KR20170143478A - Ferment collagen for preventing skin aging and improving skin wrinkle and preparing method same - Google Patents

Abstract

The present invention relates to production of a fermented collagen using microorganisms. Disclosed is a fermented collagen which is prepared by using microorganisms that have excellent effects of preventing skin-aging and reducing skin wrinkles while dramatically improving skin penetration rate. The present invention provides a fermented collagen for preventing skin-aging and reducing skin wrinkles, which is fermented by using Lactobacillus paracasei or Lactobacillus rhamnosus.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a fermented collagen for preventing skin aging and improving skin wrinkles, and a method for producing the same. BACKGROUND ART [0002]

The present invention relates to a fermented collagen and a method for producing the same, and more particularly, to a fermented collagen for preventing skin aging and improving skin wrinkles and a method for producing the same.

One of the greatest areas of interest in cosmetics is wrinkling, pigmentation, loss of elasticity, skin dryness, hair loss, and lack of shine in the hair associated with aging. The skin is subject to various changes through aging. First, the thickness of the epidermis, dermis, and subcutaneous tissues, which are components of the skin, is thinned and the extracellular matrix (ECM) component that gives skin elasticity changes. Among them, collagen which accounts for 70 ~ 80% As the age increases, the production thereof is drastically lowered to have a close relationship with wrinkle formation, and collagen, elastin, proteoglycans, glucosaminoglycan, , Laminin, fibronectin, etc. are oxidized and lose their function, causing the skin to lose its elasticity and wrinkles becoming excessively formed and changing into senile skin. There are collagen fibers, collagen fibers, reticular fibers, elastic fibers (elastic fibers, elastic fibers) in the connective tissue of the extracellular matrix. Collagen, which accounts for 70% Are mostly formed in fibroblasts of the skin. As you get older, the collagen content in the connective tissue of the skin decreases, which is due to degradation of collagen synthesis and promotion of degradation. Therefore, degradation of collagen biosynthesis and promotion of collagen degradation are the major causes of wrinkles in the skin. The collagen biosynthesis process is regulated by many factors involved in transcription level and post-translation level, and collagen degradation is triggered by collagenase degrading collagen by ultraviolet rays Promotion of matrix metalloproteases (MMP), such as collagenase, promotes collagen degradation and reduces collagen content. In addition, as the collagen's deformation accelerates due to the external environment, the skin becomes wrinkled and deepened. As a result, skin aging is caused by non-homogenization of cells, loss of elastin, destruction of collagen, reduction and delay of collagen synthesis, and the like. Therefore, the skin aging phenomenon occurs in the epidermis of the skin but can be seen as a phenomenon that occurs in the dermis.

Various cosmetic compositions have been studied for solving the problem of skin aging, and the wrinkle improvement effect of the skin is visible in some parts. For example, there have been various reports on retinoids widely used for the improvement of skin wrinkles, spots, and pigment deposition, and in particular, clinical results of removing skin wrinkles on retinol. Among them, retinol- And skin deflection and elasticity reduction.

In order to utilize the skin moisturizing effect and the wound healing effect of the collagen, there have been introduced products containing collagen in the composition for external application for skin such as cosmetics and ointment. However, these products apply collagen itself to the surface of the skin, There is a problem that the transdermal absorption is difficult and a moisturizing action or a wound healing effect can not be expected.

On the other hand, Japanese Laid-Open Patent Publication Nos. 2008-0081454 and 1254403 disclose a fermentation collagen production method using various strains. Particularly, in Patent Publication No. 2012-0081454, low molecular weight fermentation collagen is produced Although mention is made on the production of fermentation collagen that can be absorbed more quickly into the dermal layer of the skin, it is not explicitly mentioned about changes in collagen low molecular weight and physiological activity obtained through fermentation and utilization of specific strains for improving skin absorption rate have.

Accordingly, it is an object of the present invention to provide a fermented collagen produced by using a specific microorganism having excellent skin aging resistance and skin wrinkle-reducing effect, particularly in skin fermentation collagen production using microorganisms.

The present invention also provides a cosmetic composition for preventing skin aging and improving skin wrinkles containing fermented collagen produced by using a specific microorganism.

The present invention also provides a method for producing optimal fermentation collagen using specific microorganisms.

In order to solve the above problems, the present invention provides a fermented collagen for preventing skin aging and improving skin wrinkles fermented with Lactobacillus paracasei or Lactobacillus rhamnosus .

Also, the fermentation collagen has a collagenase activity inhibition rate of 20% or more.

Also, the fermentation collagen is characterized by having a skin permeability (based on 24 hours of the experiment) of at least 0.01 mg / ml using a Franz diffusion cell.

In order to solve the above-mentioned problems, the present invention provides a cosmetic composition for preventing skin aging and improving skin wrinkles containing the fermented collagen as an active ingredient.

The formulations of the cosmetic composition may further comprise at least one selected from the group consisting of softening agents, convergent lotion, nutritional lotion, eye cream, serum, nutritional cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, A cosmetic composition, a cosmetic composition, a cosmetic composition, a shampoo, and a rinse.

In order to solve the above-mentioned problem, the present invention provides a method for preventing and treating low-molecular-weight skin aging and skin wrinkles by adding Lactobacillus paracasei or Lactobacillus rhamnosus to a medium containing collagen, The present invention also provides a method for producing fermented collagen.

The Lactobacillus paracasei or Lactobacillus rhamnosus is inoculated in a culture medium of 10 ⁷ to 10 ¹⁰ cfu / ml in an amount of 0.5 to 5 parts by weight per 100 parts by weight of the culture medium. ≪ / RTI >

According to the present invention, Lactobacillus paracasei or Lactobacillus rhamnosus is selected as a microorganism used for fermentation collagen production, and has excellent skin aging prevention and skin wrinkle reducing effect, It is possible to provide a fermentation collagen which is dramatically improved compared to the prior art.

Also, by using the fermentation collagen prepared by using the specific microorganism as the cosmetic composition, it is possible to provide a cosmetic composition having excellent anti-aging effect and skin wrinkle-reducing effect.

In addition, when utilizing Lactobacillus paracasei or Lactobacillus rhamnosus for fermentation collagen production, it is possible to provide optimal conditions for fermentation collagen production that maximizes skin permeation performance.

1 is a photograph showing a comparison of collagen fermentation patterns in four fermentation broths of lactic acid bacteria through SDS-PAGE in Example 1 of the present invention,
FIG. 2 is a photograph showing a comparison of patterns of acid-treated and base-treated collagen with the fermentation broth of lactic acid bacteria through SDS-PAGE in Example 2 of the present invention,
3 is a graph showing the results of confirming the effect of improving the skin wrinkles of fermented collagen in Example 3 of the present invention,
FIG. 4 is a graph showing the skin permeability of collagen, fermented collagen, and caffeine Franz diffusion cells in Example 4 of the present invention.

Hereinafter, the present invention will be described in detail with reference to examples. Prior to this, terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms, and the inventor should appropriately interpret the concepts of the terms appropriately The present invention should be construed in accordance with the meaning and concept consistent with the technical idea of the present invention. Accordingly, it is to be understood that the constituent features of the embodiments described herein are merely the most preferred embodiments of the present invention, and are not intended to represent all of the inventive concepts of the present invention, so that various equivalents, And the like.

The inventors of the present invention have conducted intensive studies on a cosmetic composition having anti-aging effect and skin wrinkle effect containing low-molecular-weight collagen as an active ingredient. As a result, it has been found that when lactobacillus microorganisms are used, , It was found that optimal performance can be achieved only when Lactobacillus paracasei or Lactobacillus rhamnosus is used as a specific microorganism in various Lactobacillus microorganisms in order to improve skin permeability And came to the present invention.

Accordingly, the present invention discloses , as an embodiment, a fermentation collagen for preventing skin aging and improving skin wrinkles fermented with Lactobacillus paracasei or Lactobacillus rhamnosus .

The fermentation collagen according to the present invention is prepared by adding and fermenting Lactobacillus paracasei or Lactobacillus rhamnosus to a medium in which collagen is mixed, whereby the low-molecular-weight fermentation collagen is prevented from aging skin And improve the wrinkle-reducing effect of the skin.

In the present invention, the collagen to be used for fermentation collagen production may have a weight average molecular weight of 50,000 to 500,000 and a weight average molecular weight of 100,000 to 300,000 in consideration of the microorganism used.

A method for culturing fermented collagen is, for example, a method comprising culturing 100 parts by weight of Lactobacillus paracasei or Lactobacillus rhamnosus in 0.5 to 5 parts by weight of a culture medium containing 10 ⁷ to 10 ¹⁰ cfu / And incubated at 35 to 39 ° C for 1 to 5 days, preferably 0.5 to 2 parts by weight in a culture medium of 10 ⁸ to 10 ⁹ cfu / ml, and cultured at 36 to 38 ° C for 2 to 4 days . At this time, the culture medium used for the culture may include peptone, sodium acetate hydrate, magnesium sulfate hydrate, manganese sulfate hydrate, Tween 80, ammonium citrate, potassium phosphate and distilled water. 1 to 10 parts by weight, preferably 2 to 5 parts by weight of collagen may be mixed and used. After completion of the cultivation, the culture broth is centrifuged to separate the lactic acid bacteria and the culture broth to produce fermented collagen.

The fermented collagen thus prepared is highly suitable for collagen fermentation by using Lactobacillus paracasei or Lactobacillus rhamnosus as a microorganism and having excellent collagen low molecular weight as compared with other lactic acid bacteria strains According to one embodiment of the present invention, the fermentation collagen having a collagenase activity inhibition rate of 20% or more, preferably 25% or more, more preferably 35% or more can be provided.

Also, according to one embodiment of the present invention, skin permeability (based on 24 hours of the experiment) using Franz diffusion cells, which promotes skin absorption, is 0.01 mg / ml or more, preferably 0.03 mg / ml Or more, more preferably 0.05 mg / ml or more.

According to another aspect of the present invention, a cosmetic composition for preventing skin aging and improving skin wrinkles containing fermented collagen as an active ingredient is disclosed.

The cosmetic composition may contain 0.1 to 90% by weight of the fermentation collagen as an active ingredient. In addition to the fermented collagen, components commonly used in cosmetic compositions may be included in the cosmetic composition. Examples thereof include stabilizers, dissolving agents, vitamins, , And a carrier.

The cosmetic composition for preventing skin aging and improving skin wrinkles according to the present invention may be prepared in any formulations conventionally produced in the art and may be in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, , Surfactant-containing cleansing,

Oils, powder foundations, emulsion foundations, wax foundations, sprays, and the like, but are not limited thereto. More specifically, the present invention relates to a cosmetic composition for cosmetics, such as softening longevity, convergent lotion, nutritional lotion, eye cream, serum, nutritional cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, , Rinse, and the like.

When the formulation is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracer, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Butane, dimethyl ether, and the like.

When the formulation is a solution or an emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan can be used.

When the formulation is a suspension, the carrier component may be a liquid diluent such as water, ethanol, propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar, trachant and the like may be used.

In addition, when the formulation is an interface-active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, and the like can be used.

Hereinafter, the present invention will be described in detail with reference to examples.

Example  1: Confirmation of collagen fermentation and fermentation products

After the fermentation of collagen (Mw 100,000 ~ 300,000) using Lactobacillus sp. Lactic acid bacteria, fermentation conditions of collagen were examined. The minimum culture conditions were investigated to find Lactobacillus sp.

Lactobacillus acidophilus , Lactobacillus plantanum , Lactobacillus rhamnosus and Lactobacillus paracasei were used as the strains of the lactic acid bacteria, It was purchased from a bank and used.

All four lactic acid bacterial strains were inoculated in a selective medium and inoculated at a concentration of 1 8 wt% in a culture medium in which the number of bacterial strains was about 10 ⁸ to 10 ⁹ cfu / ml, cultured in an incubator at 37 ° C for 72 hours Respectively. 5 g of peptone, 2.5 g of sodium acetate trihydrate, 0.5 ml of magnesium sulfate heptahydrate, 0.5 ml of manganese sulfate tetrahydrate, 5 ml of tween 80, 1 g of diammonium citrate, 1 g of dipotassium phosphate phosphate and 480 ml of distilled water was used as a culture medium in which 15 g of collagen was mixed.

After the cultivation was completed, the culture broth was centrifuged at 4,000 rpm for 10 minutes to separate the lactic acid bacteria and the culture broth. The separated culture broth was confirmed to have low molecular weight collagen using SDS-PAGE, . Lanes 1 to 6 in Fig. 1 show the fermentation of Lactobacillus acidophilus , Lactobacillus plantanum fermentation, Lactobacillus rhamnosus fermentation, Lactobacillus paracase , ( Lactobacillus paracasei ) fermentation and protein size marker.

1, when fermentation was performed with Lactobacillus rhamnosus and Lactobacillus paracasei among four lactic acid bacteria strains, a lot of low-molecular-weight fermentation proceeded, and Lactobacillus paracasei ( Lactobacillus paracasei ) And it was confirmed that most of the low - molecular - weight fermentation was most suitable for collagen fermentation.

Example  2: Collagen through fermentation, acid and base treatment Low-molecular-weight  compare

The fermentation broth fermented with Lactobacillus paracasei and Lactobacillus rhamnosus obtained in Example 1 and acid treatment (neutralized with 1M HCl for 1 hour and collagen at pH 6.5) and base treatment (1 mM NaOH for 1 hour and then neutralized to pH 6.5). The results were compared in FIG. 2 by comparing the results of the SDS-PAGE of the four samples . In FIG. 2, lanes 1 to 5 represent acid treatment, base treatment, protein size marker, Lactobacillus paracasei fermentation and Lactobacillus rhamnosus fermentation, respectively.

2, it was confirmed through SDS-PAGE that acid treatment, base treatment, and Lactobacillus rhamnosus fermentation collagen were also effective for low molecular weight collagen. However, the fermentation broth of Lactobacillus paracasei fermentation broth It was confirmed that the collagen of the present invention was most homogeneously and low molecular weight.

Example  3: Confirmation of improvement of skin wrinkles of fermented collagen

In order to confirm the effect of improving the skin wrinkle of the fermentation collagen obtained in Example 1, the in vitro collagenase activity inhibition test was carried out as described below, and the results are shown in FIG.

[Collagenase activity inhibition test method]

1. Prepare collagen by dissolving in phosphate buffer (100, 50, 25ppm). At this time, EGCG (Epigallocatechin gallate) is prepared at the same concentration as a positive control group.

2. Prepare a test sample at 300 μl / 48 well by diluting to a suitable concentration range using a solvent (DMSO) at 48 wells.

3. Referring to the following Table 1, a 15 ml glass tube is prepared, and the solution is dispensed in the order shown in Table 2 below.

4. Add 25 mM citric acid (400 μl / tube) to terminate the reaction.

5. Add 1.5 ml / tube of ethyl acetate, vortex and leave for 10 minutes.

6. Take supernatant, transfer to sodium sulfate 150 mg / ep-tube, vortex and centrifuge at 10,000 rpm for 3 minutes.

7. Take supernatant 300 μl / ep-tube and transfer to quartz 96-well plate. Measure absorbance at 320 nm.

3, the in vitro collagenase activity inhibition test showed that the inhibition rate of collagenase activity was excellent when fermentation of Lactobacillus paracasei was performed compared to pre-fermentation collagen and acid-treated and base-treated collagen. .

Example  4: Confirmation of improved skin permeability of fermented collagen

In order to confirm the enhancement of the skin permeability of the fermentation collagen, a skin permeability comparative experiment using a Franz diffusion cell was performed on the epidermis of the human body as described below, and the results are shown in FIG. 4 .

[Experimental method of skin permeability]

1. Fill the receptor chamber of the Franz diffusion cell with 6.5 ml of phosphate buffer solution and place the horny layer face up on the skin of the human body between the donor chamber and the receptor chamber. (cadaver). The epidermis skin of Hans Biomed was used as the cadaver used.

2. In the donor chamber, an aqueous solution having a concentration of 1 mg / ml of fermentation collagen and normal collagen is prepared as a comparative group, and an aqueous solution having a concentration of 1 mg / ml of caffeine is prepared as a positive control.

3. Keep the temperature at 35 ± 1 ° C during the experiment and continue stirring the magnet.

4. At 0.5, 1, 3, 5, 8, 12, and 24 hours, each sample in the receptor chamber is sampled at a sampling port.

5. The amount of collagen and caffeine in the sample by time is checked by HPLC analysis, and the skin permeation amount and skin permeability are determined, and the comparison group and the control group are compared.

4, the skin permeability comparison experiment using Franz diffusion cell showed that collagen before fermentation hardly penetrated into skin with a large molecular weight, but Lactobacillus paracasei fermentation collagen had a tendency to increase in time Was found in the receptor chamber through the cadaver between the receptor chamber and the donor chamber of the Franz diffusion cell in proportion to the size of the donor chamber. Through this example, it was confirmed that collagen fermented using Lactobacillus paracasei significantly promotes skin absorption of collagen.

Hereinafter, a production example of applying a cosmetic composition for preventing skin aging and improving skin wrinkles containing fermented collagen according to the present invention as an active ingredient will be described, but the present invention is not limited thereto.

Manufacturing example  1: Production of softened longevity

Lactobacillus paracasei fermented collagen prepared according to Example 1 and having the composition shown in the following Table 3 was prepared according to the general method of producing lotion.

Manufacturing example  2: Nutritional cream manufacture

A nutritional cream containing Lactobacillus paracasei fermented collagen prepared according to Example 1 and having the composition shown in Table 4 below was prepared according to the general nutrition cream preparation method.

Manufacturing example  3: Nutritional lotion manufacturing

Nutritional lotion comprising Lactobacillus paracasei fermented collagen prepared according to Example 1 and having the composition shown in the following Table 5 was prepared according to the general method of producing lotion.

Manufacturing example  4: Essence  Produce

Lactobacillus paracasei fermented collagen prepared according to Example 1 and comprising the composition of the following Table 6 was prepared according to the general essence production method.

While the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, Such modifications and changes are to be considered as falling within the scope of the following claims.

Claims (7)

Fermented collagen for preventing skin aging and improving skin wrinkles fermented with Lactobacillus paracasei or Lactobacillus rhamnosus . The method according to claim 1,
Wherein the fermentation collagen has a collagenase activity inhibition rate of 20% or more. 3. The method of claim 2,
Wherein the fermentation collagen has a skin permeability (based on 24 hours as an experiment) of at least 0.01 mg / ml using a Franz diffusion cell. A cosmetic composition for preventing skin aging and improving skin wrinkles containing the fermented collagen of any one of claims 1 to 3 as an active ingredient. 5. The method of claim 4,
The formulation of the cosmetic composition may be selected from the group consisting of softening agents, convergent lotion, nutritional lotion, eye cream, serum, nutritional cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, , Shampoo, and rinse. ≪ RTI ID = 0.0 > 11. < / RTI > ( Lactobacillus paracasei or Lactobacillus rhamnosus ) is added to fermented collagen-containing medium and fermented to prepare low molecular weight fermented collagen for preventing skin aging and improving skin wrinkles. The method according to claim 6,
Wherein said Lactobacillus paracasei or Lactobacillus rhamnosus is inoculated with 0.5 to 5 parts by weight per 100 parts by weight of said medium in a culture medium containing 10 ⁷ to 10 ¹⁰ cfu / .

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