KR101930342B1 - A cosmetic composition - Google Patents

Abstract

The present invention relates to a composition for external application for skin comprising a novel Lactobacillus curvatus (Accession number: KCTC 13494BP) J2K-01 strain isolated from aged kimchi in Jeju area, a crushed product of the strain, a culture or extract of the strain as an active ingredient. The composition exhibits excellent effects on skin condition improvement such as antioxidant effects, collagen synthesis promotion effects, skin wrinkle amelioration effects, skin elasticity enhancement effects and the like.

Description

A cosmetic composition for improving skin elasticity and wrinkles {A COSMETIC COMPOSITION}

The present invention relates to a cosmetic composition for improving skin elasticity and wrinkles.

Skin caused by ultraviolet rays generates reactive oxygen species, which causes skin cancer, phototoxicity, autoimmune disorders, photosensitivity and skin aging (Cadenas, Ann. Rev. Biochem., 58:79, 1989).

Synthesis and degradation of collagen-like extracellular matrix in vivo are appropriately controlled, but as the aging progresses, the synthesis gradually decreases and the expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, is promoted, And wrinkles are formed.

These MMPs are also activated by ultraviolet irradiation.

Therefore, it is required to develop a substance capable of controlling the activation of MMP expression in cells or inhibiting its activity.

Kimchi is a typical traditional fermented food, and various functional antioxidants, anti-aging, anti-obesity, anti-inflammatory, antibacterial, anti-allergic and immunity enhancing effects are known.

The in vivo efficacy of kimchi is derived from the nutritional components and metabolites of fermented lactic acid bacteria. Lactobacillus sp., Leuconostoc sp. And Pediococcus sp. Are involved in the fermentation and nutrient production of kimchi, and lactic acid bacteria isolated from kimchi are used for various probiotics (Probiotics) health food products are being released.

Probiotics is a beneficial microorganism that enters the body in a living state and gives immunity and various functions to help health.

Most probiotics are lactic acid bacteria and have been used in a variety of fermented foods such as meat, vegetables, milk, cheese, and yogurt.

To be recognized as a probiotic, the strain must survive in the body's stomach and bile acid to reach the small intestine and attach to the epithelial cells to grow.

It should also be non-toxic and non-pathogenic, and exhibit beneficial effects such as antibacterial, anti-inflammatory, anti-cancer and antioxidant properties in the intestinal tract.

As a related prior patent, Korean Patent Laid-Open No. 10-2018-0046831 (Patent Document 1) discloses a technique for culturing a lactic acid bacterium for cosmetic use having skin barrier recovery and skin aging-improving ability, Korean Patent Laid- 0064185 (Patent Document 2) discloses a method for producing a fermentation broth as a main component of rice germinated fermented extract and a technique relating to a cosmetic composition.

KR 10-2018-0046831 (2018.05.09) KR 10-2018-0064185 (2018.06.14)

The object of the present invention is to provide a novel strain of Lactobacillus curvatus J2K-01 (accession number: KCTC 13494BP).

Another object of the present invention is to provide a cosmetic composition for improving the wrinkles and elasticity of skin by using a composition containing a novel Lactobacillus carbatis J2K-01 (accession number: KCTC 13494BP) as an active ingredient.

In order to solve the above-mentioned problems, the present invention provides a novel strain of Lactobacillus Carbatus J2K-01 (accession number: KCTC 13494BP) derived from an old Kimchi.

In the present invention, the strain preferably has the 16S rRNA sequence of SEQ ID NO: 1, but is not limited thereto.

The culture of the present invention is prepared by inoculating and cultivating the above-mentioned strain into a culture medium, wherein the pH of the culture medium is adjusted to 5.0 to 6.0, the culture time is 12 to 48 hours, the culture temperature is 27 to 37 DEG C , 0.5 to 3% (W / V) of glucose as the carbon source of the culture medium, 0.5 to 2% (W / V) of the yeast extract as the nitrogen source of the culture medium, Manganese sulfate is contained in an amount of 0.05 to 0.1% (W / V) relative to the total volume of the medium, magnesium sulfate is contained in an amount of 0.02 to 0.05% (W / V) relative to the total volume of the medium, 0.02 to 0.05% (W / V).

The cosmetic composition of the present invention contains, as an active ingredient, at least one member selected from the group consisting of the above strain, a lysate of the strain, a culture solution of the strain or an extract thereof.

In this case, the active ingredient is contained in an amount of 0.001 to 30.0% by weight based on the total amount of the cosmetic composition.

The composition for improving skin elasticity and wrinkles of the present invention contains the cosmetic composition as an active ingredient.

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As can be seen from the present invention, the composition comprising the Lactobacillus Carbatus J2K-01 (accession number: KCTC 13494BP) strain of the present invention, the lysate of the strain, the culture solution or the extract of the strain as an active ingredient, It showed excellent antioxidant effect such as active oxygen scavenging effect, MMP inhibitory effect and UV irradiation - induced MMP expression control effect than conventional simple extract even without skin irritation.

In addition, cosmetic compositions such as lotion, cream, lotion, pack, and powder containing a culture of Lactobacillus carbatus J2K-01 (Accession No .: KCTC 13494BP) strain have an active oxygen scavenging effect and a collagenase activity modulating effect, Skin wrinkles, and skin elasticity.

1 is a 16S rRNA sequence diagram (actobacillus curvatus J2K-01, 16S rRNA gene complete cds, 1521 bp) as a result of identification of Lactobacillus curvatus J2K-01 (accession number: KCTC 13494BP).
Fig. 2 is a phylogenetic tree of Lactobacillus curvatus J2K-01 showing the result of identification of the strain Lactobacillus curvatus J2K-01 (accession number: KCTC 13494BP).
Fig. 3 is a donation of the Lactobacillus carbatoth J2K-01 strain of the present invention.

Hereinafter, the present invention will be described.

In order to develop a cosmetic raw material that is safe for skin, excellent in efficacy and has no effect on cosmetic safety in searching for microorganisms useful for human body, the present inventors have found that a large number of new Lactobacillus sp. .

Also, it was confirmed that the content of the active ingredient was increased in the culture cultured in the above strain, and Lacobacillus curvatus strain having an excellent effect as a cosmetic composition was confirmed without causing skin irritation, it to March 14, 2018 South Korea Lactobacillus Pico Yerba biotechnology Research Institute biological resource Center (Korean Collection for Type Culture, KCTC ) Tooth (Lacotobacillus curvatus J2K-01 (Accession No .: KCTC 13494BP).

The culture method according to the present invention comprises culturing Lactobacillus curvatus J2K-01 (accession number: KCTC 13494BP); After the above cultivation, centrifugal separation is performed to remove the cells from the culture solution, and the supernatant is recovered; A method for producing a composition comprising the step of filtering a supernatant recovered in the above step to remove a microbial cell or a microbial cell obtained from the culture or the microbial cell recovered by the above step is filtered to remove the microbial cells, to provide.

The Lactobacillus carbatis J2K-01 (accession number: KCTC 13494BP) strain, the disruption product of the strain, and the culture or extract of the strain finally obtained in the present invention are preferably contained in an amount of 0.001 to 30.0% by weight based on the total cosmetic composition If the content is less than 0.001% by weight, the effect of improving the skin is insignificant. If the content is more than 30.0% by weight, the efficiency with respect to the amount of the material is poor, which is not economical.

In the present invention, the culture of Lactobacillus carbatis J2K-01 (accession number: KCTC 13494BP) is preferably carried out by adjusting the pH of the culture medium to 5.0 to 6.0, optimally culturing for 12 to 48 hours, (W / V) relative to the total volume of the culture medium as the optimum carbon source of the culture medium at 27 to 37 캜, and the yeast extract as an optimal nitrogen source of the culture medium containing 0.5 to 3% (W / V) Manganese sulfate as an optimal trace element source of the culture medium, 0.05 to 0.1% (W / V) of the total volume of the medium, 0.02 to 0.05% (W / V) of the magnesium sulfate to the total volume of the medium, (W / V) relative to 0.02 to 0.05%.

In the present invention, the term "included as an active ingredient" means that the culture filtrate is added to the composition to such an extent as to exhibit an improvement effect on the skin condition from the composition of the present invention. It is meant to include formulating various components by adding various components as subcomponents.

In addition, the present invention relates to a pharmaceutical composition comprising a lactobacillus carvatus J2K-01 (accession number: KCTC 13494BP) strain containing as an active ingredient of a cosmetic, a lysate thereof, a culture or extract of the strain, An effect of improving fine wrinkles, an effect of improving skin elasticity, and an excellent water hygroscopicity.

The present invention also relates to a cosmetic composition in which the cosmetic composition is in the form of a shampoo, a lotion, a gel, a water-soluble liquid, a cream, an essence, an oil-in-water (O / W) type and a water-in-oil (W / O) type; Wherein the composition is a cosmetic composition selected from the group consisting of a makeup base, a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a cheek color and an eyebrow pencil.

In the present invention, the composition preferably refers to a composition for external application for skin, and the composition for external application for skin refers generally to all the compositions applied to the skin, and includes medicines, quasi drugs, and cosmetics.

The blending amount of the effective ingredient of the strain of the present invention in the external preparation for skin is preferably 0.0001 to 10%, more preferably 0.01 to 1%, based on the total amount of the composition.

Hereinafter, the present invention will be described in more detail by way of examples. It should be noted, however, that these examples are only illustrative examples of the present invention, and the scope of the present invention is not limited to these examples.

<Separation of strain from kimchi>

Kimchi from Jeju Island was used as a lactic acid bacteria isolated kimchi.

Each of the kimchi samples was suspended and diluted in sterilized physiological saline, and 0.1 mL of each of the samples was plated on a Difco Lactobacili MRS Agar medium.

Thereafter, the plate was incubated in a constant temperature incubator at 35 DEG C for 2 days.

Each of the resulting colonies was inoculated on MRS agar plates and cultured at 35 DEG C for 2 days to obtain a colonies showing white colonies.

Lactobacillus carvatus J2K-01 was isolated from kimchi in the same manner as above.

&Lt; Preparation of Lactobacillus carbatus J2K-01 culture >

A seed culture of the Lactobacillus carbatis J2K-01 strain isolated pure from Example 1 was carried out by inoculating a single colony into MRS medium and incubating in a constant temperature incubator at 35 ° C for 24 hours.

The optimized culture medium of Lactobacillus carbbatus J2K-01 strain was optimized medium containing 2% of glucose, 1% of yeast extract, 0.1% of manganese sulfate, 0.05% of magnesium sulfate and 0.025% of calcium chloride and pH 5.5 The seed culture was inoculated to the conditioned medium in a concentration of 1% (V / V), and then cultured at a temperature of 35 ° C and a rotation speed of 120 rpm for 24 hours.

After culturing, the cells were removed from the culture by centrifugation, and the supernatant was filtered to obtain a Lactobacillus carbamazep J2K-01 culture in which the cells were completely removed.

Experimental Example 1: Measurement of free radical scavenging activity

To determine the free radical scavenging activity of the culture of the Lactobacillus carbatis J2K-01 strain obtained in Example 2, extracts having excellent antioxidative activity, such as green tea extract under laboratory conditions, were used as comparative samples and free The radical scavenging activity was measured.

The DPPH method used free radical scavenging activity by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-1picrylhydrazyl free radical).

Free radical scavenging was measured at a wavelength of 560 nm by comparing the degree of reduction of the absorbance by DPPH reduction by the test substance with the absorbance of the blank test solution.

For the measurement of DPPH free radical scavenging activity, cultures were prepared at concentrations of 0.2%, 0.1%, 0.05% and 0.005%.

The culture at the above concentration was placed in a 96-well plate, and DPPH prepared from 100 uM methanol solution was added thereto to make the total volume of the solution 200 μl.

After incubation at 37 ° C for 30 minutes, absorbance was measured at 560 nm.

&Quot; (1) &quot;

Free radical scavenging activity (%) = {100- (B / A)} X100

A: Absorbance of the control well without treatment of the sample of the present invention,

B: absorbance of the experimental group well treated with the sample of the present invention,

As shown in Table 1, the culture of the Lactobacillus carbamazep J2K-01 strain of Example 2 shows an antioxidative effect superior to that of the green tea extract having excellent antioxidative activity.

Name of sample Antioxidative effect(%) Example 2 (0.05% by weight) 85 Example 2 (0.1% by weight) 92 Example 2 (0.2% by weight) 98 Green tea extract (0.2% by weight) 90

Experimental Example 2: Cytotoxicity of a sample

The cytotoxicity of the cultured product of Lactobacillus carbatis J2K-01 strain obtained in Example 2 on skin cells was evaluated.

To the 96-well test plate, 1 x 10 ⁵ cells / mL of diluted fibroblast was added to the cell culture medium (DMEM supplemented with 10% FBS) and adhered for 24 hours.

The sample obtained in Example 2 was diluted to an appropriate concentration in each well and treated for 24 hours.

After 24 hours, the medium was removed, and 200 μL of cell culture medium containing MTT (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyl tetrazolium bromide) solution (2.5 mg / And then cultured in a 37 ° C CO ₂ incubator for 2 hours.

The medium was removed and 100 μL of DMSO (dimethyl sulfoxide) was added.

Cells were lysed by shaking for 5 minutes and absorbance at 565 nm was measured in a microplate reader. The cell viability (%) was determined as in the mathematics 2 and the concentration of the sample that did not affect cell survival was determined.

&Quot; (2) &quot;

Cell survival rate (%) = {(St-Bo) / (Bt-Bo)} X 100

Bo: 565 nm absorbance value of wells reacting with cell culture medium,

Bt: 565 nm absorbance value of a well color-reacted without sample treatment,

St: 565 nm absorbance of the well treated and chromogenic.

Name of sample 100% cell viability concentration Example 2 5% Example 2 3% Example 2 One% Example 2 0.5%

As shown in Table 2, the 100% survival treatment concentration of the dermal fibroblasts was 5% or less in the culture of the Lactobacillus carbamazep J2K-01 strain of Example 2, It is a safe sample with less possibility of skin irritation.

Experimental Example 3: Inhibitory activity of MMP-1 in vitro

Fluorescence analysis was used to measure the inhibitory effect of MMP-1 activity of the culture of the Lactobacillus coumarbatus J2K-01 strain obtained in Example 2 above.

The substrate used was DQ-Collagen labeled with fluorescein and the collagenase was purchased from Molecular probe (Eugene, OR, USA) and incubated in reaction buffer (0.5 M Tris-HCl, 1.5 M NaCl , 50 mM CaCl2, 2 mM sodium azide, pH 7.6) was used after 10-fold dilution.

20 μl of DQ collagen dissolved in a reaction buffer at 0.25 mg / ml in the reaction buffer and 40 μl each of 0.2%, 0.1%, and 0.05% concentrations of the sample obtained in Example 2 were added, and 0.5 μl And 40 [mu] l of diluted collagenase.

After 20 minutes at room temperature in a dark place, the fluorescence value was measured using a fluorescence spectrophotometer (Perkin Elmer, UK) with an absorption wavelength of 495 nm and an emission wavelength of 515 nm. As a control, a fluorescence value The fluorescence value of the sample itself was also measured and corrected for the enzyme activity.

The results are shown in Table 3. The metal protease (MMP-1) inhibitory activity was excellent at 0.2% treatment of the sample of Example 2, and the inhibitory activity was better than that of green tea extract.

From the above results, it was confirmed that the effect of inhibiting metalloproteinase (MMP-1) activity of the culture of Lactobacillus carbapthus J2K-01 strain was excellent. Based on this fact, the skin external- It is possible to infer that the elasticity of the skin and the formation of wrinkles can be prevented or improved.

Name of sample MMP-1 activity inhibition rate (%) Example 2 (0.05% by weight) 38 Example 2 (0.1% by weight) 52 Example 2 (0.2% by weight) 74 Green tea extract (0.2% by weight) 67

Experimental Example 4: Evaluation of inhibition of MMP-1 expression after UV irradiation

Enzyme immunoassay (ELISA) was performed to measure the concentration of MMP-1 after addition of the culture of Lactobacillus carbamazu strain J2K-01 obtained in Example 2 after UV irradiation.

UVA was irradiated in human dermal fibroblasts at an energy of 6.3 J / cm &lt; 2 &gt; using a UV chamber.

Ultraviolet irradiation dose and incubation time were established by preliminary experiment to maximize MMP expression level in fibroblasts.

The negative control was wrapped in a silver foil and the cells in the UVA environment were transferred to the medium containing the sample after irradiated with UVA, and cultured for 24 hours. Then, the medium was recovered and coated on 96 wells. The primary antibody (MMP-1 (Ab-5) monoclonal antibody) was treated and reacted at 37 ° C for 60 minutes.

The cells were incubated with a secondary antibody, anti-mouse IgG (alkaline phosphatase conjugated), for 60 minutes, and incubated with an alkaline phosphatase substrate solution (diethanolamine buffer solution containing 1 mg / ml p- The reaction was carried out at room temperature for 30 minutes and the absorbance at 405 nm was measured with a microplate reader. As a control group, a sample to which no sample was added was used.

As shown in Table 4, when the sample of Example 2 was added to MMP-1 in which expression was induced by ultraviolet irradiation, the MMP-1 inhibition rate was increased and the inhibition rate was over 75% as compared with the control without the sample.

This is an excellent result compared to the inhibition rate of retinol used as a control.

Test group MMP-1 expression inhibition rate (%) Negative control group - Example 2 (0.05% by weight) 75 Example 2 (0.02% by weight) 60 Example 2 (0.01% by weight) 31 Retinol 41

Experimental Example 5: Cytotoxicity mitigation effect by ultraviolet irradiation

The cytotoxic effect of the culture of the Lactobacillus coumarbatus J2K-01 strain obtained in Example 2 on UV irradiation was evaluated.

Fibroblasts were placed in 24-well test plates at 1 × 10 ⁵ cells and adhered for 24 hours.

Each well was washed once with PBS and 500 ul of PBS was added to each well. The cells were irradiated with 10 mJ / cm 2 of ultraviolet light using ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan) 1 ml) was added.

Here, the sample to be evaluated was treated and cultured for 24 hours.

After 24 hours, the medium was removed, and cell culture medium (500 μl) and MTT solution (2.5 mg / ml) (60 μl) were added to each well and incubated for 2 hours at 37 ° C in a CO2 incubator.

The medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added.

The cells were shaken for 5 minutes to dissolve the cells, and 100 쨉 l of the supernatant was transferred to a 96-well test plate, and absorbance at 565 nm was measured in a microplate reader.

The cell viability (%) was measured as shown in Equation (3), and the cytotoxic mitigation rate by ultraviolet was calculated by Equation (4).

&Quot; (3) &quot;

Cell survival rate (%) = {(St-Bo) / (Bt-Bo)} X 100

Bo: absorbance at 565 nm of the well in which only the cell culture medium was color-

Bt: absorbance at 565 nm of a well that underwent chromogenic reaction in a well not treated with the sample,

St: Absorbance at 565 nm of a well that underwent chromogenic reaction in a sample-treated well.

&Quot; (4) &quot;

Cytotoxicity mitigation rate by ultraviolet ray (%) = 1- (St-Bo) / (Bt-Bo) X 100

Bo: the cell survival rate of wells not irradiated with ultraviolet rays and not treated,

Bt: cell survival rate of wells irradiated with ultraviolet rays and not treated with samples,

St: Cell survival rate of well irradiated and treated samples.

Sample treatment concentration (%) Cytotoxic Relaxation Rate (%) Example 2 (0.05% by weight) 47 Example 2 (0.02% by weight) 31 Example 2 (0.01% by weight) 15

As shown in Table 5, the culture of the Lactobacillus carbamazep J2K-01 strain of Example 2 effectively inhibited cytotoxicity by ultraviolet light by mitigating cytotoxicity by 47% of ultraviolet rays at a concentration of 0.05% And it was found.

From the above results, it is possible to deduce the skin irritation effect of the culture of Lactobacillus carbatus strain J2K-01 by ultraviolet rays.

Experimental Example 6: Promotion of type 1 procollagen biosynthesis

To investigate the effect of promoting the biosynthesis of type 1 procollagen, which is a skin matrix component, after the culture of the Lactobacillus coumarbatus J2K-01 strain obtained in Example 2, a procollagen assay was conducted under the following conditions The same method was used.

Human dermal fibroblasts isolated from neonatal foreskin tissues were purchased from Modern Tissue Technology (MTT, Korea) and supplemented with 10% fetal bovine serum (FBS) in DMEM / F-12 (3: And cultured at a concentration of 1 × 10 ⁴ cells / cm 2.

When 70-80% of the cells were grown, the cells were subcultured at a ratio of 1: 3, and cells in the third to fourth subculture were used for the experiment.

For the experiment to measure the amount of procollagen, the fibroblasts were cultured in a 48-well plate at 90% or more, and then the sample of Example 2 was added. After 24 hours, the amount of procollagen liberated in the medium was measured using a procollagen- 1C-peptide EIA kit (MK101, Takara, Japan).

The results are shown in Table 6. In the 0.05% treatment of the sample of Example 2, the biosynthesis of procollagen promoted 58%, which was similar to that of TGF-beta, a cell signaling substance.

Name of sample Promoting effect of procollagen biosynthesis Example 2 (0.05% by weight) 58 Example 2 (0.02% by weight) 34 Example 2 (0.01% by weight) 20 TGF-beta 55

Experimental Example 7: Skin elasticity improvement effect and wrinkle improvement

A cosmetic preparation containing the culture of the Lactobacillus carbamazep J2K-01 strain obtained in Example 2 was prepared and compared with Comparative Example 1 to humans, and skin elasticity improving effect and wrinkle improving effect were evaluated.

The cosmetics used in the comparative experiments were in the form of a cream, and cream (Example 3, Comparative Example 1) was prepared according to the composition shown in Table 7 below in a conventional manner.

The cream prepared in Example 3 was applied to the right side of the face and the cream prepared in Comparative Example 1 was applied to the left side of the face for two consecutive months for 20 consecutive months for 20 subjects (20 to 35 year old female) .

The skin elasticity was measured by using a skin elasticity meter (SEM 575, C + K Electronic Co., Germany) before and after using the product for 2 months.

The experimental results are shown in the following Table 8 as the ΔR7 value of Cutometer SEM 575, where the R7 value represents the viscoelasticity of the skin. As shown in Table 8, it can be seen that the skin elasticity improving effect of the experimenter coated with the cream containing the culture of Lactobacillus carbatus strain J2K-01 is excellent.

Raw material Example 3 Comparative Example 1 Purified water Balance Balance glycerin 8 8 Beta Glucan 5 5 Carbomer 0.1 0.1 Stearic acid cholesterol 2 2 Squalane 4 4 Cetearyl alcohol 6 6 Polyoxyethylene alcohol ester 3 3 Caprylic / capric triglyceride 8 8 Example 2 One - Propylene glycol 5 5 incense Suitable amount Suitable amount Pigment Suitable amount Suitable amount antiseptic Suitable amount Suitable amount Sum 100 100

Note) Unit: wt%

Experimental product Skin elasticity effect (△ R7) Example 3 0.32 Comparative Example 1 0.11

n = 20, p < 0.05

The cream prepared in Example 3 was applied to the right side of the face and the cream prepared in Comparative Example 1 was applied to the left side of the face for two consecutive months for 20 consecutive months for 20 subjects (20 to 35 year old female) .

To evaluate the wrinkle improvement effect of the skin before and after the use of the product after the completion of the experiment, a silicone replica was made and the wrinkle state of the designated area was measured with a visiometer SV60, C + K Electronic Co., Germany).

The results are shown in Table 9, which shows the average value of each parameter after 2 months minus the parameter value two months ago.

That is, the negative value indicates that the wrinkle improving effect is higher.

Therefore, as shown in Table 9, it was confirmed that the effect of improving the wrinkles of the skin of Example 3 containing the culture of Lactobacillus coumarbatus J2K-01 strain of Example 2 was greatly enhanced.

Experimental product R1 R2 R3 R4 R5 Example 3 -0.21 -0.15 -0.11 -0.08 -0.09 Comparative Example 1 -0.11 -0.06 -0.05 -0.04 -0.02 R1: Difference between the maximum value and the minimum value of the wrinkle contour line
R2: The wrinkle contour line is arbitrarily divided into 5 squares, and the average of R1 values
R3: Maximum value of R1 divided by 5
R4: Mean value of the baseline of the wrinkle contour minus the value of each apex and valley
R5: Mean value of the value obtained by subtracting the outline of each wrinkle from the base line of the wrinkle contour line

Experimental Example 8: Effect of skin irritation mitigation

The effect of stimuli relaxation of the cosmetic containing the culture of the Lactobacillus carbapthus strain J2K-01 obtained in Example 2 was evaluated by a human skin patch test.

1% of sodium lauryl sulfate (SLS), which stimulates general cosmetic prescriptions (cream, lotion, skin, essence), and the product prepared in Example 3 were mixed for 24 hours, 48 hours and 72 hours, And the effect of stimulation relaxation was evaluated.

Each product was sprayed with 0.3 mg of FINN CHAMBER (FINLAND) on the upper arm of 50 healthy men and women aged 20 to 50 years, and the acute irritation index was evaluated after 24 hours.

After the evaluation, the same amount of the product was applied again to the same site again to evaluate the delayed irritation index after 48 hours and 72 hours.

As a result of the test, when the product containing the Lactobacillus carbamazep J2K-01 culture of the present invention was applied on the skin, There was no skin attack after 72 hours.

This result shows that there is a significant effect of reducing skin irritation caused by a stimulant (surfactant, fragrance, alcohol) when the Lactobacillus carbatis J2K-01 culture of the present invention is mixed with cosmetics .

Other examples are shown below.

That is, the shampoo, lotion, milky lotion and serum of the present invention obtained in Example 2 containing the lactobacillus carbamazu J2K-01 culture of the present invention were prepared as in Examples 4 to 7.

The shampoos, lotions, emulsions and cosmetic liquids containing the cultures of Lactobacillus carbamazep J2K-01 (Accession No .: KCTC 13494BP) of the present invention have a reactive oxygen scavenging effect and a collagenase activity controlling effect, The effect of improving elasticity, the effect of alleviating skin irritation, and the like.

Example 4: Production of Shampoo Containing Culture of Lactobacillus Carbatus J2K-01 of the Present Invention

To 5 g of 95% ethanol, 5 g of ammonium laureth sulfate, 5 g of cosyl betaine, 0.5 g of alkyl ether cellate, 0.5 g of silicone, 0.2 g of polyquater, 0.2 g of fragrance and 0.02 g of preservative were mixed. 5 g of the lactobacillus carbamazep J2K-01 culture of the present invention obtained in Example 2 and 5 g of glycerin were dissolved in 85.33 g of purified water, and the mixture was added to the mixture, followed by stirring to obtain a shampoo.

Example 5 Production of Cosmetic Lotion Containing Culture of Lactobacillus Carbatus J2K-01 of the Present Invention

0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of perfume, 0.1 g of p-hydroxybenzoic acid methyl ester, a small amount of antioxidant and a small amount of dye were mixed in 8 g of 95% ethanol.

10 g of the lactobacillus carbamazu J2K-01 culture of the present invention obtained in Example 2 and 5 g of glycerin were dissolved in 85.33 g of purified water, and the mixture was added to the mixture, followed by stirring to obtain a lotion.

Example 6: Preparation of an emulsion containing the lactobacillus carbatous J2K-01 culture of the present invention

0.2 g of cetyl alcohol, 10 g of squalane, 2 g of vaseline, 0.2 g of p-hydroxybenzoic acid ethyl ester, 1 g of glycerin monoestearylate, 1 g of polyoxyethylene (20 mol addition) monooleate and 0.1 g of coryneform were heated and mixed at 70 캜 , 0.5 g of the lactobacillus carbamazu J2K-01 culture of the present invention obtained in Example 2, 5 g of dipropylene glycol, 2 g of polyethylene glycol-1,500, 0.2 g of triethanolamine and 76.2 g of purified water were heated to 75 캜 Lt; / RTI &gt; The two were mixed and emulsified and then cooled to obtain oil-in-water (O / W) type emulsion.

Example 7: Preparation of serum of the present invention containing a culture of Lactobacillus carbamazu J2K-01

To 5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of chitoolose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium permanganate, 0.1 g of p-hydroxybenzoic acid ethyl ester 1 g of the lactobacillus carbamazu J2K-01 culture of the present invention obtained in Example 2 and an appropriate amount of pigment were mixed to obtain a serum.

Korea Biotechnology Research Institute KCTC13494BP 20180314

<110> J2KBIO CO., LTD. <120> LACTOBACILLUS CURVATUS J2K-01 STRAIN AND A COSMETIC COMPOSITION          USING THE SAME FROM KIMCHI <130> j2k-01 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1521 <212> RNA <213> Lactobacillus curvatus <400> 1 ctcaggacga acgctggcgg cgtgcctaat acatgcaagt cgaacgcact ctcgttagat 60 tgaagaagct tgcttctgat tgataacatt tgagtgagtg gcggacgggt gagtaacacg 120 tgggtaacct gccctaaagt gggggataac atttggaaac agatgctaat accgcataaa 180 acctagcacc gcatggtgca aggttgaaag atggtttcgg ctatcacttt aggatggacc 240 cgcggtgcat tagttagttg gtgaggtaaa ggctcaccaa gaccgtgatg catagccgac 300 ctgagagggt aatcggccac actgggactg agacacggcc cagactccta cgggaggcag 360 cagtagggaa tcttccacaa tggacgaaag tctgatggag caacgccgcg tgagtgaaga 420 aggttttcgg atcgtaaaac tctgttgttg gagaagaacg tatttgatag taactgatca 480 ggtagtgacg gtatccaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat 540 acgtaggtgg caagcgttgt ccggatttat tgggcgtaaa gcgagcgcag gcggtttctt 600 aagtctgatg tgaaagcctt cggctcaacc gaagaagtgc atcggaaact gggaaacttg 660 agtgcagaag aggacagtgg aactccatgt gtagcggtga aatgcgtaga tatatggaag 720 aacaccagtg gcgaaggcgg ctgtctggtc tgtaactgac gctgaggctc gaaagcatgg 780 gtagcaaaca ggattagata ccctggtagt ccatgccgta aacgatgagt gctaggtgtt 840 ggagggtttc cgcccttcag tgccgcagct aacgcattaa gcactccgcc tggggagtac 900 gaccgcaagg ttgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg 960 gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcctttg accactctag 1020 agatagagct ttcccttcgg ggacaaagtg acaggtggtg catggttgtc gtcagctcgt 1080 gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttattacta gttgccagca 1140 tttagttggg cactctagtg agactgccgg tgacaaaccg gaggaaggtg gggacgacgt 1200 caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggat ggtacaacga 1260 gtcgcgagac cgcgaggttt agctaatctc ttaaaaccat tctcagttcg gattgtaggc 1320 tgcaactcgc ctacatgaag ccggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380 atacgttccc gggccttgta cacaccgccc gtcacaccat gagagtttgt aacacccaaa 1440 gccggtgagg taaccttcgg gagccagccg tctaaggtgg gacagatgat tagggtgaag 1500 tcgtaacagg gtaacccgta a 1521

Claims (7)

A cosmetic composition for improving skin elasticity and wrinkles,
1% by weight of strain culture, 8% by weight of glycerin, 5% by weight of betaglucan, 0.1% by weight of carbomer, 2% by weight of stearic acid cholesterol, 4% by weight squalane, 6% by weight of cetearyl alcohol, 3% by weight of polyoxyethylene alcohol ester, 8% by weight of caprylic / capric triglyceride and 5% by weight of propylene glycol,
The culture of the strain may contain,
The Lactobacillus curvatus strain J2K-01 isolated from the old kimchi, which is assigned to deposit number KCTC 13494BP, is inoculated into the medium and cultured,
The pH of the culture was adjusted to 5.5, the incubation time was 24 hours, the incubation temperature was 35 ° C,
As a carbon source of the culture medium, glucose was contained in an amount of 2% (W / V) based on the volume of the culture medium,
As the nitrogen source of the culture medium, the yeast extract contained 1% (W / V) of the total volume of the culture medium,
Manganese sulfate was added to the culture medium as a trace element source in an amount of 0.1% (W / V) relative to the total volume of the culture medium. Magnesium sulfate was contained in an amount of 0.05% (W / V) (W / V)
Culturing, centrifuging to remove cells from the culture, and filtering the supernatant.
A cosmetic composition for skin elasticity and wrinkle improvement. delete delete delete delete delete delete

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