KR20160110829A - The milk fermented product using Lactobacillus paracasei HY7301 for skin whitening and products containing thereof as effective component - Google Patents

Abstract

The present invention relates to Lactobacillus paracasei HY7301 milk fermented product having skin whitening effect and a product containing it as an active ingredient, wherein the Lactobacillus paracasei HY7301 milk fermentation product of the present invention is a milk fermented product of Lactobacillus paracasei HY7301 And can be used as a pharmaceutical composition for skin whitening, a fermented food, a beverage, a health functional food, a cosmetic composition, and the like for whitening skin containing the active ingredient as it has a skin whitening effect.

Description

Lactobacillus paracase HY7301 milk fermented product having skin whitening effect and product containing it as an active ingredient {The milk fermented product using Lactobacillus paracasei HY7301 for skin whitening and products containing as effective component}

The present invention relates to a milk fermented product of Lactobacillus paracasei HY7301 having skin whitening effect and a product containing it as an active ingredient. More particularly, the present invention relates to a lactobacillus paracasei ( Lactobacillus paracasei ) Fermented milk obtained by fermenting milk with HY7301 and a product containing it as an active ingredient.

The skin color of a person is mainly determined by the content of pigment called melanin in the skin. Melanin is biosynthesized by the melanocytes (melanocytes) in the basal layer of the skin and is transferred from the basal layer to the stratum corneum by keratinocyte keratinocyte through cytoplasmic processes. Melanin is produced by several oxidative processes in melanin-producing cells after conversion of tyrosine, an amino acid, into DOPA and DOPAquinone by an enzyme called tyrosinase. Melanin absorbs ultraviolet light and protects various cell tissues inside the epidermis from damage by ultraviolet rays. When melanin synthesis is excessive, it not only causes spots and freckles but also darkens the overall skin color. Therefore, excessive inhibition of melanin pigment synthesis may not only brighten skin color but also alleviate symptoms of hypercholesterolemia of spots and freckles.

As interest in cosmetics has recently increased, interest in wrinkle-improving and whitening cosmetics is steadily increasing. In addition, according to the wellbeing trend, the development of cosmetic materials derived from natural sources has been increasing, and the development of cosmetics using microorganisms such as enzymes and lactic acid bacteria is continuing. In particular, the lactic acid bacterium Lactobacillus microorganism that is used as the main pillar's ash also (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), ronggeom Bifidobacterium (Bifidobacterium longum ), and these strains have been reported to have antibacterial activity, antioxidative activity, and anti-inflammatory activity. There have been cases where a fermentation broth obtained by fermenting these lactic acid bacterium cells or mushroom, rice, fruit juice, plant extract or the like with lactic acid bacterium has been blended in cosmetics. However, the study on the whitening function of Lactobacillus paracasei and fermented milk using the same was not yet known.

Accordingly, the inventors of the present invention have found that a fermented product obtained by fermenting milk with Lactobacillus paracasei HY7301 among the lactic acid bacteria isolated from makgeolli is superior in skin whitening efficacy.

Korean Patent Registration No. 1140572 (Name of the invention: fermented black garlic nano fraction having effect on skin cell growth and wrinkle improvement and whitening, Registration date: 2012.07.02) Korean Patent Application Laid-Open No. 10-2013-0115943 (entitled "Anti-inflammatory, skin whitening and anti-aging composition comprising a supernatant of lactic acid bacterium, filed on Apr. 20, 2013) Korean Patent Application Laid-Open No. 10-2014-0039548 (Title of the Invention: Composition for skin whitening comprising a culture liquid of Lactobacillus pentosus strain and method for producing the same, filed on Apr. 20, 2014)

The present invention aims to provide a fermented milk product of Lactobacillus paracasei HY7301 having skin whitening effect and a pharmaceutical composition containing it as an active ingredient, a fermented milk, a functional beverage, a health functional food, and a cosmetic composition .

In order to achieve the above object, the present invention provides a fermented milk product of Lactobacillus paracasei HY7301 having skin whitening effect and a pharmaceutical composition containing it as an active ingredient, a fermented milk, a functional beverage, a health functional food, a cosmetic composition .

Hereinafter, the present invention will be described in detail.

In order to isolate the strain according to the present invention, the raw rice wine samples within the shelf life were decanted and inoculated into the lactic acid bacteria selective medium (BCP agar) and cultured at 37 ° C for 72 hours. Colonies estimated to be lactic acid bacteria were repeated twice in MRS agar culture medium, and single colonies were obtained. Among these strains, strains capable of fermenting milk were selected. The milk fermentation product of the strain was freeze-dried to be pulverized, and then the tyrosinase enzyme inhibitory activity and melanin production inhibitory activity were confirmed, and the fermented milk fermented product having excellent whitening effect was produced. The results are shown in Table 1 below. The results of the 16S rDNA analysis are shown in Table 1 below.

As shown in Table 1 above, the 16S rDNA gene of the new strain of the present invention was found to be 100% identical to the 16S rDNA gene of Lactobacillus paracasei , and the present inventors confirmed that the 16S rDNA gene of Lactobacillus paracase Lactobacillus paracasei HY7301, and deposited on October 8, 2013 with the Korea Research Institute of Bioscience and Biotechnology (Accession No .: KCTC 12502BP).

The characteristics of Lactobacillus paracasei HY7301 according to the present invention are as follows.

1) Types of bacteria

The characteristics of the bacteria when cultured on an MRS agar plate medium at 37 ° C for 2 days

① Cell shape: Bacillus

② Mobility: None

③ Spore forming ability: None

④ Gram staining: positive

2) Morphology

When cultured on MRS agar plate medium at 37 ° C for 2 days,

① Shape: Circular

② Bump: convex

③ Surface: Smooth

3) Physiological properties

① Growth temperature: Growth possible Growth temperature 15 ~ 40 ℃

            Optimum growth temperature 36 ~ 38 ℃

② Growth pH: Growable growth pH 5.0 ~ 7.5

           Optimum pH 6.0 to 6.5

③ Effect on oxygen: Tumor anaerobic

4) Catalase: -

5) Whether gas is formed: -

6) Growth at 15 ° C: +

7) Growth at 45 ° C: +

8) Indole production: -

9) Lactic acid production: +

10) Fermentation experiment and identification

The results of sugar fermentation experiments using the api 50CH kit from Biomerieux are shown in Table 2 below.

0-24 per Whether or not to use Per 25-49 Whether or not to use 0 Control - 25 esculin + 1 glycerol - 26 Living + 2 Erythritol - 27 Cellobios + 3 D arabinose - 28 Maltose + 4 L arabinose - 29 lactose + 5 ribos + 30 Melibiose - 6 D xylose - 31 sucrose + 7 L xylose - 32 Trehalose + 8 Adonitor + 33 inulin - 9 beta methyl-D-xyloside - 34 Melitos + 10 galactose + 35 raffinos - 11 glucose + 36 starches - 12 fructose + 37 glycogen - 13 Manno + 38 xylitol - 14 sorbos - 39 Gentiobios + 15 Lambos - 40 D Toranos + 16 Dorsitol - 41 License Source - 17 Synoshitol + 42 D Tagatos + 18 Mannitol + 43 D Fuchos - 19 sorbitol + 44 L Fuchos - 20 [alpha] -methyl-D-mannoside - 45 D arabitol - 21? -Methyl-D-glucoside - 46 L arabitol - 22 N-acetyl-glucosamine + 47 Gluconate - 23 Amigalline + 48 2-keto-gluconate - 24 Arbutin + 49 5-keto-gluconate -

On the other hand, the pharmaceutical composition containing the fermented milk of Lactobacillus paracasei HY7301 having the skin whitening effect of the present invention as an active ingredient can be used as a pharmaceutical, alone or in combination with pharmaceutical excipients, And may be formulated in the form of tablets, capsules and the like.

For humans, a typical daily dose may range from 1 to 30 mg / kg body weight, and may be administered in single or divided doses. However, the actual dosage should be determined in light of various relevant factors such as the route of administration, the patient's age, sex and weight, health status and severity of the disease.

Of course, the pharmaceutical composition having a skin whitening effect containing Lactobacillus paracasei HY7301 milk fermented product of the present invention as an active ingredient has little toxicity and side effects, so that it can be safely used for prophylactic long-term use It is medicine.

In addition, the composition comprising Lactobacillus paracasei HY7301 milk fermented product having the skin whitening effect of the present invention as an active ingredient can be used as food, food additive, beverage, beverage additive, fermented milk, have. When used as a food, a food additive, a beverage, a beverage additive or a health functional food, it can be used in various foodstuffs, fermented milk, meat, beverages, chocolate, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, , Alcoholic beverages, and other health functional foods.

Particularly, the fermented milk containing the fermented milk of Lactobacillus paracasei HY7301 having the skin whitening effect of the present invention as an active ingredient can be obtained by mixing fermented milk of Lactobacillus paracasei HY7301 with milk fermented milk And then homogenized at 150 bar, cooled to below 10 ° C, and packaged in a container to prepare fermented milk.

In addition, the functional beverage containing Lactobacillus paracasei HY7301 milk fermented product of the present invention as an active ingredient can be prepared by mixing fruit juice syrup, Lactobacillus paracasei HY7301 milk fermented product and water at a predetermined ratio The mixture is homogenized at 150 bar, cooled to below 10 ° C, and packaged in small containers such as glass bottles and plastic bottles to produce functional beverages.

In addition, the health functional food containing the fermented product of Lactobacillus paracasei HY7301 milk of the present invention as an active ingredient contains not only Lactobacillus paracasei HY7301 milk fermented product but also nutritional supplement component Vitamin B ₁ , B ₂ , B ₅ , B ₆ , E and acetic acid ester, nicotinic acid amide, oligosaccharide and the like may be added and other food additives may be added.

The present invention can also be used as a cosmetic composition containing Lactobacillus paracasei HY7301 milk fermented product as an active ingredient. In the case of using the cosmetic composition as described above, in addition to the fermented milk of Lactobacillus paracasei HY7301 of the present invention, a substance arbitrarily selected for the purpose of formulation or use of other cosmetics may be added. For example, purified water, an oil, a surfactant, a moisturizer, a high alcohol, a thickener, a chelating agent, a pigment, a fatty acid, an antioxidant, an antiseptic, a wax, a pH adjusting agent and a perfume may be added.

The formulation of the cosmetic composition containing Lactobacillus paracasei HY7301 milk fermented product of the present invention as an active ingredient is not particularly limited, and examples thereof include nutritive creams, convergent lotions, soft lotions, lotions, Essence, nutritional gel, massage cream, mask pack, and the like.

The Lactobacillus paracasei HY7301 milk fermented product of the present invention has a skin whitening effect, so that it can be used as a skin whitening agent for skin whitening containing Lactobacillus paracasei HY7301 milk fermented product as an active ingredient Composition, fermented food, beverage, health functional food, cosmetic composition and the like.

FIG. 1 is a graph showing inhibitory activity of tyrosinase enzyme activity of Lactobacillus paracasei HY7301 milk fermented product. FIG.
FIG. 2 is a graph showing the effect of fermented milk of L. paracsei HY7301 on melanin synthesis in melanoma cells. FIG.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following embodiments are not intended to limit the scope of the present invention, and ordinary variations by those skilled in the art within the scope of the technical idea of the present invention are possible.

≪ Example 1 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk Fermentation product  And preparation of lyophilized powder

Crude oil: Skim milk: Purified water was mixed at a weight ratio of 66.7: 3.3: 30 and sufficiently dissolved. The mixture was heated at 130 to 135 ° C for 2 to 3 seconds and cooled to 35 to 40 ° C. Lactobacillus paracasei HY7301 was inoculated thereto and cultured at 37 DEG C for about 24 hours to prepare a milk fermented product. Then, the fermented milk was freeze-dried to obtain a lyophilized powder.

Meanwhile, the Lactobacillus paracasei HY7301 milk fermented product of the present invention may be provided in the form of a culture in addition to the lyophilized powder form as described above, and it may be prepared in other blending ratios using milk.

≪ Example 2 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk The fermented product  Preparation of pharmaceutical composition containing active ingredient

The preparation examples of the pharmaceutical composition containing Lactobacillus paracasei HY7301 milk fermented product of the present invention as an active ingredient will be described, but the present invention is not intended to be limited thereto but is specifically explained.

Manufacture of tablets

100 mg of lyophilized powder of fermented milk of Lactobacillus paracasei HY7301 of Example 1, 100 mg of corn starch, 100 mg of lactose and 97 mg of polyvinyl pyrrolidone were mixed homogeneously and granulated by a wet granulation method 3 mg of magnesium stearate was added to the mixture, and the resulting mixture was compressed to 400 mg per tablet.

Preparation of capsules

100 mg of lyophilized powder of fermented milk of Lactobacillus paracasei HY7301 of Example 1, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed thoroughly, Thus, the capsules were prepared by filling into hard gelatine capsules.

≪ Example 3 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk The fermented product  Preparation of fermented milk containing active ingredient

The process for producing the fermented milk comprising the fermented milk of Lactobacillus paracasei HY7301 milk of the present invention and the mixed fruit syrup is as follows.

First, the fermented milk of Lactobacillus paracasei HY7301 of Example 1 was homogenized at 150 to 200 bar to grind the curd and then cooled to 10 ° C or less.

The mixed fruit juice syrup was prepared by mixing 13 wt% of liquid fructose, 5 wt% of white sugar, 10.9 wt% of concentrated juice concentrate 56Brix ^o , 1.0 wt% of pectin, 0.1 wt% of fresh fruit mix essence and 70 wt% of purified water at 30 to 35 캜 And sterilized at 130 to 135 ° C for 2 to 3 seconds, followed by cooling.

75% by weight of Lactobacillus paracasei HY7301 milk fermentation product and 25% by weight of the above-mentioned mixed fruit syrup were mixed to prepare a fermented milk containing Lactobacillus paracasei HY7301 milk fermented product as an active ingredient .

<Example 4>

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk The fermented product  Preparation of functional beverage containing active ingredient

The method for producing a functional beverage composed of Lactobacillus paracasei HY7301 milk fermented product and a mixed fruit syrup of the present invention is as follows.

First, a mixed juice syrup was 13% by weight of liquid fructose, white sugar, 2.5 wt%, brown sugar 2.5% by weight, mixed juice concentrate 56Brix ^o 10.9% by weight of pectin, 1.0% by weight, fresh 0.1% by weight fruit mix essence and purified water 70% The mixture was stirred at 30 to 35 ° C, mixed, and heat-treated by UHT (sterilized at 135 ° C for 2 seconds) and cooled.

99% by weight of the mixed fruit juice syrup prepared by the above method and 1% by weight of the lyophilized powder of the fermented milk of Lactobacillus paracasei HY7301 of Example 1 were combined and homogenized at 150 bar, Cooled and packaged in small containers such as glass bottles and PET bottles to produce functional beverages containing Lactobacillus paracasei HY7301 milk fermented product as an active ingredient.

&Lt; Example 5 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk The fermented product  Production of Health Functional Foods Containing Active Ingredients

(Vitamin B ₁ , B ₂ , B ₅ , B ₆ , E and acetic acid ester, nicotinic acid amide) were added to 0.1 wt% of the lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example ₁ , And oligosaccharide were added in an amount of 10 parts by weight based on 100 parts by weight of the lyophilized powder of fermented milk of Lactobacillus paracasei HY7301 of Example 1 and mixed in a high-speed rotary mixer. To 100 parts by weight of the mixture, 10 parts by weight of sterilized purified water was added and mixed to form granules having a diameter of 1 to 2 mm. The molded granules were dried in a vacuum drier at 40 to 50 DEG C and then passed through 12 to 14 mesh to prepare uniform granules. The granules thus prepared were extruded in suitable amounts to be purified or powdered or filled into hard capsules to prepare hard capsule products.

&Lt; Example 6 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk The fermented product  Active ingredient Cosmetic  Produce

Softening longevity

[0064] The lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example 1, 0.01 wt% of the lyophilized powder, 3 wt% of glycerin, 2 wt% of butylene glycol, 2 wt% of propylene glycol, 0.1 wt% of carboxyvinyl polymer, 10% by weight of ethanol, 0.1% by weight of triethanolamine, a small amount of preservative, a trace amount of coloring matter, a trace amount of perfume, and a remaining amount of purified water were mixed and prepared according to a conventional method for producing softening water.

Nutritional lotion

[0070] The lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example 1, 0.01 wt% of beeswax, 4 wt% of beeswax, 1.5 wt% of polysorbate 60, 0.5 wt% of sorbitan sesquioleate, By weight of glycerin, 3% by weight of glycerin, 3% by weight of butylene glycol, 3% by weight of propylene glycol, 0.1% by weight of carboxyvinyl polymer, 0.2% by weight of triethanolamine, 5% by weight of squalane, 5% by weight of caprylic / capric triglyceride, , A trace amount of preservative, a trace amount of coloring matter, a trace amount of flavor, and a remaining amount of purified water.

Nourishing cream

[0060] The lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example 1, containing 0.01 wt% of lyophilized powder, 10 wt% of wax, 1.5 wt% of polysorbate 60, 0.5 wt% of sorbitan sesquioleate, 5% by weight of squalane, 5% by weight of caprylic / capric triglyceride, 5% by weight of glycerin, 3% by weight of butylene glycol, 3% by weight of propylene glycol, 0.2% by weight of triethanolamine, A coloring matter, a trace amount of flavor and a remaining amount of purified water were mixed to prepare a conventional nutrition cream.

Massage  cream

[0064] The lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example 1, 0.01 wt% of wax, 10 wt% of wax, 1.5 wt% of polysorbate 60, 0.8 wt% of sorbitan sesquioleate, , Propylene glycol 3 wt%, triethanolamine 0.2 wt%, a small amount of preservative, a small amount of an antiseptic agent, and the like, in an amount of 5 wt%, squalane 5 wt%, caprylic / capric triglyceride 4 wt%, glycerin 5 wt%, butylene glycol 3 wt% Pigment, trace amount of fragrance, and remaining amount of purified water were mixed to prepare a conventional massage cream.

pack

[0060] The lyophilized powder of the Lactobacillus paracasei HY7301 milk fermentation product of Example 1, 0.01 wt% of the lyophilized powder, 13 wt% of polyvinyl alcohol, 0.2 wt% of sodium carboxymethylcellulose, 0.1 wt% of allantoin, 5 wt% 0.3% by weight of nonyl phenyl ether, a small amount of preservative, a trace amount of coloring matter, a trace amount of perfume, and a remaining amount of purified water were mixed and prepared according to a conventional pack preparation method.

&Lt; Comparative Example 1 &

Lactobacillus Paracase ( Lactobacillus paracasei ) 71-2 Milk Fermented  Produce

Yeoju rice wine was decanted and inoculated on BCP agar and cultured at 37 ℃ for 72 hours. Colonies estimated to be lactic acid bacteria were repeated twice in MRS agar culture medium, and single colonies were obtained. Among the thus prepared strains, strains capable of fermenting milk were selected. Molecular genetic analysis using 16S rDNA analysis was performed to identify the selected lactic acid bacteria. Lactobacillus paracasei 71-2 was named as the isolate based on the identification results.

The milk fermented by Lactobacillus paracasei 71-2 isolated from raw rice wine was prepared in the same manner as in Example 1 above.

&Lt; Comparative Example 2 &

Lactobacillus Flora Room ( Lactobacillus plantarum ) 78-4 Milk Fermented  Produce

The strain was isolated in the same manner as in Comparative Example 1, except that raw rice wine in Gwangju area was used as a source. The selected lactic acid bacteria were identified by 16S rDNA analysis and named as Lactobacillus plantarum 78-4.

The fermented milk fermented with Lactobacillus plantarum 78-4 isolated from raw rice wine was prepared in the same manner as in Example 1.

&Lt; Comparative Example 3 &

Lactobacillus Lambrosse ( Lactobacillus rhanmnosus ) 67-1 Milk Fermented  Produce

The strains were isolated in the same manner as in Comparative Example 1, except that raw rice wine in the Jinju area was used as a separation source. The selected lactic acid bacteria were identified by 16S rDNA analysis and named as Lactobacillus rhamnosus 67-1.

The fermented milk fermented with Lactobacillus rhamnosus 67-1 isolated from raw rice wine was prepared in the same manner as in Example 1 above.

&Lt; Comparative Example 4 &

Lactobacillus Kajay ( Lactobacillus casei ) 65-3 Milk Fermented  Produce

The strains were isolated in the same manner as in Comparative Example 1, except that raw rice wine in the Jinju area was used as a separation source. The selected lactic acid bacteria were identified through 16S rDNA analysis and named as Lactobacillus casei 65-3.

The fermented milk fermented with Lactobacillus casei 65-3 isolated from raw rice wine was prepared in the same manner as in Example 1 above.

&Lt; Test Example 1 >

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk Fermented Tyrosinez  Enzyme activity Inhibition  Measure

Tyrosinase is an enzyme involved in determining the most important initial rate in the melanin biosynthetic pathway in the human body. Many whitening agents have an action mechanism that inhibits this enzyme. The efficacy of the Lactobacillus paracasei HY7301 milk fermented product of the present invention was evaluated by measuring the degree of inhibition of the activity of the tyrosinase enzyme, and the activity of the tyrosinase enzyme was determined by the method of No et al. (No JK , et al., Inhibition of tyrosinase by green tea components, Life Sci ., 1991).

(1) The tyrosinase used in the test was prepared with a mushroom tyrosinase concentration of 200 units / mL, and 5 mM L-DOPA (L-3,4-dihydroxy- phenylalanine) was prepared, and lyophilized powder of Lactobacillus paracasei HY7301 milk fermented product of Example 1 was prepared at a concentration of 100 μg / mL.

A lyophilized powder of Lactobacillus paracasei HY7301 milk fermentation of Example 1 was added to 0.1 mL of the mushroom tyrosinase and 0.2 mL of the reaction substrate (5 mM L-DOPA) Lt; / RTI &gt; After reacting at 37 ° C for 10 minutes, 200 μl of the reaction solution was transferred to a 96-well plate, and the absorbance at 490 nm was measured to calculate the inhibition of tyrosinase activity. The formula is as follows.

      Tyrosinase inhibition rate (%) = (1 - (S - B) / (C - B)

          S: absorbance of sample, test sample (fermentation product or arbutin)

          B: blank, the absorbance of the reaction substrate alone

C: control, absorbance when reacted with reaction substrate and tyrosine alone

(2) Tests were carried out in the same manner as in (1) above except that arbutin, hypoactive whitening functional ingredient, was used as a positive control in place of Lactobacillus paracasei HY7301 milk fermentation of Example 1 Respectively.

(3) In order to confirm whether specific whitening efficacy only Lactobacillus para casei (Lactobacillus paracasei) HY7301 milk fermented product of the present invention Lactobacillus bacteria (Lactobacillus) in the other lactic acid bacteria in Comparative Example 1 to carry out milk fermentation in 4. Example 1 (1) except that Lactobacillus paracasei HY7301 milk fermentation product was used instead of Lactobacillus paracasei HY7301 milk fermentation product.

The results are shown in Fig.

As can be seen from FIG. 1, the positive control group, arbutin, showed about 29% inhibition of tyrosinase, while the milk fermentation products of Comparative Lactobacillus strains of Comparative Example 1 to 4, which are other lactic acid bacteria in Lactobacillus , Lactobacillus paracasei 71-2 showed low inhibitory effect on tyrosinase , despite the fact that Lactobacillus paracasei was not the same as Lactobacillus paracasei . On the other hand, the Lactobacillus paracasei HY7301 milk fermentation of the present invention showed about 20% inhibition of tyrosinase, indicating that it has a similar function to that of positive control group arbutin.

&Lt; Test Example 2 &

Lactobacillus Paracase ( Lactobacillus paracasei ) HY7301  milk Fermented  Melanin production Inhibition  Measure

Melanin in skin tissue is produced in melanocytes and secreted inside and outside the cells, which leads to pigmentation in the skin and the like. Therefore, it was evaluated whether the fermented milk of Lactobacillus paracasei HY7301 of the present invention inhibited the melanin synthesis of melanoma cells.

(1) As melanoma cells, Murine melanocyte, B16F10 cell was used. Cells were cultured in 96-well plates in DMEM medium containing 10% fetal bovine serum (FBS) at a concentration of 6 × 10 ⁴ cells / well, and melanin stimulation hormone (α-MSH) To induce melanogenesis. At the same time, the Lactobacillus paracasei HY7301 milk fermentation product of Example 1 was treated at a level of 100 μg / mL and reacted at 37 ° C. and 5% CO ₂ for 48 hours. After the reaction was completed, the medium was removed, and the cells were washed with phosphate buffered saline (PBS), and then treated with trypsin to recover the cells. The recovered cells were dissolved in 1 N sodium hydroxide (NaOH) containing 10% dimethyl sulfoxide (DMSO) and reacted in a constant temperature water bath at 60 ° C for 30 minutes to elute intracellular melanin. The absorbance of the eluted solution was measured at 450 nm.

(2) In order to confirm whether the inhibition of melanin production is due to Lactobacillus paracasei HY7301 strain or Lactobacillus paracasei HY7301 milk fermentation, additional experiments were conducted as follows.

First, the Lactobacillus paracsei HY7301 of the present invention was cultured in a lactic acid bacteria culture medium (MRS broth) for 24 hours, and the cells were collected and washed with physiological saline. The cells were concentrated to physiological saline at a concentration of ¹ × ^{10 10} CFU / mL and heat-treated at 100 ° C. for 10 minutes to prepare dead bacteria. The Lactobacillus paracsei HY7301 killed bacterium was treated with melanoma cells at a level of 1 × 10 ⁷ CFU / well instead of the Lactobacillus paracasei HY7301 milk fermentation of the above (1) The test was conducted in the same manner as in the above (1), except that the reaction was carried out.

(3) Further, in order to confirm whether the inhibition of melanin production is caused by the whey of Lactobacillus paracasei HY7301 milk fermentation, further experiments were conducted as follows.

The whey fermented product of Lactobacillus paracasei HY7301 of Example 1 was centrifuged (500 X g, 20 minutes) to separate whey, and the separated whey and Lactobacillus paracasei ) HY7301 milk fermentation broth were each lyophilized and treated to a level of 100 μg / mL, respectively, in the same manner as in (1) above.

(4) The negative control group was a non-treated group that did not treat melanin stimulation hormone (α-MSH) and fermented milk, and did not induce melanin production. Respectively.

The positive control group was tested in the same manner as in the above (1) by inducing melanin production by treating α-melanin stimulating hormone (α-MSH) alone at a concentration of 2 μM.

As can be seen in FIG. 2, the α-melanin stimulating hormone (α-MSH) induces melanin accumulation in cells, but when the Lactobacillus paracasei HY7301 milk fermentation of the present invention is treated simultaneously, And the melanin production of the cells was reduced.

In addition, melanoma production of melanoma cells was reduced by the lactobacillus paracasei HY7301 strain itself and the Lactobacillus paracasei HY7301 milk fermented product, ( Lactobacillus paracasei ) HY7301 milk fermented product itself.

Taken together these test results, it was confirmed that the fermented milk of Lactobacillus paracasei HY7301 of the present invention inhibited the activity of tyrosinase, inhibited melanin production, and had a skin whitening effect .

Korea Biotechnology Research Institute KCTC12502BP 20131008

Claims (7)

Lactobacillus paracasei HY7301 (Accession No .: KCTC 12502BP) milk fermentation product having skin whitening efficacy.
A pharmaceutical composition for skin whitening comprising the fermented milk of Lactobacillus paracasei HY7301 of claim 1 as an active ingredient.
A fermented milk containing the fermented milk of Lactobacillus paracasei HY7301 of claim 1 as an active ingredient.
A functional whitening beverage for skin whitening comprising the fermented milk of Lactobacillus paracasei HY7301 of claim 1 as an active ingredient.
A health food for skin whitening comprising the fermented product of Lactobacillus paracasei HY7301 milk of claim 1 as an active ingredient.
A cosmetic composition comprising the fermented milk of Lactobacillus paracasei HY7301 of claim 1 as an active ingredient.
The method according to claim 6,
Wherein the cosmetic composition has one of the formulations selected from a nutritional cream, a convergent lotion, a soft lotion, a lotion, an essence, a nutritional gel, a lipstick, a foundation, an eye shadow, a patch, a mast sheet, a massage cream and a mask pack. A cosmetic composition comprising Lactobacillus paracasei HY7301 milk fermented product as an active ingredient.

Images