KR102329297B1 - Cosmetic composition for anti-inflammatory or anti-atopic comprising fermented permission leaf extract - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a DSM 20314 persimmon leaf extract fermented by Lactobacillus pentosus as an active ingredient and to a preparing method thereof. The cosmetic composition exhibits excellent anti-oxidant, anti-inflammation, and atopic dermatitis alleviation effects, and has excellent skin safety and excellent moisturizing power.

Description

Cosmetic composition for anti-inflammatory or anti-atopic improvement comprising fermented persimmon leaf extract

The present invention relates to a cosmetic composition for improving anti-inflammatory or atopic dermatitis using persimmon leaves, and more particularly, an extract obtained by fermenting persimmon leaves with Lactobacillus pentosus DSM 20314, KACC 12428 as an active ingredient, It relates to a cosmetic composition with enhanced anti-inflammatory effect.

Persimmon (Diospyros kaki Thunberg) leaves have been widely used in the private sector for the prevention and treatment of hypertension, stroke, and hyperlipidemia since ancient times (Seu OS et al., J Korean Agri Chem Soc 19:93). Persimmon leaves contain flavonoid glycosides, tannins, phenols, resins, carmine compounds, reducing sugars, essential oil, chlorophyll, etc., and contain a large amount of tannins to show pharmacological action with various biological activities (Choi, JS et al., Arch Pharm., 1989; Matsuo et al., Agric. Biol. Chem, 1978). In addition, in a study on the functionality of persimmon leaves, the effect of inhibiting the activity of angiotensin-converting enzyme of persimmon leaf flavonoids (Choi, JS et al., Arch. Pharm., 1989), hot water extract and tannins from persimmon leaves Numerous studies have been conducted on the physiologically active functions of persimmon leaves, such as the mutation inhibitory effect, caries prevention effect, and the heavy metal removal effect of persimmon leaves and persimmon leaf tea (Choi, HJ et al., Korean J.Food Sci.Technol, 1998). .

In Korea, persimmons are produced in a wide area mainly in the southern region, with a total cultivation area of about 25,060ha, which is the second largest after apples. Among them, the area of persimmon cultivation in Sangju, Gyeongsangbuk-do is about 1,537ha (as of 2018), which is increasing every year. Accordingly, the use of agricultural products corresponding to the 6th industry that creates new added value by linking primary industries, such as agriculture, forestry and fisheries, secondary industries such as food and special product manufacturing and processing, and tertiary industries such as cultural tourism service industry, development of processing technology, etc. effort is required

On the other hand, fermentation is a process of decomposing organic matter using enzymes possessed by microorganisms. Fermentation method is being studied in food, medicine, cosmetics, etc. for the purpose of increasing the extraction rate of natural products and increasing physiological functionality. Polyphenols and flavonoids in natural products do not reach the effective concentration to give actual functionality to the human body, so various processing methods such as extraction, concentration, or bioconversion through fermentation are required industrially. Fermentation method can efficiently separate and extract only active ingredients from glycosides using enzymes produced by microorganisms, and higher physiological functionality can be expected if bioconversion proceeds further.

The present inventors maximized antioxidant active substances such as polyphenols and flavonoids of persimmon leaves through fermentation of persimmon leaves while contemplating the use of persimmon leaves that are not utilized and discarded although the area of persimmon cultivation in Sangju is increasing every year. An optimal fermentation strain and fermentation method have been found, and this study was conducted to confirm the possibility of application as a cosmetic composition with anti-inflammatory or atopy improving function by utilizing it.

There are Patent Documents 1 to 3 as related prior art for fermentation of persimmon. Patent Document 1 relates to a persimmon paste fermented with Lactobacillus planetarium and a food manufacturing method using the same, but does not disclose fermentation of persimmon leaves. In addition, Patent Document 2 relates to a cosmetic composition for improving wrinkles using a ferment extract obtained by fermenting persimmon leaves with Lactobacillus planetarium, and Lactobacillus pentosus fermentation of persimmon leaves is not disclosed. not. In addition, Patent Document 3, the result of previous research by the present inventors, relates to a food composition in which persimmon leaves are fermented with Lactobacillus pentosus DSM 20314, and a cosmetic composition, anti-inflammatory or atopic improvement effect thereof is not disclosed. not,

Patent Literature 1: Republic of Korea Patent No. 10-1965144 Patent Document 2: Republic of Korea Patent No. 10-1484887 Patent Document 3: Republic of Korea Patent No. 10-2233721

The present inventors have found that when persimmon leaves that are not used and discarded while having a high content of antioxidants such as vitamins, polyphenols, and flavonoids are fermented with Lactobacillus pentosus strains, higher antioxidants compared to fermentation of persimmon leaves by other strains As a result of confirming the antioxidant, cytotoxic, and anti-inflammatory effects of persimmon leaf, the ethanol extract of the Lactobacillus pentosus fermented product of persimmon leaf showed the most excellent effect while maintaining skin safety, By confirming the excellent moisturizing power, the present invention was completed.

Therefore, the problem to be solved by the present invention is a cosmetic composition for improving anti-inflammatory and atopy containing lactic acid bacteria fermented extract of persimmon leaf as an active ingredient, wherein the lactic acid bacteria fermented extract is 16s lye containing the nucleotide sequence of SEQ ID NO: 1 It is to provide a cosmetic composition for improving anti-inflammatory and atopy, which is to extract a solid fermented fermented product by inoculating the Lactobacillus pentosus DSM 20314 strain having bosomal DNA into dried persimmon leaves.

Another problem to be solved by the present invention is (a) drying and sterilizing persimmon leaves; (b) preparing a fermented product by inoculating and fermenting the Lactobacillus pentosus DSM 20314 strain having 16s ribosomal DNA including the nucleotide sequence of SEQ ID NO: 1 in the resultant (a); and (c) extracting the fermented product with C1-C6 alcohol, water, or a mixture thereof.

The problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, in the present invention, there is provided a cosmetic composition comprising a fermented extract of persimmon leaf lactic acid bacteria as an active ingredient.

One aspect of the present invention is a cosmetic composition containing a lactic acid bacteria fermented extract of persimmon leaves as an active ingredient, wherein the lactic acid bacteria ferment extract is a dried persimmon strain having 16s ribosomal DNA comprising the nucleotide sequence of SEQ ID NO: 1 It relates to a cosmetic composition for improving anti-inflammatory and atopic dermatitis, which is obtained by extracting a fermented product obtained by inoculating leaves and fermenting solid.

According to one embodiment of the present invention, the lactic acid bacteria strain is Lactobacillus pentosus (Lactobacillus pentosus) DSM 20314.

Inflammation is a physiological response that protects the living body from intrusion of foreign substances such as bacteria and mechanical damage. This inflammatory phenomenon results in a large increase in several types of polynuclear leukocytes (PMNs) and immune substances. do. Inflammation causes damage to cells and connective tissue by proteolytic enzymes, and damage to connective tissue reduces skin elasticity and causes wrinkles, as well as adversely affects cell regeneration and proliferation, leading to rapid skin aging This results in localized erythema and swelling of the skin. The production of mediators that promote the level of white blood cells along with inflammation is also achieved by the process through arachidonic acid. This pathway involves a process by cyclooxygenase, and prostaglandins produced by these enzymes act as pro-inflammatory factors. Therefore, cyclooxygenase inhibitors can exhibit anti-inflammatory action as a substance that not only inhibits the production of inflammatory substances, but also inhibits cell membrane destruction and lipid peroxidation by free radicals generated during inflammation. In addition, by inhibiting proteins and lipolytic enzymes generated by excessive inflammation, it is possible to inhibit skin aging by protecting cells damaged thereby and to improve atopic skin.

In the present invention, the fermentation method may be applied as long as it is a fermentation method that can be used by a person skilled in the art, and preferably solid fermentation may be applied. Solid fermentation may refer to fermentation by inoculating microorganisms into a raw material in a solid state with low moisture. Solid fermentation can compensate for several shortcomings of conventional liquid fermentation (Silvia Martins et al., Biotechnology Advances, 2011). First, the conventional liquid fermentation has a risk of spoilage and contamination by E. coli, anaerobic bacteria, etc., which are harmful to the human body due to fermentation conditions with a high water content. In both cases, there is a problem in that the process cost increases because stirring and sterilization processes are required during the presentation process due to fermentation conditions with a high water content. Third, a large amount of chemicals harmful to the environment are required for the submerged fermentation process. The composition according to one embodiment overcomes the problems of conventional fermentation technology by applying a solid fermentation method to persimmon leaves, so that it is not subject to contamination of harmful bacteria, can reduce process costs, and has the advantage of easy product recovery . As an example of the present invention, after sterilizing dried persimmon leaves, the persimmon leaves were fermented by inoculating a microbial strain. The amount of strain applied to the dried persimmon leaves may be 10 to 30 ml of the strain per 20 mg of persimmon leaves, but is not limited thereto. In addition, in order to increase fermentation efficiency, nutrient components that may be commonly added in microbial fermentation, for example, glucose, yeast extract, vitamins, minerals, and the like may be further added.

According to one embodiment of the present invention, the fermented product is Lactobacillus pentosus (Lactobacillus pentosus DSM 20314, KACC 12428) inoculated into dried persimmon leaves and fermented for 48 to 72 hours.

In the present invention, the "extraction" may be obtained by extraction, separation and fractionation from nature using methods known in the art for extraction, separation and fractionation. An "extract" as defined in the present invention is one obtained by extraction treatment from a fermented persimmon leaf using a suitable solvent.

In the present invention, the extraction of the fermented persimmon leaf may be extracted according to various extraction solvents and extraction methods, and suitable solvents for extracting the extract from the fermented persimmon leaf include, for example, water, methanol, ethanol, propanol. , C1-C6 alcohols such as isopropanol and butanol or a mixed solvent thereof may be used.

Ethanol, 50 to 90% (v/v) ethanol, or 60% to 80% (v/v) ethanol may be used as the extraction solvent of the extract of the present invention, but is not limited thereto. In the case of extracting the lactic acid bacteria fermented product of persimmon leaves using ethanol, both fat-soluble and water-soluble active ingredients in the lactic acid bacteria fermented product extract of persimmon leaves can be sufficiently dissolved, thereby preparing an extract having an optimal effect.

According to one embodiment of the present invention, the fermented persimmon leaf extract is obtained by drying and pulverizing the fermented persimmon leaf and then extracting it with an extraction solvent in a weight ratio of 5 to 15 times the weight of the dry pulverized fermented product. As an example of the present invention, the weight ratio of the dry pulverized fermented product and the extraction solvent was mixed at 1 : 9 and extracted.

The extraction temperature is preferably room temperature, but is not limited thereto. In addition, as the extraction method, methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method, compression method, etc. can be used.

After obtaining the extract using water or an organic solvent as described above, the liquid can be obtained by cooling, heating and filtration at room temperature by a conventional method known in the art, or further evaporating the solvent, spray drying or freeze drying. You may.

In one embodiment of the present invention, it is possible to prepare a cosmetic composition for improving anti-inflammatory and atopy by further comprising the step of obtaining a concentrate by concentration under reduced pressure after the extraction.

In the present invention, the prepared persimmon leaf lactic acid bacteria fermented extract can be obtained by fractionation after additional distillation, specifically silica gel column chromatography, thin layer chromatography

It can be obtained as a fraction further purified using various chromatography methods such as layer chromatography, high performance liquid chromatography, etc., and any method that can obtain an additional fraction using an appropriate solvent can be used. can

The cosmetic composition is selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, packs, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. It may have a formulation.

According to one embodiment of the present invention, the persimmon leaf lactic acid bacteria ferment extract is based on the total weight of the cosmetic composition. It is characterized in that it is contained in a concentration of 0.03 to 10% by weight. If it is less than 0.03%, the effect cannot be expected, and if it exceeds 10%, it may affect skin stability and formulation stability in cosmetic formulations, and economic efficiency is deteriorated.

The cosmetic composition containing the persimmon leaf lactic acid bacteria fermented surgical product has a weak acidity with a pH of 5.5 to 6.5 and thus has skin stability.

According to one embodiment of the present invention, the method for preparing the cosmetic composition comprises the steps of (a) drying and sterilizing persimmon leaves; (b) preparing a fermented product by inoculating and fermenting a lactic acid bacteria strain having ribosomal DNA including the nucleotide sequence of SEQ ID NO: 1 in the resultant (a); and (c) extracting the fermented product with C1-C6 alcohol, water, or a mixture thereof.

According to one embodiment of the present invention, the lactic acid bacteria strain is Lactobacillus pentosus (Lactobacillus pentosus) DSM 20314.

According to one embodiment of the present invention, the fermentation is to ferment for 48 to 72 hours after inoculating the dried persimmon leaves with Lactobacillus pentosus DSM 20314.

The cosmetic composition comprising the Lactobacillus pentosus DSM 20314 ferment extract of persimmon leaves according to the present invention has excellent antioxidant effect, anti-inflammatory effect or atopic improvement effect.

1 shows the 16S rDNA sequence of the Lactobacillus pentosus DSM 20314 strain.
2 is a phylogenetic diagram of the strain of the present invention. (b) shows that the strain of the present invention is 100% consistent with the 16S rRNA reference Lactobacillus pentosus DSM 20314,
3 shows the results of DPPH removal activity related to the antioxidant activity of fermented persimmon leaf extract (PLW: persimmon leaf hot water extract, PLE: persimmon leaf ethanol extract, LPLW: persimmon leaf lactic acid bacteria fermented hot water extract, LPLE: persimmon leaf lactic acid bacteria fermented ethanol extract) .
4 shows the results of MTT analysis related to cytotoxicity of fermented persimmon leaf extract (PLW: persimmon leaf hot water extract, PLE: persimmon leaf ethanol extract, LPLW: persimmon leaf lactic acid bacteria fermented hot water extract, LPLE: persimmon leaf lactic acid bacteria fermented ethanol extract) .
5 shows the results of Nitric Oxide (NO) analysis related to the anti-inflammatory activity of fermented persimmon leaf extract.
6 shows iNOS and COX-2 measurement results of fermented persimmon leaf extract.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and that the scope of the present invention is not limited by these examples according to the gist of the present invention. It is obvious to those of ordinary skill in the art,

Example 1-2. Preparation of Lactobacillus pentosus fermented persimmon leaf extract

As the strain, lactic acid bacteria, Lactobacillus pentosus (Lactobacillus pentosus DSM 20314, KACC 12428) were used, and after inoculation with the MRS culture medium in a 1 L Erlenmeyer flask, it was cultured at 30 ° C. for 1 day.

The persimmon leaves used in the present invention were used after washing the persimmon leaves picked in May and June, drying them to a moisture content of 10%, and sterilizing them at 100° C. for 10 minutes. 20 ml of the cultured lactic acid bacteria per 20 g of dried persimmon leaves were inoculated into dried persimmon leaves, fermented at 33° C. for 72 hours, dried until the moisture content reached 10%, and then ground in a blender to make powder. A leaf ferment was prepared. Hot water extraction was carried out with a weight ratio of the fermented persimmon leaf to the extraction solvent (distilled water) of 1: 9, and concentrated with a vacuum rotary concentrator to prepare a fermented persimmon leaf hot water extract (LPLW, Example 1). In addition, under the same conditions, using 70% (v/v) ethanol as an extraction solvent, an ethanol extract of fermented persimmon leaves (LPLE, Example 2) was prepared.

Comparative Example 1-2. Preparation of Persimmon Leaf Extract

As the strain, lactic acid bacteria, Lactobacillus pentosus (Lactobacillus pentosus DSM 20314, KACC 12428) were used, and the MRS culture medium was used in a 1L Erlenmeyer flask to inoculate the bacteria and cultured at 30° C. for 1 day.

The persimmon leaves used in the present invention were used after washing the persimmon leaves picked in May-June, drying them to a moisture content of 10%, and sterilizing them at 100° C. for 10 minutes. Hot water extraction was performed at a weight ratio of the dried persimmon leaf to the extraction solvent (distilled water) of 1 : 9, and concentrated with a vacuum rotary concentrator to prepare a persimmon leaf hot water extract (PLW, Comparative Example 1). In addition, an ethanol extract of persimmon leaves (PLE, Comparative Example 2) was prepared using 70% (v/v) ethanol as an extraction solvent under the same conditions.

Experimental Example 1. Measurement of antioxidant activity by DPPH scavenging activity

Persimmon leaf fermented hot water extract (LPLW, Example 1), Persimmon leaf fermented ethanol extract (LPLE, Example 2), Persimmon leaf hot water extract (PLW, Comparative Example 1), Persimmon leaf ethanol extract (PLE, Comparative Example 2) Antioxidant activity through DPPH radical scavenging ability was tested.

DPPH (α,α-diphenyl-β-picrylhydrazyl) is a water-soluble substance with a nitrogen-centered, stabilized radical in the compound, and is a purple compound with maximum absorbance at about 515 to 525 nm. When reduced, dark purple is discolored to yellow, so it is used to measure the electron donating ability of the antioxidant material, and the antioxidant activity can be measured according to the degree of scavenging ability of this stabilized radical. In the present invention, the DPPH radical scavenging ability of the fermented persimmon leaf extract was measured by partially modifying the method of Biosis et al. (1958). 800 μL of DPPH solution was added to 200 μL of each sample obtained in Examples 1 and 2 and Comparative Examples 1 and 2, mixed, and then reacted at room temperature and dark for 10 minutes, and absorbance was measured at 525 nm to obtain Equation 1 below. The inhibition rate was calculated by As a positive control, butylated hydroxyanisole (BHA), known as an antioxidant, was used.

Equation 1

×100DPPH radical scavenging ability (%) =

×100

As a result of the experiment, the antioxidant activity by DPPH scavenging activity was highest in fermented persimmon leaf ethanol extract (LPLE) at 94%, persimmon leaf ethanol extract (PLE) at 93.3%, persimmon leaf hot water extract (PLW) at 86.9%, and persimmon leaf Fermentation hot water extract (LPLW) was 85.3% (FIG. 3).

Experimental Example 2. Cytotoxicity measurement

An experiment was conducted to determine the degree of toxicity of persimmon leaf fermented hot water extract (LPLW), persimmon leaf fermented ethanol extract (LPLE), persimmon leaf hot water extract (PLW), and persimmon leaf ethanol extract (PLE).

When cells are treated with yellow water-soluble MTT tetrazolium, the metabolic process is reduced to blue-violet water-insoluble MTT formazan crystals as the ring structure of tetrazolum is broken by dehydrogenase in mitochondria of intact living cells. Therefore, the more live cells there are, the more formazan is produced, so it is possible to quantitatively evaluate the living cells according to the measured absorbance of the formazan staining.

0.18 mL of RAW264.7 cells were aliquoted to 3 × 10 ⁴ cells/well in a 96-well plate, and the samples were prepared at concentrations of 10, 25, 50, 75, and 100 ug/mL, respectively, and 0.02 mL was added after 37 C, 5% CO ₂ Incubated in an incubator for 24 hours. After adding 0.02 mL of MTT solution prepared at a concentration of 5 mg/mL to this and incubating for 4 hours, remove the culture medium, add 0.15 mL of DMSO to each well, react at room temperature for 15 minutes, and absorb absorbance at 540 nm with an ELISA reader was measured. The cytotoxicity measurement was expressed as the absorbance reduction rate of the group with and without the sample solution.

Equation 2

Cell viability (%) = (1 - absorbance of sample addition group / absorbance of no addition group) x 100

As a result, persimmon leaf hot water extract, persimmon leaf ethanol extract (PLW, PLE), persimmon leaf fermented hot water extract, and persimmon leaf fermented ethanol extract (LPLW, LPLE) all showed a cell viability of over 80% up to a concentration of 100 μg/mL. Boa (Fig. 4), it was confirmed that all were non-toxic.

Experimental Example 3. Measurement of anti-inflammatory effect by Nitric Oxide (NO) analysis

In order to investigate the anti-inflammatory action of ethanol extract of persimmon leaf fermented (LPLE), the inhibitory ability of LPS-stimulated NO production in RAW 264.7 cells was investigated.

RAW 264.7 murine macrophage cells were prepared using DMEM medium (dulbecco's modified eagle's medium, GIBCO, 1% penicillin-streptomycin) and 10% fetal bovine serum (FBS). Grand Island, YY, USA) using 37°C, 5% CO ₂ incubator conditions, and then ^{aliquoted to a cell number of 2×10 5} cells/mL in a 24-well plate, 37° C., 5% CO ₂ Incubator conditions incubated for 12 to 18 hours. Since there is a risk of cell death if the growth of RAW264.7 is excessive, serum from the medium was removed to control this, and then the cells were waited for 30 minutes for adaptation. After incubation, the medium was removed, washed with phosphate buffered saline (PBS), and exchanged with a medium containing 1 μg/mL of lipopolysaccharides (LPS). When exchanged with a medium containing LPS, RAW264.7 cells generate nitric oxide (NO) and cytokines, and for this, 18 to 24 hours of waiting. 100 ul of the supernatant and 100 ul of Greiss reagent (1% sulfanilamide, 0.1% naphthylethylenediamine in 2.5% phosphoric acid) were mixed, put into 96 wells, and absorbance was observed.

As a result of tabulating NO production according to the concentration of ethanol extract fermented from persimmon leaves, 85.7% at 6.25 μg/ml, 84.4% at 12.5 μg/ml, 83.6% at 25 μg/ml, and 70.1% at 25 μg/ml of Dexamethasone as a positive control. It was confirmed that the amount of NO production was decreased in a concentration-dependent manner of the ethanol extract fermented with persimmon leaves (FIG. 5).

Experimental Example 4. iNOS, COX-2 measurement

To investigate the anti-inflammatory effect of persimmon leaf fermented ethanol extract (LPLE) Expression of inflammation-related proteins iNOS and COX-2 was confirmed in RAW 264.7 cells.

RAW 264.7 cells ^{were dispensed and cultured in a 6-well plate at 5.0×10 5} cell/ml, and when the cell density reached 80%, it was replaced with a serum-free medium and cultured for 24 hours. , 25, 50) ug/mL. After 30 minutes of sample treatment, LPS was put into all wells except for the Normal group, stimulated, and incubated for 24 hours. Thereafter, the supernatant was removed, washed twice with PBS, and then the cells were harvested with a scraper. Lysis buffer was added to each harvested cell at 100 ug per well, lysed by refrigeration for 1 hour, and then centrifuged at 12,000 rpm, 4° C., for 10 minutes to remove cell membrane components. Proteins obtained by centrifugation were quantified by Bradford assay, and 10ul of protein was electrophoresed using 10% sodium dodecyl sulfare polyarcylamide gelelectrophoresis, and then transferred to PVDF membrane. After the transfer, it was left in 5% skim milk for 1 to 2 hours. After washing 3 times with 1xTBST, the secondary antibody (1:1000) was reacted for 2 hours and then transferred to a film using an ECL kit (Amersham pharmacia, England) for measurement. The density of the band was confirmed with Image Quant LAS-4000 (GE life sciences, Taiwan).

As a result, it was confirmed that the expression levels of iNOS and COX2 were significantly reduced in proportion to the concentration of the persimmon leaf fermented ethanol extract ( FIG. 6 ). Through this, it was confirmed that the fermented persimmon leaf ethanol extract inhibited the expression of inflammation-related proteins, and it was confirmed that it could be used as a cosmetic composition for improving anti-inflammatory or atopy.

Experimental Example 5. Measurement of pH of Cosmetic Composition Containing Fermented Persimmon Leaf Extract of the Present Invention

Healthy skin has a weak acidity of about pH 5.5, and this acidity serves to protect the skin from harmful bacteria and microorganisms. When the pH of the skin is shifted by external stimuli, this protection becomes weak. Therefore, it is appropriate that the pH concentration of cosmetics is 5.5 ~ 6.5, and pH measurement was performed to confirm this. A cosmetic composition containing the fermented persimmon leaf extract of the present invention was prepared according to the following preparation example, and the pH was measured and compared with other products.

[Production Example]

In order to prepare the cosmetic composition of the preparation example, the aqueous phase is heated between 80 ~ 90 ˚C, the oil phase is heated at 75 ~ 85 ˚C, and the aqueous phase is put into the oil phase without cooling, and then a thickener is added to emulsify with a homomixer. . After the emulsification was completed, the mixture was cooled while stirring using a stirrer, and finally, the fragrance was added and stirred. The pH of the cosmetic composition prepared above was measured.

Third-party product A Third-party product B the present invention 6.55 5.96 5.85

As a result, the competitor's product A had a pH of 6.55, the competitor's product B had a pH of 5.96, and Example 4 containing the fermented persimmon leaf ethanol extract of the present invention had a pH of 5.85, confirming that it had a stable pH level for the skin.

Experimental Example 6. Sensory evaluation of cosmetic composition containing fermented persimmon leaf extract of the present invention

A sensory evaluation was performed on the eye cream prepared according to the above preparation example. After applying the eye cream of the preparation example and the fermented eye cream of another company by a blind method to 98 second-year students of Hapcham High School, a questionnaire evaluation was conducted for fragrance and moisture, and the results are shown in Table 2 below.

division preference item incense moisture degree Fermented persimmon leaf eye cream 62 28 34 Normal Fermented Eye Cream 36 20 16

As a result, it was confirmed that the fermented persimmon leaf eye cream prepared by the present invention obtained high scores in terms of flavor and moisture compared to other fermented eye creams, indicating a high preference.

<110> SHIN, YEONHEE <120> functional food composition for antioxidant effect comprising granted permission leaf <130> app2019-0476kr <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1527 <212> DNA <213> Lactobacillus pentosus <400> 1 tcagacgaac gctggcggcg tgcctaatac atgcaagtcg aacgaactct ggtattgatt 60 ggtgcttgca tcatgattta catttgagtg agtggcgaac tggtgagtaa cacgtgggaa 120 acctgcccag aagcggggga taacacctgg aaacagatgc taataccgca taacaacttg 180 gaccgcatgg tccgagtttg aaagatggct tcggctatca cttttggatg gtcccgcggc 240 gtattagcta gatggtgggg taacggctca ccatggcaat gatacgtagc cgacctgaga 300 gggtaatcgg ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtag 360 ggaatcttcc acaatggacg aaagtctgat ggagcaacgc cgcgtgagtg aagaagggtt 420 tcggctcgta aaactctgtt gttaaagaag aacatatctg agagtaactg ttcaggtatt 480 gacggtattt aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg taatacgtag 540 gtggcaagcg ttgtccggat ttattgggcg taaagcgagc gcaggcggtt ttttaagtct 600 gatgtgaaag ccttcggctc aaccgaagaa gtgcatcgga aactgggaaa cttgagtgca 660 gaagaggaca gtggaactcc atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc 720 agtggcgaag gcggctgtct ggtctgtaac tgacgctgag gctcgaaagt atgggtagca 780 aacaggatta gataccctgg tagtccatac cgtaaacgat gaatgctaag tgttggaggg 840 tttccgccct tcagtgctgc agctaacgca ttaagcattc cgcctgggga gtacggccgc 900 aaggctgaaa ctcaaaggaa ttgagggggg cccgcacaag cggtggagca tgtggtttaa 960 ttcgaagcta cgcgaagaac cttaccaggt cttgacatac tatgcaaatc taagagatta 1020 gacgttccct tcggggacat ggatacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg 1080 agatgttggg ttaagtcccg caacgagcgc aacccttatt atcagttgcc agcattaagt 1140 tgggcactct ggtgagactg ccggtgacaa accggaggaa ggtggggatg acgtcaaatc 1200 atcatgcccc ttatgacctg ggctacacac gtgctacaat ggatggtaca acgagttgcg 1260 aactcgcgag agtaagctaa tctcttaaag ccattctcag ttcggattgt aggctgcaac 1320 tcgcctacat gaagtcggaa tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt 1380 tcccgggcct tgtacacacc gcccgtcaca ccatgagagt ttgtaacacc caaagtcggt 1440 ggggtaacct tttaggaacc agccgcctaa ggtgggacag atgattaggg tgaagtcgta 1500 acaaggtagc cgtaggagaa cctgcgg 1527

Claims (8)

A cosmetic composition containing 6.25 to 25 μg/mL of a lactic acid bacteria fermented extract of persimmon leaves as an active ingredient, wherein the lactic acid bacteria ferment extract is Lactobacillus pentosus DSM 20314 having 16s ribosomal DNA comprising the nucleotide sequence of SEQ ID NO: 1 A cosmetic composition for inhibiting nitric oxide (NO) production, which is obtained by inoculating a strain on dried persimmon leaves and extracting a solid fermented product, which improves anti-inflammatory or atopy. The cosmetic composition for inhibiting nitric oxide (NO) production according to claim 1, wherein the fermented product is fermented for 48 to 72 hours after inoculating the Lactobacillus pentosus DSM 20314 strain into dried persimmon leaves. The cosmetic composition for inhibiting nitric oxide (NO) production according to claim 1, wherein the extraction is performed by extracting the fermented persimmon leaf lactic acid bacteria by centrifugation with C1-C6 alcohol, water, or a mixture thereof. The cosmetic composition for inhibiting nitric oxide (NO) production according to claim 1, wherein the extraction is performed by extracting the fermented persimmon leaf lactic acid bacteria with 50 to 90% (v/v) ethanol after centrifugation. The cosmetic composition for inhibiting nitric oxide (NO) production according to claim 1, which is prepared by further comprising the step of obtaining a concentrate by filtration and then concentration under reduced pressure after the extraction. According to claim 1, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, pack, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. A cosmetic composition for inhibiting nitric oxide (NO) production, which has a formulation selected from the group consisting of. In the method for preparing the cosmetic composition according to any one of claims 1 to 6,
(a) drying and sterilizing persimmon leaves;
(b) preparing a fermented product by inoculating a Lactobacillus pentosus DSM 20314 strain having 16s ribosomal DNA including the nucleotide sequence of SEQ ID NO: 1 in the resultant (a) and solid-fermenting; and
(c) extracting the fermented product with C1-C6 alcohol, water, or a mixture thereof;
The cosmetic composition contains 6.25 to 25 μg/mL of lactic acid bacteria fermented extract of persimmon leaves as an active ingredient,
A method for manufacturing a cosmetic composition, characterized in that it is for inhibiting nitric oxide (NO) production. The method according to claim 7, wherein the fermentation is performed by inoculating Lactobacillus pentosus DSM 20314 into dried persimmon leaves and then fermenting for 48 to 72 hours.

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