KR101625163B1 - Cosmetic composition comprising fermented extract of inonotus obliquus under natural cave - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a tea mushroom fermentation extract and a process for producing the tea mushroom fermentation extract. The method for preparing the mushroom fermented extract of the present invention can improve the physiological activity promoting ability of the mushroom and chaga mushroom extracts by cultivating the mushroom in the solid culture of the mushroom mycelium, By biotransformation, various physiologically active substances with increased physiological activity can be obtained. In addition, the mushroom fermented extract of the present invention is excellent in whitening, wrinkle reduction, antioxidant and anti-inflammatory effects, and thus can be usefully applied to research and industry in related cosmetics field.

Description

Technical Field [0001] The present invention relates to a cosmetic composition comprising a mushroom fermented extract fermented under natural cave conditions,

The present invention relates to a cosmetic composition comprising a tea mushroom fermentation extract and a method for producing the tea mushroom fermentation extract. In particular, the present invention relates to a cosmetic composition comprising a fermentation extract fermented in a natural limestone cave, and a process for producing the tea mushroom fermentation extract.

Chaga is a medicinal mushroom parasitized on birch, and its scientific name is Inonotus Obliquus. It is a mushroom parasitic on birch trees in Siberia, North America, northern Europe and more than 45 degrees north latitude. It is effective in treating adult diseases such as cancer. It grows by eating viruses and eating sap. It usually grows for 15 to 20 years. In Russia, from the 16th century, it has been reported as a leap for the treatment of incurable diseases. In 1951, the Soviet Academy of Sciences, Komarov Institute of Science, started to research in earnest.

Various studies on chaga mushroom have been carried out. For example, Korean Patent Laid-Open Publication No. 2013-0133477 discloses a composition for improving the enlargement of the prostate gland, improving acne and promoting hair growth, which comprises extracts of mushroom and mushroom. However, the prior art does not disclose a cosmetic composition related to anti-inflammation, antioxidation, whitening, wrinkle improvement, etc., and the anti-inflammatory, antioxidative, whitening, and wrinkle-improving effects using chaga mushrooms are not known.

On the other hand, the white mushroom belonging to the basidiomycete belongs to the mushroom mycelium which is inoculated without artificial agitation and aeration in the liquid fermentation process does not contact the fermentation object smoothly, Is different. In addition, if additional nutrients are not supplied, the fermentation process by the mushroom becomes late, and the growth of the inoculated inoculum is delayed, and the possibility of contamination by other bacteria is greatly increased.

Korea Patent Publication No. 2013-0133477

In order to solve the problems of the prior art as described above, the present invention provides a method for preparing a fermented mushroom extract, which is obtained by inoculating and cultivating mushroom into a solid culture of a mushroom mycelium, It is an object of the present invention to provide a cosmetic composition.

In order to achieve the above object, the present invention provides a cosmetic composition comprising an extract of Fermented Mushroom Fermentation as an active ingredient.

One embodiment of the present invention may be a cosmetic composition for anti-inflammation, antioxidant, whitening, or wrinkle improvement.

Another embodiment may be one in which the mushroom fermentation extract is obtained by inoculating and cultivating a mushroom or a powder thereof into a mushroom mycelial solid culture.

In another embodiment, the solid culture of the mycelium of mycelium of the above-mentioned condition comprises at least one member selected from the group consisting of senile trees, pine trees, pine trees, oak trees, cedar trees, fir trees, spruce trees, secretory trees, oak trees, oak trees, mulberry trees, Or at least one selected from the group consisting of wood, acacia tree, bloom, alder tree, ash tree, and bamboo tree can be inoculated and cultivated with a mushroom or a homogenate thereof.

In another embodiment, the solid culture of the mycelium of mycelium of the above-mentioned condition comprises at least one member selected from the group consisting of senile trees, pine trees, pine trees, oak trees, cedar trees, fir trees, spruce trees, secretory trees, oak trees, oak trees, mulberry trees, And may be one in which at least one member selected from the group consisting of wood, acacia tree, bark, alder tree, ash tree, and bark tree is inoculated with the mushroom or a homogeneous cargo thereof and cultured for 30 to 360 days.

In another embodiment, the chaga mushroom fermentation extract may be obtained by inoculating chaga mushroom or a powder thereof into a mycelia mycelial solid culture and culturing at 5 to 30 ° C.

In another embodiment, the mushroom fermentation extract may be obtained by inoculating mushroom or powder thereof into a mushroom mycelial solid culture and culturing at pH 4.5 to 7.5.

In another embodiment, the fermented mushroom extract may be contained in the cosmetic composition in an amount of 0.001 to 10% by weight.

In another embodiment, the above-mentioned mushroom fermentation extract is inoculated into a mushroom mycelial solid culture with at least one selected from the group consisting of chaga mushrooms or powder thereof, and goryeo mushroom, cordyceps, agaricus, blooming mushroom, and powder thereof And cultured.

In another embodiment, the cosmetic composition may be a formulation of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

The present invention also provides a method for producing an extract from Fermented mushroom fermentation.

An embodiment of the present invention relates to a method for producing a pine tree, a pine tree, a pine tree, a fir tree, a cedar, a fir, a spruce, a secretory tree, an oak tree, an oak tree, an oak tree, a mulberry tree, Preparing a mushroom mycelial solid culture by inoculating and cultivating a mushroom or a homogenate thereof in at least one selected from the group consisting of almond, ash, and barley; And obtaining a mushroom mycelial solid culture by inoculating and cultivating a mushroom or a powder thereof.

In another embodiment, the solid culture of the mycelia of the mycelia of mushroom is selected from the group consisting of a waxy tree, a pine tree, a pine tree, an oak tree, a cedar, a fir, a spruce tree, a secretory tree, an oak tree, an oak tree, an oak tree, , Acacia tree, bloom, alder tree, ash tree, and barley tree may be inoculated with at least one mushroom or a homogenate thereof and cultured for 30 to 360 days.

In another embodiment, the obtaining step may be to inoculate the mycelial solid culture with chaga mushroom or a powder thereof and cultivate at 5 to 30 ° C.

In another embodiment, the obtaining step is to inoculate the mycelial solid culture with chaga mushroom or a powder thereof and cultivate at pH 4.5 to 7.5.

The present invention also provides a cosmetic method, characterized by having at least one effect selected from the group consisting of anti-inflammation, antioxidation, whitening, and wrinkle improvement, including the step of applying the cosmetic composition to the skin.

The method for preparing the mushroom fermented extract of the present invention can improve the physiological activity promoting ability of the mushroom and chaga mushroom extracts by cultivating the mushroom in the solid culture of the mushroom mycelium, By biotransformation, various physiologically active substances with increased physiological activity can be obtained. In addition, the mushroom fermented extract of the present invention is excellent in whitening, wrinkle reduction, antioxidant and anti-inflammatory effects, and thus can be usefully applied to research and industry in related cosmetics field.

FIG. 1 is a photograph of a solid culture of a mushroom mycelium produced using a mulberry round piece according to Example 1.2.
FIG. 2 shows the concentration of free glucose (mg / L) measured by extracting from a mulberry solid culture of Mycelia mushroom according to the incubation time (week) according to Example 1.3.

The inventors of the present invention completed the present invention by confirming that the extract obtained by inoculating and cultivating a mushroom into a solid culture of the mycelia of mushroom mycelium has an excellent antioxidative, whitening, anti-inflammatory or wrinkle-reducing effect.

Hereinafter, the present invention will be described in detail.

The present invention provides a cosmetic composition comprising an extract of Fermented Mushroom Fermentation as an active ingredient.

The mushroom fermented extract may be one obtained by inoculating and cultivating a mushroom or a powder thereof into a solid culture of a mushroom mycelium. According to the present invention, a substance other than mushroom is added or inoculated to a mushroom mycelial solid culture And then cultured. For example, the chaga mushroom fermentation extract may be obtained by inoculating chaga mushroom or its powder, a medicinal mushroom or powder thereof into a solid culture of Mycelia mushroom mycelium. The medicinal mushroom may be Ganoderma lucidum, Cordyceps sinensis, Agaricus, and Mushroom mushroom.

In the present invention, the mushroom including the mushroom and the mushroom may be wild or cultured, and can be used in a dried or non-dried state.

In the cultures obtained above, the cultivation can be carried out at an appropriate temperature to show the effective efficacy of the skin. For example, the cultivation may be performed at 5 to 30 캜, preferably 5 to 15 캜.

In culturing, the cultivation may be carried out at an appropriate pH to show the effective efficacy of the skin. For example, the culture may be carried out at a pH of 4.5 to 7.5, preferably at a pH of 5.5 to 6.5.

In the culture obtained by culturing, the culture period can be appropriately adjusted to show the effective efficacy of the skin. For example, the culture may be performed for 30 to 360 days.

Further, if necessary, filtration and / or concentration and / or lyophilization methods known in the art can be additionally used after the above-mentioned, if necessary.

The solid culture of the mycelium of mycelia of the above-mentioned mushroom may be selected from the group consisting of coniferous trees or oak trees such as white planthoptera, pine, pine, oak, cedar, fir, spruce, (Phellinus linteus) or a homogeneous cargo thereof may be inoculated to a broad-leaved tree such as a tree, a forest tree, an almond tree, an ash tree, or a barley tree. The conifer or hardwood may be used as a piece of wood, and the wood piece may be cut and used according to a suitable size. For example, the above-mentioned solid culture of mycelium of mycelia of mushroom may be cultured by inoculation with mushroom on a mulberry round piece. The piece of wood may also be sterilized for the desired culture. In culturing by inoculation with the above-described mushroom or its homogeneous carbohydrate, the culture period can be appropriately adjusted in order to increase the concentration of free glucose in the nutrient cause. For example, the culture may be performed for 30 to 360 days, preferably 60 to 90 days.

The content of the mushroom fermentation extract is preferably 0.001 to 90% by weight based on the total weight of the composition. If the content of the fermentation extract is less than 0.001% by weight, skin improvement effect such as anti-inflammation, antioxidation, wrinkle improvement or whitening effect may not be exhibited. If it exceeds 90% by weight, There is a problem in the safety and stability of the shape, and a problem that the cost is not economical may occur.

The cosmetic composition may further comprise ingredients that can ordinarily be contained in the cosmetic composition and may include one or more selected from the group consisting of, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances and carriers However, the present invention is not limited thereto.

The cosmetic composition may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, surfactant-free cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray , But is not limited thereto. More specifically, the cosmetic composition is preferably a formulation such as a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder. It is not.

When the formulation of the cosmetic composition is paste, cream or gel, the carrier component includes an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .

When the formulation of the cosmetic composition is a powder or a spray, it may contain lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder as a carrier component. In the case of a spray, it may further contain chlorofluorohydrocarbons, Propane, such as butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or an emulsion, the carrier component may include a solvent, a solubilizing agent or an emulsifying agent. More specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the cosmetic composition is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like.

When the formulation of the cosmetic composition is a surfactant-containing cleansing, it is preferred that the carrier component is selected from the group consisting of aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

If the formulation of the cosmetic composition is a soap, a surfactant-containing cleansing agent or a surfactant-free cleansing agent, it may be applied to the skin and then wiped off, washed or rinsed with water. The surfactant-containing cleansing formulation may be a cleansing foam, a cleansing water, a cleansing towel or a cleansing pack. The surfactant-free cleansing formulation may be a cleansing cream, , A cleansing lotion, a cleansing water or a cleansing gel, but is not limited thereto.

The present invention provides a method for producing fermented mushroom extract.

The method for producing the mushroom fermented extract is a method for producing the fermented mushroom extract according to any one of the above (1) to (4), wherein the extract is selected from the group consisting of perilla, pine, pine, oak, cedar, fir, spruce, Producing a mycelia of mycelium of mushroom mycelia by inoculating and cultivating the mushroom or a homogenate thereof in at least one selected from the group consisting of almonds, almonds, ash trees, and barley trees; And obtaining the mushroom mycelial solid culture by inoculating and cultivating the mushroom or a powder thereof.

The step of preparing the mycelial solid culture comprises the steps of producing a mycelial solid culture by cultivating a mycelial solid culture in a culture medium containing at least one selected from the group consisting of perilla, pine, pine, oak, cedar, fir, spruce, , Pine tree, noodle tree, ash tree, and barley tree, and cultivating them for 30 to 360 days, preferably 60 to 90 days. In addition, substances can be added or inoculated in addition to the mushroom or its homogeneous cargo, as described above.

The obtaining step may be a step of inoculating a mushroom mycelial solid culture with a mushroom or a powder thereof and culturing at 5 to 30 캜, preferably 5 to 15 캜. In addition, the step of obtaining may be a step of inoculating a mushroom mycelial solid culture with a mushroom or a powder thereof and culturing at a pH of 4.5 to 7.5, preferably at a pH of 5.5 to 6.5. The period of inoculating and cultivating the mushroom or the powder thereof into the above-described mushroom mycelial solid culture can be preferably 30 days to 360 days.

The present invention provides a makeup method characterized by having at least one effect selected from the group consisting of anti-inflammation, antioxidation, whitening, and wrinkle improvement, including the step of applying the cosmetic composition to skin.

The cosmetic process of the present invention refers to all the cosmetic processes for applying the cosmetic composition of the present invention to human skin. That is, all methods known in the art for applying the cosmetic composition to the skin belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in combination, or may be used by overlapping with other cosmetic compositions other than the present invention. In addition, the cosmetic composition having an excellent exfoliating effect on the skin according to the present invention can be used according to a conventional method of use, and the use frequency of the cosmetic composition can be varied according to a user's skin condition or taste.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are provided to illustrate the present invention, and the present invention is not limited by the following examples, but may be variously modified and changed.

Example  1. Using mushroom mycelium Mushroom  Preparation of fermented extract

1.1 Situ mushroom mycelium Inoculation  Ready

Mycelia of Phellinus linteus, which is a white rot fungus, were subcultured in a PDA (Potato Dextrose Agar) plate medium at 5 ℃ at the Department of Biology, Department of Biology, Kangwon National University. The liquid culture for the mycelial growth was 25 And cultured in shake culture medium at 150 rpm, YMG (Yeast extract: 4 g / l, Malt extract: 10 g / l, Glucose: 4 g / l)

10 to 20 agar plugs (diameter: 1 cm) were placed in a 250 ml Erlenmeyer flask containing 100 ml of sterilized distilled water and homogenized in a homogenizer (X120, CAT, Germany) for 20 days in a PDA plate medium And homogenized for 30 to 60 seconds. After homogenization, the mycelia of mycelia were centrifuged at 6,140 g for 30 minutes, and the supernatant was removed. Then, 300 ml of sterilized tertiary distilled water was added to the centrifuged mycelium and resuspended in a reciprocal shaker (vertically reciprocating extractor) for 20 minutes. The supernatant was centrifuged, and glucose, phosphorus, and nitrogen were not detected. The wet weight (hereinafter referred to as "wt wt") of the mycelia of the mushroom from which the residual YMG medium component was removed was measured, and the homogenized 40% (w wt / v) mushroom mycelium An inoculum was prepared.

1.2 Production of mycelium mycelium solid culture

The mulberry collected from the mountain was cut with a scroll saw and cut in the shape of a circular piece (Diameter: 8 ~ 12 cm, Thickness: 1 ~ 1.5 cm). The cut mulberry round wood pieces were sterilized by wet sterilization at 121 ° C and dried in a dry oven at 80 ° C for 48 hours to remove moisture and used as a substrate of mycelia of the mushroom.

To the sterilized 3 L beaker was added 1 L of the homogenized 40% (w wt / v) mushroom mycelium prepared in Example 1.1 and 1 L of sterilized third distilled water to prepare a 20% (w wt / v) mushroom mycelium inoculum , And the sterilized dried mulberry round piece was immersed for 10 minutes. The immersed mulberry roundwood pieces were taken out and filled in a sterilized 20 L open glass water bath (D: 30 cm, H: 50 cm) and incubated at 25 ° C for 8 weeks. In a glass water tank used for the production of the mushroom mycelial solid culture, 1 L of sterilized distilled water was added to maintain the free address humidity during the incubation period, and the results are shown in FIG.

FIG. 1 is a photograph showing a solid culture of a mycelia of mushroom cultured using a mulberry round piece.

As shown in Fig. 1, the mycelium of the mushroom mycelium was grown on the surface of the round wood piece, and the solid culture of the mushroom mycelium causing the inoculation was obtained.

1.3 Condition of mushroom mycelial solid culture Glass glucose  Concentration measurement

According to Example 1.2, the mycelia of mulberry leaves were recovered, and mycelial bodies were removed with tweezers. Then, 50 g of sodium acetate buffer (pH 5.0) was added to 10 g of mulberry round wood pieces and the mixture was allowed to stand at 4 ° C for 24 hours. The extracted glucose was extracted from the slices. The extracted glucose was passed through a cotton filter to remove the mycelia of the residual mushroom. The supernatant was recovered after centrifugation at 6,140 g for 40 minutes. The recovered supernatant was filtered through a 0.45 탆 HPLC filter and purified by HPLC (column A: RS tech, NH ₂ column (250 mm x 46 mm), column no .: 091264AE, column temperature: 40 ° C, mobile phase: 75:25 acetonitrile: water), flow rate: 1.2 ml / min, detector: RI]. The results are shown in FIG.

FIG. 2 shows the concentration of free glucose (mg / L) measured by extraction from the mulberry cultured mushroom mycelium according to the incubation time (week). '○' means the concentration of free glucose extracted from the mulberry round slices cultured for 8 weeks inoculated with the mycelium of mushrooms and '○' means the free glucose concentration extracted from the mulberry round slices without the mushroom mycelium.

As shown in Fig. 2, the free glucose concentration of the mulberry cultured mycelia of mushroom mycelium was 305 mg / L at the 8th week, and the free glucose concentration was about 18 times higher than that of the non-mycelial control 17 mg / L. Therefore, mushroom mycelia inoculated on the mulberry round slab can biodegrade mulberry and receive continuous carbon source during the fermentation process.

Also, mycelium removed with tweezers was collected and added with ultrapure distilled water, homogenized at 13,000 rpm for 1 minute, centrifuged at 15,000 rpm, and w wt was measured after removing the supernatant. As a result, Respectively. Therefore, inoculation of the general fermentation broth was carried out by liquid culture of mycelia of the mushroom, centrifuging and washing the recovered mycelia with a homogenizer and securing 83 g (w wt) of mycelium.

1.4 Mushroom  Preparation of fermented extract

The dried chaga mushroom (available from Dae Kwang Yakushin Co., Ltd.) was finely divided and then pulverized again to obtain a powder. The obtained mushroom powder was filled with the mushroom solid culture prepared according to Example 1.2 above and incubated for 180 days in a cave (Gangwon-do Jeongseong Hwaam Cave) at a pH of 5.5 to 6.5 and a temperature of 5 to 15 ° C, The culture was prepared. The polymer precipitate and mycelium were removed using a filter having a size of 0.2 mu m to extract an active ingredient in the fermentation broth.

Comparative Example  1. Using yeast Mushroom  Preparation of fermented extract

The dried tea was finely ground and then ground again to give a powder. The obtained chaga mushroom powder was added to 1.2 L of a basic medium (YM Broth) so as to have a concentration of 30% by weight, the pH was adjusted to 5.5 to 6.5, and the mixture was allowed to stand at room temperature for 2 hours.

Yeast (Saccharomyces cerevisiae) distributed from the Biological Resource Center of Korea Research Institute of Bioscience and Biotechnology was liquid cultured at 35 ℃ for 24 hours to prepare fermented mushroom culture.

The yeasts were diluted in the medium containing the mushroom powder and inoculated with 10 ⁶ CFU / ml, cultured in a cave at a pH of 5.5 to 6.5 at 5 to 15 ° C for 180 days, and cultured in a fermentation broth using yeast Prepared. In order to extract the active ingredient in the fermentation culture, the polymer precipitate and cells were removed using a filter having a size of 0.2 mu m to obtain an extract of Fermented Mushroom fermentation using yeast.

Experimental Example  1. Cytotoxicity Assessment - MTT  analysis

The HaCaT cells received from the Korean Cell Line Bank were inoculated into 96-well plates at a concentration of 1 x ¹⁰ cells / ml and cultured at 37 ° C in 5% CO ₂ And cultured in an incubator for 24 hours. After the incubation, the medium was removed and the medium without serum was cultured for 24 hours in the case of the positive control. In the case of the mushroom fermentation extract of Example 1 and the mushroom fermentation extract using yeast of Comparative Example 1 The cells were diluted to 25, 50, 100, and 200 ug / ml in a serum-free medium, and treated with 100 ul per well. Then, the cells were cultured for 24 hours. After incubation for 24 hours, 20 μl of the MTT reagent dissolved in PBS at a concentration of 5 mg / ml was added and cultured for 4 hours. The medium containing MTT reagent and extract was removed and 100 μl of acid isopropanol (0.04 N HCl in iso-propanol) was added to each well. The mixture was stirred in a shaker for 10 to 15 minutes, and the absorbance at 570 nm And the cell viability (%) was calculated using the absorbance (Equation 1). The results are shown in Table 1 below.

[Equation 1]

Cell viability (%) = (absorbance of test group) / (absorbance of positive control) x 100

Experimental group absorbance: The absorbance measured when the extract prepared in Example 1 or Comparative Example 1 was treated

Positive Control Absorbance: Absorbance of untreated group

division Concentration (ug / ml) Cell viability (%) Positive control group (no treatment group) - 100 Yeast extract from fermented mushroom
(Comparative Example 1) 25 101.2 50 98.5 100 97.7 200 93.4 Mushroom fermented extract treated group
(Example 1) 25 101.4 50 93.9 100 92.4 200 91.6

As shown in Table 1, when HaCaT, which is a keratinocyte, was treated at a high concentration of 200 ug / ml in both the fermentation extract of Fermenta mushroom prepared in Example 1 and the fermented extract of Fermenta mushroom using the yeast prepared in Comparative Example 1, The viability of the cells was more than 80%, indicating no cytotoxicity.

Experimental Example  2. Determination of anti- NO ( nitric oxide ) assay

Nitric oxide (NO) is a biologically active molecule with high reactivity and is produced from L-arginine by NOS (NO synthase), which is involved in neurotransmission, vascular relaxation and cell mediated immune response However, when a living body is in an inflammatory state such as atopic dermatitis, iNOS (inducible NOS) is expressed to produce a large amount of nitrogen monoxide. The inflammation promoting medium such as nitric oxide produced in this way is known to further exacerbate atopic dermatitis. No assay was performed to confirm anti-inflammatory activity.

Specifically, excessive production of RNS (reactive nitric species) induces an inflammatory response. To measure the effect of RNS, cells are stimulated with LPS (lipopolysaccharide) to induce the production of nitrogen monoxide, The anti-inflammatory effect of the mugwort fermented extract prepared in Example 1 and the fermented mugwort mugwort extract using the yeast prepared in Comparative Example 1 was measured using the principle of measuring the inhibitory effect by the griessreagent. Raw 264.7 cells (mouse macrophages) supplied by Korean Cell Line Bank were inoculated into 24-well plates at a concentration of 1 × 10 ⁵ cells / ml and cultured at 37 ° C. in 5% CO ₂ incubator for 24 hours. 100 [mu] g / ml of LPS, while using a medium containing no serum, the final concentration of the composition of Example 1 and Comparative Example 1 was diluted to 25, 50, 100, 200 ug / ml, And cultured for 24 hours. After culturing, the culture medium was recovered and centrifuged at 12,000 rpm at 4 ° C for 10 minutes. Only the supernatant was recovered, and 50 ul of the supernatant was taken into a 96-well plate and incubated with an equal amount of a grease reagent (Griessreagent) After the reaction, the absorbance was measured at 540 nm using an ELISA reader, and the inhibitory effect (%) on the production of nitrogen monoxide was determined using the absorbance. The results are shown in Table 2 below.

&Quot; (2) "

(%) = (Absorbance of sample / LPS-treated absorbance) X 100

division Treatment concentration ([mu] g / ml) NO production inhibition (%) Untreated group (positive control group) - 100 Fermented mushroom extract with yeast extract
(Comparative Example 1) 25 5.2 50 12.5 100 49.5 200 68.7 Chaga mushroom fermentation extract
(Example 1) 25 14.2 50 48.9 100 75.1 200 79.7

As shown in Table 2, when the amount of nitrogen monoxide produced in the untreated control was taken as 100%, the mushroom fermented extract prepared in Example 1 had better anti-inflammatory activity than the mushroom fermented extract prepared in Comparative Example 1 Respectively.

Experimental Example  3. Check antioxidant efficacy - DPPH Radical  Erasure analysis

While ordinary radicals are highly reactive and unstable, DPPH (1,1-diphenyl-2-picryl-hydrazyl) radicals are relatively stable free radicals and can be measured through absorption at 517 nm in the presence of radicals The free radical scavenging ability of the mugwort fermented extract prepared in Example 1 and the yeast prepared in Comparative Example 1 was measured by DPPH (Blois, MS nature 181, 1190, 1958).

The fermented extracts of the mugwort mushroom prepared in Example 1 and the fermented mugwort fermented extract using yeast prepared in Comparative Example 1 were diluted to 25, 50, 100 and 200 ug / ml, respectively, and 0.005% (w / v) BHT (butylated hydroxytoluene) was added to each well of a 24-well plate in an amount of 500 μl each. 100 μl of 0.1 mM DPPH in a methyl alcohol solution was added to each well, and the mixture was allowed to stand at room temperature for 30 minutes and then diluted with an ELISA reader (VICTOR3, Perkin Elmer) The absorbance at 565 nm was measured, and the free radical scavenging activity (%) was calculated using the absorbance (Equation 3). The results are shown in Table 3 below. As the control group, the methyl alcohol solution was used.

&Quot; (3) "

Free radical scavenging efficacy (%) = (1-experimental group absorbance / control absorbance) X100

Experimental group absorbance: The fermentation extract prepared in Example 1, the fermented extract prepared using yeast prepared in Comparative Example 1, or the absorbance upon treatment with BHP

Control absorbance: absorbance in methyl alcohol solution in which 0.1 mM DPPH was dissolved

division Free radical scavenging efficacy (%) BHT 98.12 Fermented mushroom extract with yeast extract
(Comparative Example 1) 55.12 Chaga mushroom fermentation extract
(Example 1) 75.34

As shown in Table 3, it was confirmed that the mushroom fermentation extract prepared in Example 1 had significantly better free radical scavenging activity than the fermented mushroom fermentation extract prepared in Comparative Example 1.

Experimental Example  4. Whitening efficacy  Confirm - Tyrosinase  Enzyme inhibition assay

In order to determine the inhibitory effect of the fermented mushroom extract prepared in Example 1 and the fermented mushroom fermentation extract using the yeast prepared in Comparative Example 1, the mushroom-derived thyrosinase activity was measured using Sigma (Sigma). ) Were purchased and tested.

100 μl of a 1.5 mM tyrosine solution in which tyrosine was used as a substrate in a 96-well plate was added and the fermentation extracts prepared from the mushroom fermentation extract prepared in Example 1 and the fermented mushroom fermentation extracts prepared using the yeasts prepared in Comparative Example 1 were mixed at 25, 50, 10, 200 [mu] g / ml, and then 80 [mu] l of 100 U / ml tyrosinase was inoculated. After inoculation, they were mixed and reacted at 37 ° C for 15 minutes. A 0.1 M phosphate buffer solution was used as a buffer solution in the reaction. After treating with 20 μl of kojic acid 1 and 5 μg / ml in the same manner as above, 80 μl of 100 U / ml tyrosinase was inoculated in each case, and the mixture was reacted under the same conditions. The absorbance of the resulting plate was measured at 490 nm, and the inhibitory activity (%) of tyrosinase activity was determined using the measured absorbance. The results are shown in Table 4 below.

&Quot; (4) "

(%) = 100- (absorbance of sample - absorbance of control group) x 100

division Concentration (/ / ml) Activity inhibition of tyrosinase activity (%) Kojic acid (positive control) One 45 5 91 Fermented mushroom extract with yeast extract
(Comparative Example 1) 25 11 50 14 100 18 200 21 Chaga mushroom fermentation extract
(Example 1) 25 18 50 21 100 29 200 35

As shown in Table 4, the inhibitory activity of tyrosinase activity of the myrtle fermentation extract prepared in Example 1 was higher than that of the fermented myrtle fermentation extract prepared in Comparative Example 1. Inhibition of tyrosinase inhibited the production of melanocytes. Thus, it was confirmed that the mushroom fermented extract prepared in Example 1 had a whitening effect at the enzyme level.

Experimental Example  5. Check wrinkle improving efficacy - PIP -One assay

Procollagen type 1 C-peptide ELISA kit (TaKaRa, Japan) was used to measure the amount of collagen released to the culture medium from fibroblasts. 100 μl of antibody-POD conjugate solution was added to each well of a 96-well plate included in the kit, and the fermentation extracts of the mushroom fermentation extract prepared in Example 1 and the fermented mushroom fermentation extract prepared in Comparative Example 1 were respectively 25 and 50 20 μl of PIP-1 standard solution diluted to 0, 10, 20, 40, 80, 160, 320 or 640 ng / ml was added to each well at a concentration of 100 μg / After mixing, it was sealed, incubated at 37 ° C for 3 hours, suctioned, and washed 4 times with 400ul washing buffer. After removing all of the washing buffer, 100 μl of TBMZ (3,3 ', 5,5'-tetramethylbenzidine) substrate solution contained in the kit was incubated for 15 minutes at room temperature. Then stop solution (1N H ₂ SO ₄ ) An additional 100 ul was added and mixed, and absorbance was measured at 450 nm within 1 hour after completion. The collagen biosynthesis performance (%) was determined using the measured absorbance, and the results are shown in Table 5 below.

&Quot; (5) "

Collagen biosynthesis performance (%) = 100 X (absorbance of sample / absorbance of control group)

division Concentration (/ / mL) Collagen biosynthesis performance (%) Untreated group (positive control group) - 100.00 Fermented mushroom extract with yeast extract
(Comparative Example 1) 25 105.00 50 107.00 100 107.50 200 110.00 Chaga mushroom fermentation extract
(Example 1) 25 103.00 50 107.00 100 113.00 200 121.00

As shown in Table 5, the chaga mushroom fermented extract prepared in Example 1 showed wrinkle improving effect by increasing the collagen biosynthesis ability as compared with the fermented chaga mushroom extract prepared in the yeast prepared in Comparative Example 1.

Claims (14)

A mushroom fermented extract obtained by inoculating a mushroom mycelial solid culture with a mushroom or a powder thereof and culturing the mushroom at a pH of 5.5 to 6.5 at a temperature of 5 to 15 DEG C for 180 days,
The cosmetic composition for improving anti-inflammation, antioxidation, whitening, or wrinkle, wherein the solid culture of the mycelia of mycelium of mushroom is cultured by inoculation with mushroom or a homogenate thereof. delete delete delete delete delete The method according to claim 1,
The cosmetic composition according to claim 1, wherein the mushroom fermented extract is contained in the cosmetic composition in an amount of 0.001 to 90% by weight. The method according to claim 1,
The above-mentioned mushroom fermentation extract is prepared by mixing the mushroom or its powder with the mushroom mycelial solid culture
Wherein the composition is inoculated with at least one member selected from the group consisting of Ganoderma lucidum, Cordyceps, Agaricus, Rosaceae, and powder thereof. The method according to claim 1,
Wherein the cosmetic composition is a formulation of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder. Inoculating and cultivating the mulberry into a mushroom mycelial solid culture; And
Comprising the step of inoculating chitosan or powder thereof into a solid culture of the mycelia of mycelium of mushroom and culturing at a pH of 5.5 to 6.5 and a temperature of 5 to 15 DEG C for 150 days to obtain a mushroom fermented extract. delete delete delete delete

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