CN110267643B

CN110267643B - Composition for skin beauty comprising flocculation protein

Info

Publication number: CN110267643B
Application number: CN201980000340.5A
Authority: CN
Inventors: 柳正镇; 金旻智; 金荷荣; 姜承贤; 金延俊; 朴明三
Original assignee: Cosmax Inc
Current assignee: Cosmax Inc
Priority date: 2018-01-12
Filing date: 2019-01-07
Publication date: 2022-12-30
Anticipated expiration: 2039-01-07
Also published as: WO2019139307A1; KR20190086280A; KR102037039B1; CN110267643A

Abstract

In one aspect, the present invention provides a composition for skin beauty comprising a flocculation protein. Since the composition including the flocculation protein has effects of improving skin barrier, moisturizing, whitening and cell proliferation, and promoting collagen synthesis and elastin synthesis, it can be effectively used for skin beauty.

Description

Composition for skin beauty comprising flocculation protein

Technical Field

The present invention relates to a composition for making up the skin.

Background

Flocculation protein (Flocculin) is a lectin like protein (lectin like protein) in the cell wall of Lagger yeast (Lager yeast) for precipitating Lagger yeast. In particular, the flocculation protein has an excellent binding force with mannose, binds to mannose of other yeast walls and generates precipitates of yeast cells, and also has a function of protecting yeast from stress factors such as nutrient depletion, temperature, and ultraviolet rays (UV) during culture.

On the other hand, yeast is used as a cosmetic ingredient for the purpose of improving skin (e.g., improving wrinkles, whitening effect, etc.). After disrupting cell walls by proteolytic enzyme, organic solvent or autolysis using yeast cells as a raw material, insoluble portions are removed and concentrated to produce a yeast extract, and the yeast extract is used as a cosmetic ingredient. The yeast extract thus treated is composed of a complex of various proteins, peptides, amino acids, minerals and B vitamins, and is used as a cosmetic additive. In addition, U.S. Pat. No. 5,776,441 and U.S. Pat. No. 6,177,105 disclose that yeast extract has skin wound treatment and collagen synthesis effects, and is used as various cosmetic raw materials. However, until now, there has been no example of the use of flocculating proteins in cosmetic compositions for cosmetic purposes of the skin.

Disclosure of Invention

Technical problem

In one aspect, the present invention provides a cosmetic composition for skin beauty including a flocculation protein.

In another aspect, the present invention provides a composition for health functional food for skin beauty including flocculation protein.

In another aspect, the present invention provides a pharmaceutical composition for skin beauty comprising a flocculation protein.

In another aspect, the invention provides a method of treating or ameliorating skin damage or skin aging comprising the step of administering to an individual an effective amount of a composition comprising a flocculating protein.

In another aspect, the invention provides the use of a composition comprising a flocculating protein for the preparation of a therapeutic agent for skin damage or skin ageing.

Technical scheme

One aspect of the present invention provides a cosmetic composition for skin beauty including a flocculation protein. In another aspect, the present invention provides a composition for health functional food for skin beauty including flocculation protein. Another aspect of the present invention provides a pharmaceutical composition for preventing or treating skin injury or skin aging, comprising the flocculation protein. In another aspect, the present invention provides a composition for external application to the skin for skin beauty.

"flocculation protein" usually refers to the expression in Saccharomyces microorganisms in the cell wall of lectin like protein, and can be according to Marika H. (1994. From beer yeast flocculation protein Purification and partial characteristics) the method disclosed in the literature isolated from Saccharomyces microorganisms or commercially purchased from yeast microorganisms. <xnotran> , (Saccharomyces bayanus), (Saccharomyces boulardii), (Saccharomyces bulderi), (Saccharomyces cariocanus), (Saccharomyces cariocus), (Saccharomyces cerevisiae), (Saccharomyces chevalieri), (Saccharomyces dairenensis), (Saccharomyces ellipsoideus), (Saccharomyces eubayanus), (Saccharomyces exiguus), (Saccharomyces florentinus), (Saccharomyces kluyveri), (Saccharomyces martiniae), (Saccharomyces monacensis), (Saccharomyces norbensis), (Saccharomyces paradoxus), (Saccharomyces pastorianus), (Saccharomyces spencerorum), (Saccharomyces turicensis), (Saccharomyces unisporus), (Saccharomyces uvarum) (Saccharomyces zonatus). </xnotran> Known amino acid sequences of flocculation proteins include, for example, NCBI accession Nos. AIC64115.1, AFJ20718.1, BAG49462.1 or ABS87371.1, etc.

The composition may further comprise a culture or extract of the saccharomyces microorganism. The culture or extract can be easily obtained by those skilled in the art according to methods known in the art. For example, the culture of the microorganism can be carried out using a commercially available medium, for example, LB medium, ham's F10 culture (Sigma), minimal essential medium (MEM, sigma), RPMI-1640 medium (Sigma), and Dulbecco's modified eagle's medium (DMEM, sigma). Hormones or other growth factors, salts, buffers, nucleotides, antibiotics, trace elements, glucose, and the like may be supplemented at known suitable concentrations, if desired. The culture conditions such as temperature and pH, etc., can be appropriately determined by those skilled in the art according to the selected microorganism. In addition, water, acetone, alcohols (e.g., C1-C6 alcohols), or mixtures thereof may be used as a solvent to extract the extract. The C1-C6 alcohol may be methanol, ethanol, propanol, isopropanol, 1, 3-propanediol, butanol, pentanol or hexane. The solvent may be, for example, a mixture of water and alcohol, i.e., an alcohol aqueous solution. The alcohol concentration of the alcohol aqueous solution may be 1 (v/v)% to 100 (v/v)%, for example, 1 (v/v)% to 99.5 (v/v)%, 10 (v/v)% to 100 (v/v)%, 20 (v/v)% to 100 (v/v)%, 30 (v/v)% to 100 (v/v)%, 40 (v/v)% to 100 (v/v)%, 50 (v/v)% to 100 (v/v)%, 60 (v/v)% to 100 (v/v)%, 70 (v/v)% to 100 (v/v)%, 75 (v/v)% to 100 (v/v)%, 60 (v/v)% -90 (v/v)%, 60 (v/v)% to 80 (v/v)%, 65 (v/v)% to 75 (v/v)% or 75 (v/v)%, or. The alcohol aqueous solution may be a methanol aqueous solution, an ethanol aqueous solution, or a butanol aqueous solution. The composition may further comprise a fraction of the extract. The term "fraction" refers to a substance of a component into which the extract is divided, i.e., a substance that has been fractionated. The fraction may be obtained by solvent fractionation (fractionation). The solvent fractionation may refer to a step of mixing the extract with a solvent and separating substances present in the solvent.

The term "skin beauty" refers to restoring skin damaged due to skin damage or skin aging to a pre-damaged state by a method of improving skin wrinkles, enhancing skin plumpness, improving elasticity, or regenerating skin. The skin includes skin of all parts of the body including the face, hands, arms, legs, feet, chest, abdomen, back, buttocks and scalp.

The term "skin lesion" includes lesions of the clinical and cosmetic viewpoints of skin tissues or cells, including external physical lesions, penetration of chemical substances, bacteria, fungi, viruses and the like, exposure to ultraviolet rays, wrinkles caused by the loss of moisture and aging of the skin, etc., oxidation caused by free radicals (active oxygen), etc., skin pigmentation, skin inflammation, seborrheic dermatitis, redness (redness, erythema), edema, lichenification, eczema, itching (itching), atopic dermatitis and the like.

The term "skin aging" includes endogenous aging over time and exogenous aging due to the external environment. The skin aging may include skin wrinkles, blemishes, and brown spots. The skin wrinkles may be fine lines due to skin deterioration. The cause of skin wrinkles may be photoaging, age, facial expression, lack of moisture, or a combination thereof. The photoaging may age the skin by exposure to ultraviolet light (including UVA, UVB, and UVC). The skin wrinkle improvement may be inhibition or inhibition of the generation of skin wrinkles or alleviation of already generated wrinkles.

The formulation type of the composition may be a non-oral administration dosage form. The non-oral administration dosage form may be an injection or a skin external preparation. The skin external agent may be cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, medicated bandage, lotion, or their combination. The external preparation for skin may be appropriately mixed with ingredients generally used as external preparations for skin such as cosmetics and medicines, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, perfumes, colorants, various skin nutrients, or a combination thereof, as necessary. The skin external preparation can be mixed with disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, metal chelating agent such as gluconic acid, caffeine, tannin, verapamil, glycyrrhrizae radix extract, glabridin, hot water extract of Cirsium japonicum fruit, various crude drugs, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts, and saccharides such as vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

When the composition is a cosmetic composition, the cosmetic composition may be prepared in a dosage form comprising: lotions (moisturizers), emollients, toners, astringents (extracts), lotions, milk lotions (milk lotions), moisturizing lotions, nutritional lotions, massage creams, nutritional creams, moisturizing creams, hand creams, foundations, essences, films, soaps, cleansing foams, cleansing milks, cleansing creams, body lotions, body cleansers, suspensions, gels, powders, pastes (pastes), air pads, face masks or sheet pack masks or spray compositions. Compositions of such dosage forms may be prepared according to methods conventional in the art. The cosmetic composition may further include preservatives, stabilizers, surfactants, thickeners, solvents, moisturizers, emollients, ultraviolet absorbers, preservatives, bactericides, emulsifiers, antioxidants, pH adjusters, organic and inorganic pigments, fragrances, cold sensates or antiperspirants and the like which are commonly used in the art. The mixing amount of the additional ingredients of the above preservative and the like can be easily selected by those skilled in the art within the range that does not impair the object and effect of the present invention, and can be 0.001 to 40 weight percent based on the total weight of the composition.

When the composition is a pharmaceutical composition, the pharmaceutical composition may comprise an effective amount of a flocculation protein or comprise a flocculation protein as an active ingredient. The effective amount may be appropriately selected depending on the individual. The effective amount may be determined based on factors including the following, as well as other factors well known in the medical arts: the severity of the disease, the age, weight, health, sex of the patient, the sensitivity of the patient to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, the drug mixed with or used simultaneously with the composition. Administration can be performed by any method known in the art. Administration can be carried out directly to the individual in any way, by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. The administration may be performed systemically or locally.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat or cat. The subject may be one in need of improved skin cosmetics (e.g. skin moisturization, enhancement of skin barrier or skin rejuvenation).

The administration may be daily administration of the composition for skin beauty comprising the flocculating protein to each individual in the following amounts: 0.1mg to 1000mg, for example, 0.1mg to 500mg, 0.1mg to 100mg, 0.1mg to 50mg, 0.1mg to 25mg, 1mg to 1,000mg, 1mg to 500mg, 1mg to 100mg, 1mg to 50mg, 1mg to 25mg, 5mg to 1000mg, 5mg to 500mg, 5mg to 100mg, 5mg to 50mg, 5mg to 25mg, 10mg to 1,000mg, 10mg to 500mg, 10mg to 100mg, 10mg to 50mg, or 10mg to 25mg. However, the administration amount may be variously prescribed depending on factors such as formulation method, administration method, age, body weight, sex, pathological condition, diet, administration time, administration route, excretion rate, and response sensitivity of the patient, and the like, and those skilled in the art will understand that the administration amount may be appropriately adjusted in consideration of these factors. The number of administrations may be 1 per day or 2 or more within a clinically acceptable side effect range, with respect to the administration site, administration may be at one or two or more, and the total number of administration days per day or every 2 to 5 days as intervals may be 1 to 30 days at the time of one treatment. If desired, the same treatment can be repeated after an appropriate time. For animals other than humans, the administration may be performed in the same amount as humans per kg of the administration amount, or in an amount converted to the administration amount by a volume ratio (for example, average value) of organs (heart and the like) of the target animal and humans, or the like.

When the composition is a composition for health functional foods, the formulation may be of a conventional health food formulation known in the art. The health food can be prepared into general dosage forms such as powder, granule, tablet, pill, capsule, suspension, oil, syrup, impregnant, liquid, extractant, etc., and any health food forms such as meat, sausage, bread, chocolate, candy, snack, biscuit, pizza, stretched noodles, other noodles, chewing gum, jelly, dairy products including ice cream, various soups, beverages, tea, drinkable preparation, alcoholic beverage, vitamin complex, etc. In order to formulate the health food, a dietetically acceptable carrier or additive may be used, and any carrier or additive known in the art may be used to prepare a formulation desired to be prepared. The additives may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonating agents for carbonated beverages, and the like. In addition, the additive may contain pulp for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These additive ingredients may be used alone or in combination, and although the proportion of additives is not critical, may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition. The content of the flocculated protein extract in the composition for health functional food may be appropriately determined depending on the purpose of use (prevention or improvement). Generally, 0.01 to 15 weight percent of the total weight of the food may be included, and when manufactured as a beverage, a ratio of 0.02 to 10g, more preferably a ratio of 0.3 to 1g, may be included on the basis of 100 mL.

The inventors of the present invention confirmed significant improvement in expression of filaggrin, improvement in expression of claudin 1, improvement in expression of hyaluronidase, improvement in expression of exfoliating enzyme, and an effect of inhibiting melanin production, as compared to a general yeast culture or an untreated group, in an experiment using a composition including flocculation protein. Therefore, the composition comprising the flocculation protein can be effectively used for skin beauty or for treating skin-related diseases and the like.

Advantageous effects

Since the composition including the flocculated protein has effects of improving skin barrier, moisturizing, whitening and cell proliferation, and promoting collagen synthesis and elastin synthesis, it can be effectively used for skin beauty.

Drawings

FIG. 1 is a graph showing the effect of increasing filaggrin expression of flocculating proteins in immortalized epidermal (HaCat) cells.

FIG. 2 is a graph showing the effect of increasing the claudin 1 expression of flocculation proteins in immortalized epidermal cells.

Fig. 3 is a graph showing the effect of increasing hyaluronic acid synthase 3 (HAS 3) expression of flocculation proteins in immortalized epidermal cells.

FIG. 4 is a graph showing the effect of increasing the kallikrein 7 (KLK 7) expression of flocculating proteins in immortalized epidermal cells.

Fig. 5 is a graph showing the effect of flocculating proteins in inhibiting melanin production in immortalized epidermal cells.

Detailed Description

Examples and test examples are described in detail as examples to specifically illustrate the present invention. However, the examples and test examples of the present invention may be varied in various forms, and the scope of the present invention should not be construed as being limited to the examples and test examples described hereinafter. These examples and test examples are provided to enable those skilled in the art to more completely and completely understand the present invention.

Example 1 preparation of flocculation protein

Saccharomyces pastorianus (Saccharomyces pastorianus) was cultured according to the method of Marika (Marika) H.1994 Purification and partial characterization of flocculation proteins from brewer's yeast, and a solution including flocculation proteins was separated. The separated flocculated protein solution was freeze-dried at-70 ℃ and 5.0 Torr (Torr) to prepare a powder.

Comparative example 1 preparation of general Yeast culture

Pasteur yeasts were cultivated according to the method of Marika (Marika) H.1994 Purification and partial characterization of flocculation proteins from Saccharomyces cerevisiae. The yeast culture was centrifuged at 3500rpm, and the supernatant was filtered using a 0.45 μm filter membrane for sterilization. The sterilized yeast culture was freeze-dried at-70 ℃ and 5.0 torr to prepare a powder.

Test example 1 confirmation of the expression of the skin barrier factor Filaggrin (Filaggrin)

Immortalized epidermal (HaCat) cells as human keratinocyte cell lines were purchased from american type culture collection (ATCC, rockville, maryland, usa) and cultured in an incubator at 37 ℃ and 5% carbon dioxide using DMEM (hyclone, luogen, usa) supplemented with 10% fetal bovine serum (FBS, ATCC, rockville, maryland, usa) and 1% antibiotic/antifungal agent.

To confirm the cell barrier enhancing effect on human keratinocytes, human keratinocytes were seeded at a density of 5x 105 cells/well into 6-well cell culture dishes, and cultured for 24 hours in an incubator at 37 ℃ and 5% carbon dioxide using a medium supplemented with 10% fetal bovine serum and 1% antibiotic/antifungal agent. Comparative example 1 or example 1 was added to the cultured cells, followed by culture for 24 hours, and then the cells were recovered and 1ml of TRIZOL reagent (RNA extraction reagent (RNA iso), takara, japan) was added to isolate ribonucleic acid (RNA). After quantifying RNA using an instrument (Nanodrop 2000, sermer (Thermo), usa), complementary deoxyribonucleic acid (cDNA) was synthesized by reaction at 42 ℃ for 55 minutes and at 70 ℃ for 15 minutes (Reverse Transcriptase Mix), ELPIS biotechnology, korea). The RT-PCR technique used a PCR instrument (Step One Plus, applied Biosystems, USA), added with silk fibroin primers and cDNA to Sebocerin (SYBR Green supermix, applied Biosystems, USA), and polymerization was performed in 40 cycles as follows: polymerase activation was performed at 94 ℃ for 5 minutes, at 95 ℃ for 30 seconds, at 54 ℃ for 1 minute, and at 72 ℃ for 1 minute. The base sequences of the primers are shown in Table 1 below. The expression level of the gene was finally analyzed by correcting the β -actin (β -actin) gene.

As a result, it was observed that the expression of filaggrin was 2.84 times higher in the example 1-added group compared to the untreated group (fig. 1). Therefore, the composition of example 1 can be expected to have an excellent skin barrier strengthening effect.

[ Table 1]

Base sequence of filaggrin primer

Test example 2 confirmation of expression of the skin cell binding factor Claudin 1 (Claudin 1)

Immortalized epidermal cells as human keratinocyte cell lines were purchased from american type culture collection (ATCC, rockville, maryland, usa) and cultured in a DMEM medium (Hyclone, luogen, uk) supplemented with 10% fetal bovine serum (FBS, ATCC, rockville, maryland, usa) and 1% antibiotic/antifungal agent (AA, ATCC, rockville, maryland, usa) at 37 ℃ and 5% carbon dioxide.

Human keratinocytes were seeded at a density of 5x 105 cells/well into 6-well cell culture dishes, and then cultured for 24 hours in an incubator at 37 ℃ and 5% carbon dioxide using a medium supplemented with 10% fetal bovine serum and 1% antibiotic/antifungal agent. Comparative example 1 or example 1 was added to the cultured cells, followed by culture for 24 hours, and then the cells were recovered and 1ml of TRIZOL reagent (RNA extraction reagent (RNA iso), baobao (TAKARA), japan) was added to isolate RNA. After quantifying RNA using an instrument (Nanodrop 2000, saimer (Thermo), USA), cDNA was synthesized by reacting at 42 ℃ for 55 minutes and at 70 ℃ for 15 minutes (reverse transcriptase cocktail, ELPIS Biotechnology, korea). RT-PCR technology used a PCR instrument (Step One Plus, applied Biosystems, USA), with the addition of Siborgolin (SYBR Green supermix, applied Biosystems, USA) along with the Claudin 1 primer and cDNA, and the polymerization was performed in 40 cycles as follows: the reaction was carried out at 50 ℃ for 2 minutes, at 95 ℃ for 10 seconds, and at 60 ℃ for 1 minute. The base sequences of the primers are shown in Table 2 below. The expression level of the gene was finally analyzed by correcting the β -actin (β -actin) gene.

As a result, it was confirmed that the expression of claudin 1 in the group to which example 1 was added was increased 1.45 times as compared with the untreated group. (fig. 2) therefore, the composition of example 1 can be expected to have excellent skin moisturizing effect.

[ Table 2]

Base sequence of tight junction protein 1 primer

Test example 3 confirmation of expression of hyaluronic acid synthase HAS3

Immortalized epidermal cells as human keratinocyte cell lines were purchased from american type culture collection (ATCC, rockville, maryland, usa) and cultured in a DMEM medium (Hyclone, luogen, uk) supplemented with 10% fetal bovine serum (FBS, ATCC, rockville, maryland, usa) and 1% antibiotic/antifungal agent (AA, ATCC, rockville, maryland, usa) at 37 ℃ and 5% carbon dioxide in an incubator.

Human keratinocytes were seeded at a density of 5x 105 cells/well into 6-well cell culture dishes and cultured for 24 hours in an incubator at 37 ℃ and 5% carbon dioxide using a medium supplemented with 10% fetal bovine serum and 1% antibiotic/antifungal agent. Comparative example 1 or example 1 was added to the cultured cells, followed by culture for 24 hours, and then the cells were recovered and 1ml of TRIZOL reagent (RNA extraction reagent (rnaisso), baobao (TAKARA), japan) was added to isolate ribonucleic acid (RNA). Complementary deoxyribonucleic acid (cDNA) (Reverse Transcriptase Mix, ELPIS biotechnology, korea) was synthesized by reacting at 42 ℃ for 55 minutes and 70 ℃ for 15 minutes after quantifying RNA using an instrument (Nanodrop 2000, schirmer (Thermo), usa). The RT-PCR technique used a PCR instrument (Step One Plus, applied Biosystems, USA), and added with hyaluronic acid synthetase 3 primer and cDNA ceboglin (SYBR Green supermix, applied Biosystems, USA), and polymerization was performed according to 40 cycles as follows: polymerase activation was performed at 94 ℃ for 5 minutes, at 95 ℃ for 30 seconds, at 54 ℃ for 1 minute, and at 72 ℃ for 1 minute. The base sequences of the primers are shown in Table 3 below. The expression level of the gene was finally analyzed by correcting the β -actin (β -actin) gene.

As a result, it was confirmed that the flocculation protein of lager brewing yeast (example 1) expresses more hyaluronic acid synthase than the general yeast culture (comparative example 1) (fig. 3). Therefore, since the increased hyaluronic acid synthase can enhance the capturing ability of moisture within the skin, the composition of example 1 can be expected to have an excellent skin moisturizing effect.

[ Table 3]

Hyaluronic acid synthetase 3 primer base sequence

Test example 4 confirmation of expression of Decutinase kallikrein 7 (KLK 7)

Immortalized epidermal cells as human keratinocyte cell lines were purchased from american type culture collection (ATCC, rockville, maryland, usa) and cultured in an incubator at 37 ℃ and 5% carbon dioxide using DMEM medium (Hyclone, rogen, usa) supplemented with 10% fetal bovine serum (FBS, ATCC, rockville, maryland, usa) and 1% antibiotic/antifungal agent.

Human keratinocytes were seeded at a density of 5x 105 cells/well into 6-well cell culture dishes and cultured for 24 hours in an incubator at 37 ℃ and 5% carbon dioxide using a medium supplemented with 10% fetal bovine serum and 1% antibiotic/antifungal agent. Comparative example 1 or example 1 was added to the cells, followed by culture for 24 hours, and then the cells were recovered and 1ml of TRIZOL reagent (RNA extraction reagent (RNA iso), baobao (TAKARA), japan) was added to isolate ribonucleic acid (RNA). Complementary deoxyribonucleic acid (cDNA) (Reverse Transcriptase Mix, ELPIS biotechnology, korea) was synthesized by reacting at 42 ℃ for 55 minutes and 70 ℃ for 15 minutes after RNA quantification using an instrument (Nanodrop 2000, schirmer (Thermo), usa). The RT-PCR technique used a PCR instrument (Step One Plus, applied Biosystems, USA), added with kallikrein 7 primer and cDNA to Cyberrin (SYBR Green supermix, applied Biosystems, USA), and the polymerization was performed in 40 cycles as follows: at 50 ℃ for 2 minutes, at 95 ℃ for 10 seconds, and at 60 ℃ for 1 minute. The base sequences of the primers are shown in Table 4 below. The expression level of the gene was finally analyzed by correcting the β -actin (β -actin) gene.

It was confirmed that the flocculation protein of lager brewing yeast (example 1) expresses more of the cutinase (kallikrein 7) than the general yeast culture (comparative example 1). (FIG. 4) therefore, the composition of example 1 can be expected to have a mending effect of making skin color smooth and bright.

[ Table 4]

Kallikrein 7 primer base sequence

Test example 5 confirmation of the Effect of inhibiting melanogenesis

B16F10 cells, which were murine pigmented cells (mouse melanoma), were suspended in DMEM medium including 10% fetal bovine serum and seeded at a density of 1 × 105 cells/well in a 6-well plate until the B16F10 cells adhered to the bottom of the well. To induce melanin production, 0.1 μ M α -Melanocyte Stimulating Hormone (MSH) was treated and then cultured for 3 days after addition to each sample. As a positive control group for the test of melanin production ability, 100ppm of arbutin was treated. The cultured cells were washed with Phosphate Buffered Saline (PBS) and recovered with trypsin (trypsin). The cells were obtained only by counting the recovered cells using a hemocytometer (hemacytometer), collecting the same number of cells at 1 × 106 cells/mL for each treatment group, and centrifuging at 1000rpm for 10 minutes. The melanin in the cells was obtained by drying the recovered cells at 60 deg.C for 1 hour and adding 100. Mu.l of 1M sodium hydroxide solution including 10% dimethyl sulfoxide (DMSO). The inhibition rate of melanin production was evaluated by measuring the absorbance of the melanin solution at 490nm using a microplate reader.

As a result, it was confirmed that the flocculation protein of lager brewing yeast (example 1) inhibited melanin production (FIG. 5) to a greater extent than the general yeast culture (comparative example 1). Therefore, the composition of example 1 can have an excellent whitening effect because it inhibits melanin production in skin cells.

<110> kesi meishi co

<120> composition for skin beauty comprising flocculation protein

<130> PX057992PCT

<150> KR 10-2018-0004614

<151> 2018-01-12

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Claims

1. Use of a composition comprising a flocculating protein for the preparation of a therapeutic agent for enhancing the barrier of skin, wherein the flocculating protein is isolated from Saccharomyces pastorianus.

2. Use of a composition comprising a flocculating protein for the preparation of a therapeutic agent for skin moisturization or whitening, wherein the flocculating protein is isolated from Saccharomyces pastorianus.