, so that the thickness of the skin decreases, the wrinkles increase, the elasticity decreases Skin color is grayish, skin troubles occur frequently, and spots, freckles and black blots also increase.

As the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid and glycoprotein, which constitute the skin, are changed or decreased, and oxidative stress due to free radicals and active noxious oxygen is experienced. In addition, most of the cells constituting the skin are biosynthesized by Cox-2 (cyclooxygenase), an enzyme that produces proinflammatory cytokines, known to cause aging or inflammation by ultraviolet light , The biosynthesis of matrix metalloproteinase (MMP), an enzyme that degrades skin tissue by these inflammatory factors, is increased, and nitric oxide (NO) production by iNOS (inducible nitric oxide synthase) is increased . In other words, the decrease of cell activity due to natural endogenous aging and the decrease of biosynthesis of substrate materials due to microinflammation, the increase of stress caused by various harmful environments, and the increase of active oxygen species by sunlight The skin is degraded and thinned and the symptoms of skin aging are revealed.

Therefore, it is a reality that many researches have been conducted on active ingredients that can prevent and improve such aging phenomena. Several factors are involved in determining the skin color of a person. Among them, factors such as the activity of melanocytes that make melanin pigment, the distribution of blood vessels, the thickness of skin, and the presence or absence of pigment in and out of human body such as carotenoid and bilirubin Are important. Especially, the most important factor is melanin, a melanin pigment produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of melanin pigment affects genetic factors, hormonal secretion, physiological factors such as stress, and environmental factors such as ultraviolet irradiation. The melanin pigment produced in melanocytes of body skin is a phenolic polymer substance having a complex form of black pigment and protein. It protects the subcutaneous skin organs by blocking ultraviolet rays irradiated from the sun, and at the same time, free radicals And protects proteins and genes in the skin.

Melanin, which is produced by stress stimuli inside and outside the skin, is a stable substance that does not disappear until the skin is excreted through skin keratinization even if the stress disappears. However, when melanin is produced more than necessary, it induces hypercholesterolemia such as spots, freckles, and dots, resulting in poor cosmetic results.

Today, oriental women prefer white, clean skin like white whites, and this is an important criterion of beauty, so there has been a growing demand for treatment and cosmetic problems for hyperpigmentation.

In response to such demands, substances having ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione, derivatives thereof, or tyrosinase inhibiting activity have been conventionally used in cosmetics or pharmaceuticals, The use thereof is limited due to a whitening effect, a safety problem to the skin, a problem of formulation and stability in combination with cosmetics, and the like.

The most important function of the epidermis on the outermost skin is to protect against various external stimuli (chemical, air pollutants, dry environment, physical and chemical stimulants such as ultraviolet rays) It is a protective function to prevent excessive divergence of water in the body. This protective function can maintain its function only when the stratum corneum composed of keratinocytes is normally formed. The stratum corneum (horney layer), which is the outermost part of the epidermis, is formed from keratinocytes and consists of keratinocytes with complete differentiation and a surrounding lipid layer (J. Invest. Dermatol. 1983; 80: 49). The keratinocyte is a characteristic cell in which basal cells continuously proliferating in the lowest epidermis undergo a stepwise change in morphology and function and are elevated to the surface of the skin. After a certain period of time, And the new keratinocyte replaces its function. This repetitive sequence of changes is called "epidermal cell differentiation" or "keratinization". During keratinization, keratinocytes form the stratum corneum while producing natural moisturizing factors (NMF) and intercellular lipids (ceramides, cholesterol, fatty acids), which make the stratum corneum firm and flexible, As shown in Fig. However, such a stratum corneum can easily lose its function due to excessive skin cleansing, lifestyle factors such as bathing, environmental factors such as dry air and pollutants, and endogenous diseases such as atopic skin and senile skin, Due to the increasing number of factors in the modern age, there has been an increasing tendency in recent years to appeal to people with dry skin symptoms and disorders. Therefore, in order to maintain proper skin moisture, studies have been conducted to supply moisture from the outside or to prevent water loss from the body, and a variety of moisturizers having moisture retention ability have been developed, It is widely used in the area.

However, as the risk factors for humans increase in the living environment and the aging population rapidly increases, the turnover rate of the stratum corneum is delayed, the lipid synthesis ability of the keratinocytes decreases, The cells are not smoothly divided, maturated and differentiated, and the amount of moisturizing factors and lipids in the stratum corneum is decreased, so that the normal stratum corneum does not maintain its function. Namely, . Due to the abnormal division and differentiation of the epidermal cells, the skin causes various skin diseases such as dry skin, atopy and psoriasis. Such a disease can slightly alleviate the symptoms only by a conventional moisturizing agent having only moisture holding function , But it is difficult to expect fundamental healing. In general, sebaceous glands in the skin, including the scalp and face, play a role in preventing moisturization and microbial invasion of the skin. However, excessive secretion of sebum promotes hair loss, worsens acne, promotes enlargement of pores caused by acne, And many other diseases are caused. The hypersecretion of sebaceous glands is caused by various causes, and the most important one is the activation of sebaceous glands by the amount of dihydrotestosterone (DHT), which is one of the hormones involved in promoting sebaceous secretion in the activity of sebaceous glands, Is overdone. That is, testosterone (T) is converted into dihydrotestosterone, one of the male hormones, by 5-alpha-reductase type 2 in cells in the hair loss, (5-α-reductase type 1) in the skin or the sebaceous glands. The testosterone is converted to dihydrotestosterone by the action of 5-α-reductase type 1, (J. Invest Dermatol 105: 209-214 Diane et al.). Acne and hair loss, which are skin troubles other than simple sebum hypersecretion, are further exacerbated by the minute inflammatory reaction of the skin. If you look at the development of acne, excess sebum accumulates in the hair follicle, activating acne bacteria and causing inflammation. It has antioxidant effect with excellent antioxidant effect, inhibiting the production of prostaglandin, especially prostaglandin E2 (PGE2) in cell inflammation reaction, inhibiting the production of nitric oxide (NO) by iNOS, It is possible to alleviate the skin troubles. However, there are not many raw materials that can reduce sebum secretion as a skin-safe natural material through fundamental causes and at the same time to relieve inflammation.

The inventors of the present invention have made intensive efforts to overcome the problems of the prior art. As a result, the inventors of the present invention have found that the ginseng fruit is a fermented product of ginseng fruit bran obtained by fermentation by Botrytis sp. The cosmetic composition containing the fermented product of ginseng fruit obtained by fermenting the ginseng fruit with the prey bacterium and the lactic acid bacterium promotes cell proliferation, restores damaged cells, inhibits MMPs, inhibits skin aging by promoting collagen synthesis And it is possible to improve the wrinkles and inhibit the melanin production, thereby confirming that the skin whitening effect can be exhibited. Thus, the present invention has been completed.

US 6524626 B2

Attele AS et al, Biochem Pharmacol, 58; 1685-1693, 1999 Dey L. et al., Phytomedicine, 10; 600-605, 2003 Latest KOREA Ginseng 'Cultivation', Korea Ginseng & Tobacco Research Institute, 130-131, 1996

Therefore, the main object of the present invention is to provide a method for inhibiting skin aging and wrinkle by inhibiting melanin production by promoting cell proliferation, repairing damaged cells, inhibiting MMPs, promoting collagen synthesis, And a fermented product of the ginseng fruit of the present invention.

It is another object of the present invention to provide a method for producing a cosmetic composition containing the fermented product of ginseng berry extract using the ginseng fermented product.

According to one aspect of the present invention, there is provided a cosmetic composition comprising a fermented product of ginseng fruit bran obtained by fermenting ginseng fruit with Botrytis sp.

Recently, ginseng has been studied as one of the cosmetic materials as consumer interest in natural cosmetics has been increased. However, studies on inhibition of skin aging of the ginseng fruit ingredient, wrinkle improving effect and skin whitening effect are still insignificant. Thus, the inventors of the present invention have found that when a fermented product obtained by fermenting ginseng fruit with a prey bacterium as a cosmetic composition is effective in inhibiting skin aging and improving wrinkles, the present invention has been completed.

In the present invention, the ear bacillus is a bacterium belonging to the genus Borotis, preferably Botrytis cinerea.

The bacterium belonging to the genus Bautritis can be found in soil and decay fruit, and is usually present as a pathogen of fruits and vegetables. It is the same in morphology, but differs in that it does not have polluted mold and mold odor of strawberry. Colony is gray or dark brown, and it forms irregularly stellate endospores from bulbous, long-lined hyphae. Conidia are oval to spherical, 1 cell, and B. allii is also known as gray onion and leek germs. It is known to be used in the manufacture of wines in Europe, but has never been used to ferment the ginseng fruit.

In the present invention, the ginseng fermented bacterium fermented product is characterized in that the ginseng fruit is placed on a solid medium and inoculated with the ear bacterium to perform solid phase fermentation.

In the experimental example of the present invention, when the fermented product was to be fermented only by the ear bacterium, solid phase fermentation proceeded without proceeding with liquid fermentation (see Example 2).

In the present invention, the term "solid-state fermentation" means fermentation in a solid state using air as a medium by placing the fermentation material on a solid medium or completely exposing the fermentation material to air without solid medium. The solid medium may be any solid medium known in the art, but it is preferable to use agar medium. Solid phase fermentation is known to maximize the diversity and yield of functional materials when compared with liquid fermentation, and the yield is also excellent. On the other hand, liquid fermentation means that fermentation material is put into a liquid medium and fermented in a liquid state using water as a medium.

Conventionally, it is known that fermentation of ginseng fruit with yeast or lactic acid bacteria is carried out in liquid phase, but it is not yet known how to ferment the ginseng fruit using the ginseng fungus. In addition, it was confirmed that when the ginseng fruit was solid - phase fermented using ear buds, the effect was better than that of liquid fermentation with yeast or lactic acid bacteria.

The method of the present invention may further comprise a second fermentation step in which the solid fermented ginseng fruit is put into a liquid medium and the lactic acid bacteria are further inoculated and subjected to liquid fermentation after the solid phase fermentation step. The lactic acid bacteria may be Lactobacillus sp , Leuconostoc sp., And Bifidobacteria sp.). In the present invention,

In the case of solid fermentation using only the ear bacterium, the effect was more excellent than that in the case of liquid fermentation by inoculation with yeast or lactic acid bacteria, but the second liquor fermentation with lactic acid bacteria after the first solid phase fermentation as the ear bacterium was more effective I did.

In the present invention, the fermented product of the ginseng berry strain may be contained in an amount of 0.001 to 30 wt%, preferably 0.001 to 30.0 wt%, based on the total weight of the composition. If the content of the active ingredient is less than 0.001% by weight, the effect or effect is insufficient. If the content of the active ingredient is more than 90% by weight, there is a problem in stabilizing the formulation and the increase in the effect is insignificant.

In the present invention, the composition is characterized by having an effect of suppressing skin aging and improving wrinkles. In addition, the composition promotes collagen production, exhibits an MMP-1 inhibitory effect, and exhibits cell regeneration and cell proliferation effects.

According to the Experimental Example of the present invention, the fermented product obtained by solid-phase fermentation of the ginseng fruit into the fungus bacterium (Example 2) was superior to the fermented product obtained by liquid fermentation with yeast or lactic acid bacteria (Comparative Example 2-3) -1 inhibition effect, and cell regeneration and cell proliferation effect. Moreover, the fermented product obtained by fermenting liquid fermented with lactic acid bacterium after solid phase fermentation (Example 3) showed better collagen production promotion effect, MMP-1 inhibitory effect and cell regeneration And cell proliferation effect. As a result, the fermented product obtained by solid-phase fermentation of ginseng fruit as a prey bacterium and the fermented product fermented by liquid fermentation with lactic acid bacterium after solid phase fermentation promotes collagen production, inhibits MMP-1, promotes cell regeneration and cell proliferation Skin aging inhibition and wrinkle-improving effect (see Experimental Example 1-4).

In the present invention, the composition is characterized by having a skin whitening effect. Further, the composition is characterized by exhibiting an effect of inhibiting melanin production.

According to the Experimental Example of the present invention, the fermented product obtained by solid-phase fermentation of the ginseng fruit into the fungus bacterium (Example 2) exhibited a better melanin production inhibitory effect than the fermented product obtained by liquid fermentation with the yeast or lactic acid bacterium (Comparative Example 2-3) Furthermore, the fermented product fermented by liquid fermentation with lactic acid bacterium after solid phase fermentation (Example 3) showed more excellent inhibitory effect on melanin production. From these results, it can be seen that the fermented product obtained by solid-phase fermentation of the ginseng fruit as a prey bacterium and the fermented product fermented by the liquid fermentation with the lactic acid bacterium after the solid phase fermentation can suppress the melanin production, 5).

In the present invention, the composition may be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, , A soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser. For example, softening longevity, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body essence , And may also be a cosmetic composition having transdermal delivery formulations such as lotions, ointments, gels, creams, patches or sprays.

In accordance with the present invention, there is provided a process for producing a fermented product of a ginseng fruit bacterium for cosmetic composition comprising the following two steps.

a) primary fermentation stage in which ginseng fruit is placed on a solid medium and inoculated with ear buds to perform solid phase fermentation; And

b) Secondary fermentation step in which the first fermented ginseng fruit is put into a liquid medium and the lactic acid bacteria are further inoculated to perform liquid fermentation.

The method for producing the fermented extract of the ginseng fruit used in the composition of the present invention is particularly limited and can be carried out by a general method known in the art. Specifically, the ginseng can be obtained by adding the ginseng to the ginseng fruit, have.

More specifically, when using Botrytis cinerea during fermentation, the ginseng fruit sterilized at 121 ° C was sprayed onto a Potato Dextrose Agar solid medium, Botrytis cinerea seedlings were inoculated in the middle of the sprayed tray, and cultured for 7 days do. At this time, the cells to be used are inoculated (about 105 to 107 cells / mL) and cultured. After 7 days of fermentation, the first fermented ginseng fruit is re-mixed and mixed with MRS media (BD) at a ratio of 1: 3 and prepared by high-pressure sterilization at 121 ° C. After completion of sterilization, lactic acid bacteria are inoculated to perform secondary fermentation. At this time, the inoculated cells (about 107 ~ 109) are inoculated. The inoculated ginseng fruit is fermented for about 5 days. After the culture, the fermentation broth is centrifuged to obtain a final fermented extract of ginseng fruit from which the polymer precipitate and the lactic acid bacterium have been removed.

As described above, the cosmetic composition containing the fermented product of the ginseng fruit bacterium using the ear bacterium provides an effect of improving the overall skin condition while safely protecting the skin through skin whitening effect as well as skin aging inhibition and wrinkle improvement effect can do.

Figure 1 is a diagram showing the solid phase fermentation of ginseng fruit.
2 is a graph showing the cell proliferation effect of the fermented product of ginseng fruit. (GB: ginseng fruit extract (Example 1), GBF: ginseng fruit + lactobacillus ginseng fruit extract (Example 3)
FIG. 3 is a graph showing the cell regeneration effect of the fermented product of ginseng fruit. (GB: ginseng fruit extract (Example 1), GBF: ginseng fruit + lactobacillus ginseng fruit extract (Example 3)
4 is a graph showing the inhibitory effect of MMP-1 on the fermented product of ginseng fruit. (GB: ginseng fruit extract (Example 1), GBF: ginseng fruit + lactobacillus ginseng fruit extract (Example 3)
Fig. 5 is a graph showing the inhibitory effect of fermenting bacteria and ginseng fermented product on MMP-1. (Comparative Example 1-1), G-2: Lactic Acid Bacteria Extract (Comparative Example 1-2), G-6: Ear Bud Extract (Comparative Example 1-3), G-8: Ginseng Extract Fermented product of lactic acid bacteria (Example 3))
6 is a graph showing the inhibitory effect of MMP-1 on the fermented product of ginseng fruit. (Comparative Example 2), G-3: Ginseng fermented yeast fermented product (Comparative Example 3), G-4: Ginseng fermented lactic acid fermentation product (Comparative Example 2)
FIG. 7 is a graph showing the effect of promoting collagen synthesis of the ginseng fruit fermented product. FIG. (GB: ginseng fruit extract (Example 1), GBF: ginseng fruit + lactobacillus ginseng fruit extract (Example 3)
FIG. 8 is a diagram showing the promoting effect of collagen synthesis of fermenting bacteria and ginseng fruit fermented product. FIG. (Comparative Example 1-1), G-2: Lactic Acid Bacteria Extract (Comparative Example 1-2), G-6: Ear Bud Extract (Comparative Example 1-3), G-8: Ginseng Extract Fermented product of lactic acid bacteria (Example 3))
FIG. 9 is a graph showing the promoting effect of collagen synthesis of the ginseng fruit fermented product. FIG. (Comparative Example 2), G-3: Ginseng fermented yeast fermented product (Comparative Example 3), G-4: Ginseng fermented lactic acid fermentation product (Comparative Example 2)
10 is a graph showing the melanin production inhibitory effect of fermenting bacteria and ginseng fruit fermented products. (Comparative Example 1-1), G-2: Lactic Acid Bacteria Extract (Comparative Example 1-2), G-6: Ear Bud Extract (Comparative Example 1-3) , G-8: fermented product of ginseng fruit extract + lactic acid bacteria (Example 3))
Fig. 11 is a graph showing the melanin production inhibitory effect of the fermented product of ginseng fruit. (AB: alpha-arbutin), G-3: fermented product of ginseng fruit yeast (Comparative Example 3), G-4: fermented product of lactic acid fermented with ginseng (Comparative Example 2), G-5: fermented product of ginseng fruit Example 2))

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Pretreatment of primary fermentation of ginseng, preparation and preparation of ginseng fruit extract

1. Ginseng fruit pretreatment

The raw ginseng fruit was harvested and the seeds were separated and removed. Then, the ginseng fruit and the skin of the ginseng fruit were pretreated through daylight drying or flowing water washing.

2. Ginseng fruit sterilization treatment and first preparation

After the raw ginseng fruit is washed, it is subjected to high pressure wet sterilization at 121 ° C, and it is naturally dried under sterile conditions. Potato Dextrose Agar media is solidified in a tray of size 100 * 100 * 10 cm and prepared by spraying well dried dried berry on it.

3. Preparation of ginseng fruit extract

To prepare ginseng fruit extract, 300 g of ginseng fruit was washed with flowing water, dried naturally, partially crushed with a crusher, and subjected to compression extraction using a juicer. Extracted ginseng fruit extracts were sterilized by using 0.2um syringe filter for sterilization.

Example 2: Fermented Extract of Ginseng Fruit Using Ear Bud

The ginseng fruit of Example 1 was sprayed on the tray, and pre-cultured pre-cultured bacterium (Botrytis cinerea) was inoculated in the middle of the tray and cultured for 7 days (168 hours). The amount of cells used for inoculation is 10 ⁷ -10 ⁹ , and incubation temperature is 28 ° C. After 7 days, the fermented ginseng fruit on the tray is re-recovered and mixed with MRS media at a ratio of 1: 3 and sterilized at 121 ° C.

Example 3: Fermented Extract of Ginseng Fruit Using Mycorrhizae + Lactic Acid Bacteria

The ginseng fruit of Example 1 was sprayed on the tray, and pre-cultured ear bacillus was inoculated in the middle of the tray and solid cultured for 7 days (168 hr). The amount of cells used for inoculation is 10 ⁷ -10 ⁹ , and incubation temperature is 28 ° C. After 7 days, the fermented ginseng fruit on the tray is re-recovered and mixed with MRS media at a ratio of 1: 3 and sterilized at 121 ° C. Lactobacillus sp. CXHB-4 was inoculated at 10 ⁷ -10 ⁹ cells / mL in the final sterilized MRS media and ginseng fruit culture medium, cultured for 5 days (120 hr) , A temperature of 35 to 38 DEG C, and a ventilation amount of 1 VVM. The fermentation broth was centrifuged at 10000 xg for 30 minutes to obtain a final ginseng fermentation extract in which the polymer precipitate and cells were removed.

Comparative Example 1 Yeast Extract, Lactic Acid Bacteria Extract, Ear Bud Extract

1. Yeast extract

In order to cultivate Saccharomyces sp., CXHB-2 isolated from ginseng, Potato Dextrose Broth (PDB) medium is subjected to 1 liter wet autoclaving (121degree). After sterilization, the temperature is cooled to 25 ° C and the seeds are inoculated. The inoculated PDB medium was agitated at 28 ° C and 100 rpm for 5 days. When the cells were cultured for 5 days, the amount of the cells was found to be about 10 ⁹ / ml. The culture broth was centrifuged at 4000 × g for 1 hour using a centrifuge, and the supernatant was removed and washed twice with sterilized distilled water. The washed cells were dried at room temperature (25 ° C) for 4 days, re-dissolved in 50 ml of sterilized distilled water, and the cells were disrupted for 1 minute using an ultrasonic disrupter and centrifuged (4000 xg) The supernatant was used for comparative experiments.

2. Lactic acid bacteria extract

In order to cultivate Lactobacillus sp. CXHB-4 isolated from ginseng, the MRS medium is subjected to 1 liter wet autoclaving (121 ° C). After sterilization, the temperature is cooled to 25 ° C and the seeds are inoculated. The inoculated MRS medium was incubated at 37 ° C for 5 days. When the cells were cultured for 5 days, the amount of the cells was found to be about 10 ⁹ / ml. The culture broth was centrifuged at 4000 × g for 1 hour using a centrifuge, and the supernatant was removed and washed twice with sterilized distilled water. The washed cells were dried at room temperature (25 ° C) for 4 days, re-dissolved in 50 ml of sterilized distilled water, and the cells were disrupted for 1 minute using an ultrasonic disrupter and centrifuged (4000 xg) The supernatant was used for comparative experiments.

3. Ear buds extract

In order to cultivate Botrytis cinerea (CXHB-3) isolated from ginseng, PDA medium is subjected to 1 liter wet autoclaving (121 degrees). After sterilization, the medium is poured into a 150-pie petri dish and solidified. Inoculate the seeds in the hardened PDA medium and incubate in a 28 degree incubator for about 7 days. After the cells were cultured, the cells were cut using a sterilized surgical knife, and 5 g of the cells were dissolved in 50 ml of sterilized distilled water. The cells were disrupted for 1 minute using an ultrasonic disrupter and then centrifuged (4000 X g) Were used for comparison experiments.

Comparative Example 2: Fermented extract of ginseng fruit using lactic acid bacteria

Lactobacillus sp. CXHB-4 was inoculated at 10 ⁷ -10 ⁹ cells / mL into the final sterilized MRS media and ginseng fruit culture medium in Example 2, and cultured for 5 days (120 hr) , pH 6.5 to 7.5, temperature 35 to 38 占 폚, and aeration amount 1 VVM. The fermentation broth was centrifuged at 10000 xg for 30 minutes to obtain a final ginseng fermentation extract in which the polymer precipitate and cells were removed.

Comparative Example 3: Yeast extract of fermented ginseng fruit

Cells were inoculated at 10 ⁷ -10 ⁹ cells / mL in sterile PD media and ginseng fruit mixed medium and incubated for 5 days (120 hr) To 38 캜, and a ventilation amount of 1 VVM. After the fermentation, the fermentation broth was centrifuged at 10,000 xg for 30 minutes to obtain a final fermented extract of ginseng fruit in which the polymer precipitate and cells were removed.

Test Example 1: Assessment of cell proliferation promoting ability of fermented product of ginseng fruit

The effect of promoting cell proliferation in human fibroblast Hs68 fibroblast of ginseng fruit (Example 1) and fermented product of ginseng fruit (Example 3) was evaluated by MTT (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyltetrazolium bromide reagent.

Human fibroblast Hs68 fibroblast was dispensed into 96-well plates at a concentration of 2 × 10 ⁴ cells / well and cultured in an incubator at 37 ° C and 5% CO ₂ for 24 hours. After that, the medium was replaced with the medium supplemented with the extract of Comparative Example 1 or the fermented material of Example 2, and further cultured for 24 hours. After incubation, the medium was removed from each well, DPBS washing step was performed once, MTT reagent with 0.5 mg / ml concentration was added and reacted at 37 ° C for 4 hours. Formazan crystals formed during the reaction were dissolved in DMSO after removing the MTT reagent, and then measured at 590 nm using an ELISA reader. Cell viability was expressed as a percentage of the average absorbance value of the control group without any sample.

As a result, as shown in Fig. 2, it was confirmed that the ginseng fermented product (Example 3) exhibited higher cell proliferation potency than the ginseng fruit extract (Example 1) compared to the same concentration.

Test Example 2: Ginseng fruit Fermented Cell damage Recoverability evaluation

Exposure to ultraviolet light has direct action on the cell and inhibits cell proliferation. In order to compare the ability of the ginseng fruit extract (Example 1) and the fermentation product (Example 3) to recover cell damage, experiments were conducted using a human fibroblast cell line (Hs68) irradiated with ultraviolet light.

After removing the medium from the fibroblasts inoculated and cultured at the same concentration as in Test Example 1, DPBS was added, and the remaining cells except for the control were irradiated with UVB of 15 mJ / cm ^2. The extracts of Example 1 and Example 3 The fermented product was replaced with a medium supplemented with each concentration and cultured for an additional 24 hours. The subsequent test using the MTT reagent was carried out in the same manner as in Test Example 1.

As a result, as shown in FIG. 3, the ginseng fruit extract and the fermented product showed excellent cell damaging restoration effect, but in the case of the high concentration treated group, the fermented product (Example 3) exhibited remarkably excellent efficacy.

Test Example 3: Evaluation of MMP-1 Inhibition

In order to compare the inhibitory effect against the collagenase (MMP-1) increased by ultraviolet rays of Examples 1-3 and 1-3, MMP-1 expression level was measured using human fibroblast cell line Hs68 Respectively.

The fibroblast cell line Hs68 was divided into 4 × 10 ⁵ cells in a 6-well plate, and cultured in an incubator at 37 ° C. and 5% CO ₂ for 24 hours. After the medium was removed, DPBS was added, and UVB of 15 mJ / cm ² was irradiated to the remaining cells except the UVB non-irradiated group. Then, the ginseng fruit extract or the fermented substance was added to each concentration and further cultured for 24 hours. Then, RNA was isolated from the cells treated with each sample using triazole (RNA iso, DAKARA, Japan), and RNA was quantitated at 260 nm using nanodrop. Then, 2 μg of each RNA was used for amplification cDNA was synthesized (C1000 Thermal Cycler, Bio-Rad, USA). Real-time PCR was performed in a real-time PCR machine using a mixture of a target protein, MMP-1, and a cyanogen dye, SYBR Green supermis (Applied Biosystems, USA) The expression level of the MMP-1 gene was evaluated. The sequence of the template and the reaction conditions were as shown in Table 1 below, and the expression amount of the gene was finally analyzed by correcting for the β-actin gene.

Primer order Reaction conditions MMP-1 F 5'-CGAATTTGCCGACAGAGATGA-3 ' After activation of the polymerase at 94 ° C for 5 minutes, polymerization was carried out at 40 ° C for 30 seconds at 95 ° C for 30 seconds, at 55 ° C for 30 seconds, and at 72 ° C for 30 seconds R 5'-GTCCCTGAACAGCCCAGTACTT-3 ' COL1A1 F 5'-CACCAATCACCTGCGGTACAGAA-3 ' R 5'-CAGATCACGTCATCGCACAAC-3 ' COL3A1 F 5'-TGGAGGATGGTTGCACGAAA-3 ' R 5'-ACAGCCTTGCGTGTTCGATA-3 ' β-Actin F 5'-GGCCATCTCTTGCTCGAAGT-3 ' R 5'-GAGACCTTCAACACCCCAGC-3 '

As a result, as shown in Fig. 4 to Fig. 6, the ginseng fruit extract (Example 1) and the ginseng fruit fermented product (Example 2-3) were significantly increased in the treatment group while the MMP-1 expression level was rapidly increased in the UVB irradiation group (Comparative Example 2) or yeast (Comparative Example 3) among the ginseng fruit fermented products showed a decrease in the expression level of the ginseng fermented product in the fermented product of the ginseng fruit (Example 2) (Example 3) exhibited remarkably excellent MMP-1 expression inhibiting activity.

Test Example 4: Assessment of collagen synthesis promoting ability of fermented product of ginseng fruit

Experiments were carried out under the same conditions using the same cells as in Test Example 3 in order to compare and evaluate the collagen synthesis promoting effects of Examples 1-3 and 1-3 of the present invention. In the process of real-time polymerase chain reaction, COL1A1 template, which is a collagen precursor synthesis gene, was used instead of MMP-1 template. The sequence of the template and the reaction conditions were as shown in Table 1, and the expression level of the gene was finally analyzed by correcting for the β-actin gene.

As a result, as shown in FIG. 7-9, the expression of COL1A1 reduced by UVB irradiation did not increase or the increase rate was small when the extract of Ginseng Fruit (Example 1, Comparative Example 1-5) was treated, whereas the fermentation product The results are shown in Table 2.

Experimental Example 5: Assessment of inhibition of melanin formation

In order to comparatively evaluate the melanogenesis inhibitory effects of Examples 1-3 and Comparative Examples 1-3 of the present invention, mouse mel-Ab cells (Dooley, TP et al, Skin pharmacol, 7 , pp 188-200) were cultured in DMEM supplemented with 10% fetal calf serum, 100 nM 12-O-tetradecanoylphorbol-13-acetate and 1 nM cholera toxin at 37 ° C and 5% CO2. The cultured Mel-Ab cells were removed with 0.25% trypsin-EDTA, and the cells were cultured at a concentration of 10 ⁵ cells / well (cells / well) in a 24-well plate. And cultured. As test substances, arbutin and the extracts of Examples 1-3 and Comparative Examples 1-3 were used at a concentration of 100 ppm. At this time, the hydroquinone was used as a positive control. Then, the culture solution was removed, washed with PBS, and the cells were dissolved with 1N sodium hydroxide. The absorbance at 400 nm was measured, and the inhibition rate of melanin production was calculated.

As a result, as shown in FIG. 10-11, compared with the control group in which melanin production was rapidly increased by the alpha-MSH treatment, the melanin production was not suppressed or the effect was small in Example 1 and Comparative Example 1-3, Of Example 2, which is a fermented product of ginseng fruit fermented with a prey bacterium or a prefecture bacterium and a lactic acid bacterium, is excellent in the melanin production inhibitory effect.

Claims

The present invention relates to a cosmetic composition comprising ginseng fruit obtained by fermenting ginseng fruit with Botrytis sp. The present invention relates to a cosmetic composition comprising a fermented product of ginseng fruit ear buds obtained by fermenting ginseng fruit with Botrytis sp. Wherein the composition is solid-phase fermented.

The cosmetic composition according to claim 1, wherein the ear bacterium is Botrytis cinerea.

delete

The cosmetic composition according to claim 1, further comprising a second fermentation step in which the solid-phase fermented ginseng fruit is put into a liquid medium after the solid-phase fermentation step and the lactic acid bacteria are further inoculated to perform liquid fermentation.

The composition according to claim 4, wherein the lactic acid bacterium is at least one selected from the group consisting of Lactobacillus sp., Leuconostoc sp., And Bifidobacteria sp.

[Claim 3] The cosmetic composition according to claim 1, wherein the fermented product of the ginseng root extract is contained in an amount of 0.001 to 30.0% by weight based on the total weight of the composition.

The cosmetic composition according to claim 1, wherein the composition has an effect of inhibiting skin aging and improving wrinkles.

The cosmetic composition according to claim 1, wherein the composition has a skin whitening effect.

The composition according to claim 1, wherein the composition is at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, Wherein the cosmetic composition is at least one selected from the group consisting of a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

A method for producing a fermented product of a ginseng berry extract for cosmetic composition comprising the following two steps:
a) primary fermentation stage in which ginseng fruit is placed on a solid medium and inoculated with ear buds to perform solid phase fermentation; And
b) Secondary fermentation step in which the first fermented ginseng fruit is put into a liquid medium and the lactic acid bacteria are further inoculated to perform liquid fermentation.