Composition comprising culture medium of fructobacillus fructosus PRC-1 for antioxidation, wound healing, skin whitening or wrinkle alleviation

Abstract

The present invention relates to a composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP, a culture medium thereof, or extracellular vesicles thereof. The composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP, a culture medium thereof, or extracellular vesicles thereof, according to the present invention, exhibits excellent antioxidant activity, and thus can be used as an antioxidant composition, exhibits an excellent effect on skin whitening by inhibiting melanin production, can exhibit improved efficacy against skin pigmentation diseases, facilitates wound healing and skin regeneration through wound recovery and the like, reduces skin wrinkles by inhibiting elastase activity, and exhibits therapeutic efficacy on skin wrinkle-related diseases. Therefore, the present invention is remarkably effective as a food, a pharmaceutical composition, and a cosmetic composition.

Description

Composition for antioxidant, wound healing, skin whitening or wrinkle improvement comprising Fructobacillus fructosus PRC-1 culture solution

[0001] The present invention relates to a composition for antioxidant, wound healing, skin whitening or wrinkle improvement, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0002] As modern people's economic well-being improves and their participation in social life increases, interest in skin beauty and health is on the rise across all age groups. Numerous beauty-related cosmetics and foods are being developed to prevent skin aging and maintain clear, healthy skin by suppressing skin stress.

[0003] In addition, the development of compositions that have the function of suppressing the production of free radicals or active oxygen is ongoing, but due to consumers' low preference for synthetic compounds, recently developed products are made from natural materials.

[0004] Meanwhile, probiotics are live bacteria that are distributed as dominant bacteria in the human intestines, where various microorganisms exist, and promote the growth of beneficial bacteria in the body. They live symbiotically in the human digestive system, breaking down fiber and complex proteins to transform them into important nutrients. They also inhibit the growth of harmful bacteria such as Escherichia coli and Clostridium difficile, improve diarrhea and constipation, and play a role in vitamin synthesis and lowering blood cholesterol. Among these probiotics, lactic acid bacteria are bacteria that obtain energy by fermenting sugars and produce large amounts of lactic acid. They are widely distributed in nature, including agricultural products, foods, and the bodies of humans and animals. They are widely used in the fermentation of cheese, fermented milk, kimchi, and bread. In general, it is known that consuming lactic acid bacteria not only suppresses harmful bacteria among the intestinal microflora, but also increases beneficial bacteria that aid in the digestion, absorption, and breakdown of food. Therefore, it has been reported that consuming lactic acid bacteria has various benefits, including reducing blood cholesterol, boosting immunity, suppressing endogenous infections, improving liver cirrhosis, and having anticancer effects. Furthermore, recent research is also being conducted to enhance the efficacy of natural products fermented with lactic acid bacteria.

[0005] To date, approximately 400 species of lactic acid bacteria have been discovered, and they are being used in the manufacture of pharmaceuticals, health functional foods, and cosmetics. Fermented solutions obtained by culturing these lactic acid bacteria in common culture media, such as those containing skim milk, whey, and sugars, have been reported to have effects such as whitening, moisturizing, skin softening by increasing the turnover rate of the stratum corneum, and wrinkle reduction. However, in reality, these functional effects of compositions containing these fermented solutions are often due to other raw materials added to the composition rather than the active ingredients contained in the fermented solution.

[0006] Accordingly, the inventors of the present invention have completed the present invention by confirming that Fructobacillus fructosus PRC-1 KCTC15271BP isolated from honey; its culture solution; or its extracellular vesicles; has excellent antioxidant, wound healing, skin whitening, or wrinkle improvement effects.

[0007] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating wounds, skin pigmentation diseases, or skin wrinkle-related diseases, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0008] The purpose of the present invention is to provide a food composition for antioxidant, wound healing, skin whitening or wrinkle improvement, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0009] The purpose of the present invention is to provide a cosmetic composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0010] The purpose of the present invention is to provide a feed composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof.

[0011] To avoid confusion due to overlapping content, the description of redundant content will be omitted below. In other words, the content of the invention is not limited to the content described below, and the content of the invention should be interpreted based on the overall content of the invention.

[0012] The present invention provides Fructobacillus fructosus PRC-1 KCTC15271BP (Deposit institution: Korea Research Institute of Bioscience and Biotechnology, Deposit date: January 5, 2023, Accession number: KCTC15271BP).

[0013] The Fructobacillus fructosus PRC-1 KCTC15271BP of the present invention is characterized as a novel probiotic isolated and identified from honey.

[0014] The 16S rDNA base sequence for identification and classification of Fructobacillus fructosus PRC-1 KCTC15271BP of the present invention is as shown in SEQ ID NO: 1 attached to this specification. Therefore, Fructobacillus fructosus PRC-1 KCTC15271BP of the present invention may have the 16S rDNA of SEQ ID NO: 1.

[0015] The present invention provides a pharmaceutical composition for preventing or treating wounds, skin pigmentation diseases, or skin wrinkle-related diseases, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0016] The pharmaceutical composition according to the present invention has an excellent effect in inhibiting intracellular ROS production and melanin production, facilitating wound healing and regeneration, and inhibiting the activity of elastase.

[0017] In the present invention, the strain may be deposited under the accession number KCTC15271BP.

[0018] In the present invention, the strain may be a live cell or a dead cell.

[0019] The above live cells refer to living bacteria, and the above dead cells refer to live cells cultured under certain conditions, collected, and then dried, pressurized, treated with drugs, or the like to prevent further growth of the bacteria.

[0020] The pharmaceutical composition according to the present invention may include a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or its extracellular vesicles. 12

[0021] In the present invention, the culture solution (or culture) refers to a product obtained by culturing Fructobacillus fructosus PRC-1 in a known liquid medium, and is a concept that includes the strain according to the present invention or various factors secreted therefrom. The product may include Fructobacillus fructosus PRC-1.

[0022] The above medium may be selected from known liquid media, and may be, for example, MRS liquid medium, GAM liquid medium, but is not limited thereto.

[0023] Additionally, depending on the need, live cells, dead cells or cultures thereof may be used after being crushed or after further extraction.

[0024] Shredded material refers to a material that has been separated and processed into a shredded form through mechanical or chemical methods. For example, shredded material can be produced through bead mills, presses, sonicators, microfluidizers, or enzyme treatment.

[0025] The term "extract" refers to a "water (liquid)" form obtained by further extracting living cells, dead cells, and / or fragments using a commonly known extraction method. For example, it refers to a product obtained by extracting living cells, dead cells, and / or fragments using a commonly known extraction method (a known extraction solvent (e.g., water, C1 to C4 alcohol (methanol, ethanol, etc.)).

[0026] Preferably, the culture medium may be obtained by culturing the strain for 12 hours or more, and preferably, may be obtained by culturing for 12 to 30 hours, 12 to 28 hours, 12 to 26 hours, 12 to 24 hours, 18 to 30 hours, 18 to 28 hours, 18 to 26 hours, 18 to 24 hours, 24 to 30 hours, or 24 to 28 hours, and for example, may be obtained by culturing for 24 to 26 hours, but is not limited thereto.

[0027] In the present invention, the culture solution may have a pH of 6.5 to 8.5, preferably a pH of 6.5 to 8.0, a pH of 6.5 to 7.5, a pH of 7.0 to 8.5, or a pH of 7.0 to 8.0, and for example, a pH of 7.0 to 7.5, but is not limited thereto.

[0028] In culture media, Fructobacillus fructosus PRC-1 grows and produces metabolites such as lactic acid, which can lead to acidification. This pH range is not suitable for cell culture. Based on this, the most appropriate pH for cell culture is 7.4.

[0029] In the present invention, the culture solution may be included in an amount of 10.0 to 100.0% (v / v) of the total composition, preferably 10.0 to 90.0%, 20.0 to 80.0%, but is not limited thereto. In addition, for example, the culture solution may be included in an amount of 1 x 10 5 1 x 10 11 It may contain live or dead cells at a concentration of CFU / ml, preferably 1 x 10 5 1 x 10 10 CFU / ml, 1 x 10 5 1 x 10 9 CFU / ml, or 1 x 10 7 1 x 10 10 It may be included at a concentration of CFU / ml, for example, 1 x 10 8 1 x 10 10 It may be included at a concentration of CFU / ml, but is not limited thereto.

[0030] In the present invention, the culture solution may be a culture supernatant obtained by culturing the strain and then removing the strain, a concentrate thereof, a fraction thereof, or a lyophilized product thereof, and may be, for example, a culture supernatant, but is not limited thereto.

[0031] In the present invention, "extracellular vesicles (EV)" refers to particles secreted from cells and released into the extracellular space, and are called exosomes, ectosomes, microvesicles, microparticles, exosome-like vesicles, etc. Extracellular vesicles were observed in electron microscopic studies to originate from specific intracellular compartments called multivesicular bodies (MVBs) and to be released and secreted outside of the cell, rather than being directly separated from the plasma membrane. That is, when fusion of multivesicular bodies and the plasma membrane occurs, the vesicles are released into the extracellular environment, which are called extracellular vesicles.

[0032] Extracellular vesicles (EVs) facilitate the exchange of materials (proteins, lipids, and genetic material) between cells and function as mediators for physiological and pathological signaling. Extracellular vesicles are broadly categorized into exosomes and microvesicles. Exosomes vary in size depending on their biological origin. They are intraluminal vesicles created when the endosomal membrane inwardly inflates during the maturation of multi-vesicular endosomes, and are secreted when multi-vesicular endosomes fuse with the cell surface. Microvesicles, 50-1000 nm in size, are vesicles secreted outside the cell by protruding from the plasma membrane. Each cell produces different types of EVs depending on its physiological state and secretes EVs with specific lipid / protein / nucleic acid compositions. The above “extracellular vesicles” is used to mean exosomes and microvesicles.

[0033] The above extracellular vesicles may have an average diameter of 20 nm to 400 nm, preferably 50 nm to 300 nm, more preferably 100 nm to 250 nm.

[0034] The above extracellular vesicles can be obtained using an extracellular vesicle (EV) extraction method known in the art, and are not limited thereto, but can be obtained by an extraction method including, for example, the following steps:

[0035] 1) Step of culturing Fructobacillus fructosus PRC-1;

[0036] 2) A step of recovering the above Fructobacillus fructosus PRC-1 culture solution;

[0037] 3) A step of centrifuging the recovered culture solution to remove cell residue; and

[0038] 4) A step of obtaining extracellular vesicles from the culture medium from which the cell debris has been removed using at least one selected from the group consisting of tangential flow filtration (TFF), ultracentrifugation, size exclusion chromatography, and an exosome isolation kit.

[0039] In one embodiment, the present invention can purify extracellular vesicles of Fructobacillus fructosus PRC-1 with high purity through filtration and ultracentrifugation.

[0040] In the process of isolating extracellular vesicles in the present invention, any method commonly used in the art can be used without limitation, for example, utilizing the commercially available exosome isolation kit (e.g., EXO-BB, ExoQuick ® -ULTRA, ExoQuick ® -TC, Capturem TM Exosome Isolation Kit, Total Exosome Isolation Kit, ExoTrap TM Exosome Isolation Spin Column Kit, Exo2DTM, etc.), or including separation based on the difference in specific gravity between components in a solution (e.g., centrifugation), separation based on size (e.g., ultrafiltration or vacuum filter), separation based on affinity for a specific substrate (e.g., affinity chromatography), but is not limited thereto, and any method commonly used in the art as a separation method based on the unique physical properties of a target substance in a heterogeneous sample can be used without limitation.

[0041] In the present invention, the extracellular vesicles are 1 x 10 compared to the entire composition. 5 1 x 10 11 particles / ml concentration, preferably 1 x 10 5 1 x 10 10 particles / ml, 1 x 10 5 1 x 10 9 particles / ml, or 1 x 10 6 1 x 10 10 particles / ml, for example, 1 x 10 7 1 x 10 9 It may be included at a concentration of particles / ml, but is not limited thereto.

[0042] The present invention provides a pharmaceutical composition for preventing or treating wounds, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0043] A wound is, for example, a "wound," which means a physical disruption of the continuity or integrity of a tissue structure.

[0044] Although not limited thereto, the wound may be at least one selected from the group consisting of burns, lacerations, epidermal wounds, ulcers, trauma, post-surgical, childbirth, chronic wounds, damage due to dermatitis, corneal ulcers, corneal epithelial detachment, keratitis, and damage due to dry eye.

[0045] The present invention provides a pharmaceutical composition for preventing or treating skin pigmentation disease, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0046] In the present invention, the skin pigmentation disease may be at least one selected from the group consisting of, for example, lentigines, freckles, hypopigmentation, and hyperpigmentation.

[0047] The present invention provides a pharmaceutical composition for preventing or treating skin wrinkle-related diseases, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0048] The above skin wrinkle-related disease may be at least one selected from the group consisting of elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, and premature skin aging.

[0049] Specifically, the skin wrinkle-related disease may be a skin wrinkle-related disease caused by skin photodamage. "Skin photodamage" refers to skin damage caused by skin exposure to sunlight or UV light (especially UV-B).

[0050] Protection against the aforementioned skin wrinkle-related conditions may be, for example, "photoprotective." This may refer to blocking or mitigating adverse clinical, histological, and immunological effects caused by the sun or ultraviolet rays. These effects include, in particular, chronic effects (e.g., elastosis and wrinkle formation).

[0051] The pharmaceutical composition of the present invention can be used as a pharmaceutical composition comprising a pharmaceutically effective amount of Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; and / or a pharmaceutically acceptable carrier.

[0052] The term “pharmaceutically effective amount” as used herein means an amount sufficient to achieve the efficacy or activity of the Fructobacillus fructosus PRC-1 strain described above, a culture medium thereof, or an extracellular vesicle thereof.

[0053] Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in formulation, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

[0054] The pharmaceutical composition according to the present invention can be administered to mammals, including humans, via various routes. Any commonly used route of administration may be used, including oral, transdermal, intravenous, intramuscular, and subcutaneous administration. Preferably, the composition is administered orally or applied to the skin.

[0055] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, patient age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. An ordinary skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention. For a desirable effect, in the case of oral administration, the composition of the present invention may generally be administered to adults at 0.0001 to 100 mg / kg per 1 kg of body weight per day, preferably 0.001 to 100 mg / kg. Administration may be administered once a day or divided into several times. The above dosage does not limit the scope of the present invention in any way.

[0056] The present invention provides a method for preventing or treating a wound, a skin pigmentation disease, or a skin wrinkle-related disease, comprising administering to a subject in need thereof Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0057] The pharmaceutical composition of the present invention can be manufactured in the form of a unit dose or can be manufactured by inserting it into a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablets, capsules or gel (e.g., hydrogel), and may additionally include a dispersant or stabilizer.

[0058] The present invention provides the use of Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; in the manufacture of a medicament for treating at least one disease selected from the group consisting of wounds, skin pigmentation diseases, and skin wrinkle-related diseases.

[0059] The present invention provides Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; for use in the prevention or treatment of at least one disease selected from the group consisting of wounds, skin pigmentation diseases, and skin wrinkle-related diseases.

[0060] The present invention provides a food composition for preventing or improving skin pigmentation disease, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0061] The present invention provides a food composition for antioxidant, wound healing, skin whitening or wrinkle improvement, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0062] A food composition according to the present invention may include a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

[0063] The food composition according to the present invention has an excellent effect in suppressing the production of reactive oxygen species (ROS), inhibiting melanin production, facilitating wound healing and regeneration, and inhibiting the activity of elastase.

[0064] In the present invention, "antioxidant" means preventing the production of active oxygen in the body and preventing oxidation phenomena that cause irreparable damage to cells. In a narrow sense, it means the action of inhibiting, reducing or controlling the production or reaction of free radicals produced in the body, hydrogen peroxide or peroxide produced from free radicals, and hydroxyl radicals produced from hydrogen peroxide. In a broad sense, it means the action of inhibiting, reducing or controlling the production of oxidation reactions that occur in nature.

[0065] "Wound treatment" refers to the restoration of tissue integrity, which may include partial or complete restoration of tissue integrity, and may be used interchangeably with "cure" or "heal." The treatment may involve healing a wound in a shorter period of time compared to natural healing. The treatment may involve improving and / or alleviating the wound. Wound improvement refers to reducing the severity of the wound. Furthermore, the treatment may encompass the healing of both the wound and / or a disease associated with the wound. The treatment may involve the healing and / or regeneration of damaged tissue resulting from the wound. The treatment may encompass skin regeneration. Furthermore, the treatment may involve maintaining the original composition of the damaged tissue. Furthermore, the treatment may involve promoting the healing and / or regeneration of the damaged tissue while minimizing complications and / or scarring of the wound-related disease. Therefore, wound treatment refers to the promotion, improvement, advancement, acceleration, or progress of one or more steps or processes involved in the wound healing process.

[0066] In the present invention, "skin whitening" generally refers to any action that inhibits or prevents melanin deposition on the skin by inhibiting melanin production. Examples include, but are not limited to, improving freckles, blemishes, and age-related dark spots, reducing the melanin index on the skin surface, and improving skin tone.

[0067] In the present invention, “wrinkle improvement” means suppressing or inhibiting the formation of wrinkles on the skin or alleviating wrinkles that have already formed.

[0068] Additionally, it can be used to prevent and improve the above-mentioned wounds, skin pigmentation diseases or skin wrinkle-related diseases.

[0069] When the food composition of the present invention is used as a food additive, the food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. Generally, when manufacturing a food or beverage, the food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, relative to the raw materials.

[0070] There are no specific restrictions on the types of the above foods. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and all foods in the conventional sense are included.

[0071] The above beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates may be appropriately determined by those skilled in the art.

[0072] In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination. The proportions of these additives may also be appropriately selected by those skilled in the art.

[0073] The food composition of the present invention can also be used as a health functional food. A health functional food is a food that emphasizes the bioregulatory function of food, and is a food that has added value by using physical, biochemical, and biotechnological methods to act and express for a specific purpose. The food composition of the present invention can be used as a health functional food. The ingredients of such a health functional food are designed and processed to sufficiently exert the body's body regulatory functions related to biodefense, regulation of body rhythm, and prevention and recovery from disease. The food composition may contain food-acceptable food additives, sweeteners, or functional raw materials. When the strain of the present invention is used as a health functional food (or health functional beverage additive), the novel strain may be added as is or used in combination with other foods or food ingredients, and may be used appropriately according to conventional methods. The mixing amount of the strains may be appropriately determined depending on the intended use (prevention, health or improvement, therapeutic treatment).

[0074] The present invention provides a cosmetic composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0075] Preferably, a cosmetic composition for antioxidant, wound healing, skin wrinkle improvement, skin whitening, skin soothing, or anti-aging, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof, is provided.

[0076] The cosmetic composition according to the present invention can exhibit antioxidant, wound healing, wrinkle improvement, skin whitening, skin soothing, and anti-aging effects.

[0077] The cosmetic composition may be used for any one purpose selected from the group consisting of antioxidant, wound healing, skin wrinkle improvement, skin whitening, skin soothing, and anti-aging, and more specifically, may be used for any one purpose selected from the group consisting of antioxidant, wound healing, skin whitening, and wrinkle improvement.

[0078] More specifically, the present invention provides a cosmetic composition for antioxidant, wound healing, skin whitening or wrinkle improvement, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

[0079] The cosmetic composition according to the present invention has an excellent effect in suppressing the production of reactive oxygen species (ROS), inhibiting melanin production, facilitating wound healing and regeneration, and inhibiting the activity of elastase.

[0080] That is, Fructobacillus fructosus PRC-1 KCTC15271BP; its culture medium; or its extracellular vesicles; may exhibit at least one effect selected from the group consisting of inhibition of reactive oxygen species (ROS) production, inhibition of melanin production, wound healing and regeneration, and inhibition of elastase activity.

[0081] The cosmetic composition according to the present invention may include a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

[0082] The cosmetic composition of the present invention can be manufactured into any formulation commonly manufactured in the art, and for example, can be manufactured into a formulation including a toner (skin lotion), skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, hand sanitizer, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, suspension, gel, powder, paste, mask pack, and sheet. The composition of such a formulation can be manufactured according to a method conventional in the art. The mixing amount of additional ingredients such as the moisturizer can be easily selected by a person skilled in the art within a range that does not impair the purpose and effect of the present invention.

[0083] In addition to the effective ingredients disclosed herein, the above cosmetic composition may further include functional additives and components included in general cosmetic compositions, and may further include purified water, thickeners, preservatives, stabilizers, solubilizers, surfactants, carriers, fragrances, or combinations thereof that are commonly used. The functional additives may include components selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular peptides, high molecular polysaccharides, sphingolipids, and seaweed extracts. The carriers may include, for example, alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosity modifiers, emulsifiers, stabilizers, ultraviolet scatterers, ultraviolet absorbers, colorants, fragrances, and the like. Compounds / compositions that can be used as the above alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosity modifiers, emulsifiers, stabilizers, ultraviolet scatterers, ultraviolet absorbers, colorants, fragrances, etc. are already known in the art, so a person skilled in the art can select and use the appropriate corresponding substances / compositions.

[0084] The strain may be contained in the composition in an amount of, for example, 0.001% to 30% by weight.

[0085] In addition, the cosmetic composition may further contain, as necessary, ingredients such as sunscreens, antioxidants (butylated hydroxyanisole, propyl gallate, ellisorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinyl urea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, sodium malate, formaldehyde, sodium formaldehyde, succinic acid, sodium succinate, sodium hydroxide, sodium hydrogen phosphate, etc.), moisturizers (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycereth-26, methylgluceth-20, etc.), and lubricants.

[0086] Additionally, for each type of cosmetic composition, appropriate ingredients can be selected and blended according to the cosmetic formulation or intended use. Since the blending ingredients and method can be achieved using conventional techniques, a detailed description thereof is omitted herein.

[0087] The present invention provides a feed composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof.

[0088] Preferably, a feed composition for antioxidant, wound healing, skin wrinkle improvement, skin whitening, skin soothing, or anti-aging, comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof, is provided.

[0089] The feed composition according to the present invention can exhibit antioxidant, wound healing, skin wrinkle improvement, skin whitening, skin soothing, and anti-aging effects.

[0090] In the present invention, the feed may be feed for fish, birds, or mammals, and preferably, feed for livestock or aquatic organisms that have been acclimated to the wild and are suitable for breeding, as defined in Article 2, Paragraph 1 of the Livestock Industry Act and Article 2, each clause of the Enforcement Decree of the same Act. The livestock may include cats, dogs, cows, horses, mules, donkeys, goats, goats, sheep, deer, pigs, rabbits, poultry, etc., and the poultry may include chickens, turkeys, ducks, ostriches, geese, quail, etc.

[0091] In addition, “livestock products” means meat, milk, eggs, honey and processed products thereof, raw hides (including raw fur), raw wool, and other livestock products produced from livestock, as defined in Article 2, Paragraph 3 of the Livestock Industry Act, as determined by the Ordinance of the Ministry of Agriculture and Forestry.

[0092] It may also include, but is not limited to, pets such as dogs and cats.

[0093] The term "feed" of the invention means any natural or artificial diet, meal, etc., or a component of said meal, which is intended for or suitable for an animal to eat, ingest, or digest. In one embodiment, the feed composition containing Fructobacillus fructosus PRC-1 KCTC15271BP of the present invention; a culture medium thereof; or an extracellular vesicle thereof; may include a concentrate feed, a forage, and / or a special feed.

[0094] The composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP of the present invention; a culture medium thereof; or an extracellular vesicle thereof; exhibits excellent antioxidant activity and can be utilized as an antioxidant composition, exhibits an excellent skin whitening effect by inhibiting melanin production, can exhibit improved efficacy on skin pigmentation diseases, facilitates wound healing and skin regeneration through wound healing, etc., and improves skin wrinkles and exhibits therapeutic efficacy on skin wrinkle-related diseases by inhibiting the activity of elastase. Accordingly, it has excellent effects as a food, pharmaceutical composition, cosmetic, and feed composition.

[0095] Hereinafter, in Figures 1 to 7, CSF refers to the Fructobacillus fructosus PRC-1 culture medium, and PC refers to the positive control.

[0096] Figure 1 is a diagram showing the 16S rRNA base sequence of Fructobacillus fructosus PRC-1 KCTC15271BP.

[0097] Figure 2 is a diagram showing the cytotoxicity of Fructobacillus fructosus PRC-1 culture solution.

[0098] Figure 3 is a diagram showing the effect of Fructobacillus fructosus PRC-1 culture solution on enhancing the migration ability of keratinocytes.

[0099] Figure 4 is a diagram showing the ROS production inhibitory effect of Fructobacillus fructosus PRC-1 culture solution.

[0100] Figure 5 is a diagram showing the inhibitory effect of Fructobacillus fructosus PRC-1 culture solution on melanin production.

[0101] Figure 6 is a diagram showing the elastase activity inhibitory effect of Fructobacillus fructosus PRC-1 culture solution.

[0102] Figure 7 is a diagram showing the inhibitory effect of extracellular vesicles derived from Fructobacillus fructosus PRC-1 on melanin production.

[0103] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention and are not intended to limit the scope of the present invention.

[0104] <Manufacturing Example 1> Manufacture of Fructobacillus fructosus PRC-1 KCTC15271BP strain, its culture solution, extracellular vesicles, dead cells, and lysate

[0105] 1. Isolation and identification of Fructobacillus fructosus PRC-1

[0106] Honey used in the experiment was collected from Jangseong-gun, South Korea. 2 g of the collected honey was dissolved in 10 ml of sterile phosphate buffered saline (PBS) and further diluted 10-fold. The diluted honey was spread on MRS solid medium and cultured at 30°C for 2 days, after which the strain was isolated. The isolated lactic acid bacteria strain was inoculated into MRS liquid medium, cultured at 30°C for 24 hours, and glycerol was added to a concentration of 25% (v / v) to create a glycerol stock, which was stored at -70°C and used.

[0107] The isolated strain was identified through 16S rRNA sequence analysis. The isolated strain was sent to Macrogen in the form of an MRS plate, and 16S rRNA sequence analysis was performed using the universal primer set of 27F (5'-AGAGTTTGATCMTGGCTCAG-3', SEQ ID NO: 2) and 1492R (5'-TACGGYTACCTTGTTACGACTT-3', SEQ ID NO: 3). A total of 1,475 bp of the 16S rRNA sequence of the isolated strain Fructobacillus fructosus PRC-1 was determined, and the results are shown in Fig. 1.

[0108] As can be seen in Figure 1, it showed 99% homology with the Fructobacillus fructosus strain, and was finally identified as Fructobacillus fructosus.

[0109] The isolated strain was named Fructobacillus fructosus PRC-1 and was deposited on January 5, 2023, at the Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology, 823 Sinjeong-dong, Jeongeup-si, Jeollabuk-do, Republic of Korea, and was assigned the accession number KCTC15271BP.

[0110] 2. Preparation of Fructobacillus fructosus PRC-1 culture solution

[0111] For the preparation of culture medium, live Fructobacillus fructosus PRC-1, a lactic acid bacterium isolated from honey, was added to DMEM medium (1*10 9 CFU / ml) was cultured for 24 hours, centrifuged, and the supernatant was separated. The supernatant was adjusted to pH 7.4, sterilized using a syringe filter (0.2 ㎛), and then stored.

[0112] 3. Preparation of extracellular vesicles derived from Fructobacillus fructosus PRC-1

[0113] Extracellular vesicles derived from Fructobacillus fructosus PRC-1 were isolated according to the following steps.

[0114] 1) The culture solution of Fructobacillus fructosus PRC-1 according to the above manufacturing example 1-2 was centrifuged at 2000 g for 10 minutes to separate and remove the strain from the culture solution.

[0115] 2) The culture solution centrifuged in step 1 was filtered through a 0.2㎛ bottle top filter to increase the purity of the culture solution.

[0116] 3) It was concentrated 10-fold through tangential flow filtration (TFF).

[0117] 4) The culture solution concentrated in step 3 above was filtered through a 0.22 ㎛ syringe filter.

[0118] 5) The culture solution filtered in step 4 above was centrifuged at 4°C, 150,000g for 3 hours to separate extracellular vesicles.

[0119] 6) After ultra-high-speed centrifugation in step 5 above, the supernatant was removed and the remaining extracellular vesicle pellet was resuspended in a buffer solution (PBS).

[0120] 4. Preparation of Fructobacillus fructosus PRC-1 dead cells

[0121] Fructobacillus fructosus PRC-1 KCTC15271BP was heat treated three times at 90°C for 10 minutes to produce dead cells.

[0122] 5. Preparation of Fructobacillus fructosus PRC-1 lysate (soluble and insoluble)

[0123] Fructobacillus fructosus PRC-1 (Fructobacillus fructosus PRC-1) KCTC15271BP was centrifuged and 1x10 6 Suspend in sterile distillation to CFU / mL, 4o Sonication at C (1 min treatment, 1 min rest, repeated 5 times) and centrifugation (10,000 g, 4 o C, 15 minutes) to obtain the supernatant (soluble matter) and precipitate (insoluble matter). This was freeze-dried and used.

[0124] <Example 1> Measurement of skin cell protection ability of Fructobacillus fructosus PRC-1 culture solution

[0125] 1. Cytotoxicity evaluation of Fructobacillus fructosus PRC-1 culture solution

[0126] Keratinocytes (HaCaT) were seeded in 96-well plates at 5X10 4 The cells were cultured for 16 hours in DMEM medium containing 10% (v / v) FBS at a concentration of 10 cells / ㎖. Thereafter, DMEM medium containing 1% (v / v) FBS with 2.5%, 5%, or 10% of the Fructobacillus fructosus PRC-1 culture solution according to Manufacturing Example 1-2 was treated, and DMEM medium containing 1% (v / v) FBS without the Fructobacillus fructosus PRC-1 culture solution was used as the control group.

[0127] After culturing for 24 h, 4 mg / ml MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reagent was diluted 10-fold in DMEM medium and treated for 4 h. Afterwards, formazan crystals were dissolved using DMSO, and the absorbance was measured at 540 nm using a spectrophotometer. Cytotoxicity was expressed as a percentage based on the absorbance measured in the control group.

[0128] When cells are treated with MTT, MTT is converted into formazan crystals due to a reduction reaction in the mitochondria within the cells, and cytotoxicity can be determined through the amount of formazan crystals produced.

[0129] The results of this experiment confirmed that the Fructobacillus fructosus PRC-1 culture solution did not affect cytotoxicity (see Figure 2). Based on these results, subsequent experiments were conducted using the Fructobacillus fructosus PRC-1 culture solution at a maximum concentration of 10%.

[0130] 2. Confirmation of the keratinocyte recovery ability of Fructobacillus fructosus PRC-1 culture solution.

[0131] Keratinocytes (HaCaT) were seeded at 3X10 in a 12-well plate with 2-well silicone attached. 5 The cells were cultured for 16 hours in DMEM medium containing 10% (v / v) FBS at a concentration of 10 cells / ㎖. The medium was replaced with a medium not containing FBS, and the silicone was removed to create a cell-free gap. Thereafter, the medium was replaced with DMEM medium containing 1% (v / v) FBS supplemented with 2.5%, 5%, or 10% of the Fructobacillus fructosus PRC-1 culture medium according to Manufacturing Example 1-2. After 1 hour of culture, hydrogen peroxide was treated at a concentration of 500 μM, and the cells were imaged after 0 h, 12 h, and 24 h. In addition, the area of ​​the cell-free gap in the image was quantified using the Image J program.

[0132] This experiment aimed to determine the effect of Fructobacillus fructosus PRC-1 culture on the migratory ability of keratinocytes, which are essential for wound healing and regeneration. First, when keratinocytes were stressed by treating them with hydrogen peroxide, it was confirmed that the migratory ability of the cells was reduced and the cell-free gap was maintained. However, when treated with Fructobacillus fructosus PRC-1 culture, the cell migratory ability increased despite the stress caused by hydrogen peroxide treatment, and it was confirmed that the cell-free gap disappeared at all concentrations after 24 h of treatment (see Figures 3A and 3B).

[0133] That is, it was confirmed that Fructobacillus fructosus PRC-1 culture solution increases the migratory ability of keratinocytes and contributes to wound healing and regeneration.

[0134] <Example 2> Measurement of the ability of Fructobacillus fructosus PRC-1 culture solution to inhibit intracellular ROS production

[0135] 1. ROS measurement (DCF-DA assay) method

[0136] Keratinocytes (HaCaT) were seeded in 12-well plates at 5X10 4 Cells were cultured in DMEM medium containing 10% (v / v) FBS at a concentration of 10 cells / ㎖ for 16 hours, and then treated with DMEM medium containing 1% (v / v) FBS supplemented with 2.5%, 5%, or 10% Fructobacillus fructosus PRC-1 culture. After 1 hour, hydrogen peroxide was treated at a concentration of 500 μM and cultured for 3 hours. Afterwards, the cells were stained with 10 μM DCF-DA for 20 minutes, and the generated ROS were imaged using a fluorescence microscope. In addition, the relative level of the generated ROS was quantified using the Image J program.

[0137] 2. Evaluation of the ability of Fructobacillus fructosus PRC-1 culture solution to inhibit intracellular ROS production

[0138] DCF-DA, a non-fluorescent substance, is oxidized by ROS to the fluorescent DCF. Using this principle, the DCF-DA assay was used to confirm the inhibitory ability of Fructobacillus fructosus PRC-1 culture to inhibit intracellular ROS production.

[0139] In an experiment where Fructobacillus fructosus PRC-1 culture was treated together with hydrogen peroxide, it was confirmed that Fructobacillus fructosus PRC-1 culture inhibited ROS production as fluorescence was not observed (see Figures 4A and 4B). In particular, compared to the hydrogen peroxide control group which showed strong fluorescence, it was confirmed that the Fructobacillus fructosus PRC-1 culture strongly inhibited ROS production starting from the low concentration treatment of 2.5%.

[0140] <Example 3> Evaluation of the whitening improvement efficacy of Fructobacillus fructosus PRC-1 culture solution

[0141] 1. Method for measuring the inhibition rate of melanin biosynthesis

[0142] B16F10 cell line, a human-derived malignant melanoma cell line, was purchased from ATCC and used in the experiment. B16F10 cells were cultured and 1X10 cells were seeded per well in a 12-well plate. 5 After dispensing at cells / ml, the cells were cultured for 24 hours for attachment and stabilization. After the culture was completed, DMEM medium containing 1% (v / v) FBS supplemented with 2.5%, 5%, or 10% Fructobacillus fructosus PRC-1 culture medium or 2% Arbutin, known as a whitening agent, was treated for 1 hour. Afterwards, α-melanocyte-stimulating hormone (α-MSH), a melanin-stimulating hormone, was treated, and the melanin production concentration was expressed as a percentage compared to the control group. After 48 hours, each well was washed with PBS, 500 ㎕ of 1 N NaOH solution was added, and the cells were dissolved at 60°C for 3 hours. The absorbance was measured at 475 nm using a microplate reader.

[0143] 2. Evaluation of the inhibition rate of melanin biosynthesis by lactic acid bacteria culture solution

[0144] Excessive melanin synthesis and accumulation on the skin surface leads to hyperpigmentation symptoms such as freckles and melasma. Experimental results showed that α-MSH treatment in B16F10 cells induced melanin production by 193.6%, while treatment with 2% arbutin, the positive control, inhibited melanin production by 115.8%.

[0145] Meanwhile, Fructobacillus fructosus PRC-1 culture inhibited melanin production in a concentration-dependent manner. In particular, treatment with 10% Fructobacillus fructosus PRC-1 culture inhibited melanin production by up to 70%, resulting in a decrease in melanin production compared to the control group not exposed to α-MSH. (See Fig. 5).

[0146] <Example 4> Evaluation of the wrinkle improvement efficacy of Fructobacillus fructosus PRC-1 culture solution

[0147] 1. Method for measuring elastase inhibition

[0148] 30 μL of the test substance was added to 170 μL of 0.2 M Tris-HCl buffer (pH 8.0), and 50 μL of porcine pancreas elastase diluted to 0.5 units / mg was added, and then left at room temperature for 10 minutes. The test substance was 2.5%, 5%, or 10% Fructobacillus fructosus PRC-1 culture medium, or DMEM medium containing 1% (v / v) FBS supplemented with 2 mg / mL of vitamin C as a positive control. Afterwards, 50 μL of N-succinyl-(L-Ala)3-p-nitroanilide, a substrate, was diluted to 2 mg / mL and added, and the amount of p-nitroanilide produced was measured using a microplate reader at a wavelength of 410 nm. The elastase activity inhibition ability was calculated using the following equation 1.

[0149] [Formula 1]

[0150] Elastase inhibition (%) = (Control absorbance - Experimental absorbance) / Control absorbance * 100

[0151] 2. Evaluation of the Elastase Activity Inhibition Ability of Fructobacillus fructosus PRC-1 Culture Solution

[0152] Elastase, an elastin-degrading enzyme known to be the cause of wrinkles, reacts with its substrate, N-succinyl-(L-Ala)3-p-nitroanilide, to produce p-nitroanilide. In this experiment, the elastase activity inhibitory ability of Fructobacillus fructosus PRC-1 culture was evaluated through changes in the amount of p-nitroanilide produced.

[0153] As a result, it was confirmed that the positive control group treated with Vitamin C inhibited the activity of Elastase by approximately 95%. In the case of Fructobacillus fructosus PRC-1 culture, the higher the concentration, the higher the inhibition effect of Elastase activity, and in particular, it was confirmed that the group treated with 10% of Fructobacillus fructosus PRC-1 culture showed inhibition of up to 85% (see Figure 6).

[0154] <Example 5> Evaluation of the whitening improvement efficacy of extracellular vesicles derived from Fructobacillus fructosus PRC-1

[0155] 1. Evaluation of the inhibition rate of melanin biosynthesis by extracellular vesicles derived from Fructobacillus fructosus PRC-1

[0156] The inhibition rate of melanin biosynthesis of extracellular vesicles derived from Fructobacillus fructosus PRC-1 was measured in the same manner as in Example 3-1 above, but 0.5x10 of extracellular vesicles derived from Fructobacillus fructosus PRC-1 was used instead of the Fructobacillus fructosus PRC-1 culture medium. 8 particles / ml, 5x10 8 particles / ml or 50x10 8 particles / ml were used.

[0157] As a result of the experiment, it was confirmed that melanin production was induced by 219.1% by α-MSH treatment in B16F10 cells, and melanin production was inhibited by 142.5% by treatment with 2% Arbutin as a positive control.

[0158] Meanwhile, extracellular vesicles derived from Fructobacillus fructosus PRC-1 inhibited melanin production in a concentration-dependent manner. In particular, 50x10 extracellular vesicles 8 When treated with particles / ml, melanin production was inhibited by up to 125.2%, confirming that melanin production was inhibited more strongly than the positive control group (see Figure 7).

[0159] Through the above results, it was confirmed that Fructobacillus fructosus PRC-1, its culture solution, or its extracellular vesicles according to the present invention have excellent efficacy in inhibiting reactive oxygen species (ROS) production, inhibiting melanin production, promoting wound healing and regeneration, and inhibiting elastase activity, and thus it is expected to have high utility as a cosmetic, pharmaceutical, and food agent.

[0160] [Accession number]

[0161] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0162] Accession number: KCTC15271BP

[0163] Date of acceptance: 20230105

[0164]

[0165]

Claims

1. A cosmetic composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

2. A cosmetic composition according to claim 1, wherein the cosmetic composition is used for any one purpose selected from the group consisting of anti-oxidation, wound healing, skin whitening, and wrinkle improvement.

3. A cosmetic composition according to claim 1, wherein Fructobacillus fructosus PRC-1 KCTC15271BP; its culture medium; or its extracellular vesicles exhibit at least one effect selected from the group consisting of inhibition of reactive oxygen species (ROS) production, inhibition of melanin production, wound healing and regeneration, and inhibition of elastase activity.

4. A cosmetic composition according to claim 1, wherein the cosmetic composition comprises a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

5. A cosmetic composition according to claim 1, wherein the Fructobacillus fructosus PRC-1 KCTC15271BP has 16S rRNA of sequence number 1.

6. Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or a food composition for antioxidant, wound healing, skin whitening, or wrinkle improvement comprising an extracellular vesicle thereof.

7. A food composition according to claim 6, wherein the Fructobacillus fructosus PRC-1 KCTC15271BP has 16S rRNA of sequence number 1.

8. In the 6th paragraph, the food composition comprises a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

9. Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or a pharmaceutical composition for preventing or treating a wound, skin pigmentation disease, or skin wrinkle-related disease, comprising an extracellular vesicle thereof.

10. A pharmaceutical composition according to claim 9, wherein the Fructobacillus fructosus PRC-1 KCTC15271BP has 16S rRNA of sequence number 1.

11. A pharmaceutical composition according to claim 9, wherein the pharmaceutical composition comprises a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

12. A pharmaceutical composition according to claim 9, wherein the wound is at least one selected from the group consisting of burns, thermal injuries, epidermal wounds, ulcers, trauma, post-surgical, childbirth, chronic wounds, damage due to dermatitis, corneal ulcers, corneal epithelial detachment, keratitis, and damage due to dry eye.

13. A pharmaceutical composition according to claim 9, wherein the skin pigmentation disease is at least one selected from the group consisting of lentigines, freckles, hypopigmentation, and hyperpigmentation.

14. A pharmaceutical composition according to claim 9, wherein the skin wrinkle-related disease is at least one selected from the group consisting of elastic fibrosis, thinning of the skin, skin atrophy, reduction of collagen fibers and elastic fibers, loss of skin elasticity, dryness, wrinkle formation, and premature skin aging.

15. Feed composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof.

16. A feed composition according to claim 15, wherein the feed composition is used for any one purpose selected from the group consisting of antioxidant, wound healing, skin whitening, and wrinkle improvement.

17. In claim 15, the feed composition comprises a culture solution of Fructobacillus fructosus PRC-1 KCTC15271BP or extracellular vesicles thereof.

18. A pharmaceutical composition comprising Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; and a pharmaceutically acceptable carrier.

19. A method for preventing or treating a wound, skin pigmentation disease, or skin wrinkle-related disease, comprising administering to a subject in need thereof Fructobacillus fructosus PRC-1 KCTC15271BP; a culture solution thereof; or an extracellular vesicle thereof.

20. Use of Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; in the manufacture of a medicament for treating at least one disease selected from the group consisting of wounds, skin pigmentation diseases, and skin wrinkle-related diseases.

21. Fructobacillus fructosus PRC-1 KCTC15271BP; a culture medium thereof; or an extracellular vesicle thereof; for use in the prevention or treatment of any one or more diseases selected from the group consisting of wounds, skin pigmentation diseases, and skin wrinkle-related diseases.

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