KR20180103374A - A cosmetic composition containing Dendropanax morbifera ferment extract as an active ingredient and Functional cosmetics having anti-inflammation and sensitive skin improvement containing the same - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a fermented extract of Dendropanax morbiferus as an active ingredient, and more specifically, can provide a cosmetic composition containing a fermented extract of Dendropanax morbiferus leaves/branches having anti-inflammatory, anti-allergic and antibacterial effects, cosmetics containing the same, a method for manufacturing a fermented extract of Dendropanax morbiferus and an extracted Dendropanax morbiferus solution.

Description

[0001] The present invention relates to a cosmetic composition containing a fermented extract of Huangchu tree as an active ingredient and a functional cosmetic composition containing the fermented product. [0002]

The present invention relates to a cosmetic composition containing a fermented product obtained by fermenting a leafy goat's leaf extract and a yellowtail millet extract as an active ingredient. More particularly, the present invention relates to a cosmetic composition containing a fermented product extracted from a leafy yellowtail extract as an active ingredient, A cosmetic composition having skin improving effect, and a cosmetic using the same.

Dendropanax morbifera ) is a Korean species and mainly grows in southern islands, reaching a height of 15m. The young branches are green and have no hairs. The leaves are ovate or elliptical, with leaf edge smooth, but split in 3 to 5 in young trees and have serrations. The flower is pale yellowish green, blooming in June, bisexual, and hangs on a mountain-shaped inflorescence. Flower stalk is 3 ~ 5cm long, flower stalk is 5 ~ 10mm long, calyx is divided into 5, petals and stamen are 5, and there is honey gland in the petiole. The stigma is divided into five, the nucleus is elliptical, and it is mature in October and stigmas remain. Yellowish lacquer is a yellowish lacquer, which has been used as a paint for furniture for a long time. However, recent studies have shown that anti-cancer effect, hepatocyte regeneration, diabetes treatment, hard tissue cell regeneration It turned out to be effective.

Korean Patent Laid-Open No. 10-2011-0078527 discloses a cosmetic composition containing an extract of Hwangchujang as an active ingredient, wherein the extract of Hwangchujang extract has an effect of inhibiting MMP-1 production, enhancing collagen synthesis, Cytotoxic relaxation effect by ultraviolet irradiation, and suppression of inflammatory cytokine expression by ultraviolet irradiation. Korean Patent No. 10-1544176 also discloses a cosmetic composition having a whitening or anti-inflammatory function comprising a method for stabilizing Jeju Horticulturist's extract and a Horticultural extract which is stabilized by the method.

However, studies on the hypersensitive skin improvement, antiallergic effect, and anti-inflammatory effect of the leaf extract or fermented extract of saponin leaves have not yet been made yet, and the related patent has not yet been disclosed.

Accordingly, the present invention provides a cosmetic composition having excellent hypersensitive skin improvement, anti-allergic and anti-inflammatory functions, cosmetics and cosmetics using the extracts of Huchilki leaf and Huchilchia yezoensis extract which are almost no toxic to skin cells, Manufacturing methods, and further, health functional foods.

Accordingly, the present inventors have completed the present invention by developing a fermented product obtained by fermenting an extract of Hwangchujangguk (Leucaceae) Leaf and Hwangchujanggwa (Leaves), and a Leucigenia japonica.

It is therefore an object of the present invention hwangchil tree (Dendropanax The present invention relates to a cosmetic composition having an irritant skin improving, anti-allergic and / or anti-inflammatory effect containing an extract of fermented morbifera as an active ingredient, and a cosmetic and / or health functional food using the composition.

It is still another object of the present invention to provide a method for preparing a fermented extract of Huangchu tree which can be used as a cosmetic and / or health functional food material.

It is still another object of the present invention to provide a fermented broccoli extract having an irritant skin improving effect, an anti-allergic effect and an anti-inflammatory effect.

In order to achieve the above object, the present invention hwangchil tree (Dendropanax The present invention provides a cosmetic composition and / or a health functional food composition having an irritant skin improving, anti-allergic and / or anti-inflammatory effect, comprising a fermented product obtained by fermenting morbifera leaf and eggplant extract and an extract of saponin extract as an active ingredient.

In order to accomplish still another object of the present invention, the present invention relates to a method for preparing a plant extract, b) wherein a) the extract, or hwangchil hwangchil tree sap prepared in step fermentation process the Lactobacillus plantarum strains ilchiwhangcil1785 represented by Lactobacillus plantarum strain or ilchiwhangchil2020 accession No. KACC92131P represented by accession No. KACC92132P obtain a fermented product; And c) filtering the fermentation product obtained in step b) into a filter,

The method for preparing the extract of Hwangchil-myeon, which is a mixed extract prepared by extracting a mixture of Hwangchu-myeon leaf and Hwangchu-myeon branch, which comprises the extract of Hwangchu-myeon leaf and the extract of Hwangchu-myeon branch,

In order to accomplish still another object of the present invention, the present invention provides a fermented milk powder obtained by the above method.

In order to accomplish still another object of the present invention, the present invention provides a cosmetic and / or health functional food having the fermentation liquid as an active ingredient.

The fermented extract of Huchulchindo extract of the present invention has excellent radical scavenging ability of DPPH (1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity and ABTS (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) And it has excellent antioxidant effect. In addition, it has an anti-inflammatory and / or whitening effect as well as an excellent anti-allergic effect because of its excellent inhibitory effect on β-hexosaminidase enzyme production , Has excellent antimicrobial effect and low cytotoxicity, and thus can provide a new complex functional cosmetic composition, a cosmetic product and / or a health functional food.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing a process for producing a leafy leaf / eggplant extract and its fermentation process. FIG.
Fig. 2 is a photograph showing the MRS medium and the fermentations prepared in Examples 3-1 and 3-2. Fig.
FIG. 3 is a graph showing changes in pH of fermented water extracts and fermented water bodies of perilla leaf / branch extract according to microbial fermentation time (A: Lactobacillus K2020, B: Lactobacillus K1785).
FIG. 4 is a graph showing changes in the number of viable cells in the fermented water extract and fermented broth of U. pergolas leaf / root according to the fermentation time of microorganism (A: Lactobacillus K2020, B: Lactobacillus K1785).
FIG. 5 shows the results of measurement of cell viability (A) and reduction rate of NO production (B) according to the concentration of the extract.
6A to 6B show the results of measuring the inhibitory effect on the cell viability and NO production of RAW264.7 macrophage cell line according to the contents of the extract of the leafy goat / leaf extract (K2020-20%, K1785-20%).
FIG. 6C shows the results of measuring the inhibitory effect of RAW264.7 macrophage lineage on NO production according to the contents of the fermented fish oil (K2020-1%, K1785-1%).
FIG. 7 shows the results of the RBL-2H3 mast cell survival rate (A) and the inhibitory effect of β-hexosaminidase enzyme production (B) on H. japonicus extract.
8A to 8B are the results of measuring the RBL-2H3 mast cell line cell viability and β-hexosaminidase enzyme inhibitory effect of the extract of fermented Hwangbokgol (leaf) / branchy (K2020-20%, K1785-20%).
8C to 8D are the results of measuring the RBL-2H3 mast cell line cell viability and the inhibitory effect of β-hexosaminidase enzyme production on the fermented product of the perilla leaf juice (K2020-1%, K1785-1%).
Fig. 9 shows the results of antimicrobial measurement on E. coli bifidus leaf / bifidobacterium extract, Fusarium oxyspora leaf / bifidobacterium fermented product, and Fusarium spp.
Fig. 10 shows the results of antimicrobial measurement of yellow stucco leaf / egg plant extract, yellowtail leaf / branch extract fermented product, and yellowtail oil fermented product against Staphylococcus aureus.
Fig. 11 shows the results of antimicrobial measurement of P. gingko leaf / egg plant extract, P. fructus leaf / branch extract fermented, and P. vivax sap of fermented product against P. aeruginosa.
Fig. 12 shows the result of antimicrobial measurement against Candida vaginitis on the leaves and leaves of Huaxia tree, the leaves of Huaxia tree leaf / fermented products and the fermented water of Huaxia wood.

Hereinafter, the present invention will be described in more detail.

As described above, there have been no studies on the antioxidant, anti-allergic, sensitive skin improving or anti-inflammatory effects of the fermented product of Leucigenia sp.

The extract fermented product of the present invention has excellent DPPH radical scavenging ability and ABTS radical scavenging ability, and thus has excellent antioxidative effect, thereby improving sensitive skin.

The extract fermented product of the present invention has a low cytotoxicity and inhibits NO production, thereby inhibiting inflammation and inhibiting melanin production, thereby exhibiting a whitening effect.

In addition, the fermented extract of the present invention has an anti-allergic effect because of its excellent inhibitory effect on the production of? -Hexosaminidase enzyme.

Accordingly, the present invention provides a cosmetic composition comprising an extract fermented with Dendropanax morbifera as an active ingredient.

The woody trees reach 15m in height and the young branches are green and have no hairs. Leaves are alternate, ovate or oval. In addition, the edge of the leaf is flat, but in young trees it divides into 3 to 5 and has sawteeth. The flower blooms in light yellowish green in June, is bisexual, and hangs in a mountain-shaped inflorescence. Flower stem is 3 ~ 5cm long and small flower stem is 5 ~ 10mm long. Calyx is divided into 5, petals and stamens are 5, and there is nectar in the petiole. The stigma is divided into five, the nucleus is elliptical, and it is mature in October and stigmas remain. When the bark is cut, a yellow liquid comes out. It grows in southern islands as a Korean species.

In the present invention, the Huangchuang extract fermented product is obtained by extraction and fermentation from various organs or parts (e.g., leaves, flowers, roots, stems, branches, bark, sap, seeds and the like) Is an extracted fermentation product obtained from a leaf, a stem, a branch, a shell, a sap or a root, more preferably an extract fermentation product obtained from a leaf, a branch and / or a sap, and most preferably, Fermented product after fermentation, or fermented product obtained by fermenting an extract obtained from a mixture of leaves and branches.

The cosmetic composition of the present invention is not limited to be produced in any formulations conventionally produced in the art, and may be, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, Active-agent-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, but are not limited thereto. More specifically, the present invention is applicable to the production of functional cosmetics which are basic cosmetics such as lotion, nutritional cream, moisture cream, essence, eye cream, lotion, pack, essence.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the above extract or fermented extract, such as antioxidants, stabilizers, solubilizers, vitamins, Such as conventional adjuvants, and carriers.

The fermented extract of Huangchu tree of the present invention may be fermented into a bacterium of the genus Lactobacillus , more preferably fermenting into a bacterium of the genus Lactobacillus plantarum . And more preferably may be to fermentation with Lactobacillus plantarum ilchiwhangcil1785 bacteria represented by Lactobacillus plantarum strain or ilchiwhangchil2020 accession No. KACC92131P represented by accession No. KACC92132P.

According to one embodiment of the invention, Oriolus versicolor , Stereum hirustum , Phellinus linteus , Aspergillus oryzae , Bacillus sp. 　 Fomes fomentarius , Coriolus sinensis And The Lactobacillus sp. Strain was used to produce fermented extracts of Huangchuang tree, and it was confirmed that Lactobacillus sp. Strain exhibited the highest anti - inflammatory and whitening effect. Thereafter, nine out of 122 Lactobacillus spp. L. acidophilus, L. delbrueckii, L. helveticus , L. curvatus , L. plantarum , L. sakei , L. buchneri , L. fermentum And the result to prepare the L. reuteri confirm the effects of anti-allergy, anti-oxidation, anti-inflammatory, when fermented by Lactobacillus plantarum strain confirmed the highest anti-allergy, anti-oxidation, anti-inflammatory effects. That were selected strains of the two species showed the highest effect among were named respectively accession No. KACC92132P Lactobacillus plantarum ilchiwhangcil1785 bacteria represented by Lactobacillus plantarum ilchiwhangchil2020 bacteria and accession No. KACC92131P represented by.

In the present invention, the fermentation may be performed for 26 to 60 hours. If the fermentation time is less than 26 hours, the secondary metabolites may not be sufficiently generated, which may not be effective for whitening or antiinflammation. If the fermentation is performed for 60 hours or more, the acidity decreases and the number of fermentation bacteria decreases rapidly have. Particularly, it is preferable that the fermentation is carried out for about 26 hours to 40 hours in the fermentation of the leaves and the eggplant extract, and fermentation is carried out for the fermentation of the perennials in the fermentation for 36 hours to 60 hours.

In the present invention, the fermented extract of Huangchuang tree may be contained in an amount of 0.001 to 30.0% by weight based on the total weight of the cosmetic composition. The extract may be contained in an amount of 0.001 to 25.0% by weight, more preferably 0.01 to 20.0% by weight, based on the total weight of the cosmetic composition, .

According to an embodiment of the present invention, it is known that cytotoxicity is exhibited as the content of fermented Hwanyeolmum extract is increased. Therefore, it is preferable to use it in an amount of less than 20% by weight when it is used as a cosmetic raw material.

The present invention provides a functional cosmetic having an anti-inflammatory or sensitive skin soothing effect containing a fermented extract of Huangchuang tree.

The cosmetic composition of the present invention is not limited to be produced in any formulations conventionally produced in the art, and may be, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, Active-agent-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, but are not limited thereto. More specifically, the present invention is applicable to the production of functional cosmetics which are basic cosmetics such as lotion, nutritional cream, moisture cream, essence, eye cream, lotion, pack, essence.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the fermented extract or the fermented extract as described above. For example, antioxidants, stabilizers, solubilizers, vitamins, Such as conventional adjuvants, and carriers.

The present invention relates to a method for the production of Lactobacillus plantarum ilchiwhangchil2020 or Lactobacillus plantarum ilchiwhangcil1785, which is represented by Accession No. KACC92132P or Accession No. KACC92131P, To obtain an extracted fermented product; And c) filtering the extracted fermented product obtained in the step b) into a filter.

In the step a), the Hwigyeolmu tree extract may include a Hwangbukki leaf extract and a Hwangchujangguk branch extract, or a mixed extract prepared by extracting a mixture of Hwangchujanggwa leaf and Hwangchujang branch.

The step (a) of the present invention is a method for producing the extract of Hwigulrugae leaf and / or P. japonicus, which comprises extracting 8 to 12 times, preferably 9 to 11 times (wt%) And adding purified water, followed by extraction at a temperature of 70 to 90 캜 for 2 to 4 hours.

If the extract is extracted at a temperature lower than the temperature of 70 ° C or extracted within 2 hours, the active ingredient may not be extracted sufficiently from the leaves and / or branches of the leaves. When extracted at a temperature higher than 90 ° C, , The anti-inflammatory or whitening effect may not be sufficient. In addition, if the extraction is performed for a time longer than 4 hours, a large amount of ancillary components other than the active ingredient may be extracted and the anti-inflammatory or whitening effect may not be effective.

The fermentation of step b) of the present invention may be carried out by inoculating the microorganism with a number of bacteria of 6 × 10 ⁸ to 8 × 10 ⁸ cfu / ml and fermenting at a temperature of 35 to 40 ° C. for 26 to 60 hours.

If the fermentation is carried out with less than 6 × 10 ⁸ cfu / ml, secondary metabolites may not be sufficiently produced. If the fermentation proceeds more than 8 × 10 ⁸ cfu / ml, the acidity in the solution will increase There may be a side effect that the fermentation time is reduced and the production of beneficial fermentation product is decreased.

According to one embodiment of the present invention, the fermentation of the fermented extract of Huchilchia mushroom with two strains showed the highest pH change at 28-32 hrs. . In addition, the number of viable cells was confirmed at 28 to 32 hours or 2 days. It was confirmed that the cultivation of Hwangchu - wood leaf / eggplant extract for 26 ~ 36 hours and the cultivation of Hwangchu - tree sap for 46 ~ 54 hours is the optimum time to produce the Hwangchol - wood extract fermented product. Therefore, the fermentation in the step b) may be performed for 26 to 60 hours, more preferably for 28 to 54 hours, and the optimal fermentation time may vary depending on the fermentation target.

The present invention provides a fermented Lycopersiconum lentivirus having an anti-inflammatory or sensitive skin improving effect.

The leucocephalic fermentation broth may be a leaf and sap extract, or a fermentation broth obtained by fermentation of leek shark sap.

The present invention provides a cosmetic product having an anti-inflammatory or sensitive skin improving effect containing the fermentation broth as an active ingredient.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

[ Example ]

Example  1. Preparation of Leaf / Leaf Extracts and Preparation of Sapwood

Example  1-1. Leaves Heat number  Preparation of extract (powder)

In March 2016, the leaves and branches of Hwacheonchu were obtained from Jocheon-eup Jeju-si, Jeju Special Self-Governing Province and dried at 30 ℃ for 24 hours. Next, a mixture of chopped leaves and branches (leaf: branch = 1: 1 weight ratio) was added with 10 times of purified water and extracted at 80 ° C for 3 hours. Then, the extract was filtered under reduced pressure to obtain a hotchpot leaf / gang hot water extract. Next, it was concentrated and then lyophilized to be powdered.

Example  1-2. Preparing the Sapporo Saplings

In March, 2015, the sap of the Japanese whitewood tree obtained from Seogwipo City, Jeju Special Self-Governing Province was prepared.

Example  2. Selection of fermentation strains

Example  2-1: Selection of fermentation strains by anti-inflammatory or whitening effect of extracts and sap of Hwangchurak

The following experiment was conducted to select the optimum fermentation strains for fermentation of the leafy leaves / egg plant extracts or Hwangcholgwa sap that were prepared in Example 1 above.

Oriolus versicolor , Stereum hirustum , Phellinus linteus , Aspergillus oryzae, Bacillus sp. 　 Fomes fomentarius , Coriolus sinensis And Lactobacillus spp. Were prepared, and the extracts of Leucumber leaf / eggplant and Hwangjak tree were fermented. At the time of fermentation, the extracts were inoculated at 7 × 10 ⁸ cfu / ml. Mulberry leaf and eggplant extracts were prepared at a ratio of 20% by weight, and 1% of bacteria (7 × 10 < ⁸ > cfu / ml). It was then cultured and fermented in an incubator for 48 hours.

After that, NO assay was carried out using RAW264.7 to measure the anti-inflammatory effect of the fermented product of leafworm / leaf extract and fermented water. To measure the inhibitory activity of melanin production, melanin of melanocytic cell line B16-F0 Production inhibitory ability was measured.

As a result, it showed the best anti - inflammatory and whitening effect in the fermented Lactobacillus spp.

Example  2-2. Through the anti-inflammatory or whitening effect Lactobacillus  Selection of fermentation strains among the genus strains

In Example 2-1, the anti-inflammatory or whitening effect of the fermented product of the leafy / branchy fermented product and the fermented product obtained from the fermented Lactobacillus sp. Strain was excellent. Therefore, we tried to identify the most optimal Lactobacillus species.

L. acidophilus, L. delbrueckii , L. helveticus , L. curvatus, L. plantarum , L. sakei , L. buchneri , L. fermentum and L. reuteri were prepared. Then, the extracts of Leucumber leaf / eggplant and Leucillin seedlings were fermented with the strains prepared in the same manner as in Example 2-1. The extracts were prepared by inoculation of 7 × 10 ⁸ cfu / ml at the fermentation of Hwacheonchulchu extract, 20% by weight of Hwangchuhei leaf / eggplant extract and sap in the MRS medium, and 1% of bacteria (7 × 10 ⁸ cfu / ml). And then incubated in an incubator for 48 hours.

After that, NO assay was performed using RAW264.7 to measure the anti-inflammatory effect of the extract of fermented Hwangcholgusa leaf / fermented broth or liquid fermentation. To measure the inhibitory activity of melanin production, rat melanocyte B16-F0 melanin (See Example 5 hereinafter for detailed experimental methods).

As a result, it showed the best anti - inflammatory and whitening effect in the fermented Lactobacillus plantarum fermented leafy / branchy fermented or fermented liquid.

The present inventors selected two strains which can be fermented to exhibit the best anti-inflammatory and whitening effect among Lactobacillus plantarum strains. Lactobacillus plantarum ilchiwhangchil2020 (hereinafter abbreviated as K2020, microorganism accession number: KACC 92132P), Lactobacillus plantarum ilchiwhangchil 1785 (Hereinafter abbreviated as K1785, Microorganism Accession No .: KACC 92131P) and entrusted to the microorganism depository.

Example  3. Yellow-eyed Tree Extract Fermented  Produce

Example  3-1. Hwangchulgyeo leaf / eggplant extract Fermented  Produce

The extracts of hot pepper leaves / branches obtained in Example 1-1 were fermented using K2020 and K1785, respectively. Culture of K2020 and K1785 strain was used as an MRS medium ^{(Difco TM Lactobacilli MRS Broth, Becton} , Dickinson and Company, USA). The extracts of K2020 and K1785 were inoculated with 7 × 10 ⁸ cfu / ml.

Mulberry leaves and wood extracts were prepared in the ratio of 0, 10, 20, 30, and 40% by weight, respectively, and inoculated with 1% of the culture medium (7 × 10 ⁸ cfu / ml) Lt; / RTI > in a shaking incubator. Thereafter, the culture was centrifuged (4,000 x g) for 10 minutes to collect the bacterium-free supernatant. The collected supernatant was filtered (0.22 ㎛ pore size) to finally obtain a leafy / branchy fermented product. And, a rough process diagram of the fermentation of the leaf extract of Tochigi koreana leaf / gangue and its fermentation is shown in Fig.

And, actobacillus the MRS broth, MRS broth before the entry into force of applying the control of MRS broth, hwangchil leaves / branches extracts Leaf / branch extract fermented (20% by weight of extract) inoculated with plantarum K2020, MRS medium was transferred to Lactobacillus plantarum (A) to (D) of FIG. 2 are photographs of each of the fermented product of the leafy / branched extract fermented with K1785 (20 wt% of the extract).

Example  3-2. Perennial sap Fermented  Produce

The perilla seedlings obtained in Example 1-2 were fermented using K2020 and K1785, respectively. MRS medium was used as the culture medium of K2020 and K1785. The extracts of K2020 and K1785 were inoculated with 7 × 10 ⁸ cfu / ml.

MRS medium was prepared in the ratio of 0, 0.5, 1.0 and 2.0 wt%, respectively. Then, the bacteria were inoculated and then cultured in a shaking incubator for 48 hours. Thereafter, the culture was centrifuged (4,000 x g) for 10 minutes to collect the bacterium-free supernatant. The collected supernatant was filtered (0.22 μm pore size) to finally obtain a fermented extract of Hwangryuki sap. Then, the MRS medium was treated with actobacillus Fermented sap roup (fermented with 1% by weight of sap) fermented with plantarum K2020, MRS medium was cultured in Lactobacillus plantarum (E) to (F) of FIG. 2 are photographs of each of the fermented sap rosemary (fermented with 1% by weight of sap) fermented with K1785.

Example  3-3. Leaf leaf / sap extract Fermented  denomination

The leaf-leaf / branch-extracted fermented product or the liquid fermented product obtained in Examples 3-1 and 3-2 were named as shown in Table 1 below.

Sample Woodwort
Leaf / eggplant extract content
(%) Amount of sapwood in the Hwacheon tree (%) Lactobacillus  K2020 Lactobacillus  K1785 denomination A - - - - Control (0%) B 10 - O - K2020-10% C 20 - O - K2020-20% D 30 - O - K2020-30% E 40 - O - K2020-40% F 10 - - O K1785-10% G 20 - - O K1785-20% H 30 - - O K1785-30% I 40 - - O K1785-40% J - 0.5 O - K2020-0.5% K - One O - K2020-1% L - 2 O - K2020-2% M - 0.5 - O K1785-0.5% N - One - O K1785-1% O - 2 - O K1785-2%

Example  4. Setting the fermentation condition of Leaf leaf / eggplant extract or sap

Example  4-1. Set the appropriate date of fermentation according to pH change

K2020 and K1785 strains were inoculated on MRS medium containing extracts of Yulchicolorus vulgaris L. and extracts, and K2020-20%, K1785-20%, K2020 -1% and K1785-1%, respectively, were measured using a pH meter, and the results are shown in FIGS. 3A and 3B.

Fig. 3 (A) is a measurement result for K2020-20% and K2020-1%, and (B) is a measurement result for K1785-20% and K1785-1%.

The pH change of K2020 strain showed the greatest change from pH 7 to 3.0 when the culture time was 32 hours, and the pH change of K1785 strain showed the greatest change from pH 7 to 3.5 when the culture time was 32 hours Respectively.

Therefore, as shown in Fig. 3, the pH of the two strains was determined as the incubation time, and the fermentation time of the extract and the sap was set to 32 hours or more.

Example 4 -2. Depending on incubation time Viable cell count  Measure

In order to determine the viable cell counts according to the incubation time of the leavened leaf / branchy fermented product or the fermented water, the strain K2020 or K1785 was inoculated on the MRS medium containing the leafy goat / leaf extract and the sap solution (K2020-20% K1785-20%, K2020-1% and K1785-1%). Thereafter, the strain was diluted 10 ⁴ times and spread on the MRS medium at intervals of one day. After spreading, the number of colonies generated after incubation at 37 ° C for 15 hours in an incubator was counted to confirm viable cell count.

As a result, in the case of K2020-20% and K2020-1%, the viable cell count increased to the maximum number of bacteria after culturing for 30 hours, and thereafter the number of bacteria was decreased (see FIG. Also, in the case of K1785-20% and K1785-1%, the number of viable cells increased after 28 hours of culture, and thereafter the number of bacteria was decreased (see FIG. 4 (A)).

This is because the pH of Lactobacillus plantarum is increased with increasing lactic acid content. Therefore, it is considered that Lactobacillus plantarum is killed by the change of culture composition at pH 3.2 ~ 3.5.

Example  4-3. Hwangchulgyeo leaf / eggplant extract Fermentation product  And sap Fermented  Lactic acid analysis

Lactic acid production was determined by fermentation of the extracts of Leaf / Leaf Leaf / Leaf Extract and Leaf Extract to determine whether the change in pH and viable cell count were due to lactic acid production. Quantification of lactic acid was performed by measuring lactic acid dehydrogenase (LDH) at 340 nm of NADH formed in NAD ⁺ during pyruvic acid formation. The measured NADH means the presence of lactic acid, which means that the equivalence ratios are the same.

The activity of lactate dehydrogenase was determined by measuring the amount of lactic acid that was already known, and after reacting the sample with lactic dehydrogenase, the absorbance was measured at 340 nm to determine the presence or absence of lactic acid. The principle of the measurement of lactic acid is shown in Scheme 1 below.

[Reaction Scheme 1]

As a result, the fermentation of leaf - leaf - leavening fermented products showed the highest amount of lactic acid after about 28 ~ 32 hours of fermentation. This result is consistent with the result of viable cell count and the change of pH. It can be concluded that the decrease of viable cell count after about 28 ~ 32 hours is due to the change of growth environment due to the formation of lactic acid. In the case of fermented extract of Hwangchu - tree, fermentation of both strains started and lactic acid was detected most 28 ~ 32 hours.

Example  5. Perennial leaf / branch extract Fermentation product  And sap Fermented  Effect analysis

Example  5-1: Antioxidant effect analysis

A control group was prepared by mixing 6.20 g frozen powder of Hwangchujigae leaf / eggplant extract into 55 g of MRS medium.

Then, fermentations of K2020-20%, K1785-20%, K2020-1% and K1785-1% in Table 1 were prepared.

The DPPH radical scavenging activity and the ABTS radical scavenging activity were measured, and the results are shown in Table 2 below.

At this time, the DPPH radical scavenging ability was prepared by diluting the fermented water extract and fermented broth of Hwangchu-wood leaf / branch and 160 μL of each diluted solution was dispensed into a 96-well plate, and then 40 μL of 0.15 M DPPH solution was added thereto The reaction was dark room temperature for 30 minutes. After completion of the dark room reaction, the absorbance was measured using an ELISA reader (517 nm).

In addition, the ABTS radical scavenging activity was measured by mixing 14 mM ABTS reagent and 4.9 mM potassium persulfate solution in 1: 1 volume ratio, and reacting at room temperature for 24 hours. Next, in a 1.5-mL tube, 980 μL of ABTS reagent, 20 μL of leaf juice / leaf extract and 20 μL of fermentation broth were reacted. After incubation at room temperature for 10 minutes, the absorbance was measured using an ELISA reader (734 nm).

division DPPH radical scavenging ability
(IC50, 占 퐂 / ml) ABTS radical scavenging ability
(IC50, 占 퐂 / ml) Control group 133.47 ± 14.59 64.35 + - 2.22 K2020-20% 2561.96 + - 326.94 1611.05 + 415.21 K1785-20% 2301.54 ± 353.21 1633.24 + - 290.98 K2020-1% 729.55 + 152.50 345.15 +/- 12.85 K1785-1% 806.94 + - 116.47 257.24 + - 6.02

As shown in Table 2, it can be seen that the fermented product and the fermented product obtained by fermenting Kwangwoongolgwa leaf / eggplant extract with K2020 or K1785 strain had better DPPH radical scavenging ability and ABTS radical scavenging ability than the control. And, the antioxidative effect of the extracts of Ganoderma lucidum Leaf / Streptococcus sp. From the results, it was confirmed that the extract of U. perilla leaf / branch extract of the present invention and the extract of U. perch had excellent antioxidative activity.

Example  5-2: Analysis of cell survival rate and anti-inflammatory effect

In order to investigate the anti - inflammatory effects of the extracts of Hwangchu - wood and leaf - thrips, the NO assay was carried out to determine the survival rate and anti - inflammatory effect of Hwangchu -

The macrophage cell line RAW264.7 was cultured in DMEM medium (Welgene, Gyeongsan, Korea) supplemented with 10% FBS and 1% antibiotic (penicillin / streptomycin) from Korean Cell Line Bank (KCLB, Seoul, Korea) ₂ was incubated at 37 ℃ for 2 ~ 3 times. RAW264.7 cells were plated at a density of 1 × 10 ⁶ cells / well in a 96-well plate, pre-incubated in a 5% CO ₂ incubator for 20 to 24 hours, and cultured for 24 hours.

At this time, to induce inflammation of the macrophage RAW264.7 cell, 10 μl of 1 mg / ml lipopolysaccharide (Sigma-Aldrich, USA) was injected into each well. Then, 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of a griess reagent (2.5% phosphoric acid, 1% sulfanilamide, 0.1% N- (1-Naphtyl) ethylenediamine) was added to each well for reaction. The optical density was then measured with an ELISA reader at a wavelength of 540 nm. The measured optical density was expressed as a percentage (%) in comparison with the optical density value in the cell to which the lipopolysaccharide was added.

L-NAME-L-arginine methyl ester (L-NAME, Sigma-Aldrich, USA), which acts as an NO inhibitor, was used as a negative control value in RAW264.7 cell without lipid polysaccharide as a positive control. It is the optical density value in the added cells.

In addition, MTT assay was performed to measure the cell survival rate of RAW264.7. RAW264.7 cells were plated in 96-well plates at a density of 1 × 10 ⁶ cells / well, pre-cultured in a 5% CO ₂ incubator for 20 to 24 hours, treated with the concentration of each sample for 24 hours, Ten μl of MTT (5 mg / ml) solution was added to each well and reacted for 4 hours in an incubator. The supernatant was then removed and 100 μl of DMSO was added to each well to dissolve the formazan crystal and the absorbance was measured with an ELISA reader at a wavelength of 550 nm. The measured degree of formazan production was expressed as percentage (%) compared with the value of normal cells.

(1) Results of cell viability and reduction of NO production according to the concentration of Wortech extract are shown in (A) and (B) of FIG. 5, respectively. As shown in the graph of FIG. 5, no cytotoxicity was observed in the extracts of Wuchuo leaf / eggplant extracts, and the inhibition of NO production by RAW264.7 macrophage lineages according to the contents of Wuchuo leaf / ) Showed no NO production inhibitory effect (not shown), whereas NO production was decreased in a concentration-dependent manner in the leaf and leaf extracts of U. pergolas. Here, L-NAME was used as a negative control as a reagent acting as an inhibitor of NO production through inhibition of NO enzyme activity.

(2) The results of measuring the inhibitory effect on the cell viability and NO production of the RAW264.7 macrophage cell line according to the contents of the leafy leaf / branch extract fermented with Wortook seedlings were shown in FIGS. 6A and 6B, respectively. At this time, the fermented product of leafy leaf / branch extract was tested using K2020-20% and K1785-20%.

6a, it can be confirmed that there is almost no cytotoxicity due to the concentration of K2020-20% and K1785-20% fermentation.

6b, the inhibitory effect of the two strains on the production of NO in Lactobacillus plantarum ilchiwhangchil2020 (K2020-20%) and Lactobacillus plantarum ilchiwhangchil 1785 (K1785-20%) was increased (18.2 μM at 100 μg / ml), yellowish leaf / eggplant fermentation (20.22 μM at 500 μg / ml (estimated value), yellowish leaf / eggplant fermentation (K2020-20%)) Water (K1785-20%): 17.41 [mu] M at 100 [mu] g / ml).

In control (MRS medium), NO production inhibitory effect was not observed, while K2020 and K1785 showed a tendency to decrease NO production in a concentration dependent manner.

(3) The results of measuring the inhibitory effect of RAW264.7 macrophage lineage on NO production according to the content of the fermented watery perennial tree are shown in FIG. 6C. At this time, the fermented product of the perilla seedlings was tested using K2020-1% and K1785-1%.

6C shows that the NO production inhibitory effect was not observed in the control (MRS medium) (not shown), whereas NO production in Lactobacillus plantarum ilchiwhangchil2020 (K2020-1%) and Lactobacillus plantarum ilchiwhangchil 1785 (K1785-1% And the decrease in the number of workers. (K2020-1%: 17.04 μM at 100 μg / ml, K1785-1%: 16.74 μM at 100 μg / ml) than that of K2020-1% ml).

Example  5-3: Anti-allergy  Effect analysis

The cell viability and activated mast cell counts of RBL-2H3 mast cells were investigated for the antiallergic effect of fermented water extracts of Hwangchujang, leafy / branched K2020-20%, K1785-20% The inhibitory effect of β-hexosaminidase on the production of the marker enzyme for histamine release was measured.

RBL-2H3 cells were purchased from ATCC (American Type Culture Collection, USA) for the antiallergic effect. RBL-2H3 cells were cultured in EMEM (ATCC, USA) medium containing 10% FBS and 1% penicillin / streptomycin. Both cells were grown in an incubator at 37 ° C and 5% CO ₂ .

The cytotoxicity of the samples was confirmed by MTT analysis, which analyzes active mitochondria that reduce the penetrating formazan dye to MTT. RBL-2H3 cells were plated at 4 x 10 ⁴ cells per well in 96-well plates and the compounds were treated at various concentrations at 37 ° C the following day. After 24 hours, 200 占 퐇 of MTT (5 mg / ml) was added to each well and cultured at 37 占 폚 for 4 hours. Thereafter, 200 쨉 l of DMSO was added to each well to dissolve the formazan formed. The plate was maintained at room temperature for 5 minutes and the cytotoxicity was measured by measuring the absorbance at 550 nm using a microplate reader.

The inhibitory effect of β-hexosaminidase was determined by dividing RBL-2H3 cells into 24-well plates at 1 × 10 ⁵ cells per well and stimulating with anti-DNP IgE (450 ng / mL) overnight at 37 ° C. . The cells were then washed with Siraganian buffer and incubated for 20 min at 37 ° C with the addition of 160 μl of incubate buffer. 20 μl of the extract was then treated for 10 min with cells (DNP-BSA, 100 mu] g / mL) was added to stimulate the cells to form granules at 37 [deg.] C for 10 minutes. Thereafter, the reaction was stopped by soaking in an ice water bath for 10 minutes. 50 μl of the supernatant was transferred to a 96-well plate and 50 μl of 0.1 M citrate buffer (pH 4.5) containing 1 mM p-nitrophenyl-Nacetyl-β-D-glucosaminide was added thereto. And reacted for 1 hour.

Then, 100 μL of a stop solution (0.1 M Na ₂ CO ₃ / NaHCO ₃ , pH 10.0) was added to stop the reaction and the absorbance was measured at 405 nm using a microplate reader to obtain β-hexosaminidase (hexosaminidase) release inhibitory effect was measured.

(1) RBL-2H3 mast cell survival rate and inhibitory effect on the production of? -Hexasinamidase by the extract of Wuchu chrysanthemum were shown in Fig. The results of measuring the RBL-2H3 mast cell viability and the inhibitory effect on β-hexosaminidase enzyme production of the perennial leaf / branch extract fermented are shown in FIGS. 8A and 8B, respectively.

(2) FIG. 7 (A) and (B) show that the RBL-2H3 obesity cell line tended to show cytotoxicity at a concentration of 500 μg / it was confirmed that there was an inhibitory effect on the production of? -hexasaminidase enzyme.

(3) The results of the RBL-2H3 mast cell line survival rate of the leafy / leafy fermented product (K2020-20%, K1785-20%) of FIG. 8A were not cytotoxic, , It was confirmed that the fermentation products of the present invention have an excellent inhibitory effect on the production of β-hexosaminidase enzyme even at a low concentration (leaf / egg extract: 89.37% at 625 μg / ml (estimated value Lactobacillus plantarum ilchiwangchil2020: 54.47% at 100 μg / ml, and Lactobacillus plantarum ilchiwhangchil 1785: 44.43% at 100 μg / ml).

(4) Furthermore, the results of RBL-2H3 mast cell line cell survival rate of the fermented watery sapwood (K2020-1%, K1785-1%) in FIG. 8C were not cytotoxic, and as shown in FIG. 8D, It was confirmed that the effect of inhibiting the production of? -Hexasaminidase was dependent on the concentration of the fermented water of the seedlings. (K2020-1%: 71.73% efflux, 12.5 μg / ml, K1785-1%: 99.12% efflux, and 12.5 μg / ml) at the low concentration of K2020-1% , And showed similar inhibitory effect at concentrations above.

Example  5-4: Antibacterial effect analysis

The antimicrobial activity of each of the extracts of Hwangchujanggwa (leaf) / eggplant, the extracts of Hwangchuogi (leaf) / branches (fermented K2020-20%, K1785-20% Respectively.

The strains used for the antimicrobial effect were cultured as shown in Table 3 below. The paper disc diffusion method was used for the antimicrobial activity against E. coli , S. aureus , P. aeruginosa , and C. albicans .

NB or YM agar medium, and sterilized paper discs were placed. The samples were absorbed into paper disks (6 mm, GM Healthcare, UK) and cultured in a 37 ° C. incubator for 24 hours Next, it was confirmed whether a clear zone was formed around the disk.

Photographs of the analysis results are shown in Figs. 9 to 12 (Fig. 9: Escherichia coli , Figure 10: Staphylococcus aureus , Figure 11: Pseudomonas aeruginosa , Figure 12: Candida albicans ).

Each of Figs. 9 to 12 (A) shows the results of extracting 100% of (B) Hwanyeolmu leaf / branch extract before fermentation, (C) 20-20% Water K1785-20%, (D) is K2020-1%, and (E) is the result of the measurement of K1785-1%, which is a liquid fermented product.

9 to 12, (a) is a control (MRS medium), (b) to (c) are applied at different concentrations, and (e) is a measurement result after applying antibiotics .

(1) The following Table 4 shows the results of the antimicrobial measurement on E. coli and the size of the clear zone.

division Control group
(a) 1 mg
(b) 2 mg
(c) 3 mg
(d) Antibiotic
(e) Leaf / Leaf Extract 100% (A) - - - - 3.2 cm K2020-20% (B) - 1.2 cm 1.7 cm 2.0 cm 3.2 cm K1785-20% (C) - 1.5 cm 1.7 cm 2.0 cm 3.2 cm K2020-1% (D) - - - - 3.2 cm K1785-1% (E) - - - - 3.4 cm

As shown in FIG. 9 and Table 4, it was confirmed that a growth inhibition loop was formed in the leaf / branch extract fermentation broth of E. coli strain. In comparison with before fermentation, the same amount of the fermented sample (3 mg) (Before fermentation: 0.6 cm, K2020-20% after fermentation: 2.0 cm, K1785-20%: 2.0 cm). However, the fermented product of Hwangchulchae showed no antimicrobial activity.

(2) Table 5 below shows the results of the antimicrobial measurement on Staphylococcus aureus.

division Control group
(a) 50 mg
(b) 100 mg
(c) 150 mg
(d) Antibiotic
(e) Leaf / Leaf Extract 100% (A) - 0.7 cm 0.8 cm 1 cm 3 cm K2020-20% (B) - 1.2 cm 1.3 cm 1.6 cm 2.8 cm K1785-20% (C) - 1.1 cm 1.3 cm 1.6 cm 2.7 cm K2020-1% (D) - 0.7 cm 0.8 cm 0.9 cm 2.7 cm K1785-1% (E) - - - 0.9 cm 2.8 cm

10 and Table 5, it was confirmed that the growth inhibitory ring was formed in proportion to the concentration in the yellowish leaf / branch extract fermented with Staphylococcus aureus. In comparison with before and after fermentation, (1 cm before fermentation, 1.620 cm after K2020-20%, and 1.6 cm from K1785-20% after fermentation). However, the fermented product of Hwangchulchisang was K2020-1% And K1785-1%, respectively, and the growth inhibition was 0.8 cm at the concentration of 150 mg. These results show that the extract of Fermented Leaf Blade / Leaf Extracts is more effective against Staphylococcus aureus than the sap of Fusarium oxysporum.

(3) The following Table 6 shows the results of the antimicrobial measurement on P. aeruginosa and the size of the clear zone.

division Control group
(a) 1 mg
(b) 2 mg
(c) 3 mg
(d) Antibiotic
(e) Leaf / Leaf Extract 100% (A) - - - - - K2020-20% (B) - 0.8 cm 1.2 cm 1.6 cm - K1785-20% (C) - 0.8 cm 1.3 cm 1.6 cm - K2020-1% (D) - - - 0.8 cm - K1785-1% (E) - - - 0.8 cm -

11 and Table 6, it was confirmed that the growth inhibition ring was formed in proportion to the concentration of Pseudomonas sp. Leafy / branch extract in Pseudomonas sp. E. pseudomonas. In comparison with before and after fermentation, (Before fermentation: 0 cm, K2020-20% after fermentation: 1.6 cm, K1785-20%: 1.6 cm). However, the fermented product of Hwangchulchisang was K2020-1% And K1785-1% showed no antimicrobial activity at concentrations below 2mg and 0.8cm at 3mg concentration. These results show that the extracts of Leucigenia japonica Leaf / Leaf Extracts are superior to P. aeruginosa in their antibacterial effect against P. aeruginosa.

(4) As shown in FIG. 12 which is an antibacterial measurement result against Candida vaginitis, K2020-20%, K1785-20%, K2020-1% And K1785-1% can be confirmed to have no antimicrobial activity against candida vaginitis.

Formulation Example 1. Flexible life

Formulation examples of the softening longevity were prepared according to the usual method as shown in Table 7 below.

ingredient content( Unit: wt% ) Leaf leaf (or sap) extracted fermented water
glycerin
1,3-butylene glycol
PEG 1500
Allantoin
DL-Panthenol
Lee D. TEE-2NA
Benzophenone-9
Sodium hyaruronate
ethanol
Octidodeces-16
Polysorbate 20
Preservative, fragrance, pigment
Distilled water 3.0 (or 1.0)
5.0
3.0
1.0
0.1
0.3
0.02
0.04
5.0
10.0
0.2
0.1
a very small amount
Balance Sum 100

Formulation Example 2. Essence

The formulation examples of the essence were prepared as shown in the following Table 8 according to a conventional method.

ingredient content( Unit: wt% ) Leaf leaf (or sap) extracted fermented water
glycerin
Betaine
PIZZY 1500
Allantoin
DL-Panthenol
Lee D. T.-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaruronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodec-16
ethanol
Preservative, fragrance, pigment
Distilled water 5.0 (or 1.0)
10.0
5.0
2.0
0.1
0.3
0.02
0.04
0.1
8.0
0.2
0.18
0.3
0.4
6.0
a very small amount
Balance Sum 100

Formulation Example 3. Pack

Formulation examples of the pack were prepared according to the conventional method as shown in Table 9 below.

ingredient Content (unit:% by weight) Leaf leaf (or sap) extracted fermented water
Polyvinyl alcohol
Cellulose sword
glycerin
PIZZY 1500
Cyclostristin
DL - Panthenol
Allantoin
Monoammonium glycyrrhizin acid
Nicotinamide
ethanol
FYGEN 40 Hardened castor oil
Fragrance, coloring, preservative
Distilled water 3.0 (or 1.0)
15.0
0.15
3.0
2.0
0.15
0.4
0.1
0.3
0.5
6.0
0.3
a very small amount
Balance Sum 100

In the examples of the present invention, Lactobacillus plantarum strain was used for fermentation of Hwanyeolmu leaf extract or sap, and pH was lowered because lactic acid was produced as a metabolite during fermentation. In addition, the fermentation time of the fermentation broth was determined to be 28 ~ 32 days and the cell viability was 80% %, It is confirmed that a suitable dilution drainage is required when it is used as a cosmetic raw material. In addition, the fermented extract of Hwangjak tree sap is more than 80% cell viability at a high concentration, and thus it is considered safe for use as a cosmetic raw material.

In addition, since it is known that cytotoxicity is exhibited as the content of fermented product of leafy leaf extract is increased, it is recommended that the content is less than 20% by weight when it is used as a cosmetic raw material. When the content of fermented extract of Hwangjak tree sap is more than 1% by weight, whitening activity is excellent, but cytotoxicity in melanoma cells is confirmed to be strong. Therefore, when fermented extract of Hwangcholgak extract is used as a cosmetic raw material, It is recommended to use it in the content range of.

In conclusion, this study has established the optimum process for the fermentation of leafy leaves / branches by using two new Lactobacillus plantarum strains, K2020 and K1785. The effect of fermentation on skin activity (antioxidant, anti - inflammation, (Anti-allergic effects such as asthma, allergic rhinitis and atopy), and high-value cosmetics and / or health functional foods for inner beauty (skin irritation improvement and skin health) We believe the company will be able to maximize its business scope.

National Institute of Agricultural Science KACC92132P, KACC92131P 20160630

Claims (13)

Dendropanax morbifera ) fermented product as an active ingredient,
The cosmetic composition as set forth in claim 1, wherein the fermented product is a fermented product obtained by fermenting the extract of Huangchu tree and the extract of Huangchu tree. The method according to claim 1,
The hwangchil tree extracted fermentation water is as Lactobacillus plantarum ilchiwhangchil2020 bacteria or accession No. Lactobacillus plantarum ilchiwhangcil1785 bacteria characterized in that is water fermentation fermented hwangchil leaves and hwangchil branches extracts or hwangchil sap represented by KACC92131P represented by accession No. KACC92132P Cosmetic composition. The method according to claim 1,
Wherein the fermentation is carried out for 26 to 60 hours. The method according to claim 1,
The cosmetic composition according to any one of claims 1 to 3, wherein the fermented extract of Huangchu tree is contained in an amount of 0.001 to 30.0% by weight based on the total weight of the cosmetic composition. The fermented product according to claim 1, wherein the fermentation comprises a fermentation product obtained by separately fermenting each of the extracts of Hwanyeong tree leaf and Huangchu tree branch, or a fermentation product obtained by fermenting an extract extracted from a mixture of Hwangchu- ≪ / RTI > 7. A functional cosmetic having anti-inflammatory and sensitive skin-improving effects, comprising the cosmetic composition of any one of claims 1 to 5. The method according to claim 6,
Wherein the cosmetic is manufactured from at least one selected from a lotion, a nutritional cream, an eye cream, a water cream, a lotion, a pack, and an essence. comprising the steps of: a) preparing an extract of Hwangchujang or extract of Hwangryeok tree;
b) obtaining the said extract or hwangchil hwangchil wood by treating the bacterium Lactobacillus plantarum ilchiwhangcil1785 represented by the tree Lactobacillus plantarum ilchiwhangchil2020 represented by accession No. KACC92132P the fluid bacteria or accession No. KACC92131P fermentation extract fermented product; And
c) filtering the extracted fermentation product into a filter,
The extract of Hwangchu-myeon in step (a) comprises a mixture of extracts of Hwangchu-myeon leaves and Hwangchujangguk branch, or a mixture of extracts of Hwangchu-myeon Gt; 9. The method of claim 8,
In the step (a) step, the extract of Udo leaf is prepared by adding 8 to 12 times of purified water to the weight of Udo leaf, and extracting it at 70 to 90 ° C for 2 to 4 hours.
The method for preparing a fermented yellowtail millet according to item (a), wherein the extract is obtained by adding 8 to 12 times of purified water to the weight of the woody branch tree and then extracting the fermented water at 70 to 90 ° C for 2 to 4 hours. 9. The method of claim 8,
The mixed extract of step a) is prepared by adding 8 to 12 times of purified water to the weight of the mixture of Hwangchujangguk and Hwangchujang branches and extracting it for 2 to 4 hours at 70 to 90 ℃. ≪ / RTI > 9. The method of claim 8,
wherein the fermentation in step (b) is carried out by inoculating the microorganism with a number of bacteria of 6 × 10 ⁸ to 8 × 10 ⁸ cfu / ml and fermenting the fermented product at a temperature of 35 to 40 ° C. for 26 to 60 hours Gt; 11. A fermented green tea leafy mildew containing fermented yellowtail mill produced by the method of any one of claims 8 to 11. A functional cosmetic having an anti-inflammatory and sensitive skin improving effect containing the fermentation liquid of claim 12 as an active ingredient.

Images