KR102440022B1 - A cosmetic composition for alleviating sebum secretion comprising culture of bacillus strain - Google Patents

Abstract

The present invention relates to a cosmetic for relieving sebum secretion and provides a cosmetic composition comprising: a Bacillus stratosphericus strain; a culture thereof; a lysate; an extract; or a purified product as an active ingredient.

Description

COSMETIC COMPOSITION FOR ALLEVIATING SEBUM SECRETION COMPRISING CULTURE OF BACILLUS STRAIN}

The present invention relates to the use of a culture of Bacillus stratospericus strain for alleviating sebum secretion.

Acne, which mainly occurs during puberty, is a worldwide skin disease and is observed in more than 85% of adolescents, and in some cases persists until their 30s.

Acne is a chronic inflammatory disease of the sebaceous gland unit of the hair follicle, and it is accompanied by various skin changes such as comedones, papules, pus blisters, nodules, and pseudocysts. .

Acne is caused by a combination of several causes. Typically, due to an excess of male hormones, the secretion of sebaceous glands is active, and the epithelium of the hair follicles causes heterokeratosis (abnormal keratinization), which blocks the hair follicles and forms comedones, the basic lesion of acne. .

The sebaceous glands are distributed in almost all parts of the human body, and are especially abundant in the back and chest, in addition to the head, forehead, and central parts of the face.

The sebaceous glands secrete sebum by decomposition of sebaceous cells. Sebum is excreted by holocrine, and it is known that the lifespan of sebum cells starting from cell division is about 21 to 25 days.

On the other hand, since the peroxisome proliferator activated receptor (PPAR) forms a heterodimer together with the retinoid X receptor, transcription of genes related to various intracellular reactions including lipid metabolism, cell proliferation and differentiation is known to regulate

Three subtypes α, β/δ, and γ of peroxisome proliferator-activated receptors (PPARs) are found in basal sebaceous cells, and peroxisome proliferator-activated receptor γ (PPAR γ) differentiates and lipids in sebaceous cells. It is known to be an important factor in synthesis.

As another cause, among microorganisms residing in hair follicles, especially Cutibacterium acnes ) causes acne.

The Cutibacterium acnes is the main cause of troubles while eating sebum, and in normal people, even if the microorganism is present on the skin, it does not cause a problem.

Therefore, there has been a continuous demand in the art to discover a safe material capable of effectively suppressing the production of sebum.

Conventionally, as a typical attempt to improve acne-prone skin, it removes dead skin cells that are clogging pores by removing sebum generated using an adsorption powder or applying salicylic acid, trichloric acid and trihydroxy palmitic acid to a cosmetic composition The growth of bacteria was inhibited.

In addition, a method for preventing acne by using a natural extract having antibacterial activity or suppressing sebum secretion has been proposed, but it is insufficient to show a sufficient effect.

Meanwhile, interest in non-toxic and non-toxic microorganisms has recently been increasing. .

Microorganisms that exist widely in nature secrete various kinds of physiologically active substances, and these microorganism-derived metabolites are natural products and are spotlighted as substitutes for chemical components that present various risks.

In addition, since microorganisms are easy to manipulate and can be cultured in large quantities, they have advantages in terms of economy and utility.

The present inventors completed the present invention by confirming that as a result of culturing and studying Bacillus strains, metabolites affecting the inflammatory response can be produced, thereby effectively alleviating sebum secretion.

The present invention is to solve the problems of the prior art described above, an object of the present invention is to provide a natural material with excellent biosafety and physiological activity that can replace conventional chemical components.

According to one aspect of the present invention, Bacillus stratosphericus ( Bacillus stratosphericus ) A cosmetic composition for alleviating sebum secretion comprising a strain, its culture, lysate, extract or purified product as an active ingredient is provided.

In one embodiment, the cosmetic composition may suppress the production of sebum by inhibiting the expression of NO and COX-2.

In one embodiment, the cosmetic composition may further include an anti-inflammatory use.

In one embodiment, the cosmetic composition may further include a use for improving acne.

In one embodiment, the cosmetic composition comprises a softening lotion, a nutrient lotion, a moisture cream, a nutrient cream, a massage cream, a nutrient lotion, an essence, an ampoule, a gel, an eye cream, an oil, a foundation, a cleansing cream, a cleansing foam, a cleansing water, It may be formulated with one or more selected from the group consisting of a mask pack, a spray, and a powder.

According to another aspect of the present invention, Bacillus stratosphericus ( Bacillus stratosphericus ) strain, its culture, lysate, extract or purified product; as an active ingredient; there is provided a composition for external application for improving acne comprising the active ingredient.

According to another aspect of the present invention, Bacillus stratosphericus ( Bacillus stratosphericus ) strain, its culture, lysate, extract or purified product; as an active ingredient, an anti-inflammatory composition for external application for skin is provided.

According to the present invention, the strain produces various kinds of physiologically active substances, and the physiologically active substances have low toxicity to the human body and effectively relieve sebum secretion through anti-inflammatory activity, so it can replace conventional chemical materials.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a component analysis result for a Bacillus stratospericus strain culture according to an embodiment of the present invention.
2 is a result of evaluating the cytotoxic concentration of the Bacillus stratospericus strain culture according to an embodiment and a comparative example of the present invention.
3 and 4 are results of evaluating the anti-inflammatory efficacy of the Bacillus stratospericus strain culture according to an embodiment of the present invention.
5 is a result of evaluating the antibacterial activity of the Bacillus stratospericus strain culture according to an embodiment of the present invention.

The terms used in this specification have been selected as currently widely used general terms as possible while considering the functions in the present invention, which may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, and the like.

In addition, in a specific case, there is a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than the name of a simple term.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present application. does not

Numerical ranges are inclusive of the values defined in that range. Every maximum numerical limitation given throughout this specification includes all lower numerical limitations as if the lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes all higher numerical limitations as if the higher numerical limitation were expressly written. All numerical limitations given throughout this specification will include all numerical ranges that are better within the broader numerical limits, as if the narrower numerical limitations were expressly written.

Hereinafter, examples of the present invention will be described in detail, but it is obvious that the present invention is not limited by the following examples.

One aspect of the present invention provides a cosmetic composition for alleviating sebum secretion comprising a Bacillus stratosphericus strain, its culture, lysate, extract or purified product as an active ingredient.

The Bacillus stratosphericus ( Bacillus stratosphericus ) is a microorganism commonly found in high concentrations in the stratosphere, is generally found in the atmosphere, but comes down to the earth as a result of the atmospheric circulation process, and is also found in freshwater such as rivers and streams.

The present inventors cultured the strain under optimal conditions through medium optimization technology, analyzed metabolites according to the culture, and confirmed that the metabolites according to the culture can effectively inhibit the sebum production mechanism.

The cosmetic composition may include the strain or its culture, lysate, extract or purified product, and suppress sebum production through anti-inflammatory activity, thereby exhibiting a sebum relief effect.

The culture may be the culture itself obtained by culturing the strain, or a culture supernatant obtained by removing the strain therefrom, or a concentrate or freeze-dried product of the culture supernatant.

In one embodiment, the culture comprises the steps of (a) pre-culturing the Bacillus stratosphericus strain; (b) inoculating the strain into a culture medium and fermenting; And (c) removing the strain and recovering the supernatant; can be obtained through.

The culture comprises at least one carbon source selected from the group consisting of glucose, fructose, mannose, galactose, xylose, ribose, maltose, sucrose, dextrin, glycerin, and inulin; And soy protein hydrolyzate, potassium phosphate, diammonium phosphate, potassium diphosphate, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium oxalate, and culturing the strain in a medium containing at least one nitrogen source selected from the group consisting of inulin can be obtained.

The carbon source or nitrogen source used in the culture medium is, for example, about 0.1 to 30 w / v%, 1 to 25 w / v%, 1 to 20 w / v%, 1 to 10 w / v%, 1 to 5 w / v %, or 1 to 1.5 w/v%.

The culture medium may include commonly used salts, and the salts may include, for example, one or more salts selected from the group consisting of potassium nitrate, potassium sulfate, magnesium carbonate, magnesium sulfate, and magnesium nitrate, The salt may be included in, for example, about 0.01 to 10 w/v%, 0.1 to 7 w/v%, or 0.5 to 5 w/v%.

The culture medium may contain about 0.01 to 10 w/v%, 0.05 to 5 w/v%, or 0.1 to 2 w/v% of yeast autolysate, and the culture medium may have a pH of about 2.0 to 7.0. However, the present invention is not limited thereto.

The culture is cultured under a temperature condition of, for example, about 20 to 42 ° C, about 25 to 35 ° C, or about 27 to 30 ° C, for example, about 24 to 144 hours, 48 to 120 hours, or 15 to 35 hours of culturing the strain. It may be obtained by

The culture conditions of the strain may include the step of fermenting by adding the strain to the medium, and the fermentation may use a commonly used fermentation method, for example, the method may use fed batch fermentation. can

The extract may be obtained by extracting the culture of the isolated strain with various organic solvents.

The lysate is obtained by isolating the strain from the culture of the strain, disrupting the cell wall or membrane through mechanical methods or nonmechanical methods, and releasing intracellular products. it could be

The purified product may be obtained by culturing the strain and extracting and purifying the active ingredient from the liquid medium or the fermentation process is completed.

The Bacillus stratospericus strain may further include an anti-inflammatory use of the cosmetic composition.

The “anti-inflamatory” refers to an effect of improving inflammation. The “inflammation” is a mechanism to repair and regenerate the damaged area when an invasion that causes any organic changes, such as physical action, chemical substances, or bacterial infection, is applied to a living body or tissue. Vascular active substances such as serotonin, bradykinin, prostaglandin, HETE (hydroxyeicosatetraenoic acid), and leukotriene are liberated, and vascular permeability is increased and inflammation can be induced.

The cosmetic composition can suppress the production of sebum by reducing the expression of COX-2 and NO.

The expression of COX-2 and NO is regulated by the activity of NF-kB, a transcription factor, and the cosmetic composition inhibits the expression of inflammatory response-related genes induced by the C. acnes extract and increases due to this It can reduce the production of sebum.

The cosmetic composition may further include a use for improving acne.

The “acne” is acne vulgaris, which can be caused by increased sebum production in pores, abnormal keratinization of skin epithelial cells, blocking the opening of pores, and proliferation of acne-causing bacteria to cause inflammation.

The cosmetic composition can improve acne by inhibiting the acne-causing bacteria, thereby inhibiting clogging of pores and excessive secretion of sebum.

The cosmetic composition includes a softening lotion, a nourishing lotion, a moisture cream, a nourishing cream, a massage cream, a nourishing lotion, an essence, an ampoule, a gel, an eye cream, an oil, a foundation, a cleansing cream, a cleansing foam, a cleansing water, a mask pack, a spray and a powder It may be formulated with one or more selected from the group consisting of, but is not limited thereto and may be manufactured in various forms.

Another aspect of the present invention provides a composition for external application for improving acne comprising a Bacillus stratosphericus strain, a culture, lysate, extract or purified product thereof as an active ingredient.

The skin external preparation or cosmetic composition may be formulated by a conventional method. In the formulation of external skin preparations, reference may be made to the contents disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, and in the formulation of cosmetic compositions, International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry and Fragrance Association, Inc., Washington, 1995 may be referred to.

Specifically, the cosmetic composition may be prepared in the form of a general emulsified formulation and a solubilized formulation. For example, lotion, such as a softening lotion or a nourishing lotion; emulsions such as facial lotions and body lotions; creams such as nourishing creams, moisturizing creams, and eye creams; essence; makeup ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils; Or it may be formulated as a cleaning agent such as a cleansing foam, soap, body wash, but is not limited thereto. In addition, the external preparation for skin may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.

In the skin external preparation or cosmetic composition, in each formulation, other ingredients may be appropriately blended within the range that does not impair the purpose according to the present invention, depending on the type of formulation or the purpose of use, in addition to the essential ingredients.

The external preparation for skin or cosmetic composition may include a conventionally acceptable carrier, for example, oil, water, surfactant, humectant, lower alcohol, thickener, chelating agent, pigment, preservative, fragrance, etc. may be appropriately blended, but this It is not limited.

The acceptable carrier may vary depending on the formulation. For example, when formulated into an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or their Mixtures may be used.

When the skin external preparation or cosmetic composition is formulated as a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or mixtures thereof may be used as a carrier component, and in the case of a spray, chloro It may further comprise a propellant such as fluorohydrocarbon, propane, butane or dimethyl ether.

When the skin external application or cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, fatty acid esters of polyethylene glycol or sorbitan may be used. can

When the skin external preparation or cosmetic composition is formulated as a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, such as Suspending agents, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like can be used.

When the cosmetic composition is formulated as a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars, etc. can be used

The skin external application or cosmetic composition is a fatty substance, an organic solvent, a solubilizer, a thickener, a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent commonly used in the industry according to the quality or function of the final product. ), fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, common in cosmetics It may additionally contain adjuvants commonly used in the field of cosmetics or dermatology, such as any other ingredients used as

However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.

Through the following examples, the present invention will be described in more detail, but it is obvious that the present invention is not limited by the following examples.

Example: Preparation of strain cultures

A strain culture was prepared using a single strain of Bacillus stratosphericus obtained from the Korea Research Institute of Bioscience and Biotechnology.

The Bacillus stratospericus strain was suspended in 1.0 mL of sterile physiological saline (0.9% NaCl), diluted stepwise, spread on a commercial TSB (Tryptic Soy Broth) solid medium by Difco, and cultured at 37° C. for 3 days.

A single colony formed after culturing was taken, separated and inoculated in the same medium by a stroke plate method, and then cultured at 30° C. for 3 days to obtain a pure single colony.

The isolated single colony was inoculated into 3 mL of TSB liquid medium and cultured with shaking at 37°C for 2 days. It was stored and, if necessary, revaccinated and used for later experiments.

Preparation of cultured products

Since the microorganism produces metabolites having various sebum secretion activities in the growth phase, the microorganism was cultured and a culture was obtained therefrom.

The appropriate culture conditions for the identified microorganisms are glucose or sucrose in the medium 1 to 10.0 w/v%, potassium diphosphate or potassium phosphate 1.0 to 4.0 w/v%, protein hydrolyzate or soy protein hydrolyzate 1.0 to 5.0 w/v% , each of the inorganic salts were included in an amount of 0.1 to 1.5 w/v% and sterilized at 121 ° C. for 15 minutes.

The prepared culture solution was incubated at 30 to 37° C. at 200 rpm for 48 hours to 96 hours, followed by ultrasonic extraction for 1 hour to 3 hours.

The prepared ultrasonic extract was adjusted to pH 0.5 to 2.0 using a hydrogen chloride solution having a concentration of 0.1N to 1.5N, and allowed to stand at 3 to 10°C for 12 hours.

One organic solvent selected from among ethanol, ethyl acetate, and methanol was added to the still purified product for extraction.

After repeating the above series of processes 1 to 3 times, the supernatant separated at 12,000 rpm for 15 to 30 minutes was collected after sterilization using a 0.22 μm membrane filter.

Comparative Example: Bacillus subtilis ( Bacillus subtilis ) and Lactobacillus plantarum ( Lactobacillus plantarum ) strain culture preparation

Commercially obtained single strains of Bacillus subtilis and Lactobacillus plantarum were inoculated in TSB liquid medium sterilized at 121 °C for 15 minutes.

After culturing at 30 to 37° C. at 200 rpm for 48 hours, the supernatant separated in a centrifuge at 12,000 rpm for 15 to 30 minutes was sterilized using a 0.22 μm membrane filter, and then the culture was recovered.

Then, a sample was obtained by preparing a purified culture product in the same manner as in Example (Table 1).

division strain Comparative Example 1 Bacillus subtilis Comparative Example 2 Lactobacillus plantarum

Experimental Example 1: Analysis of strain culture components

The components contained in the purified strain culture obtained in Examples were analyzed as follows.

The culture obtained in Example was freeze-dried and suspended by adding 3 mL of purified water to the freeze-dried culture.

Components included in the suspension were analyzed using thin layer chromatography (TLC).

A silica plate was used, and the mobile phase used for development was chloroform, methanol, and water in a weight ratio of 65:25:4. After development, 10% (w/w) sulfuric acid was used to analyze the components of the cultured purified product after color development.

Referring to FIG. 1 , it can be seen that the color is located on the same line as the reference material, and the reference material used is Surfactin.

Experimental Example 2: Cytotoxicity evaluation (MTT assay)

In order to evaluate the cytotoxicity of the strain culture purified products prepared in Examples and Comparative Examples, the effect on the proliferation of macrophages was tested.

After processing the samples of Examples and Comparative Examples at each concentration in the cultured Raw264.7 cells, the cell viability was measured.

It was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reagent, and MTT is absorbed into cells to form formazan by succinate anhydrase in mitochondria. Since the accumulation of substances in cells means mitochondrial activity, broadly, cellular activity, cellular toxicity can be evaluated.

Each cell was aliquoted at 2 x 10 ⁴ cells/well in a 96 well plate, and then cultured in a cell incubator for 24 hours.

After culturing, the medium was replaced with a medium containing 1% FBS to make it starvation and cultured for 24 hours.

Samples of Examples and Comparative Examples 1 and 2 were treated at each concentration and cultured for 24 hours, then treated with MTT solution and cultured for 4 hours.

After incubation, the medium was removed and the cells were completely dissolved with DMSO (dimethyl sulfoxide), and absorbance was measured at 450 nm with an absorbance measuring instrument Multiskan GO (Thermo Fisher Scientific).

Based on the measurement results, the cell viability was expressed as a percentage (%) by comparing the cell viability with the non-treated group through the formula below.

Cell viability (%) = [Absorbance of the sample added group (450 nm) / Absorbance of the non-sample group (450 nm)] x 100

In Figure 2, it was confirmed that the concentration does not show toxicity of the culture purified product prepared in Examples and Comparative Examples for Raw264.7 macrophages.

In a later experiment, the concentration of the sample was determined based on the toxicity results.

Experimental Example 3: Nitric oxide production inhibitory activity evaluation (NO assay)

The irritant was used using the culture medium of the Cutibacterium acnes strain.

Specifically, Difco's BHI (Brain Heart Infusion) medium sold on the market was sterilized at 121 ° C. for 15 minutes and inoculated at a level of 1% in the cooled medium, and anaerobically cultured at 30 to 37 ° C. for 48 hours. .

The obtained culture solution was centrifuged at 8,000 rpm to remove the supernatant, and phosphate buffered saline (PBS) was added and sonicated for 20 minutes.

After re-grinding the cells using liquid nitrogen and adding phosphate-buffered saline (PBS) to the culture solution after the grinding is completed, the absorbance (optical density value) is diluted to 2 to 3 when measured at a wavelength of 600 nm using a turbidimeter. did.

After centrifugation again at the same speed as above, only the supernatant was taken and used as a stimulation source.

The anti-inflammatory activity of the sample was evaluated by analyzing the NO production inhibitory ability in the rat macrophage RAW 264.7 cell induced by stimulation with the lysate of the Cutibacterium acnes strain.

The rat macrophages, RAW 264.7 cells (Korea Cell Line Bank) used in this experiment, were treated with DMEM medium (Gibco/BRL, USA) supplemented with 10% FBS (Gibco/BRL, USA) at 37°C, 5% CO2. Cultured in an incubator.

When the cells reached about 80% of the culture dish, the monolayer of cells was washed with PBS (pH 7.4), washed, and subcultured by adding about 3 ml of medium and scraping with a scraper.

RAW 264.7 cells were aliquoted into a 48-well plate at a volume of 6 x 10 ⁴ cells/well by 0.5 ml and cultured for 24 hours.

Samples of Examples and Comparative Examples (0.1 to 5%) and C. acnes (8%) were simultaneously treated and cultured for 24 hours.

The negative control group was treated only with C.acnes , and the positive control group was treated with NAC (N-Acetyl-L-Cysteine) and the amount of NO generated was measured.

The generated NO was measured in the form of NO ₂ ⁻ present in cell culture using Griess reagent.

100 μL of cell culture supernatant and Griess reagent (1% (w/v) sulfanilamide, 0.1% (w/v) naphtylehtylenediamine in 2.5% (v/v) phosphoric acid) were mixed in equal amounts and reacted in the dark at room temperature for 10 minutes, followed by ELISA Absorbance (540 nm) was measured using a reader.

The concentration of NO was shown in FIG. 3 by comparing the results with the control after a standard curve was prepared using NaNO2.

Referring to FIG. 3 , while the NO (nitric oxide) production inhibitory activity of the example was excellent, the comparative example was evaluated to have insufficient NO production inhibitory activity.

Experimental Example 4: Evaluation of COX-2 mRNA expression level

An experiment was conducted to determine whether the expression level of COX-2 mRNA was regulated for the purified culture products prepared in Examples and Comparative Examples.

After dispensing at 1 x 10 ⁶ cells/well in a 6 well plate, the cells were cultured in a cell incubator for 24 hours.

After culturing, the medium was replaced with a medium containing 1% FBS to make it starvation and cultured for 24 hours.

The samples of Examples and Comparative Examples were simultaneously treated with C. acnes (8%) according to the concentration. At this time, 20 μM NAC was used as a control.

After incubation for 24 hours, the medium was removed, RNA was extracted using an RNA extract kit (TaKaRa MiniBest RNA Extraction Kit), and RNA was quantified using a Qubit Fluorometer (Qubit 2.0 Fluorometer, life technologies).

cDNA was synthesized with the same amount of RNA using a High-Capacity RNA-to-cDNA kit (applied biosystems).

Taqman primer COX-2 and GAPDH were purchased from life technologies and used.

Using Taqman master mix, the desired gene was amplified using the Real-time PCR equipment Step One Plus Real-Time PCR (Applied Biosystem) to confirm the change in gene expression level.

The result of confirming the expression level of the COX-2 gene is shown in FIG. 4 .

Referring to FIG. 4 , the Example effectively inhibits the expression of COX-2 mRNA compared to the Comparative Example, thereby inhibiting or alleviating inflammation, suggesting that it is possible to block the sebum production pathway.

Referring to Figures 3 and 4, the amount of production of inflammatory cytokines COX-2 and NO is significantly reduced by the sample of the example, thereby inhibiting or alleviating inflammation, suggesting that it is possible to block the sebum production pathway. do.

Experimental Example 5: Evaluation of antibacterial activity

Using the purified strain culture obtained in Examples, the antibacterial activity against the Cutibacterium acnes strain was evaluated.

Specifically, after sterilizing RCM (Reinforced Clostridial Medium) medium with 1.5% Agar at 121°C for 15 minutes and cooling the internal temperature to about 40°C, the purified strain culture obtained in Example is added to the medium to an appropriate concentration. and solidified at room temperature to form a solid plate medium.

Example Strain Inoculation of a cutibacterium acnes strain suspended in 5.0 × 10 ⁵ CFU/mL bacterial number on RCM plate medium to which the culture purified product is added, and anaerobically cultured in an incubator at 30 to 37° C. for 48 hours, Cutibacterium acnes Antibacterial activity was confirmed by checking the number of viable cells of the strain.

The cutibacterium acnes strain was sterilized by Difco's commercially available RCM liquid medium at 121°C for 15 minutes, and anaerobic culture at 30 to 37°C in an incubator for 48 hours.

Example concentration 0% 0.5% 1.0% 2.0% 3.0% 5.0% Reduction Efficacy - - + ++ +++ +++

<Evaluation criteria> +++: more than 91% / ++: 71 to 90% / +: 51 to 70% / -: less than 50%

Referring to Table 2 and Figure 5, the sample of Example compared to the untreated group shows a strain reduction effect of about 83% with respect to the Cutibacterium acnes strain at the concentration treated with 2.0%, and when treated with 5.0%, the strain of about 95% Reduction efficacy was shown.

The results shown in Experimental Example 5 suggest that the cultured product of Example exhibited excellent antibacterial activity against the Cutibacterium acnes strain, which is the main causative agent of sebum and acne, and can effectively inhibit the growth of microorganisms at a predetermined concentration. do.

Experimental Example 6: Evaluation of artificial sebum reduction efficacy

The reduction efficacy of artificial sebum (ASTM D4265-14, Pickerking) was evaluated using the purified strain culture obtained in Examples.

Artificial sebum is in a solid state at room temperature, heated to 40 to 60 °C to make a liquid state, and then cooled again to prepare a hexagonal shape.

0.15 to 0.2 g of artificial sebum was put in a Petri dish, and 5 mL of the purified strain culture of Examples and Comparative Examples was added. Water was used as a control.

The petri dish to which artificial sebum and cultured products were added was stored in an incubator at 25 to 35° C. for up to 36 hours.

division reduction degree 0 hours 2 hours 4 hours 8 hours 16 hours 32 hours control - - - - - - Example - + ++ +++ +++ +++ Comparative Example 1 - - - ± ± + Comparative Example 2 - - - - ± +

<Evaluation criteria>+++: Very good improvement effect / ++: Significant improvement effect / +: slight improvement effect / ±: Little improvement effect / -: No improvement effect

Referring to Table 3, the control group treated only with water did not change with time, and the reduction effect of the culture purified product of the comparative example was evaluated at a very low level.

On the other hand, the sample of Example reduced artificial sebum after 2 hours, and the reduction phenomenon was accelerated over time.

The above results suggest that the purified culture product of Examples can exhibit excellent reducing efficacy against artificial sebum, and can effectively reduce sebum present in human skin.

The above description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.

Claims (7)

Bacillus stratosphericus ( Bacillus stratosphericus ) Strain, its culture, lysate, extract or purified; a cosmetic composition for reducing sebum comprising as an active ingredient. delete The method of claim 1,
Further comprising an anti-inflammatory use, a cosmetic composition. The method of claim 1,
Further comprising the use of improving acne, a cosmetic composition. 5. The method of any one of claims 1, 3 and 4,
From the group consisting of softening lotion, nourishing lotion, moisture cream, nourishing cream, massage cream, nourishing lotion, essence, ampoule, gel, eye cream, oil, foundation, cleansing cream, cleansing foam, cleansing water, mask pack, spray and powder A cosmetic composition formulated with one or more selected. delete delete

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