KR20170122431A - A cosmetic containing fermentation product of water extract of broccoli, fermentation product of water extract of spinach or fermentation product of water extract of tomato by Daedaleopsis Tricolor mycelium for skin moisturizing - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin moisturization which comprises, as active ingredients, one or more selected from a broccoli fermentation product obtained by fermenting a hot water extract of broccoli with heat-killed cells of Daedaleopsis tricolor, a spinach fermentation product obtained by fermenting a hot water extract of spinach with heat-killed cells of Daedaleopsis tricolor, and a tomato fermentation product obtained by fermenting a hot water extract of tomato with heat-killed cells of Daedaleopsis tricolor. The fermentation products contained as active ingredients in the cosmetic composition of the present invention, such as the broccoli fermentation product obtained by fermenting a hot water extract of broccoli with heat-killed cells of Daedaleopsis tricolor, the spinach fermentation product obtained by fermenting a hot water extract of spinach with heat-killed cells of Daedaleopsis tricolor, and the tomato fermentation product obtained by fermenting a hot water extract of tomato with heat-killed cells of Daedaleopsis tricolor, exhibit excellent effects in terms of antioxidant activity, skin restoration, and skin moisturization.

Description

The fermented broccoli fermented product obtained by fermenting the broccoli hot water extract with the tricolor coated mushroom mycelia and the spinach fermented product obtained by fermenting the fermented product of broccoli fermented product and spinach hot water extract with the tricolor coated mushroom mycelium and fermenting the hot water extract of tomato with the tricolor coated mushroom mycelium, The present invention relates to a cosmetic composition for skin moisturizing, which comprises as an active ingredient a cosmetic composition containing a water-extracting agent,

The present invention relates to a method for fermenting a fermented broccoli fermented product, a spinach fermented product, a spinach fermented product obtained by fermenting a hot water extract of a spinach with a mushroom of a tricolor coated mushroom, and a hot water extract of a tomato, obtained by fermenting the fermented broth with the tricolor coated mushroom mycelium And a tomato fermentation product as an active ingredient.

Skin is an organ that covers the outside of the human body and acts as a primary barrier to protect the human body from external environmental stimuli such as ultraviolet rays, temperature and humidity changes. This skin consists of three layers, from the outside, the epidermis, the dermis and the subcutaneous fat layer. The epidermis is divided into the basal layer, the superficial layer, the granular layer, and the stratum corneum as keratinocytes differentiate and keratinize. The stratum corneum is the outermost layer of the epidermis, and the dead keratinocytes are removed from the skin and replaced with new keratinocytes. The replacement cycle of keratinocytes is about 4 weeks in normal skin. However, if the keratinocyte replacement is not performed normally, the skin becomes rough.

On the other hand, the dermis consists of matrix proteins such as collagen fibers and elastic fibers, and the water supplied from the dermis gradually diffuses to the upper part of the stratum corneum and disappears to the outside. This is called Trans Epidermal Water Loss (TEWL). The lipid component of the stratum corneum and epidermal lipids play a role in maintaining this level of transdermal water loss at an appropriate level. In addition, the stratum corneum is known to have an important role in moisturizing the skin due to the existence of a natural moisturizing factor (NMF) having hydrophilic nature and water retention ability.

Therefore, the moisturizing state of the stratum corneum is directly related to the skin appearance, and the skin moisture content of the stratum corneum influences the skin membrane homeostasis related to the condition and permeability of the skin, which is an important factor for moisturizing the skin.

On the other hand, Daedaleopsis tricolor is a brown mushroom that occurs in the form of a tile in dead hardwoods, with mushroom and mushroom in the throat of Japanese mushroom. The surface of the mushroom is flat, but its shape is semicircular or flat shell. It is named Daedaleopsis tricolor because of the three colors of yellowish brown, reddish brown and dark brown, which are the colors of the gut. The glans layer on the underside of the freshly formed wrinkles is completely wrinkled, initially becoming ashy white or gradually grayish brown to black. The efficacy of tricolor coated mushrooms is known to have immunological and anticancer effects.

However, the contents of antioxidative and skin moisturizing effects of each of the fermented products obtained by using broccoli, spinach or hot water extract of tomato with the mycelium of tricolor coated mushrooms have not yet been confirmed.

Korean Patent Laid-Open No. 10-2015-0144129 (published on Dec. 24, 2015) relates to a cosmetic composition comprising an effective ingredient of a fermented product of Zanthoxylum mushroom. The cosmetic composition for wrinkle improvement according to the present invention is safe for skin But it is excellent in the effect of promoting collagen synthesis so that the effect of improving the wrinkle of the skin is excellent and it is highly suitable for wrinkle-improving skin care. Korean Patent No. 10-1375137 (Registration Date: 2014.06.11) discloses a cosmetic composition comprising a fermented product of a red hot cherry mushroom mycelium of golden hot water extract, wherein the fermented product of the red hot cherry mushroom mycelium of golden hot water extract is golden Discloses a high-functional cosmetic composition for improving skin, wherein the glycoside contained in the glycoside is effectively converted into Baicalaine, which is a non-glycoside, and contains the Baicalaine in a high concentration to thereby excel in antioxidation and skin moisturizing effect.

An object of the present invention is to provide a cosmetic composition which is safe to the skin and has excellent effects on antioxidation and skin moisturization by using the fermented product obtained by fermenting the natural hot water extract with the mushroom mycelium of three colors.

In order to achieve the above object, the present invention provides a fermented broccoli fermented product obtained by fermenting Broccoli hot-water extract with mycelia of tricolor-coated mushroom, fermenting Broccoli fermented product, spinach hot-water extract with tricolor coated mushroom mycelium, And a tomato fermented product obtained by fermenting with a mushroom mycelium as an active ingredient.

The present invention also relates to an extract of a spinach fermented product obtained by fermenting broccoli hot-water extract with a culture solution of a tricolor coated mushroom mycelia, a fermented product of broccoli fermented product, a spinach hot- And an extract of a tomato fermented product obtained by fermenting with a mushroom mycelial culture liquid as an active ingredient. The present invention also provides a cosmetic composition for skin moisturizing.

The present invention is characterized in that a fermented product obtained by fermenting broccoli hot water extract, spinach hot water extract or tomato hot water extract with ferric triclosan mushroom mycelium is used. Preferably, these hot water extracts are prepared by cutting broccoli, spinach, After rapidly freezing at 30 to 50 ° C, the water is sublimed in a vacuum state and lyophilized. The lyophilized broccoli, spinach or tomatoes are each pulverized and subjected to hot water extraction, and then filtered with 300 to 500 mesh to obtain each hot water extract Can be used.

On the other hand, the 'extract' of the broccoli fermented product obtained by fermenting the broccoli hot water extract of the present invention with the culture solution of the tricolor coated mushroom mycelia, the 'extract' of the spinach fermented product obtained by fermenting the hot water extract of the spinach with the culture solution of the mushroom cultured in the tricolor coated mushroom, The 'extract' of the tomato fermented product obtained by fermenting the extract with the culture solution of the tricolor coated mushroom mycelium refers to an extract obtained by adding a solvent to the fermented product and then extracting it secondarily.

At this time, as a solvent to be used, a solvent suitable for adding to cosmetic formulations can be used, for example, ethanol, butylene glycol and the like. That is, various extract forms can be prepared using various solvents suitable for the final product. The extract extracted secondarily using a solvent suitable for addition to cosmetic formulations can be added directly to the finished cosmetic product as an intermediate raw material and can be used by concentrating and extracting a specific ingredient according to the characteristics of the solvent.

In the present invention, the term 'fermented product' is a concept including both a state immediately after completion of fermentation, a state in which the mycelium is filtered after completion of fermentation, or a state in which other residues are filtered out in addition to the mycelium. It may also contain volatilized solids through various drying methods.

Meanwhile, in the cosmetic composition of the present invention, the 'broccoli fermentation product obtained by fermenting broccoli hot water extract with the tricolor coated mushroom mycelium' is preferably fermented for 1 to 10 days with the culture solution of the mushroom mycelium of tricolor coated mushroom, To < RTI ID = 0.0 > 600 mesh. ≪ / RTI > When the above range is satisfied, the skin moisturizing ability is remarkably excellent, which can be confirmed in Experimental Example 4 below.

In addition, in the cosmetic composition of the present invention, the 'spinach fermented product obtained by fermenting the spinach hot-water extract with the tricolor coated mushroom mycelium' is preferably fermented for 1 to 10 days in the culture solution of the mushroom mycelium of tricolor coated mushroom, To < RTI ID = 0.0 > 600 mesh. ≪ / RTI > When the above range is satisfied, the skin moisturizing ability and skin antioxidant effect are remarkably excellent, which can be confirmed in Experimental Examples 1 and 4 described below.

In addition, in the cosmetic composition of the present invention, the 'tomato fermented product obtained by fermenting tomato hot water extract with the tricolor coated mushroom mycelium' is preferably fermented for 1 to 10 days in the culture solution of mushroom mycelium of tricolor coated mushroom, To < RTI ID = 0.0 > 600 mesh. ≪ / RTI > When the above range is satisfied, the skin moisturizing ability and antioxidant efficacy are remarkably excellent, which can be confirmed in Experimental Examples 1 and 4 described below.

In the present invention, the term "included as an active ingredient" means that the extract of broccoli hot water, hot water extract of spinach or hot water extract of tomato is added to cosmetic composition to the extent that skin moisturizing effect can be exhibited from the cosmetic composition of the present invention, Means a fermented product fermented with a mycelial culture liquid.

On the other hand, the cosmetic composition of the present invention may be formulated into cosmetic formulations, for example, lotions, gels, water-soluble liquids, creams, essences, oil-in-water (O / W) and water-in-oil (W / O) formulations. Such as a makeup base, a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a teak color and an eyebrow pencil.

The broccoli fermented product obtained by fermenting broccoli hot water extract with the culture solution of the tricolor coated mushroom mycelium contained in the cosmetic composition of the present invention as an active ingredient, the spinach fermented product obtained by fermenting the spinach hot water extract with the culture solution of the tricolor coated mushroom mycelium, And 'Tomato fermented product obtained by fermenting tomato hot water extract with a culture solution of tricolor coated mushroom mycelium' have excellent effects on antioxidation, skin regeneration and skin moisturization.

Fig. 1 (A) shows the result of confirming the antioxidant activity of the broccoli hot-water extract using the DPPH method.
Fig. 1 (B) shows the antioxidant activity of the spinach extract prepared by the DPPH method.
1 (C) shows the result of confirming the antioxidant activity of the hot water extract of tomato using the DPPH method.
Fig. 1 (D) shows the result of confirming the antioxidative activity of the broccoli fermented product obtained by fermenting broccoli hot-water extract with the culture solution of the mushroom mycelium of tricolor by using the DPPH method.
Fig. 1 (E) shows the result of confirming the antioxidant activity of the fermented broccoli obtained by fermenting the spinach extract with the culture solution of the mushroom mycelium of trivalent color using the DPPH method.
FIG. 1 (F) shows the result of confirming the antioxidant activity of the fermented broccoli obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium using the DPPH method.
Fig. 2 (A) is a result of confirming the cytotoxicity of the broccoli hot-water extract in human fibroblasts using the MTT method.
Fig. 2 (B) shows the result of confirming the cytotoxicity of the spinach hot-water extract in human fibroblasts using the MTT method.
Fig. 2 (C) shows the cytotoxicity of tomato hot-water extract in human fibroblasts using MTT assay.
FIG. 2 (D) is a result of confirming the cytotoxicity of the fermented broccoli obtained by fermenting broth hot-water extract with human cultured mushroom mycelium in human fibroblasts using MTT method.
FIG. 2 (E) is the result of confirming the cytotoxicity of the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the tricolor-coated mushroom mycelium in human fibroblasts using the MTT method.
FIG. 2 (F) shows the cytotoxicity of the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium in human fibroblasts using MTT method.
Fig. 3 (A) shows the results of confirming the cell activity of the broccoli hot-water extract in human fibroblasts using the MTT method.
Fig. 3 (B) shows the result of confirming the cell activity of the spinach extract in human fibroblasts using the MTT method.
Fig. 3 (C) shows the result of confirming the cell activity of the hot-water extract of tomato in the human fibroblasts using the MTT method.
Fig. 3 (D) is a result of confirming the cell activity of the broth fermentation obtained by fermentation of the broth hot water extract in the human fibroblast with the culture medium of the tricolor coated mushroom mycelium using the MTT method.
FIG. 3 (E) shows the results of confirming the cell activity of the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the mushroom mycelium of triplicate in human fibroblasts using the MTT method.
FIG. 3 (F) is a result of confirming the cell activity of the tomato fermented product obtained by fermenting tomato hot water extract with the culture solution of the mushroom mycelium of triplicate in human fibroblasts using the MTT method.
4 (A) is a result of measuring the moisture resistance for 4 days using water as a control group.
Fig. 4 (B) shows the result of measuring the moisture resistance for 4 days using 30% butylene glycol as a positive control group.
Fig. 4 (C) shows the result of measuring the moisture resistance of the broccoli hot-water extract for 4 days.
Fig. 4 (D) shows the result of measuring the moisture resistance of the hot-water extract of spinach for 4 days.
Fig. 4 (E) shows the result of measuring the moisture resistance of the hot water extract of tomato for 4 days.
Fig. 4 (F) shows the result of measuring the moisture resistance of the broccoli fermented product obtained by fermenting broccoli hot-water extract with the culture solution of the mushroom mycelium of three colors for 4 days.
FIG. 4 (G) is a result of measuring the moisture-holding ability of the spinach fermented product obtained by fermenting the spinach extract with hot water for three days.
FIG. 4 (H) shows the result of measuring the moisture resistance of the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the mushroom mycelium of three colors for 4 days.
FIG. 4 (H) is a graph showing the results of four days measurement of the moisture resistance of broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli fermented product, spinach fermented product and tomato fermented product.

Hereinafter, the present invention will be described in more detail in the following Examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Preparation Example 1: Preparation of broccoli hot-water extract]

In this Example, broccoli hot water extract was prepared. 200 g of broccoli was cut and lyophilized at -40 ° C and then pulverized to prepare 25 g of broccoli powder. 10 g of the broccoli powder was added to 200 g of purified water, subjected to hot water extraction, and filtered through 400 mesh to prepare a broccoli hot water extract .

[Preparation Example 2: Preparation of spinach hot water extract]

In this Example, a spinach hot water extract was prepared. 200 g of spinach were lyophilized at minus 40 ° C. and pulverized to prepare 47.3 g of spinach powder. 10 g of the spinach powder was added to 200 g of purified water, subjected to hot water extraction, and then filtered through 400 mesh to prepare a spinach hot water extract.

[Preparation Example 3: Preparation of hot water extract of tomato]

In this example, a hot water extract of tomato was prepared. The tomato powder was prepared by cutting 200 g of the tomato with a slice, lyophilized at -40 ° C, and pulverized to prepare 21.5 g of the tomato powder. 10 g of the tomato powder was added to 200 g of purified water and subjected to hot water extraction. .

[Preparation Example 4: Culture of mycelia of tricolor-coated mushrooms]

In this example, the mycelia of three colors of mushroom were cultured.

The mycelium of the initial tricolor painted mushroom was cultivated at Jeonnam Forest Resources Research Institute. Cultivation and maintenance of the mycelium of tricolor coated mushrooms were stored at 4 ℃ on PDA (Potato Dextrose Agar) and subcultured at intervals of 4 weeks. The three-color coated mushroom mycelial liquid culture was used in the following Examples 1 to 3 and Experimental Examples 1 to 4 by culturing in 200 ml of PDB (Potato Dextrose Broth) medium at 150 rpm and 25 ° C for 7 days.

[ Example  1: Broccoli Hot water extract Tricolored mushroom  Preparation of fermented broccoli obtained by fermentation with a mycelial culture liquid]

In this Example, a broth fermented product obtained by fermenting broccoli hot-water extract with a culture solution of mushroom mycelium of tricolor was prepared.

To 200 g of the broccoli hot water extract prepared in Preparation Example 1 was inoculated 2.5% of the cultured medium prepared in Preparation Example 4 for 7 days, and then cultured for 7 days. Then, the culture broth was sterilized and filtered through 400 mesh to prepare a broccoli fermented product obtained by fermenting broccoli hot-water extract with a culture solution of tricolor-coated mushroom mycelium.

[ Example  2: spinach Hot water extract Tricolored mushroom  Preparation of fermented spinach obtained by fermentation with mycelial culture liquid]

In this Example, a spinach fermented product was obtained by fermenting a spinach hot water extract with a culture solution of a mushroom mycelium of three colors.

To the 200 g of the spinach extract prepared in Preparation Example 2 was added 2.5% of the culture solution prepared in Preparation Example 4 for culturing the mycelium of the tricolor coated mushroom for 7 days and then cultured for 7 days. Then, the culture broth was sterilized and filtered through 400 mesh to prepare a spinach fermented product obtained by fermenting a spinach hot-water extract with a culture solution of a mushroom mycelium of three colors.

[ Example  3: Tomato Hot water extract Tricolored mushroom  Preparation of tomato fermentation product obtained by fermentation with a mycelial culture liquid]

In this example, the tomato hot water extract was fermented with a culture solution of mushroom mycelium of tricolor to obtain a tomato fermented product.

To the 200 g of the hot water extract of tomato prepared in Preparation Example 3, 2.5% of the culture solution prepared in Preparation Example 4 and cultured for 7 days in the tricolor coated mushroom mycelium was inoculated and cultured for 7 days. Then, the culture broth was sterilized and filtered through 400 mesh to prepare a tomato fermented product obtained by fermenting tomato hot water extract with a culture solution of mushroom mycelium of three colors.

[ Experimental Example  One: Manufacturing example  1 to 3 Hot water extract  And Example  Evaluation of antioxidant efficacy of the fermented product prepared in 1 to 3]

In this experiment, the broccoli hot-water extract, spinach hot-water extract and tomato hot-water extract prepared in Production Examples 1 to 3 and 1.1-diphenyl-2-picrylhydrazyl (DPPH) Obtained by fermenting the fermented broccoli fermented product obtained by fermenting with the culture solution of the tricolor coated mushroom mycelium, the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the mushroom mycelium of three colors, and the hot water extract of tomato, Were investigated for the antioxidative effect of 'tomato fermented water'.

DPPH (1,1-diphenyl-picryl hydrazyl) used in the DPPH method is a very stable free radical, a deep purple compound that exhibits characteristic absorption at 517 nm. When DPPH is supplied from the antioxidant, electrons or hydrogen are converted into non-radicals, and decolorized. Thus, by measuring the changed absorbance, antioxidant activity is measured. The measurement method was measured as follows.

Broccoli fermented product obtained by fermenting broth with hot water extract of broccoli prepared in Preparation Examples 1 to 3, hot water extract of spinach and hot water of tomato and hot water extract of broccoli prepared in Examples 1 to 3, 0.325, 1.25, 2.35, and 5%, respectively, of the fermented spinach fermented by fermentation of the hot water extract with the culture solution of the tricolor coated mushroom mycelium, and the tomato fermented product obtained by fermenting the hot water extract of tomato with the culture solution of the mushroom mycelium of three colors. , And then 20 [mu] L of each solution was added to a 96-well plate. Then, 180 μl of 0.1 mM DPPH was added thereto and reacted at room temperature for 10 minutes.

As a positive control, 1 mM ascorbic acid was used and reacted in the same manner as described above. Thereafter, the absorbance was measured at a wavelength of 540 nm using a spectrophotometer (MULTI SKAN Thermo, Rockford, Ill.), And the inhibition rate was calculated by the following equation.

[Equation 1]

Inhibition rate (%) = [(A-B) - (C-D)] /

A: absorbance of control group

B: absorbance of blank control group

C: absorbance of sample group

D: absorbance of blank sample group

As a result, the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the mushroom mycelium of three colors exhibited excellent antioxidative activity (Fig. 1 (B) and (E)). In addition, it was confirmed that the antioxidative effect of tomato fermented water obtained by fermenting tomato hot water extract with fermentation solution of tricolor coated mushroom mycelium was 3% higher than that of hot water extract of tomato (Fig. 1 (C) and (F)).

However, in the case of broccoli, it was confirmed that the broccoli hot-water extract had an excellent antioxidative effect as compared to the broccoli fermented product obtained by fermenting broccoli hot-water extract with the culture solution of the tricolor-coated mushroom mycelium (Fig. 1 (A) and (D)).

Fig. 1 (A) shows the result of confirming the antioxidant activity of the broccoli hot-water extract using the DPPH method. Fig. 1 (B) shows the antioxidant activity of the spinach extract prepared by the DPPH method. 1 (C) shows the result of confirming the antioxidant activity of the hot water extract of tomato using the DPPH method. Fig. 1 (D) shows the result of confirming the antioxidative activity of the broccoli fermented product obtained by fermenting broccoli hot-water extract with the culture solution of the mushroom mycelium of tricolor by using the DPPH method. Fig. 1 (E) shows the result of confirming the antioxidant activity of the fermented broccoli obtained by fermenting the spinach extract with the culture solution of the mushroom mycelium of trivalent color using the DPPH method. FIG. 1 (F) shows the result of confirming the antioxidant activity of the fermented broccoli obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium using the DPPH method.

[ Experimental Example  2 : Human fibroblast  (Human dermal fibroblast neonatal, HDFn ) ≪ / RTI >

(1) Culture of human fibroblasts

In this experimental example, human fibroblasts were cultured.

DMEM medium and F12 medium were mixed at a ratio of 3: 1 and the mixture was cultured in a mixed medium supplemented with fetal bovine serum (FBS) at a concentration of 10% at 37 ° C, 5% CO _{2 and} 95% The cells were cultured using a cell culture machine and subcultured every time they grew to about 70% of the Petri dish area, and used in Experimental Example 3 below.

(2) Evaluation of cytotoxicity against human fibroblasts

In this experimental example, the broccoli hot water extract, spinach hot water extract, tomato hot water extract and hot water extract prepared in Preparative Examples 1 to 3 were prepared through MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) The broccoli fermented product obtained by fermenting with the broth of broccoli hot water extract tricolor coated mushroom prepared in Examples 1 to 3, the spinach fermented product obtained by fermenting the hot water extract of spinach with the cultured mushroom mycelium of three colors, To evaluate the cytotoxicity against human fibroblasts of the tomato fermented product obtained by fermenting the extract with the culture solution of the mushroom mycelium of tricolor.

Human fibroblasts were cultured for 24 hours under the conditions of humidity of 1 × 10 ⁴ after a concentration of the cell / well into a 96-well plate in 37 ℃ cell _{incubator, 5% CO 2, 95%} .

Thereafter, the broccoli fermented product obtained by fermenting the broccoli hot water extract, spinach hot water extract, tomato hot water extract and the broccoli hot water extract prepared in Examples 1 to 3 with the culture solution of the tricolor coated mushroom mycelium produced in Production Examples 1 to 3 , 'Spinach fermented product obtained by fermenting spinach hot-water extract with culture solution of tricolor coated mushroom mycelium', and 'tomato fermented product obtained by fermenting tomato hot-water extract with culture solution of mushroom mycelia of tricolor' were 0.312, 0.625, 1.25 and 2.35 , 5, 10%, and then reacted for 24 hours under the above culture conditions.

After the reaction, each plate was treated with 50 μl of MTT reagent at a concentration of 2 mg / ml, and cultured under the same conditions as above. After 2 hours, the medium was removed and treated with 200 μl of dimethylsulfoxide (DMSO) Absorbance was measured at a wavelength of 540 nm.

As a control group, human fibroblasts without treatment of hot water extract and 'fermented product obtained from fermentation with tricolor coated mushroom fermentation broth' were used, and the culture and measurement were the same as above.

As a result of the experiment, broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli hot water extract, broccoli fermented product obtained by fermenting with the culture solution of tricolor coated mushroom mycelia, and spinach hot water extract were fermented with a culture solution of tricolor coated mushroom mycelium Spinach fermented product 'and' tomato hot - water extract 'were not significantly changed at all concentrations of the tomato fermented product obtained by fermentation with the culture solution of the mushroom mycelium of three colors.

Therefore, the broccoli fermented product obtained by fermenting broccoli hot water extract, tomato hot water extract, spinach hot water extract, broccoli hot water extract with fermentation broth of tricolor coated mushroom mycelium, tomato hot water extract, fermented with the culture solution of mushroom mycelium of tricolor Fermented water 'and' spinach hot water extract 'were fermented with a culture solution of mushroom mycelium of tricolor (FIG. 2).

Fig. 2 (A) is a result of confirming the cytotoxicity of the broccoli hot-water extract in human fibroblasts using the MTT method. Fig. 2 (B) shows the result of confirming the cytotoxicity of the spinach hot-water extract in human fibroblasts using the MTT method. Fig. 2 (C) shows the cytotoxicity of tomato hot-water extract in human fibroblasts using MTT assay. FIG. 2 (D) is a result of confirming the cytotoxicity of the fermented broccoli obtained by fermenting broth hot-water extract with human cultured mushroom mycelium in human fibroblasts using MTT method. FIG. 2 (E) is the result of confirming the cytotoxicity of the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the tricolor-coated mushroom mycelium in human fibroblasts using the MTT method. FIG. 2 (F) shows the cytotoxicity of the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium in human fibroblasts using MTT method.

[Experimental Example 3: Evaluation of cellular activity of human fibroblasts]

In this Experimental Example, broccoli fermentation obtained by fermenting broth with hot water extract of broccoli, spinach hot water extract, hot water extract of tomato, and hot water extract of broccoli prepared in Examples 1 to 3 with a culture solution of tricolor coated mushroom mycelium And 'Tomato fermented product obtained by fermentation of the spinach fermented product obtained by fermenting the spinach hot water extract with the culture solution of the tricolor coated mushroom mycelium and the tomato hot water extract with the culture solution of the mushroom cultured in the tricolor coated mushroom' Respectively.

Human fibroblasts were added to a 96-well plate at a concentration of 5 × 10 ³ cells / well and cultured in a cell incubator at 37 ° C., 5% CO _{2, and} 95% humidity.

After 24 hours, broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli hot water extract, fermented broth fermented product obtained by fermenting with a culture solution of tricolor coated mushroom mycelium, and spinach hot water extract were fermented with a culture solution of tricolor coated mushroom mycelium 0.25% fetal bovine serum was obtained at the concentration of 0.312, 0.625, 1.25, 2.35, 5, and 10%, respectively, and the tomato fermented product obtained by fermenting a spinach fermented product and tomato hot water extract with fermentation broth of tricolor coated mushroom mycelium, And replaced with medium containing serum (FBS).

Then, the cells were cultured in a cell culture incubator for 48 hours, and 50 μl of 2 mg / ml MTT reagent was added to each well, followed by reaction for 2 hours. After 2 hours, the medium was removed, treated with 200 [mu] l of dimethylsulfoxide (DMSO) and incubated at 540 nm wavelength Absorbance was measured.

In the case of human fibroblasts treated with broccoli hot - water extract, the cell activity was increased in all concentration - dependent manner. The spinach fermented by the fermentation of the spinach hot - water extract to the tricolor coated mushroom mycelium was 0.312, 0.625, 1.25, In the case of human fibroblasts treated with the concentrations of 2.35, 5, and 10%, cell activity was increased in a concentration-dependent manner at a concentration of 0.312% to 5% (FIG. 3).

Fig. 3 (A) shows the results of confirming the cell activity of the broccoli hot-water extract in human fibroblasts using the MTT method. Fig. 3 (B) shows the result of confirming the cell activity of the spinach extract in human fibroblasts using the MTT method. Fig. 3 (C) shows the result of confirming the cell activity of the hot-water extract of tomato in the human fibroblasts using the MTT method. Fig. 3 (D) is a result of confirming the cell activity of the broth fermentation obtained by fermentation of the broth hot water extract in the human fibroblast with the culture medium of the tricolor coated mushroom mycelium using the MTT method. FIG. 3 (E) shows the results of confirming the cell activity of the spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the mushroom mycelium of triplicate in human fibroblasts using the MTT method. FIG. 3 (F) shows the results of confirming the cell activity of the tomato fermented product obtained by fermenting tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium in human fibroblasts using the MTT method.

[Experimental Example 4: Evaluation of skin moisture resistance]

In this Experimental Example, broccoli fermentation obtained by fermenting broth with hot water extract of broccoli, spinach hot water extract, hot water extract of tomato, and hot water extract of broccoli prepared in Examples 1 to 3 with a culture solution of tricolor coated mushroom mycelium Water fermented by spinach fermented with 3 - colored mushroom mycelia, and 'Tomato fermented water obtained by fermentation of tomato hot - water extract with 3 - color coated mushroom mycelia' were evaluated.

Broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli fermented product obtained by fermenting broccoli hot water extract with the culture solution of tricolor coated mushroom mycelium, spinach fermented product obtained by fermenting the hot water extract of spinach with the culture solution of mushroom mycelium of tricolor In order to measure the moisture content of the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the mushroom mycelia of three colors, broccoli hot water extract, spinach hot water extract, tomato hot water extract, The fermented broccoli fermented product obtained by fermenting broccoli hot water extract with the culture solution of the tricolor coated mushroom mycelium, the spinach fermented product obtained by fermenting the hot water extract of spinach with the culture solution of the mushroom mycelium of three colors, and the hot water extract of tomato, Fermented tomatoes obtained by fermentation ' The amount of decrease in moisture was measured.

Five milliliters of broccoli hot water extract, spinach hot water extract, tomato hot water extract, fermented product obtained by fermenting with broccoli hot water extract tricolor coated mushroom mycelia, and spinach hot water extract were added to a three-color coated mushroom mycelial culture solution And the tomato fermented product obtained by fermenting the spinach fermented product obtained by fermenting with the culture solution of the mushroom mycelium of tricolor coated with the tomato hot water extract on the bottom plate of the ceramic ware, And the results are shown in Tables 1 to 4 below.

As a control, distilled water and 30% butylen glycol (BG) were used.

Time (Day) Distilled water  30% BG Broccoli hot water extract The broccoli fermented product obtained by fermenting the broth of hot water extract with the culture solution of the mushroom mycelium of tricolor Spinach hot water extract The spinach fermented product obtained by fermenting the spinach hot water extract with the culture solution of the mushroom mycelium of tricolor Tomato hot water extract The tomato hot water extract was fermented with the culture solution of the mushroom mycelium of three colors, CaCl ₂ 333 (g) 0 100 100 100 100 100 100 100 100 One 68.47 78.79 59.96 60.69 58.66 61.60 63.86 61.52 2 55.82 76.85 54.73 55.85 53.35 56.40 59.04 56.38 3 31.53 69.26 32.80 34.27 32.10 37.00 37.75 37.04 4 12.85 57.39 7.24 30.85 8.31 35.00 13.86 30.04

As a result of the experiment, the moisture content for 1 to 2 days was obtained by extracting all the broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli hot water extract, broccoli fermented product obtained by culturing with the culture solution of tricolor coated mushroom mycelium, There was no significant difference in the fermented spinach fermented by the fermented mushroom mycelia culture and the tomato fermented by the fermentation of the mushroom cultivated with the tricolor coated mushroom cultures. In the case of the spinach fermented product obtained by fermenting the fermented broccoli fermented product obtained by fermenting with the culture solution of the tricolor coated mushroom mycelium and the hot water extract of the spinach with the culture solution of the mushroom cultured in the tricolor coated mushroom, the fermented broccoli hot water extract 'And' Spinach hydrothermal extract 'which have about 4 times more moisture Respectively. In addition, it was confirmed that the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium had a moisture resistance more than twice that of the tomato hot water extract which was not fermented by the culture solution of the mushroom mycelium of the tricolor coated mushroom.

It was confirmed that the effect of the active ingredient contained in the fermented product was not exhibited, but the difference in the moisture resistance between the fermented product and the fermented product was remarkable after 3 days because the fermented product contained excessive water in the first and second days of the experiment.

4 (A) is a result of measuring the moisture resistance for 4 days using water as a control group. Fig. 4 (B) shows the result of measuring the moisture resistance for 4 days using 30% butylene glycol as a positive control. Fig. 4 (C) shows the result of measuring the moisture resistance of the broccoli hot-water extract for 4 days. Fig. 4 (D) shows the result of measuring the moisture resistance of the hot-water extract of spinach for 4 days. Fig. 4 (E) shows the result of measuring the moisture resistance of the hot water extract of tomato for 4 days. Fig. 4 (F) shows the result of measuring the moisture resistance of the broccoli fermented product obtained by fermenting broccoli hot-water extract with the culture solution of the mushroom mycelium of three colors for 4 days. FIG. 4 (G) is a result of measuring the moisture-holding ability of the spinach fermented product obtained by fermenting the spinach extract with hot water for three days. FIG. 4 (H) shows the result of measuring the moisture resistance of the tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the mushroom mycelium of three colors for 4 days. Fig. 4 (I) shows the result of measuring the moisture-holding ability of broccoli hot water extract, spinach hot water extract, tomato hot water extract, broccoli fermented product, spinach fermented product and tomato fermented product for 4 days All of which are graphs.

[ Example  4 :  skin  For moisturizing Cosmetics  Produce]

In this example, broccoli fermented product obtained by fermentation of the tricolor coated mushroom mycelium culture solution, spinach fermented product, and spinach hot water extract prepared in Production Example 4 and Examples 1 to 3, And a tomato fermented product obtained by fermenting the spinach fermented product obtained by fermenting with the culture solution of the tricolor coated mushroom mycelium and the tomato hot water extract with the culture solution of the mushroom mycelium of tricolor.

The composition of the cosmetic for moisturizing the skin is shown in Table 2 below.

turn Name content(%) One Purified water to 100 2 Centipede extract 18,000 3 glycerin 10,000 4 Caprylic / capric triglyceride 8.000 5 Hydrogenated polydecene 4.000 6 Propanediol 4.000 7 Cetyl hexanoate 4.000 8 Macadamia seed oil 4.000 9 Glyceryl stearate S 4.000 10 1,2-hexanediol 1,000 11 Stearyl alcohol 1,000 12 Sorbitan olivate 1,000 13 Cetyl alcohol 1,000 14 Cetearyl olivate 1,000 15 Corn starch 0.100 16 Myristyl alcohol 0.100 17 Sorbitan stearate 0.100 18 Arachidyl alcohol 0.100 19 Behenyl alcohol 0.100 20 Arachidyl glucoside 0.100 21 Broccoli Fermentation ^{1 )} 0.100 22 Spinach fermented water ^{2 )} 0.100 23 Tomato fermented water ^{3 )} 0.100 24 Butylene glycol 0.100 25 Acrylate / C10-30 alkyl acrylate crosspolymer 0.100 26 Tromethamine 0.100 27 Disodium iodide 0.040 28 Zinc sulphate 0.030 29 Potassium sorbate 0.030

1) A fermented broccoli obtained by fermenting broccoli hot water extract with a culture solution of a mushroom mycelium of tricolor.

2) Spinach fermented product obtained by fermenting spinach hot water extract with culture medium of tricolor coated mushroom mycelium.

3) Tomato fermented product obtained by fermenting tomato hot water extract with culture medium of tricolor coated mushroom mycelium.

The fermented broccoli fermented product obtained by fermenting broccoli hot water extract with the culture solution of the tricolor coated mushroom mycelium, the spinach fermented product obtained by fermenting the spinach hot water extract with the culture solution of the tricolor coated mushroom mycelium, And 'Tomato fermented product obtained by fermenting the tomato hot water extract with the culture solution of the tricolor coated mushroom mycelium', the cosmetic composition for skin moisturizing having excellent moisturizing effect could be produced.

Claims (5)

Broccoli fermented product obtained by fermenting broccoli hot water extract with a culture solution of mushroom mycelium of tricolor,
The spinach fermented product obtained by fermenting the spinach hot water extract with the culture solution of the mushroom mycelium of three colors,
A cosmetic composition for skin moisturizing, characterized by comprising at least one selected from the tomato fermented products obtained by fermenting the hot water extract of tomato with the culture solution of the mushroom mycelium of three colors.
Extract of broccoli fermented product obtained by fermenting broccoli hot water extract with a culture solution of mushroom mycelium of tricolor,
Extract of spinach fermented product obtained by fermenting spinach hot-water extract with a culture solution of mushroom mycelium of tricolor,
A cosmetic composition for skin moisturizing, characterized in that it contains, as an active ingredient, at least one selected from the extracts of the tomato fermented product obtained by fermenting the hot water extract of tomato with the culture solution of the mushroom mycelium of tricolor.
3. The method according to claim 1 or 2,
The broccoli fermented product obtained by fermenting the above broccoli hot-water extract with the culture solution of the mushroom mycelium of tri-
The broccoli hot-water extract was fermented for 1 to 10 days with a culture solution of the mushroom mycelium of three colors, and the mixture was filtered through 200 to 600 mesh Broccoli fermented product Wherein the cosmetic composition is a cosmetic composition for skin moisturizing.
3. The method according to claim 1 or 2,
The spinach fermented product obtained by fermenting the spinach hot-water extract with the culture solution of the mushroom mycelium of tri-
A cosmetic composition for skin moisturizing, characterized by being a spinach fermented product obtained by fermenting a spinach hot-water extract with a culture solution of a mushroom mycelium of three colors for 1 to 10 days and filtering with 200 to 600 mesh.
3. The method according to claim 1 or 2,
The tomato fermented product obtained by fermenting the above hot water extract of tomato with the culture solution of the mushroom mycelium of three colors,
A cosmetic composition for skin moisturizing, characterized in that the hot water extract of tomato is a tomato fermented product obtained by fermenting with a culture solution of mushroom mycelium of three colors for 1 to 10 days and filtering with 200 to 600 mesh.

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