KR101627850B1 - Method of fermentation using complex strains and cosmetic composition comprising thereof - Google Patents

Abstract

The present invention relates to a human-derived Lactobacillus lambosus rhamnosus ) HK-9 and Lactobacillus paracasei paracasei HS-05 complex lactic acid bacteria and a functional cosmetic composition having antioxidative and skin whitening effect containing the fermented product. Human-derived Lactobacillus Lactobacillus rhamnosus HK-9 and Lactobacillus paracasei The hot-water extract of Lactobacillus paracasei HS-05 lactic acid bacteria and the fermented fermenting material using the strain showed high antioxidative and whitening activity, and more physiologically active substances and effective substances It is possible to obtain the maximum effect even with a small amount of elution, which is economically advantageous.

Description

[0001] The present invention relates to a method for producing a fermented product using a human strain derived complex strain, and a functional cosmetic composition containing the fermented product,

The present invention relates to a human-derived Lactobacillus lambosus rhamnosus ) HK-9 and Lactobacillus paracase paracasei HS-05 complex lactic acid bacteria and a functional cosmetic composition having antioxidative and skin whitening effect containing the fermented product.

Melanin is an important means of protecting harmful ultraviolet rays from harmful ultraviolet rays of sunlight. It is a high molecular substance of phenols widely present in animals, plants and microorganisms, and enhances survival ability against ultraviolet ray, drying, extreme temperature, etc. However, excessive melanin production , Freckles, black blotch, accelerate skin aging, and is known to be involved in skin cancer induction of malignant melanoma. Furthermore, since melanin is chemically and physically very stable, it is almost impossible to decompose the melanin in a short period of time without significant damage to the skin. Therefore, a method for preventing and treating melanin is a substance that inhibits the synthesis of melanin . In addition, there is a problem in foods that causes a change in the quality of vegetables, fruits, fish, etc. (Bell and Weeler, 1986; Chen et al., 1991; Lerner and Fitzpatrick, 1950). Melanin is biosynthesized by the continuous oxidation of tyrosinase enzymes in the intracellular melanosomes of melanocytes in the epidermal basal layer.

Tyrosinase is a major enzyme of the melanin biosynthetic process and is a copper-containing enzyme that uses a phenolic compound as a substrate. Tyrosine is oxidized by tyrosinase to L-3,4-dihydroxyl-L-phenylalanine (LDOPA) and L-DOPA to dopaquinone, which is then converted to 5,6-dihydroxy indole, , 6-quinone, and melanin is biosynthesized by polymerization (Lopez et al., 1992).

Known as tyrosinase inhibitors are ascorbic acid, kojic acid (Lee et al., 2006), hydroquinone (Fitton and Goa, 1991; Parvez et al., 2006), benzoic acid benzoic acid, retinoid, and arbutin (Maeda and Fukuda, 1996). Especially, kojic acid and arbutin have strong whitening effect, but problems such as product stability and economical efficiency (Ando et al., 1993; Masuda et al., 1996).

Therefore, it is necessary to develop cosmetics for antioxidation and skin whitening which are free from side effects and have low cost and excellent stability.

Therefore, the present inventors have found that lactic acid bacteria derived from human body, such as Lactobacillus rhamnosus ) HK-9 KCCM11349P and Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P was eluted with more physiologically active substance and effective substance than when each of these strains were fermented, and thus it was possible to obtain more effect in a smaller amount, thereby completing the present invention .

It is therefore an object of the present invention to provide a process for preparing Lactobacillus rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P strain as an active ingredient, and to provide a cosmetic composition for antioxidant or skin whitening.

Another object of the present invention is to provide a method for producing Lactobacillus < RTI ID = 0.0 > rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P strain as an active ingredient, and to provide a cosmetic composition for skin whitening.

Another object of the present invention is to provide a method for producing Lactobacillus < RTI ID = 0.0 > rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 strain KCCM11254P as an active ingredient.

Further, it is an object of the present invention to provide a method for producing a cosmetic composition for antioxidant or skin whitening.

In order to accomplish the above object, the present invention provides a pharmaceutical composition comprising Lactobacillus rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P strain as an active ingredient to provide an antioxidant or skin whitening cosmetic composition.

In one embodiment of the present invention, the cosmetic composition is prepared by adding to a golden powder (Scutellaria baicalensis GEORGE) Lactobacillus rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P strain may be inoculated and then fermented to exhibit antioxidant or skin whitening effect.

The present invention relates to Lactobacillus < RTI ID = 0.0 > rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 A cosmetic composition for antioxidant or skin whitening comprising a hot-water extract of KCCM11254P strain as an active ingredient.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 KCCM11254P strain as an active ingredient.

Further, the present invention relates to a pharmaceutical composition comprising (a) Lactobacillus rhamnosus ) HK-9 KCCM11349P and / or Lactobacillus paracase (La ctobacillus paracasei ) HS-05 100 ml of KCCM11254P strain is sonicated at 50 to 60 kHz for 20 to 50 minutes to disrupt the strain; And (b) adding 500 ml to 1 L of water at 50 to 70 캜 to the strain and the disrupted lysate obtained in step (a), followed by hot water extraction for 12 to 24 hours. And a method for producing the same.

In one embodiment of the present invention, the natural substance of step (a) is selected from the group consisting of gold, garlic, green tea, mung bean, sunflower seed, lily flower, peony root, cedar, , Angelica gigas, Lentil, Quercus, Ginseng or Wild ginseng.

The present invention relates to a pharmaceutical composition comprising Lactobacillus rhamnosus HK-9 derived from human skin and Lactobacillus parasite The present invention relates to a method for preparing a fermented product using the HS-05 complex lactic acid bacterium and to a functional cosmetic composition having antioxidative and skin whitening effect containing the fermented product. When the strain is fermented in a complex manner, It has a higher antioxidative and whitening activity. Further, it can exert the physiologically active substance and the active substance in a larger amount in a small amount, and thus has an economical advantage.

Figure 1 is a graphical representation of Lactobacillus rhamnosus HK-9 < / RTI > KCCM11349P.
Figure 2 is a graph paracasei HS-05 < / RTI > KCCM11254P.
FIG. 3 is a graph showing the activity of Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P lactic acid bacteria.
Figure 4 is a graph rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, the two fermented lactic acid bacteria of the present invention and the control group, Lactobacillus casei FIG. 5 is a graph showing the results of measuring the radical scavenging ability of the fermentation group. FIG.
Figure 5 is a graph rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, the two fermented lactic acid bacteria of the present invention and the control group, Lactobacillus casei FIG. 5 is a graph showing the results of measuring the reducing power of the fermentation group. FIG.
Figure 6 is a graph rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, the two fermented lactic acid bacteria of the present invention and the control group, Lactobacillus casei FIG. 5 is a graph showing the results of measuring the inhibition rate of melanin synthesis in the fermentation group. FIG.
Figure 7 is a graphical representation of Lactobacillus rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, the two fermented lactic acid bacteria of the present invention and the control group, Lactobacillus casei FIG. 5 is a graph showing the result of measuring the inhibition rate of tyrosinase in a fermentation group. FIG.

Best Mode for Carrying Out the Invention

The present invention relates to a pharmaceutical composition comprising Lactobacillus rhamnosus HK-9 derived from human skin and Lactobacillus parasite paracasei HS-05 complex lactic acid bacterium, and a functional cosmetic composition having antioxidative and skin whitening effect containing the fermented product.

In recent years, not only many women but also men have become increasingly interested in cosmetics, and various functional cosmetics for skin whitening, acne, atopic dermatitis and wrinkle are being released. However, most functional cosmetics sold on the market are products using synthetic compounds, and cosmetic products containing synthetic compounds can cause sensitive skin reactions depending on the skin, which causes various skin side effects.

Accordingly, the present inventors have focused on lactic acid bacteria, which have been studied for the production of a skin whitening cosmetic product having no side effects, low cost, and excellent stability.

Lactic acid bacteria are important microorganisms widely used and applied worldwide, and they have been used for a long time in fermented dairy products such as cheese, fermented milk and lactic acid bacteria drink, and food industry such as kimchi and chungkukjang. Lactobacillus has a long-lasting effect on the treatment and prevention of intestinal diseases, thus preventing the growth of harmful bacteria in the intestines and making good conditions for the growth of beneficial bacteria, thereby preventing food poisoning, preventing intestinal decline, preventing constipation, And is used in various dairy products, foods, and cosmetics industries.

The inventors of the present invention focused on the fact that the hand of a woman is softer than that of a man's hand and that lactic acid bacterium is abundantly present (PNAS 105 (46) 17994-17999) Two kinds of Lactobacillus spp. Were selected, and the microorganisms obtained by the above method were deposited in the international patent institution and assigned the deposit numbers as KCCM11349P and KCCM11254P respectively, Lactobacillus rhamnosus HK-9 KCCM11349P " and " Lactobacillus paracasei HS-05 KCCM11254P ".

According to one embodiment of the present invention, Lactobacillus rhamnosus HK-9 of the present invention and Lactobacillus < RTI ID = 0.0 > paracasei ) HS-05 was tested for antioxidative and skin whitening effects. Lactobacillus rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, a group in which the two lactic acid bacteria of the present invention were combined and fermented and a control group, Lactobacillus casei The DPPH scavenging activity, reducing power and total phenol content of the fermented group were measured. As a result, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei In the gold fermented with HS-05 KCCM11254P, the DPPH scavenging activity, the reducing power and the total phenol content were higher than that of the fermentation of the common lactic acid bacteria and the above strains, respectively, indicating high antioxidative activity (see Table 1) Showed results similar to those obtained when the lactic acid bacteria were disrupted to measure DPPH scavenging activity and reducing power (Figs. 4 to 5).

According to another embodiment, the natural gold, Lactobacillus rhamnosus HK-9 KCCM11349P group, Lactobacillus paracasei HS-05 KCCM11254P group, by treating the normal lactic acid fermentation of Lactobacillus casei group the above two lactic acid bacteria of the present invention in a group and the control group composite culture fermentation was measured melanin synthesis inhibition and tyrosinase inhibition tee, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei In the gold fermented with HS-05 KCCM11254P, it was found that melanin synthesis inhibition and tyrosinase inhibition were significantly higher than those of fermentation of the common lactic acid bacteria and the strain, respectively (see Table 2) Similar results were obtained when the lactic acid bacteria were disrupted to measure melanin synthesis inhibition and tyrosinase inhibition (Figs. 6 to 7).

Therefore, based on these results, it was found that Lactobacillus rhamnosus HK-9 and Lactobacillus paracasei HS-05 complex lactic acid bacteria are excellent in antioxidative and skin whitening effects, and thus can provide a functional cosmetic composition.

The strain according to the present invention may be added in an amount of 0.001 to 20% by weight based on the total weight of the cosmetic composition.

Lactobacillus Lactobacillus rhamnosus ) HK-9 KCCM11349P and Lactobacillus < RTI ID = 0.0 > paracasei ) When the HS-05 KCCM11254P complex is added in an amount of less than 0.001% by weight based on the total weight of the cosmetic composition, the cosmetic composition of the present invention does not exhibit various physiological activity effects. On the other hand, Or the desired effect can not be obtained in proportion to the added amount. Therefore, it is preferable to add the cosmetic composition in an amount within the range described above in consideration of economic efficiency, and the cost of the cosmetic composition according to the amount of the cosmetic composition increases, which is not economical .

When the composition of the present invention is prepared with a cosmetic composition, the composition of the present invention not only ferments the above-mentioned complex strain, but may also contain components conventionally used in cosmetic compositions. Examples thereof include antioxidants, stabilizers, , Customary adjuvants such as vitamins, pigments and flavoring agents, and carriers.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

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

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving 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, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial 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 sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

The present invention also provides a cosmetic method characterized by applying the cosmetic composition of the present invention to human skin.

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

The cosmetic composition of the present invention may be used alone or in combination, or may be used by overlapping with other cosmetic compositions other than the present invention. Further, the cosmetic composition having excellent skin wrinkle-improving effect according to the present invention can be used according to a conventional method of use, and the use frequency of the cosmetic composition can be changed according to a user's skin condition or taste.

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

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

DETAILED DESCRIPTION OF THE INVENTION

< Example  1>

A new strain Lactobacillus Lambrosse KCCM11349P  And Lactobacillus Paracase  Isolation of KCCM11254P

In order to isolate the new lactic acid bacteria, the present inventors requested 40 male and female participants in the 20s and 40s to participate in the experiment 7 days before the experiment. In order to prevent the hygiene treatment such as hand washing before the experiment, The experiment was carried out.

In the pre-prepared MRS agar medium, the palms of the participants were inoculated, and the initial bacteria in the hands were inoculated. After incubation at 30 ° C for 24 hours, the colonies were inoculated into the new MRS agar medium with platinum, . The strains isolated from the second inoculation were transferred to a new 10 ml MRS broth for each colony and subcultured. Two microorganisms with the highest activity were selected by fermentation for each colony.

The microorganisms obtained by the above method were referred to the Korean Microorganism Conservation Center to identify lactic acid bacteria by PCR, and the microorganisms were deposited with the international patent institutions.

As a result of the Korean Microorganism Conservation Center, it was confirmed that the two novel strains obtained in the present invention are strains belonging to the genus Lactobacillus , and the present inventors have found that the two strains can be used as Lactobacillus rhamnosus HK-9 and Lactobacillus &lt; RTI ID = 0.0 &gt; paracasei HS-05, and each strain was deposited at the Korean Culture Center of Microorganism and assigned accession numbers KCCM11349P and KCCM11254P. In the present invention, the two new strains identified above are referred to as &quot; Lactobacillus rhamnosus HK-9 KCCM11349P &quot; and &quot; Lactobacillus paracasei HS-05 KCCM11254P &quot;.

< Example  2>

A new strain Lactobacillus Lambrosse KCCM11349P  And Lactobacillus Paracase  Culture method of KCCM11254P

The inventors of the present invention found that two kinds of human skin-derived lactic acid bacteria Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus After culturing the paracasei HS-05 KCCM11254P on MRS agar medium set by the optimal incubation time it is maximized the number of bacteria, was added to 1ℓ of distilled water and 55% of the MRS broth powder and 0.5% L-cysteine hydrochloride to 2ℓ flask, and the medium Respectively. The cultivation temperature of the lactic acid bacteria was 36 ° C, and the experiment was carried out by anaerobic culture for 48 hours at a minimum stirring speed of 50 to 60 rpm.

Lactobacillus rhamnosus The culture of HK-9 strain KCCM11349P was measured at different time points. As a result, Lactobacillus rhamnosus When HK-9 KCCM11349P was cultured for 12 hours, the number of bacteria was 6.4 × 10 ⁶ CFU / ㎖. When cultured for 24 hours, the number of bacteria was increased to 3.2 × 10 ⁷ CFU / ㎖, The maximum number of individuals increased to about 4.3 × 10 ⁷ CFU / ㎖, but the number of individuals gradually decreased when cultured for more than 48 hours. Therefore, the Lactobacillus rhamnosus The optimum incubation time of the strain HK-9 KCCM11349P was 48 hours (see FIG. 1).

Lactobacillus paracasei HS-05 KCCM11254P was incubated with Lactobacillus paracasei When HS-05 KCCM11254P was cultured for 12 hours, the number of bacteria was 6.1 × 10 ⁶ CFU / ㎖. When cultured for 24 hours, the number of bacteria was increased to 3.0 × 10 ⁷ CFU / ㎖, and when cultured for 48 hours The maximum number of individuals was increased to 4.1 × 10 ⁷ CFU / ㎖, but it was observed that the number of individuals gradually decreased when cultured for more than 48 hours. Therefore, it was found that the optimum incubation time of the Lactobacillus paracasei strain HS-05 KCCM11254P of the present invention was 48 hours (see FIG. 2).

In addition, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P was incubated with Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei When the two strains of HS-05 KCCM11254P were cultured for 12 hours, the number of bacteria was about 7.1 × 10 ⁶ CFU / ㎖. When cultured for 24 hours, the number of bacteria increased to about 4.2 × 10 ⁷ CFU / ㎖, And the number of individuals was increased up to about 4.7 × 10 ⁷ CFU / ㎖ when cultured for 48 hours. However, when cultured for more than 48 hours, the number of individuals gradually decreased. Thus, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei The optimum incubation time for HS-05 KCCM11254P complex culture was 48 hours, and Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei It was found that more cell populations were cultured when the two cultures of the above two strains were cultured than the single culture of HS-05 KCCM11254P (see FIG. 3).

< Experimental Example  1>

Analysis of Antioxidant Activity Enhancement through Natural Product Fermentation

The Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P to enhance the antioxidant activity.

First, 100 g of gold was ground into a 2 L flask in powder form, and then 1 L of culture medium was prepared. The medium was supplemented with Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 strain KCCM11254P was added at a concentration of 5% (v / v), and the mixture was inoculated and fermented for 48 hours. After the fermentation broth was lyophilized to powder form, DPPH scavenging activity, reducing power and total phenol content were measured .

As a result, skin-derived lactic acid bacteria, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei When HS-05 KCCM11254P was fermented, DPPH scavenging activity, reducing power and total phenol content were higher than that of general lactic acid fermentation, indicating that Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P were mixed to show the DPPH activity up to 90% in the combined fermented gold. Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P alone had higher antioxidant activity than the fermentation.

In addition, the skin-derived lactic acid bacteria, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei The total phenol contents of the HS-05 KCCM11254P fermentation strain and the combination of the two strains were 6.49 mg / ml, 5.69 mg / ml and 7.08 mg / ml, respectively, Lactobacillus rhamnosus , a skin-derived lactic acid bacterium HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P showed significantly higher antioxidant activity than the intestinal lactobacillus (see Table 1).

< Experimental Example  2>

Analysis of whitening activity enhancement through natural product fermentation

To investigate the enhancement of whitening activity through natural product fermentation, the inventors of the present invention found that Lactobacillus rhamnosus HK-9 KCCM11349P, Lactobacillus paracasei HS-05 KCCM11254P and melanin synthesis inhibitory ability and tyrosinase inhibitory ability of the combined culture fermented product of the two strains were measured.

First, 100 g of gold was ground into a 2 L flask in powder form, and then 1 L of culture medium was prepared. The medium was supplemented with Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 strain KCCM11254P was added at 5% (v / v), and the fermented broth was fermented for 48 hours. After the fermentation broth was lyophilized to powder form, melanin synthesis inhibition and tyrosinase inhibition were measured the skin-derived lactic acid bacteria when compared with normal lactic acid bacteria Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P the fermentation strain and the composite of the two strains was found that high melanin synthesis inhibition and tyrosinase inhibition tee with bleaching activity, in particular Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei When combined with HS-05 KCCM11254P, melanin synthesis inhibition was 42% and tyrosinase inhibition was 45%, indicating the highest whitening activity (see Table 2).

Therefore, the Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei The complex fermentation of HS-05 KCCM11254P was carried out using a conventional lactobacillus and Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P was found to be easier to elute the useful physiologically active substances and the active ingredients than the individual fermentation, and the extraction efficiency was also good.

< Experimental Example  3>

Derived from skin  Lactobacillus Crushed  Antioxidant activity verification

The present inventors have found that the Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei Radical scavenging activity and Reducing power were tested in order to examine the efficacy of antioxidant activity of HS-05 KCCM11254P.

First, in order to disrupt the cultured lactic acid bacteria and the lactic acid bacteria complex-cultured, a total of 100 ml of each strain was obtained, and then, for 20 to 50 minutes The strains were disrupted by sonication at 50-60 kHz. The crushed strain and the crushed solution were incubated at 60 DEG C 500 ml ~ 1 L of hot water was extracted and the extract of the strain was obtained.

The radical scavenging ability test showed that the radical scavenging ability was enhanced by the ability of scavenging free radicals, and the radical scavenging activity was measured. As a result, it was confirmed that the radical scavenging ratio of the complex- It was found that when the concentration of the strain was 100% (v / v), it showed a radical scavenging ability of about 30% (see FIG. 4). These results suggest that Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 &lt; / RTI &gt; KCCM11254P.

Reducing power is a force for reducing the oxidized substance to the original substance and is an index for judging the antioxidant activity. As a result of the measurement of the reducing power, the absorbance value of each of the skin-derived complex-cultured lysates was higher than that of the single skin-derived lactic acid bacteria culture broth, and the absorbance value was 1.473 at 100% (v / v) The highest antioxidant activity was observed (see Fig. 5).

< Experimental Example  4>

Derived from skin  Lactobacillus Crushed  Verification of whitening activity

The present inventors have found that the Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei In order to test the efficacy of the whitening active substance produced from the strain itself by the combined culture of HS-05 KCCM11254P, the strain was disrupted in the same manner as in Experimental Example 3, and melanin synthesis inhibition activity and tyrosinase inhibition experiment Tyrosinase inhibition experiments were performed.

The effects of melanocyte clone-M3 cells on melanin biosynthesis and cell survival were investigated using the samples treated with different concentrations (25, 50, 75, 100% (v / v)) and melanocytes It was found that the inhibition rate of melanin synthesis was increased depending on the lysate concentration of the complex culture strain, which was higher than the inhibition rate of melanin synthesis of the single skin-derived lysophosphate bacterium. In particular, when the concentration of the skin-derived complex culture was 100% (v / v) And showed about 35% melanin inhibition (see FIG. 6).

In addition, as a result of tyrosinase inhibition, tyrosinase inhibition rate of a single skin-derived strain was 28% at maximum, but the tyrosinase inhibition rate of the complex strain culture broth was 35% at maximum (See FIG. 7).

Therefore, the Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei When HS-05 KCCM11254P was cultured in combination, Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P, respectively, and thus it is advantageous to use it as a whitening-related cosmetic product.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Korea Microorganism Conservation Center (overseas) KCCM11254P 20120217 Korea Microorganism Conservation Center (overseas) KCCM11349P 20130108

Claims (5)

delete delete delete (a) Lactobacillus rhamnosus HK-9 KCCM11349P and Lactobacillus paracasei . HS-05 100 ml of KCCM11254P strain is sonicated at 50 to 60 kHz for 20 to 50 minutes to disrupt the strain; And
(b) adding 500 ml to 1 L of water at 60 ° C. to the strain and the disrupted solution obtained in step (a), and extracting the mixture by hot water for 12 to 24 hours. 100 parts by weight of Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) HK-9 KCCM11349P and Lactobacillus paracasei HS-05 KCCM11254P strains each in an amount of 10 to 500 parts by weight, and fermenting the mixture for 45 to 50 hours, the method comprising:
Wherein the composition prepared by the above method contains 7.08 mg / ml of total phenol having antioxidant activity.

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