KR20240014021A - Streptococcus mutans strain and its use for improving skin conditions - Google Patents

Abstract

It relates to a novel Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, and their use for improving skin condition. Since the above strain has an effect of improving skin condition, it can be usefully used in cosmetics, medicine, food, etc.

Description

Streptococcus mutans strain and its use for improving skin conditions {Streptococcus mutans strain and its use for improving skin conditions}

It relates to novel microorganisms, their lysate, culture medium, and extract of the culture medium, and their use for improving skin condition.

The skin ecosystem provides various types of habitats for microorganisms, and a wide range of microorganisms live there. They form a symbiotic relationship with their human hosts, and are known to have many positive effects on the host. The skin constitutes various types of habitats, including indentations and specialized crevices, and helps a wide range of microorganisms to grow. Basically, the skin forms a physical membrane and helps defend against potential hazards and toxic substances from the outside. The skin serves as a point of contact with the external environment and is also a home for various microorganisms (e.g. fungi, bacteria, viruses, and small larvae). Depending on the selection of physical and chemical functions, microorganisms adapt to specialized niches and establish habitats. Typically, the skin remains cool, acidic, and dry. Structurally, the epidermis forms the skin barrier and plays an important role in blocking the penetration of microorganisms and toxins and maintaining moisture. The uppermost layer of the epidermis is composed of the stratum corneum. The epidermis has a form called a 'brick and mortar structure'. The skin tissue goes through a continuous self-recovery process, and the scales (squames) that have completed the differentiation process constantly repeat the process of being shed from the skin tissue.

Probiotics are a general term for microorganisms that have beneficial effects on the human body. They refer to microorganisms that benefit our bodies. Most probiotics known to date are known as lactic acid bacteria. Probiotics have been reported to produce effective effects through various beneficial effects on the human body, but research on the relationship between skin flora and skin is insufficient.

Accordingly, there is a need to develop skin flora that can be useful for skin-related conditions.

One aspect is to provide novel Streptococcus mutans strains.

Another aspect is to provide a lysate, culture medium, or extract of the culture medium of the strain.

Another aspect provides the use of a Streptococcus mutans strain, its lysate, culture, or extract of the culture.

Another aspect is to provide a composition comprising a Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.

Another aspect is to provide a method of preventing, ameliorating, or treating a condition in a subject comprising administering an effective amount of the composition described above to the subject in need thereof.

One aspect provides a Streptococcus mutans strain.

The strain may be isolated. In this specification, the term “isolated” means something that does not exist in the natural world but can be artificially separated and used. The strain may be isolated from human skin. The strain may be a strain isolated by inoculating human skin cells into TSB (Tryptic Soy Broth) or R2A (Reasoner's 2A) medium and pure isolation and culturing of the cultured colonies.

The Streptococcus mutans strain may be a strain belonging to the Streptococcus genus ( Streptococcus sp.).

The Streptococcus mutans strain may be a strain deposited under the deposit number KCCM13190P. The strain may be Streptococcus mutans CX108 strain.

The Streptococcus mutans strain may contain 16S rRNA of SEQ ID NO: 1. In one embodiment, the strain is at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, at least about 99.9%, at least 95% of SEQ ID NO. Comprising 16S rRNA having a sequence identity of 100%, 96% to 100%, 97% to 100%, 98% to 100%, 99% to 100%, 99.5% to 100%, or 99.9% to 100% You can.

As used herein, the term “sequence identity” refers to the degree of identity of amino acid residues or bases between sequences after aligning both sequences to be as consistent as possible in a specific comparison region. Sequence identity can be confirmed according to methods known in the art. The percent sequence identity can be determined using known sequence comparison programs, examples of which include BLASTN (NCBI), CLC Main Workbench (CLC bio), MegAlignTM (DNASTAR Inc), etc.

The strain may have an effect of improving skin condition, improving skin beauty, or preventing, improving, or treating skin diseases. For example, the strain may have anti-skin aging, skin wrinkle improvement, skin elasticity enhancement, skin moisturizing, skin barrier strengthening, skin whitening, anti-inflammatory, or anti-allergic effects. In addition, the strain may have the effect of preventing, treating, or improving diseases caused by impaired skin barrier function, melanin hyperpigmentation diseases, skin inflammatory diseases, or skin allergic diseases.

In one embodiment, the strain may have a skin moisturizing or skin barrier strengthening effect.

In one embodiment, the strain may increase the expression of aquaporin (Aquaporin 3, AQP3).

Another aspect provides a lysate, culture fluid, or extract of the culture fluid of the strain.

Details of the strain are as described above.

As used herein, the term “lysate” may be used interchangeably with “lysate” and may refer to a product obtained by breaking the cell wall of a strain by chemical or physical force. The lysate may include the lysate itself, its concentrate, or its freeze-dried product.

As used herein, the term “culture medium” may be used interchangeably with “culture supernatant,” “conditioned culture medium,” or “conditioned medium,” and refers to a medium that can supply nutrients to enable the strain to grow and survive in vitro. It may refer to the entire medium containing the strain, its metabolites, extra nutrients, etc. obtained by culturing the strain for a certain period of time. Additionally, the culture medium may refer to a culture medium obtained by removing the bacterial cells from the bacterial culture medium obtained by culturing the strain. On the other hand, the liquid from which the bacteria have been removed from the culture medium is also called "supernatant", and the supernatant is obtained by leaving the culture solution still for a certain period of time and taking only the upper liquid excluding the part that has settled in the lower layer, or by removing the bacteria through filtration, or by removing the culture liquid. It can be obtained by centrifuging to remove the sediment at the bottom and taking only the liquid at the top. The "bacteria" refers to the strain itself of the present invention, and includes the strain itself isolated and selected from a skin sample, etc., or a strain isolated from the culture medium by culturing the strain. The bacterial cells can be obtained by centrifuging the culture medium and taking the part that sinks to the lower layer. Alternatively, since they sink to the lower layer of the culture medium by gravity, they can be obtained by leaving them still for a certain period of time and then removing the upper liquid.

Culture media and culture conditions for cultivating the above strain can be appropriately selected or modified by those skilled in the art. For example, the culture medium is cultured by culturing the strain in an appropriate medium (e.g., R2A medium, TSA medium, or TSB medium) at an appropriate temperature (e.g., either above 10°C or below 40°C) for a certain period of time (e.g., 4 to 120 hours). ) may have been obtained by culturing for a period of time.

The culture medium may include the culture medium itself, its concentrate, or freeze-dried product obtained by culturing the strain. The culture medium may include a culture supernatant obtained by removing the strain from the culture medium, a concentrate thereof, or a freeze-dried product.

In one embodiment, the culture supernatant of the strain may be obtained by removing the strain by centrifuging or filtering the strain culture fluid.

In another embodiment, the concentrate may be obtained by concentrating the strain culture itself or the supernatant obtained after centrifuging or filtering the culture. As used herein, the term "culture extract" refers to an extract from the culture medium or its concentrate, and may include extracts, diluted or concentrated extracts, dried products obtained by drying the extracts, crude or purified products thereof, and fractions thereof. You can.

The strain, its lysate, culture medium, or extract of the culture medium may be biologically pure. The strain may be obtained by pure culture.

Another aspect provides the use of a Streptococcus mutans strain, its lysate, culture, or extract of the culture. In detail, a composition comprising a Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof is provided.

In the above use or composition, the strain may be a Streptococcus mutans strain according to the above aspect. Therefore, the strain may be the Streptococcus mutans CX108 strain.

Uses of the strain may include improving skin condition, improving skin beauty, or preventing, improving or treating skin diseases.

The improvement of skin condition or skin beauty may include preventing skin aging, improving skin wrinkles, enhancing skin elasticity, moisturizing skin, strengthening skin barrier, whitening skin, anti-inflammatory, or anti-allergy.

In one embodiment, the improvement of skin condition or skin beauty may be skin moisturizing or skin barrier strengthening.

As used herein, the term “skin aging” refers to both tangible and intangible changes that occur in the skin as one ages. Skin aging includes, for example, a thinning of the epidermis, a decrease in the number of cells or blood vessels in the dermis, a decrease in the ability to repair DNA damage, an increase in the cell turnover cycle, a decrease in the ability to heal wounds, a weakening of the skin barrier function, a decrease in the moisture retention ability of the epidermis, These include reduced sweat secretion, reduced sebum secretion, reduced vitamin D production, reduced ability to defend against physical damage, reduced ability to eliminate chemicals, reduced immune response, weakened sensory function, and reduced body temperature regulation.

The prevention of skin aging may mean improving skin aging caused by extrinsic or endogenous factors. The exogenous factors refer to various external factors, such as ultraviolet rays (light). The endogenous factors are also referred to as chronological factors and mainly refer to factors that occur due to the passage of time. In other words, the skin aging is specifically not only a symptom of premature aging induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age. It is a concept that includes wrinkles, loss of elasticity, skin sagging and dryness, etc. In addition, wrinkles include stimulation caused by changes in internal and external factors that change the components that make up skin tissue, causing wrinkles.

The aging may be photoaging. The term “photoaging” is a phenomenon caused by external environmental factors, the most representative factor being ultraviolet rays. Ultraviolet rays cause damage to biological components such as activation of proteolytic enzymes, chain cutting of matrix proteins, and abnormal cross-linking, and repetition of this mechanism causes skin aging that is evident in appearance.

The term “wrinkle” refers to a state in which the skin loses its elasticity and becomes loose, for example, the skin may be folded. “Prevention or improvement of skin wrinkles” may refer to any action that prevents or improves wrinkles by suppressing the expression of factors related to wrinkles, or increases the total amount of collagen.

The term “promoting skin elasticity” may refer to any action that improves the elasticity of loose skin.

The term “skin moisturizing” may refer to any action that maintains skin moisture or prevents moisture loss.

The “skin barrier strengthening” may refer to any action that improves the function of the skin barrier, which is located on the outermost layer of the skin and prevents moisture and nutrition loss.

The “skin whitening” may mean not only brightening skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or genetics.

The term “anti-inflammatory” may be used interchangeably with “improvement of inflammation” and “inhibition of inflammation,” and may include, for example, all actions that inhibit the expression of inflammatory cytokines.

The term “anti-allergy” may be used interchangeably with “allergy improvement” and “allergy suppression,” for example, removing or suppressing allergens (antigens that cause allergic diseases) or alleviating allergic reactions caused by allergens. It can include all actions required.

The “skin disease” may be a disease caused by damage to the skin barrier function, skin aging, melanin hyperpigmentation disease, skin inflammatory disease, or skin allergic disease.

The damage to the skin barrier function may mean any change that appears in the skin due to decreased or damaged skin barrier function. For example, it may include increased skin wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, etc.

The term “melanin hyperpigmentation” is a broad concept that includes all melanin hyperpigmentation caused by various causes, and means that certain areas of the skin, fingernails, or toenails become darker than other areas due to an excessive increase in melanin pigment. .

The melanin hyperpigmentation disease includes freckles; melasma; chloasma; liver spots; nevus; solar lentigo; melanosis; peutz-jegher's syndrome; Chloasma gravidarum; Hyperpigmentation after drug use; and postinflammatory hyperpigmentation, but is not limited thereto.

The skin inflammatory disease may be any one selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, drug rash, and acne.

The skin allergic disease may include atopic dermatitis, allergic dermatitis, urticaria, itching, rash, etc.

The term “prevention” includes suppressing the occurrence of a disease. The term “treatment” includes inhibiting, alleviating, or eliminating the development of a disease. The term “improvement” may refer to any action that results in at least reducing the severity of a parameter associated with the alleviation or treatment of a condition, such as symptoms.

In one embodiment, the strain, its lysate, culture medium, or extract of the culture medium can increase the expression of AQP3. Therefore, the strain, its lysate, culture medium, or extract of the culture medium may exhibit a skin moisturizing or skin barrier strengthening effect.

The strain can be used with other strains that have a skin-improving effect, such as strains belonging to the Streptococcus genus, to exhibit a synergistic effect. Strains belonging to the Streptococcus genus include, for example, Streptococcus equinus , Streptococcus australis, Streptococcus downii , Streptococcus halichoeri , and Streptococcus. Streptococcus intermedius , Streptococcus lutetiensis, Streptococcus massiliensis, Streptococcus pseudopneumoniae , Streptococcus oralis , Streptococcus Streptococcus parasanguinis, Streptococcus anginosus, Streptococcus pyogenes , Streptococcus rubneri, Streptococcus salivarius , Streptococcus sanguis ( Streptococcus sanguinis ), Streptococcus sobrinus , and Streptococcus vestibularis .

The composition may include the strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof as an active ingredient.

The term “included as an active ingredient” means that the strain of the present specification, its lysate, culture medium, or extract of the culture medium is added to the extent that it can exhibit the effects mentioned herein, and various ingredients are added for drug delivery and stabilization, etc. This means that it is formulated into various forms by adding as a sub-ingredient.

The strain, its lysate, culture medium, or extract of the culture medium is 0.0001% by weight to 99.9999% by weight, for example, 0.001% by weight to 80% by weight, 0.01% by weight to 60% by weight, 0.01% by weight, based on the total weight of the composition. to 40% by weight, 0.01% to 30% by weight, 0.01% to 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight. Weight %, 0.05 weight % to 30 weight %, 0.05 weight % to 20 weight %, 0.05 weight % to 10 weight %, 0.05 weight % to 5 weight %, 0.1 weight % to 60 weight %, 0.1 weight % to 40 weight % , 0.1 wt% to 30 wt%, 0.1 wt% to 20 wt%, 0.1 wt% to 10 wt%, or 0.1 wt% to 5 wt%, but is not particularly limited thereto. The content of the active ingredient can be selected by a person skilled in the art at an appropriate amount that can be formulated within a content range that can produce the effects mentioned in this specification.

The composition may be in a liquid state or a dry state. In one embodiment, the composition may be in dry powder form.

The drying method for preparing the composition in a dry state may be a method generally used in the art and is not particularly limited. Non-limiting examples of the drying method include air drying method, natural drying method, spray drying method, freeze drying method, etc. These methods can be used alone or at least two methods can be used together.

The composition may contain an effective amount of additives sufficient to reduce deterioration of the strain, its lysate, culture medium, or extract of the culture medium. The additive may be, for example, a binder, but is not limited thereto.

The composition may further include a cosmetically, pharmaceutically, or foodologically acceptable carrier. The composition can be formulated with a carrier and provided as cosmetics, drugs, food additives, etc.

The composition may be a cosmetic composition.

In addition to the active ingredients disclosed in this specification, the cosmetic composition may further include ingredients commonly used in cosmetic compositions, functional additives, etc., such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, emulsifiers, and preservatives. , conventional auxiliaries such as vitamins, pigments, fragrances, etc., and carriers.

The cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected depending on the purpose. The cosmetic composition may have, for example, a solubilized formulation, an emulsified formulation, or a dispersed formulation. The cosmetic composition includes, for example, lotion, cream, essence, cleansing foam, cleansing water, pack, ampoule, body lotion, body oil, body gel, shampoo, rinse, hair conditioner, hair gel, foundation, lipstick, mascara, and makeup. It may have a base or skin-adhesive cosmetic formulation. The skin-adhesive cosmetic formulation may be, for example, a mask pack, but is not limited thereto.

Additionally, the composition may be a composition for external skin application.

The external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The skin external preparations include ingredients commonly used in external skin preparations such as cosmetics and medicines, such as aqueous ingredients, oil-based ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, metal ion sequestrants, sugars, or combinations thereof may be appropriately mixed as needed.

The composition may be a pharmaceutical composition.

The pharmaceutical composition may additionally include 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, disintegrant, binder, 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, calcium monohydrogen phosphate anhydride, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated as an oral or parenteral dosage form. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrup, etc. Parenteral dosage forms may be injections, ointments, etc.

The composition may be a food composition. The composition may be a health functional food composition. In this case, it can be formulated in the form of a typical health functional food known in the art.

The food composition may use the strain, its lysate, culture medium, or extract of the culture medium alone, or may be used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). There are no particular restrictions on the types of health functional foods. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary beverages. The natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; and polysaccharides such as dextrins and cyclodextrins; Sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The food composition is also used in nutritional supplements, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonated beverages. It may contain a carbonating agent, or a combination thereof. The food composition may also contain pulp for the production of natural fruit juice, fruit juice beverage, vegetable beverage, or a combination thereof.

Another aspect provides a method of preventing, ameliorating, or treating a condition in a subject comprising administering an effective amount of the composition described above to the subject in need thereof.

The condition of the subject may be a condition related to the skin, and specifically, it may be a disease caused by damage to the skin barrier function, skin aging, melanin hyperpigmentation disease, skin inflammatory disease, or skin allergic disease.

The term “effective amount” refers to an amount effective enough to produce the above-mentioned effects.

As used herein, the terms “administering,” “introducing,” and “implanting” are used interchangeably and refer to an individual by a method or route that results in at least partial localization of the composition according to one embodiment to the desired site. It may refer to the arrangement of the composition according to one embodiment into the inner space.

Administration may be by methods known in the art. The administration method, such as administration route and administration frequency, can be appropriately selected by a person skilled in the art. Administration may be administered directly to the subject by any means, such as, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. You can. The administration may be administered systemically or locally. The administration may include application to the skin.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an entity in need of skin condition improvement, for example, an entity in need of anti-aging skin, improvement of skin wrinkles, enhancement of skin elasticity, skin moisturization, skin barrier strengthening, skin whitening, anti-inflammatory, or anti-allergy effect. You can.

The administration of the composition according to one embodiment is 0.1 mg to 1,000 mg per day, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1 mg. 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and those skilled in the art will Taking these factors into consideration, the dosage can be adjusted appropriately. The frequency of administration can be once a day or two or more times within the range of clinically acceptable side effects. Regarding the administration site, it can be administered at one or two or more locations, and the total number of administration days can be from 1 to 30 days for one treatment, either daily or at intervals of 2 to 5 days. If necessary, the same treatment can be repeated after an appropriate period. For animals other than humans, the dosage per kg is the same as for humans, or the above dosage is converted into, for example, the volume ratio (e.g., average value) of the organs (heart, etc.) between the target animal and human. One dose can be administered.

Another aspect provides the use of the strain, its lysate, culture, or extract of the culture for use in preparing a composition for improving skin condition.

Another aspect provides the use of said strain, its lysate, culture, or extract of the culture for use in preparing a medicament for the prevention or treatment of melanin hyperpigmentation disease, skin inflammatory disease, or skin allergic disease.

Redundant content is omitted in consideration of the complexity of the present specification, and terms not otherwise defined in the present specification have meanings commonly used in the technical field to which the present invention pertains.

According to one aspect, the new Streptococcus mutans strain has an effect of improving skin condition such as moisturizing the skin or strengthening the skin barrier, so it can be usefully used in cosmetics, medicine, food, etc.

Figure 1 is a graph showing the relative expression level of AQP3. Negative control: medium treated + untreated group with strain culture, RA: group treated with retinoic acid (positive control), Example 2: group treated with CX108 strain culture.

This will be described in more detail through examples below. However, these examples are intended to illustrate one or more embodiments and the scope of the present invention is not limited to these examples.

[Example]

Example 1. Isolation and identification of strains

Streptococcus mutans strains were isolated and identified from the skin of healthy women.

Specifically, the cheeks, tip of the nose, and alar border of a healthy woman were swabbed two to three times with a sterile cotton swab soaked in PBS (phosphate buffer saline). A cotton swab was placed in a tube containing PBS and vortexed for 10 seconds. 10-fold, 100-fold, and 1,000-fold dilutions of the sample were prepared, and 100 μL each was added to 0.1X TSB (Tryptic Soy Broth) and 0.5X R2A (Reasoner's 2A) agar medium and spread with a spreader. After culturing the plate at 30°C or 37°C, 100 colonies formed were isolated and cultured in pure form, and re-cultured in an incubator at 28°C for 48 hours. The 16S rRNA gene sequence of the colonies after completion of culture was identified. The primers used at this time were designed to react and amplify only bacteria, and the primer sequences are as follows. 27F, forward primer: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO: 2); 1492R, reverse primer: 5'-GGTTACCTTGTTACGACTT-3 (SEQ ID NO: 3). PCR amplification was performed in 30 cycles of 95°C for 1 minute, 55°C for 1 minute, and 75°C for 1 minute and 30 seconds, and was finally treated at 72°C for 8 minutes and stored at 4°C. After the PCR reaction was completed, the DNA sequences of the isolated and cultured species were determined using ABI-3730XL (ABI, USA).

Using the BLAST program provided on the US National Center for Biotechnology Information (NCBI) website, the nucleotide sequence of the 16S rRNA region determined among the isolated and cultured microbial colonies was analyzed and compared with other strains registered at NCBI. Among them, a novel microorganism Streptococcus mutans strain (hereinafter referred to as “CX108”) with 99.78% homology was selected.

The selected CX108 strain was deposited at the Korean Culture Center of Microorganisms (KCCM) on May 19, 2022, and was given the deposit number KCCM13190P. Strain CX108 has the 16S rRNA sequence of SEQ ID NO: 1 (complementary DNA).

Example 2. Preparation of strain culture medium

A culture medium of Streptococcus mutans CX108 strain was prepared.

Specifically, the CX108 strain isolated in Example 1 was cultured in TSB (Tryptic Soy Broth) medium. The cells were removed through centrifugation, and a culture medium of Streptococcus mutans CX108 strain was prepared through supernatant recovery and filtration.

[Experimental method]

Experimental method 1. Analysis of AQP3 expression level

AQP3 (Aquaporin 3) is a factor related to skin moisturization and skin barrier strengthening.

1-1. Cell culture and sample processing

HaCaT cells, a human keratinocyte cell line, were cultured in DMEM medium (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) containing 10% fetal bovin serum, and all cultures were performed at 37°C and 5% CO ₂ It was performed in an incubator. The cultured cell line was treated with 1% (w/w) of the strain culture medium and further cultured for 24 hours. As a positive control, 1 uM of retinoic acid (RA) was used.

1-2. Gene expression level analysis

The culture was recovered, RNA was isolated by adding 1 ml of Trizol (RNA iso, DAKARA, Japan), and RNA was quantified using Nanodrop 2000 (Thermo, USA), then incubated at 42°C for 55 minutes and at 70°C for 15 minutes. cDNA was synthesized by reacting for a while (Reverse Transcriptase Mix, ELPIS biotech, Korea). RT-PCR used Step One Plus (Applied Biosystems, USA), and Cybergreen (SYBR Green supermix, Applied Biosystems, USA) was added together with primers for AQP3 and cDNA to activate the polymerase at 94°C for 5 minutes. Afterwards, polymerization was performed in cycles of 95°C for 30 seconds, 54°C for 1 minute, and 72°C for 1 minute for 40 seconds. The expression level of the gene was finally analyzed through correction for the β-actin gene.

[Experimental example]

Experimental Example 1. Confirmation of skin moisturizing and skin barrier strengthening efficacy

In order to confirm the skin moisturizing and skin barrier strengthening efficacy of the CX108 strain culture medium, the expression level of AQP3 was analyzed.

Figure 1 is a graph showing the relative expression level of AQP3.

As a result, as shown in Figure 1, it was confirmed that Example 2 (strain culture) significantly increased the expression of AQP3 compared to the negative control.

Therefore, it was confirmed that the CX108 strain has skin moisturizing and skin barrier strengthening effects.

In summary, it was confirmed that Streptococcus mutans CX108 strain increased the expression of AQP3. Therefore, it was found that the CX108 strain had skin moisturizing and skin barrier strengthening effects.

Korea Center for Microbial Conservation (KCCM) KCCM13190P 20220519

Claims (10)

Streptococcus mutans strain belonging to the Streptococcus genus ( Streptococcus sp.). The strain according to claim 1, wherein the strain is deposited under deposit number KCCM13190P. The strain of claim 1, wherein the strain comprises 16S rRNA having 97% to 100% sequence identity with SEQ ID NO: 1. The method according to claim 1, wherein the strain increases the expression of AQP3 (Aquaporin 3). Culture medium of the strain of claim 1. A cosmetic composition comprising Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof. The cosmetic composition according to claim 6, which is for improving skin condition. The cosmetic composition according to claim 7, wherein the skin condition improvement is skin moisturizing or skin barrier strengthening. A composition for external application containing Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof. A food composition comprising a Streptococcus mutans strain, its lysate, culture medium, extract of the culture medium, or a mixture thereof.