Cosmetic composition comprising double fermentation product of white tea tree extract

Abstract

The present invention provides a composition comprising a white tea tree double fermentation product obtained by primary fermentation of a white tea tree extract using a microorganism of the genus Lactobacillus and secondary fermentation using a microorganism of the genus Bacillus. The composition has excellent effects of resisting oxidation, resisting inflammation, moisturizing skin, removing cutin, conditioning skin microorganisms and the like due to the inclusion of the white tea tree double fermentation product, and thus can be used as a raw material for cosmetics and the like.

Description

Technical Field

[0001] Disclosed is a cosmetic composition comprising a double-fermented product of white tea tree extract.

Background Art

[0002] Recently, with the increasing attention to cosmetic raw materials that are harmless, safe, and have skin-improving functionality to the skin, research aimed at developing natural product extracts into cosmetic raw materials has continued.

[0003] White tea tree (Kunzea ericoides), also known as kanuka, is known to have been first discovered in New Zealand. White tea tree leaf extract is used in cosmetics for purposes such as antioxidant and fragrance enhancer, and white tea tree leaf oil is used as a fragrance enhancer in cosmetics. As described above, some research is currently being conducted to use white tea tree extract as a cosmetic raw material.

[0004] However, there has been no report on the skin-improving effect of white tea tree fermentation products so far.

Summary of the Invention

[0005]

Technical Problem

[0006] On the one hand, provided is a composition comprising a fermentation product of white tea tree extract.

[0007] Specifically, the one hand provides a cosmetic composition, a topical skin composition, or a pharmaceutical composition comprising a double-fermented product of white tea tree, wherein the double-fermented product of white tea tree is obtained by first fermenting the white tea tree extract with a Lactobacillus sp. microorganism and then second fermenting with a Bacillus sp. microorganism.

[0008] On the other hand, provided is a method for preparing a fermentation product of white tea tree extract.

[0009] On the other hand, provided is the use of the double-fermented product of white tea tree in the preparation of a composition for antioxidant, anti-inflammatory, skin soothing, reducing skin irritation, skin moisturizing, strengthening skin barrier, exfoliating, or regulating skin microbiota.

[0010]

Technical Solution

[0011] On the one hand, provided is a composition comprising a fermentation product of white tea tree extract.

[0012] Specifically, the composition comprises a double-fermented product of white tea tree. The double-fermented product of white tea tree is obtained by first fermenting the extract of white tea tree (Kunzea ericoides) with a Lactobacillus sp. microorganism and then second fermenting with a Bacillus sp. microorganism.

[0013] The available parts of the tea tree (Kunzea ericoides) can be the whole plant, leaves, stems, roots, seeds, etc., and are not limited to specific parts. In a specific embodiment, the tea tree may refer to the leaves of the tea tree.

[0014] Regardless of the extraction method, extraction solvent, extraction conditions, the components extracted or the form of the extract, the term "extract" includes all substances obtained by extracting the components of natural products, and also includes substances that can be obtained by processing or treating the components of natural products using other methods after extraction. For example, the processing or treatment may include dilution, concentration, drying, purification, fractionation, filtration, etc. Therefore, the extract may include an extract solution, a diluted or concentrated solution of the extract solution, a dry product obtained by drying the extract solution, or its crude purified product or purified product, and a fractionated extract obtained by fractionated extraction thereof.

[0015] The extraction method may adopt well-known natural product extraction methods such as hot water extraction, ethanol extraction, heating extraction, cold soaking extraction, circulating extraction, circulating cooling extraction, ultrasonic extraction, subcritical extraction, etc., but is not limited thereto.

[0016] The extraction solvent may be selected from water, an organic solvent, or a mixture thereof. The water may be distilled water or pure water. The organic solvent may include one or more of alcohols having 1 to 6 carbon atoms, acetone, ether, ethyl acetate, diethyl ether, methyl ethyl ketone, and chloroform, but is not limited thereto.

[0017] In a specific embodiment, the extraction solvent of the extract may be water, an alcohol having 1 to 6 carbon atoms, or a mixture thereof. In another specific embodiment, the extraction solvent of the extraction may be water, an alcohol having 1 to 4 carbon atoms, or a mixture thereof. In another specific embodiment, the extraction solvent of the extract may be a mixture of water and an alcohol having 1 to 4 carbon atoms. In another specific embodiment, the extraction solvent of the extract may be butylene glycol. In another specific embodiment, the extraction solvent of the extract may be 10% (v / v) to 90% (v / v), 30% (v / v) to 90% (v / v), 50% (v / v) to 90% (v / v), 60% (v / v) to 80% (v / v), or 70% (v / v) butylene glycol.

[0018] The extraction conditions refer to the conditions such as extraction time and extraction temperature for extracting the components of natural products.

[0019] The extraction time can be 30 minutes to 168 hours, 30 minutes to 120 hours, 30 minutes to 100 hours, 30 minutes to 80 hours, 30 minutes to 60 hours, 30 minutes to 40 hours, 1 hour to 168 hours, 1 hour to 120 hours, 1 hour to 100 hours, 1 hour to 80 hours, 1 hour to 60 hours, 1 hour to 40 hours, 10 hours to 168 hours, 10 hours to 120 hours, 10 hours to 100 hours, 10 hours to 80 hours, 10 hours to 60 hours, 10 hours to 40 hours, 10 hours to 30 hours, 24 hours to 168 hours, 24 hours to 120 hours, 24 hours to 100 hours, or 24 hours to 80 hours. However, it can be appropriately selected according to other conditions such as the extraction solvent and extraction temperature.

[0020] The extraction temperature can be 1°C to 100°C, 10°C to 100°C, 50°C to 100°C, 70°C to 100°C, 80°C to 100°C, 1°C to 60°C, 2°C to 60°C, 2°C to 50°C, or 2°C to 40°C. However, it can be appropriately selected according to other conditions such as the extraction solvent and extraction time. In a specific embodiment, the extraction temperature can be about 90°C.

[0021] The double-fermented product of white tea tree can be obtained through the following steps: preparing an extract of white tea tree; performing a first fermentation on the extract of white tea tree using a microorganism of the genus Lactobacillus to prepare a first-fermented product of white tea tree; and performing a second fermentation on the first-fermented product of white tea tree using a microorganism of the genus Bacillus to prepare a double-fermented product of white tea tree.

[0022] Any microorganism of the genus Lactobacillus known in the art can be used as the microorganism of the genus Lactobacillus (Lactobacillus sp.). The microorganism of the genus Lactobacillus is not limited to a specific species. However, it can be selected, for example, from the group consisting of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, and Lactobacillus salivarius. In a specific embodiment, the microorganism of the genus Lactobacillus can be Lactobacillus plantarum.

[0023] The Bacillus sp. all include aerobic bacilli or facultative anaerobic bacilli that are widely distributed in nature. The Bacillus sp. microorganisms are not limited to specific species, but can be selected, for example, from the group consisting of Bacillus subtilis, Bacillus coagulans, Bacillus velezensis, Bacillus licheniformis, Bacillus polyfermenticus, Bacillus methylotrophicus, and Bacillus amyloliquefaciens. In a specific embodiment, the Bacillus sp. microorganism can be Bacillus subtilis. The Bacillus sp. microorganism can be a strain that produces keratinase.

[0024] The Lactobacillus sp. microorganism or Bacillus sp. microorganism can be used in the form of a microbial culture solution. The culture medium or culture conditions for culturing the Lactobacillus sp. microorganism or Bacillus sp. microorganism can be appropriately selected or changed by those of ordinary skill in the art for use.

[0025] For the fermentation conditions, appropriate temperature and time can be selected according to the type of microorganism. For example, the fermentation can be carried out at 20°C to 40°C, but is not particularly limited thereto. And the fermentation can be carried out for 1 hour to 168 hours, 1 hour to 120 hours, 1 hour to 72 hours, 1 hour to 48 hours, 12 hours to 168 hours, 12 hours to 120 hours, 12 hours to 72 hours, 12 hours to 48 hours, 24 hours to 168 hours, 24 hours to 120 hours, 24 hours to 72 hours, 24 hours to 48 hours, 48 hours to 168 hours, 48 hours to 120 hours, or 48 hours to 72 hours, but is not particularly limited thereto.

[0026] The fermentation medium can be appropriately selected according to the type of microorganism. For the medium, a well-known medium or a conventional medium that can be used for culturing the microorganism can be used according to the type of microorganism.

[0027] The fermentation product can include a fermentation broth obtained by fermenting white tea trees using microorganisms, a dilution or concentrate of the fermentation broth, a dry matter obtained by drying the fermentation broth, or a crude purification or purification thereof, or a fractionated extract obtained by fractionated extraction thereof, etc.

[0028] The fermentation product may be the supernatant or filtrate of the fermentation broth obtained by fermenting white tea trees using microorganisms. The supernatant or filtrate may be obtained by removing the microbial cells from the fermentation broth. Therefore, the fermentation product may be the supernatant of the fermentation broth. Alternatively, the fermentation product may be a ferment filtrate.

[0029] The content of the fermentation product relative to the total weight of the composition may be 0.0001 to 99.9999% by weight, 0.0001 to 90% by weight, 0.0001 to 50% by weight, 0.0001 to 10% by weight, 0.001 to 99.9999% by weight, 0.001 to 90% by weight, 0.001 to 50% by weight, 0.001 to 10% by weight, 0.01 to 99.9999% by weight, 0.01 to 90% by weight, 0.01 to 50% by weight, 0.01 to 20% by weight, or 0.01 to 10% by weight. However, those of ordinary skill in the art can select an appropriate content considering the desired effects and dosage forms, etc.

[0030] The composition may include a double-fermented product of white tea trees as an active ingredient.

[0031] The phrase "including... as an active ingredient" means adding the ingredient to an extent that can exhibit the effects mentioned in this specification, and also means adding various ingredients as auxiliary ingredients for drug delivery and stabilization and formulating into various forms.

[0032] The composition may be a composition for improving skin condition, improving skin beauty, improving skin health, or preventing or treating skin diseases.

[0033] The double-fermented product of white tea trees may have 2,2′-azino-bis(3-ethylbenzhothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability. Therefore, the composition including the double-fermented product of white tea trees may have an antioxidant effect.

[0034] The double-fermented product of white tea trees may have nitric oxide (NO) inhibitory ability. Therefore, the composition including the double-fermented product of white tea trees may have anti-inflammatory, skin-soothing, and skin irritation-reducing effects.

[0035] The double-fermented product of white tea trees may have the ability to increase the expression of aquaporin-3 (AQP-3). Therefore, the composition including the double-fermented product of white tea trees may have skin moisturizing and skin barrier strengthening effects.

[0036] The double-fermented product of the white tea tree may have proteolytic enzyme activity. Accordingly, a composition comprising the double-fermented product of the white tea tree may have an exfoliating effect.

[0037] The double-fermented product of the white tea tree may have an effect of regulating skin microbiota. The skin microbiota regulation may refer to reducing harmful skin bacteria while not affecting beneficial skin bacteria. The beneficial skin bacteria may be, for example, Staphylococcus epidermidis. The harmful skin bacteria may be, for example, Pseudomonas aeruginosa, Staphylococcus aureus, etc.

[0038] The ABTS radical scavenging ability, NO inhibitory ability, AQP-3 expression increasing ability, proteolytic enzyme activity, and skin microbiota regulation effect of the double-fermented product of the white tea tree may be significantly superior to those of the unfermented white tea tree extract or the Lactobacillus-fermented filtrate product.

[0039] Accordingly, a composition comprising the double-fermented product of the white tea tree may be used for antioxidant, anti-inflammatory, skin soothing, reducing skin irritation, skin moisturizing, strengthening skin barrier, exfoliating, or skin microbiota regulation.

[0040] The term "skin moisturizing" may refer to all actions for maintaining skin moisture or preventing moisture loss. The skin moisturizing effect may contribute to improving skin wrinkles and increasing elasticity.

[0041] The term "strengthening skin barrier" may refer to all actions for enhancing skin barrier function. The skin barrier is located on the outermost layer of the skin and prevents loss of moisture and nutrients.

[0042] The "anti-inflammatory" may be used interchangeably with "improving inflammation" and "inhibiting inflammation", and may include, for example, all actions for inhibiting nitric oxide (NO) production.

[0043] The "skin soothing" may include all actions for relieving and stabilizing the symptoms (examples: heat sensation, pain, redness, pruritus, etc.) of damaged or irritated skin or restoring the skin.

[0044] The "skin disease" may be a disease caused by damaged skin barrier function or a skin inflammatory disease.

[0045] The damaged skin barrier function may refer to all changes that occur on the skin due to reduced or impaired skin barrier function. For example, it may include increased skin wrinkles, dryness, dermatitis, allergic dermatitis, atopic dermatitis, acne, etc.

[0046] The skin inflammatory disease can be any one selected from the group consisting of skin wounds, dermatitis, allergic dermatitis, pruritus, eczematous skin diseases, dry eczema, erythema, urticaria, psoriasis, drug eruptions, and acne.

[0047] The term "skin health" can refer to all actions that maintain skin function and protect the skin from external stimuli. For example, it can include maintaining the skin barrier function; skin moisturization; protection from external factors such as ultraviolet rays; maintaining elasticity; calming and recovery from skin damage, etc.

[0048] The term "prevention" includes inhibiting the occurrence of a disease. The term "treatment" includes inhibiting, alleviating, or removing the progression of a disease. The term "improvement" can refer to all actions that at least reduce a parameter related to the alleviation or treatment of a condition, such as the degree of symptoms.

[0049] In one specific embodiment, the composition can be a cosmetic composition.

[0050] In addition to the active ingredients disclosed in this specification, the cosmetic composition can further include ingredients commonly used in cosmetic compositions, functional additives, etc. For example, it can include conventional auxiliaries such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, thickeners, preservatives, vitamins, pigments, fragrances, etc., as well as a cosmetically acceptable carrier.

[0051] The cosmetic composition can be prepared into any dosage form commonly prepared. For example, the cosmetic composition can have the cosmetic dosage forms of lotion, skin cream, essence, facial cleansing foam, cleansing water, facial mask, ampoule, body lotion, body oil, shampoo, rinse-off hair conditioner, hair treatment, hairspray, foundation cream, lipstick, mascara, or primer.

[0052] Optionally, the composition can be a topical skin agent composition.

[0053] The topical skin agent can be a skin cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, medicated bandage, skin lotion, or a combination thereof. The topical skin agent can be appropriately formulated with ingredients commonly used in topical skin agents such as cosmetics or pharmaceuticals, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, humectants, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or a combination thereof as needed.

[0054] Optionally, the composition can be a pharmaceutical composition.

[0055] The pharmaceutical composition may further comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, mannitol, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, anhydrous monobasic calcium phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silica, talc, or a combination thereof.

[0056] The pharmaceutical composition may be formulated into an oral or parenteral dosage form. The oral dosage form may be a granule, a powder, a liquid, a lozenge, a capsule, a dry syrup, etc. The parenteral dosage form may be an injection, a drip, an ointment, a skin lotion, a gel, a skin cream, a spray, a suspension, an emulsion, an adjuvant, a patch, etc.

[0057] On the other hand, a method for preparing a fermented product of white tea tree extract is provided.

[0058] Specifically, the method may be a method for preparing a double-fermented product of white tea tree.

[0059] The method includes: preparing a white tea tree extract; subjecting the white tea tree extract to a first fermentation using a Lactobacillus microorganism to prepare a first-fermented product of white tea tree; and subjecting the first-fermented product of white tea tree to a second fermentation using a Bacillus microorganism to prepare a double-fermented product of white tea tree.

[0060] On the other hand, the use of the double-fermented product of white tea tree in the preparation of a composition for antioxidant, anti-inflammatory, skin soothing, reducing skin irritation, skin moisturizing, strengthening skin barrier, exfoliating, or regulating skin microbiota is provided.

[0061] The composition may be a cosmetic composition, a topical skin agent composition, or a pharmaceutical composition.

[0062] Details regarding the double-fermented product of white tea tree are as described above. Therefore, the double-fermented product of white tea tree may be obtained by subjecting a white tea tree extract to a first fermentation using a Lactobacillus sp. microorganism and then subjecting it to a second fermentation using a Bacillus sp.

[0063] Considering the complexity of this specification, repetitive content will be omitted, and terms not otherwise defined in this specification have the meanings commonly used in the technical field of the present invention.

Description of the Drawings

[0064] The accompanying drawings, which are included to provide a further understanding of the present invention and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the present invention and, together with the description, serve to explain the inventive concept.

[0065] Figure 1 A graph showing the radical scavenging ability (%) of 2,2'-azino-bis(3-ethylbenzhothiazoline-6-sulfonic acid) (ABTS). L-Ascorbic acid: positive control group, white tea tree extract: white tea tree extract of Preparation Example 1, Lactobacillus plantarum: fermentation filtrate product of Lactobacillus genus of Preparation Example 3, White-Keratinase: double-fermented product of white tea tree of Example 1.

[0066] Figure 2 A graph for confirming the inhibitory ability of nitric oxide (No, Nitric Acid). L-NMMA: positive control group, white tea tree extract: white tea tree extract of Preparation Example 1, Lactobacillus plantarum: fermentation filtrate product of Lactobacillus genus of Preparation Example 3, White-Keratinase: double-fermented product of white tea tree of Example 1.

[0067] Figure 3 A graph showing the expression level of aquaporin-3 (AQP-3) gene. White tea tree extract: white tea tree extract of Preparation Example 1, Lactobacillus plantarum: fermentation filtrate product of Lactobacillus genus of Preparation Example 3, White-Keratinase: double-fermented product of white tea tree of Example 1, CaCl2: positive control group.

[0068] Figure 4 Showing the results of the paper disk experiment for confirming the proteolytic efficacy. After fermentation: double-fermented product of white tea tree of Example 1, Before fermentation: white tea tree extract of Preparation Example 1.

[0069] Figure 5 A graph showing the change in the number of bacteria in the skin microflora before and after co-culture in the double-fermented product of white tea tree of Example 1.

Detailed Description of the Invention

[0070] Hereinafter, the present invention will be described in more detail with reference to the embodiments. However, these embodiments are only used to exemplarily describe the present invention, and the scope of the present invention is not limited to these embodiments.

[0071]

Preparation Example

[0072]

Preparation Example 1: Preparation of White Tea Tree Extract

[0073] 500 mL of 70% (v / v) butylene glycol was added to 100 g of leaves of Kunzea ericoides, and extraction was carried out at 90 °C for 24 hours. Then, the crude product was removed using an 8 μm filter, and filtration was performed using a 0.45 μm membrane filter to prepare a white tea tree extract.

[0074]

Preparation Example 2: Preparation of Lactobacillus Culture Solution

[0075] 1,000 ml of purified water was added to 55 g of MRS medium, autoclaved at 121 °C for 15 minutes, and then cooled to room temperature. Then, Lactobacillus plantarum strain (accession number: KCTC14687BP) isolated from Centella asiatica was inoculated at 2% of the culture solution volume and cultured at 180 rpm and 37 °C for 48 hours.

[0076]

Preparation Example 3: Preparation of Lactobacillus Fermentation Filtrate

[0077] The Lactobacillus culture solution of Preparation Example 2 was centrifuged at 4,500 rpm for 20 minutes to remove the cells, and after obtaining the supernatant, filtration was performed using a 0.45 μm membrane filter to prepare a Lactobacillus fermentation filtrate.

[0078]

Preparation Example 4: Preparation of Bacillus Culture Solution

[0079] 1,000 mL of purified water was added to 55 g of Nutrient medium, autoclaved at 121 °C for 15 minutes and then cooled to room temperature. Then, Bacillus subtilis isolated from natto was inoculated in an amount of 2% in the medium volume and cultured at 180 rpm and 37 °C for 48 hours.

[0080]

Example 1

[0081]

Example 1: Preparation of White Tea Tree Fermentation Product

[0082]

Example 1-1: Preparation of Primary Fermentation Product of White Tea Tree

[0083] The white tea tree extract of Preparation Example 1 was fermented using the Lactobacillus culture solution of Preparation Example 2 to prepare a white tea tree fermentation product.

[0084] Specifically, the Lactobacillus culture solution of Preparation Example 2 was inoculated into the white tea tree extract of Preparation Example 1 at a concentration of 2% (v / v) of the extraction volume, and cultured at 37°C and 150 rpm for 72 hours to prepare the primary fermentation product of the white tea tree. The fermentation product was centrifuged at 4,500 rpm for 20 minutes to remove the bacteria, and after obtaining the supernatant, it was filtered through a 0.45 μm membrane filter to obtain the filtrate. The filtrate was used as the primary fermentation product of the white tea tree in the following experiments.

[0085]

Example 1-2, Preparation of the secondary fermentation product of the white tea tree

[0086] The secondary fermentation product of the white tea tree was prepared by fermenting the primary fermentation product of the white tea tree in Example 1-1 with the Bacillus culture solution of Preparation Example 4.

[0087] Specifically, the Bacillus culture solution of Preparation Example 4 was inoculated into the primary fermentation product of the white tea tree in Example 1-1 at a concentration of 2% (v / v) of the extraction volume, and cultured at 37°C and 150 rpm for 72 hours to prepare the secondary fermentation product of the white tea tree. The fermentation product was centrifuged at 4,500 rpm for 20 minutes to remove the bacteria and obtain the supernatant, and then filtered through a 0.45 μm membrane filter to obtain the filtrate. The filtrate was used as the double fermentation of the white tea tree in the following experiments. The double fermentation product of the white tea tree was named "White-Keratinase".

[0088]

Experimental Example

[0089]

Experimental Example 1, Determination of antioxidant activity

[0090] An experiment was conducted to confirm the antioxidant activity of the double fermentation product of the white tea tree according to an embodiment.

[0091] Specifically, the stable free radical 2,2′-azino-bis(3-ethylbenzhothiazoline-6-sulfonic acid) (ABTS) was used to measure the degree of reduction of ABTS free radicals due to the reaction with the sample. The white tea tree extract of Preparation Example 1, the Lactobacillus fermentation filtrate of Preparation Example 3, or the double fermentation product of the white tea tree in Example 1 was used as the sample, and each sample was used at a concentration of 1% (v / v). The L-ascorbic acid 100 μg / ml treatment group was used as the positive control group.

[0092] Figure 1 A chart showing the ABTS free radical scavenging ability (%).

[0093] As a result, it was confirmed that the white tea tree extract of Preparation Example 1, the Lactobacillus fermentation filtrate of Preparation Example 3, and the double-fermented product of white tea tree of Example 1 all have antioxidant activity, as Figure 1 shown. In particular, it was confirmed that the double-fermented product of white tea tree of Example 1 has significantly superior antioxidant activity compared to the white tea tree extract or the Lactobacillus fermentation filtrate.

[0094] Therefore, it can be seen that the double-fermented product of white tea tree of Example 1 has excellent antioxidant effects.

[0095]

Experimental Example 2, Confirmation of Anti-inflammatory and Skin Soothing Efficacy

[0096] An experiment was conducted to confirm the anti-inflammatory and skin soothing efficacy of the double-fermented product of white tea tree according to an embodiment.

[0097] Specifically, after treating mouse macrophages Raw264.7 with lipopolysaccharides (LPS), each sample was treated at a concentration of 1% (v / v) to measure the gene expression level of nitric oxide (NO). The untreated group (MO: Media Only) was used as the negative control group. The group treated with 30 μg / ml of L-NMMA was used as the positive control group.

[0098] Figure 2 A chart showing the results for confirming the nitric oxide (NO) inhibitory ability.

[0099] As a result, it was confirmed that the white tea tree extract of Preparation Example 1 and the double-fermented product of white tea tree of Example 1 significantly inhibited the expression of the NO gene that causes skin irritation, as Figure 2 shown. In particular, the double-fermented product of white tea tree of Example 1 showed the most excellent NO inhibitory ability.

[0100] Therefore, it can be seen that the double-fermented product of white tea tree of Example 1 has excellent effects on anti-inflammation, skin soothing, and reduction of skin irritation.

[0101]

Experimental Example 3, Confirmation of Skin Moisturizing and Skin Barrier Strengthening Efficacy

[0102] An experiment was conducted to confirm the skin moisturizing and skin barrier strengthening efficacy of the double-fermented product of white tea tree according to an embodiment.

[0103] Specifically, after culturing keratinocytes, each sample was treated at a concentration of 1% (v / v) to confirm the gene expression level of aquaporin-3 (AQP-3). The group treated with 2 mM of CaCl2 was used as the positive control group.

[0104] Figure 3 A graph showing the expression level of aquaporin-3 (AQP-3) gene.

[0105] As a result, it was confirmed that the double-fermented product of white tea tree in Example 1 had significantly superior efficacy in increasing AQP-3 expression compared to the white tea tree extract in Preparation Example 1 or the Lactobacillus fermented and filtered product in Preparation Example 3.

[0106] Therefore, it can be known that the double-fermented product of white tea tree in Example 1 has excellent effects on skin moisturization and skin barrier strengthening.

[0107]

Experimental Example 4, Confirmation of Keratin Removal Effect

[0108] The keratin removal effect of the double-fermented product of white tea tree according to an embodiment was confirmed based on protease activity.

[0109] Specifically, a circular paper disc was placed on a 1% skim milk agar plate for cultivation, and the protease activity was confirmed by the diameter of the clear zone.

[0110] Figure 4 The results of the paper disc experiment for confirming protease activity are shown.

[0111] As a result, it was confirmed that the double-fermented product of white tea tree in Example 1 had a more prominent clear zone and a larger diameter compared to the white tea tree extract in Preparation Example 1, and thus had greater protease activity according to the group, as Figure 4 shown.

[0112] Therefore, the double-fermented product of white tea tree in Example 1 has excellent protease activity and can be known to have excellent keratin removal effect.

[0113]

Experimental Example 5, Confirmation of Skin Microorganism Regulation Effect

[0114] An experiment was conducted to confirm the skin microorganism regulation effect of the double-fermented product of white tea tree according to an embodiment.

[0115] Specifically, Tryptic Soy Broth (TSB, BD Difco, USA) was used. For the growth of bacteria, the corresponding strain was adjusted to 1×10 6The growth rates were compared by inoculating at a concentration of CFU / mL into an appropriate medium, culturing at 37 °C for 24 hours, and measuring the absorbance at 600 nm, or by confirming the actual viable cell count by the serial dilution method. The experimental groups with the addition of the same amount of phosphate buffer saline (PBS) as the test sample were compared based on the percentage. 1 mL of 10% TBS was added to a 12-well plate for co-culturing skin harmful bacteria and skin beneficial bacteria, and the harmful bacteria (Pseudomonas aeruginosa, Staphylococcus aureus) were inoculated at a concentration of 1×10 6 CFU / mL. A 0.4-μm cell culture insert was loaded above it, and the beneficial bacteria (Staphylococcus epidermidis) were inoculated at 1×10 6 CFU / mL into 1 mL of 10% TBS. For the control group, the same amount of phosphate buffer saline (PBS) was used for treatment. Then, after culturing at 32 °C for 12 hours, the respective bacterial counts were confirmed to compare the growth extent.

[0116] Figure 5 A graph showing the change in the bacterial count in skin microorganisms before and after co-culture in the double-fermented product of white tea tree in Example 1.

[0117] As a result, it was confirmed that the bacterial counts of the skin beneficial bacteria Staphylococcus epidermidis, the skin harmful bacteria Staphylococcus aureus, and Pseudomonas aeruginosa did not change before co-culture, as Figure 5 shown. However, after co-culture with the double-fermented product of white tea tree in Example 1, it was confirmed that there was no effect on the skin beneficial bacteria Staphylococcus epidermidis, but the bacterial counts of the skin harmful bacteria Staphylococcus aureus and Pseudomonas aeruginosa decreased.

[0118] Therefore, the double-fermented product of white tea tree in Example 1 does not affect skin beneficial bacteria while reducing skin harmful bacteria, and it can be known that it has the effect of regulating skin microorganisms.

[0119] In summary, it was confirmed that the double-fermented product of white tea tree obtained by first fermenting the white tea tree extract with Lactobacillus and then second fermenting with Bacillus has the following effects:

[0120] (i) It has ABTS radical scavenging ability, so it has excellent antioxidant effects;

[0121] (ii) It has NO inhibitory ability, so it has excellent effects on anti-inflammation, skin calming, and reducing skin irritation;

[0122] (iii) It has the ability to increase AQP-3 expression, so it has excellent effects on skin moisturization and strengthening the skin barrier;

[0123] (iv) Having proteolytic enzyme activity and excellent horny layer removal effect; and

[0124] (v) Without affecting beneficial skin bacteria (e.g., Staphylococcus epidermidis) while reducing harmful skin bacteria (e.g., Staphylococcus aureus, Pseudomonas aeruginosa), thus having excellent skin microbiota regulation effect.

Claims

1. A cosmetic composition comprising a double-fermented product of white tea trees, wherein, The double-fermented product of white tea tree is obtained by first fermenting the extract of white tea tree ( Lactobacillus sp. ), using Lactobacillus Kunzea ericoides ) microorganisms, and then performing a second fermentation using Bacillus microorganisms.

2. The cosmetic composition according to claim 1, wherein, The extraction solvent of the white tea tree extract is water, an alcohol having 1 to 6 carbon atoms, or a mixture thereof.

3. The cosmetic composition according to claim 1, wherein, The Lactobacillus microorganism is selected from the group consisting of Lactobacillus acidophilus ( L.acidophilus ), Lactobacillus casei ( L.casei ), Lactobacillus gasseri ( L.gasseri ), Lactobacillus delbrueckii subsp. bulgaricus ( L. delbrueckii ssp. bulgaricus ), Lactobacillus helveticus ( L. helveticus ), Lactobacillus fermentum ( L. fermentum ), Lactobacillus paracasei ( L. paracasei ), Lactobacillus plantarum ( L. plantarum ), Lactobacillus reuteri ( L. reuteri ), Lactobacillus rhamnosus ( L. rhamnosus ), and Lactobacillus salivarius ( L. salivarius ).

4. The cosmetic composition according to claim 1, wherein, The Bacillus microorganism is selected from the group consisting of Bacillus subtilis ( B. subtilis ), Bacillus coagulans ( B. coagulans ), Bacillus velezensis ( B. velezensis ), Bacillus licheniformis ( B. licheniformis ), Bacillus polyfermenticus ( B. polyfermenticus ), Bacillus methylotrophicus ( B. methylotrophicus ), and Bacillus amyloliquefaciens ( B. amyloliquefaciens ).

5. The cosmetic composition according to claim 1, wherein, The cosmetic composition has one or more effects selected from the free radical scavenging ability of 2,2′-hydrazine-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS, 2,2'-azino-bis(3-ethylbenzhothiazoline-6-sulfonic acid)), nitric oxide (NO, Nitric Oxide) inhibitory ability, and the ability to increase the expression of aquaporin-3 (AQP-3, Aquaporin-3).

6. The cosmetic composition according to claim 1, wherein, The cosmetic composition has proteolytic enzyme activity.

7. The cosmetic composition according to claim 1, wherein, The cosmetic composition has a skin microbiota modulating effect of reducing skin harmful bacteria while not affecting skin beneficial bacteria.

8. The cosmetic composition according to claim 7, wherein, The skin beneficial bacterium is Staphylococcus epidermidis ( Staphylococcus epidermidis ), and the skin harmful bacterium is Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) or Staphylococcus aureus (Staphylococcus aureus) .

9. The cosmetic composition according to claim 1, wherein, The cosmetic composition is used for antioxidant, anti-inflammatory, calming the skin, reducing skin irritation, skin moisturization, strengthening the skin barrier, exfoliating, or modulating skin microbiota.

10. Use of the double-fermented product of white tea tree in the preparation of a composition for antioxidant, anti-inflammatory, calming the skin, reducing skin irritation, skin moisturization, strengthening the skin barrier, exfoliating, or modulating skin microbiota, wherein, The double-fermented product of the white tea tree is obtained by first fermenting an extract of the white tea tree ( Lactobacillus sp.) with a microorganism of the genus Lactobacillus and then secondarily fermenting it with a microorganism of the genus Bacillus ( Kunzeaericoides ). Bacillus sp.).

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