NOVEL LACTOCOCCUS LACTIS SUBSPECIES LACTIS STRAIN AND ITS USE

Abstract

The present invention relates to a novel Lactococcus lactis subsp. lactis LB1022 strain and uses thereof. The novel Lactococcus lactis subsp. lactis LB1022 strain, isolated from fermented dairy products, is a useful functional lactic acid bacterium with excellent acid tolerance, bile tolerance, and intestinal epithelial cell adhesion ability. It also inhibits the production of any one inflammatory factor selected from the group consisting of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β), and inhibits the NF-kB pathway (Nuclear Factor Kappa B). Since it has an excellent anti-inflammatory effect by suppressing the gene expression of p65 or p50 in the inflammatory pathway, a composition containing live or killed cells of Lactococcus lactis subsp. lactis LB1022 strain or its culture solution as an active ingredient can be used for the prevention, treatment, or amelioration of inflammatory diseases.

Description

[Technical Field]

[0001] The present invention relates to a novel Lactococcus lactis subsp. lactis LB1022 strain and uses thereof. [Background technology]

[0002] Inflammation is a local defense mechanism triggered by external or internal stimuli, such as physical trauma, harmful chemicals, microbial infection, and irritating substances in the body's metabolic products. Inflammation is triggered by various mediators produced by damaged tissue and migrating cells, and the response varies depending on the cause of inflammation. Generally, the inflammatory response removes or neutralizes the cause of the disease, restoring the normal function of damaged tissues and cells. However, an abnormally excessive inflammatory response can also lead to disease. Inflammatory bowel disease (IBD), a disease caused by an inflammatory response, is a condition that causes chronic inflammation and ulcers in the intestines, with repeated periods of remission and recurrence. Environmental factors, genetic factors, and an excessive immune response caused by intestinal bacteria are reported to be the main causes of IBD. IBD includes ulcerative colitis and Crohn's disease, and these diseases are collectively diagnosed as IBD due to similar symptoms, prognosis, and treatment processes. Recently, in Korea, Westernized eating habits and changes in lifestyle have led to an increase in the number of cases of IBD diagnosed in people in their 20s to 40s. Immunosuppressants are commonly prescribed to treat inflammatory diseases, but these can cause side effects such as nausea, heartburn, headache, dizziness, anemia, and skin rashes. Therefore, there is a need to develop new therapeutic methods that are effective using natural materials without side effects. Lactic acid bacteria are intestinal bacteria that live symbiotically in the human digestive system and play a role in breaking down fiber and complex proteins to produce important nutrients. For lactic acid bacteria to exert their beneficial effects in the intestine, they must be able to survive in a low pH environment, such as gastric acid, and must be resistant to bile, as in the small intestine. The intestinal microbial environment may improve the host's gastrointestinal tract, and recently, new functional lactic acid bacteria have been reported that exhibit various physiological activities, such as improving the symptoms of gastrointestinal diseases. Summary of the Invention [Problem to be solved by the invention]

[0003] The present invention relates to a novel Lactococcus lactis subsp. lactis LB1022 strain and uses thereof. The novel Lactococcus lactis subsp. lactis LB1022 strain, isolated from fermented dairy products, is a useful functional lactic acid bacterium with excellent acid tolerance, bile tolerance, and intestinal epithelial cell adhesion ability. It inhibits the production of any one inflammatory factor selected from the group consisting of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β), and inhibits the NF-kB pathway (Nuclear Factor Kappa B). Since the LB1022 strain exhibits excellent anti-inflammatory effects by suppressing the expression of p65 or p50 genes in the LB1022 pathway, compositions containing live or killed Lactococcus lactis subsp. lactis LB1022 strain or its culture solution as active ingredients can be used to prevent, treat, or improve inflammatory diseases. [Means for solving the problem]

[0004] The present invention provides the Lactococcus lactis subsp. lactis LB1022 strain, deposited under accession number KCTC 15324BP. The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or ameliorating inflammatory diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a health food composition for preventing or alleviating hangovers, which contains live or killed cells of the strain or a culture solution thereof as an active ingredient.

[0005] The present invention also provides a health food composition for preventing or improving liver function, which contains live or killed cells of the strain or a culture solution thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating allergic skin diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or ameliorating allergic skin diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating skin damage, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient, and a cosmetic composition for preventing or improving skin damage, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient.

[0006] The present invention also provides a pharmaceutical composition for preventing or treating obesity, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or improving obesity, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating diabetes, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or ameliorating diabetes, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating muscle loss, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient.The present invention also provides a health functional food composition for preventing or improving muscle loss, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a health food composition for gastrointestinal protection, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient. [Effects of the Invention]

[0007] According to the present invention, Lactococcus lactis subsp. lactis LB1022 strain, a novel strain isolated from fermented dairy products, is a useful functional lactic acid bacterium with excellent acid tolerance, bile tolerance, and intestinal epithelial cell adhesion ability. It also has excellent anti-inflammatory effects by suppressing the production of any one inflammatory factor selected from the group consisting of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β), and by suppressing the gene expression of p65 or p50 in the Nuclear Factor Kappa B pathway. A composition containing live or killed cells of Lactis LB1022 strain or a culture solution thereof as an active ingredient can be used for the prevention, treatment, or amelioration of inflammatory diseases. [Brief explanation of the drawings]

[0008] [Figure 1] 1 is a graph showing an analysis of bile resistance and acid resistance of a novel strain, Lactococcus lactis subsp. lactis LB1022. [Figure 2]1 is a graph showing the results of analyzing the adhesion ability of a novel bacterial strain, Lactococcus lactis subsp. lactis LB1022, to intestinal epithelial cells. [Figure 3a] This is a graph showing the analysis of nitric oxide (NO) production levels in RAW264.7 cells in which an inflammatory response was induced to evaluate the anti-inflammatory effect of the novel strain, Lactococcus lactis subsp. lactis LB1022. [Figure 3b] This is a graph showing the analysis of pro-inflammatory cytokine secretion levels in RAW264.7 cells in which an inflammatory response was induced, in order to evaluate the anti-inflammatory effect of the novel strain, Lactococcus lactis subsp. lactis LB1022. [Figure 3c] This is a graph showing the analysis of gene expression levels of NF-kB subunits p65 and p50 in RAW264.7 cells in which an inflammatory response was induced, in order to evaluate the anti-inflammatory effect of the novel strain, Lactococcus lactis subsp. lactis LB1022. [Figure 4] 1 is a graph showing the effectiveness of a novel strain, Lactococcus lactis subsp. lactis LB1022, in treating hangovers, by measuring alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities. [Figure 5a]This is a graph showing the measurement of blood AST (Aspartate Aminotransferase) and ALT (Alanine aminotransferase) levels, which are indicators of alcoholic liver damage, in alcohol-fed SD rats to evaluate the liver function protecting efficacy of the new strain, Lactococcus lactis subsp. lactis LB1022. [Figure 5b] This is a graph showing the measurement of the expression levels of factors involved in liver protection in alcohol-fed SD rats to evaluate the liver function protecting efficacy of the new strain, Lactococcus lactis subsp. lactis LB1022. [Figure 6a] 1 is a graph showing the mRNA expression levels of serum IgE and allergic dermatitis-related factors in order to evaluate the efficacy of a novel strain, Lactococcus lactis subsp. lactis LB1022, on allergic diseases. [Figure 6b] This is a graph showing serum IgG1, IgG2a, and cytokine expression levels in bronchoalveolar lavage fluid (BALF) to evaluate the efficacy of the novel strain, Lactococcus lactis subsp. lactis LB1022, against allergic diseases. [Figure 7] The results show that the skin regeneration efficacy of the new strain, Lactococcus lactis subsp. lactis LB1022, was evaluated using human skin epithelial cells to evaluate its efficacy on skin damage. [Figure 8]In order to evaluate the efficacy of the novel strain Lactococcus lactis subsp. lactis LB1022 against obesity, C57BL / 6 mice were used to evaluate its efficacy in reducing body fat. [Figure 9a] The results show that the inhibitory effect of glycolytic enzyme activity on the novel strain Lactococcus lactis subsp. lactis LB1022 was evaluated to assess its efficacy against diabetes. [Figure 9b] To evaluate the efficacy of the novel strain Lactococcus lactis subsp. lactis LB1022 against diabetes, male C57BL / 6 mice were used to analyze blood glucose lowering indicators. [Figure 10] These are the results of an analysis of indicators related to muscle differentiation using muscle cells to evaluate the efficacy of the new strain, Lactococcus lactis subsp. lactis LB1022, against muscle loss. [Figure 11] These are the results of an analysis of indicators related to gastrointestinal protection using gastrointestinal epithelial cells to evaluate the gastrointestinal protective efficacy of the novel strain, Lactococcus lactis subsp. lactis LB1022. [Figure 12] To evaluate the efficacy of the novel strain Lactococcus lactis subsp. lactis LB1022 against gout, human-derived macrophages were used to analyze indicators related to gouty inflammation. [Figure 13]To evaluate the efficacy of the novel strain Lactococcus lactis subsp. lactis LB1022 against stress-related diseases, male C57BL / 6 mice were used to evaluate its effects on reducing corticosterone and the expression of stress-related factors in adipose tissue. DETAILED DESCRIPTION OF THE INVENTION

[0009] BEST MODE FOR CARRYING OUT THE INVENTION The terms used in this specification have been selected as widely used as possible while taking into consideration the functions of the present invention, but these may change depending on the intentions of engineers in the relevant field, precedents, the emergence of new technologies, etc. In addition, in certain cases, the applicant may have arbitrarily selected terms, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this specification should be defined not simply by their names, but based on the meanings of the terms and the overall content of the present invention. Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which this invention pertains. Terms as defined in commonly used dictionaries should be interpreted to have a meaning consistent with the meaning they have in the context of the relevant art, and should not be interpreted in an idealized or overly formal sense unless expressly defined in this application.

[0010] The present invention will now be described in more detail. The present invention provides Lactococcus lactis subsp. lactis LB1022 strain, deposited under accession number KCTC15324BP. The Lactococcus lactis subsp. lactis LB1022 strain is characterized by having 16S rRNA represented by SEQ ID NO:1. The strain has excellent acid tolerance, bile tolerance, and ability to adhere to intestinal epithelial cells, and can be used as a functional lactic acid bacterium. Furthermore, the strain exhibits anti-inflammatory effects by suppressing the production of any one inflammatory factor selected from the group consisting of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β), and by suppressing the gene expression of p65 or p50 in the Nuclear Factor Kappa B pathway. Therefore, compositions containing live or killed cells of the strain or a culture solution thereof as an active ingredient can be used for the prevention, treatment, or amelioration of inflammatory diseases.

[0011] The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or ameliorating inflammatory diseases, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a health food composition for preventing or alleviating hangovers, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient. The present invention also provides a health food composition for preventing or improving liver function, which contains live or killed cells of the strain or a culture solution thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating allergic skin diseases, which contains live or killed bacteria of the above strains or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or improving allergic skin diseases, which contains live or killed bacteria of the above strains or a culture medium thereof as an active ingredient.

[0012] The present invention also provides a pharmaceutical composition for preventing or treating skin damage, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient, and a cosmetic composition for preventing or improving skin damage, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating obesity, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or improving obesity, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating diabetes, which contains live or killed bacteria of the above strains or a culture medium thereof as an active ingredient, and a health functional food composition for preventing or ameliorating diabetes, which contains live or killed bacteria of the above strains or a culture medium thereof as an active ingredient.

[0013] The present invention also provides a pharmaceutical composition for preventing or treating muscle loss, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient.The present invention also provides a health functional food composition for preventing or improving muscle loss, which contains live or killed bacteria of the strain or a culture medium thereof as an active ingredient. The present invention also provides a health food composition for gastrointestinal protection, which contains live or killed bacteria of the strain or a culture solution thereof as an active ingredient.

[0014] The pharmaceutical composition of the present invention can be prepared in unit dose form or packaged in a multi-dose container by formulating it using a pharmaceutically acceptable carrier according to a method that can be easily carried out by a person of ordinary skill in the art to which the invention pertains. The pharmaceutically acceptable carriers are those commonly used in formulations, 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, mineral oil, etc. The pharmaceutical composition of the present invention may additionally contain, in addition to the above ingredients, lubricants, wetting agents, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, etc.

[0015] In the present invention, the content of the additives contained in the pharmaceutical composition is not particularly limited, and can be appropriately adjusted within the content range used in ordinary formulations. The pharmaceutical composition may be formulated in one or more external preparation forms selected from the group consisting of injectable preparations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, plasters, lotions, liniments, pastes, and cataplasms. The pharmaceutical compositions of the present invention may further contain pharmaceutically acceptable carriers and diluents for formulation. Examples of pharmaceutically acceptable carriers and diluents include, but are not limited to, excipients such as starch, sugars, and mannitol; fillers and extenders such as calcium phosphate; cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose; binders such as gelatin, alginates, and polyvinylpyrrolidone; lubricants such as talc, calcium stearate, hydrogenated castor oil, and polyethylene glycol; disintegrants such as povidone and crospovidone; and surfactants such as polysorbates, cetyl alcohol, and glycerol. The pharmaceutically acceptable carriers and diluents may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffer solutions, solvents, and / or dispersion media.

[0016] The present invention can be used generally as a commonly used food product. The food composition of the present invention can be used as a functional health food. The term "functional health food" refers to a food manufactured and processed using raw materials or ingredients that have functional properties beneficial to the human body in accordance with the Act on Functional Health Foods. "Functional" refers to food that is consumed for the purpose of obtaining beneficial health effects, such as regulating nutrients or physiological effects, on the structure and function of the human body.

[0017] The food composition of the present invention may contain common food additives, and unless otherwise specified, the suitability as a "food additive" is determined in accordance with the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Official Specification of Food Additives approved by the Ministry of Food and Drug Safety. Examples of items listed in the "Japanese Standards of Food Additives" include chemically synthesized substances such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon color, licorice extract, crystalline cellulose, sorghum color, and guar gum; and mixed preparations such as monosodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0018] The food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. For example, among capsule-type health functional foods, hard capsules can be prepared by mixing the composition of the present invention with additives such as excipients and filling the mixture into a conventional hard capsule, while soft capsules can be prepared by mixing the composition of the present invention with additives such as excipients and filling the mixture into a capsule base such as gelatin. The soft capsules can contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., as needed. The definitions of the terms excipient, binder, disintegrant, lubricant, flavoring agent, flavoring agent, etc. are those described in documents known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and all health functional foods in the usual sense are included.

[0019] In the present invention, the term "prevention" refers to any action of administering a composition according to the present invention to suppress or delay the onset of a disease. In the present invention, the term "treatment" refers to any action of administering a composition according to the present invention to improve or beneficially alter the symptoms of a disease. In the present invention, "amelioration" refers to any action of administering or ingesting a composition according to the present invention to an individual to improve the worsening state of a disease.

[0020] MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below with reference to experimental examples and examples to aid in understanding the present invention. However, the following experimental examples and examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following experimental examples and examples. The experimental examples and examples of the present invention are provided to more completely explain the present invention to those skilled in the art. [Example]

[0021] <Preparation example> Control group and comparison group To evaluate the efficacy of the new strain, Lactococcus lactis subsp. lactis LB1022, the standard strain, Lactococcus lactis subsp. lactis KCTC 3769, was used. T (Lactococcus lactis subsp. lactis KCTC 3769 T The ) strain was selected as the comparison group. Cells that were not treated with inducers or experimental groups were defined as the control group. In all experimental results, significance is expressed as the difference between the control group and the mean value of T-test and ANOVA analysis. * means p < 0.05, ** means p < 0.005, *** means p < 0.0005, and **** means p < 0.0001.

[0022] Example 1 Isolation and identification of novel functional lactic acid bacteria In order to discover various functional lactic acid bacteria, we isolated new strains from fermented dairy products and then identified the species of the isolated new strains.

[0023] All dairy samples were finely chopped and homogenized using the agar plate dilution method, serially diluting 10-fold using 10 ml of peptone solution (0.85% mass / vol). The diluted samples were then plated onto soybean-casein digest agar (Tryptic Soy Agar-TSA; BD BBL) or Lactococcus selective medium (Alsan media) and cultured at 30°C for 2–3 days in an MIR-254-PK incubator (Panasonic, Osaka, Japan). After the culture was completed, white colonies formed were selected and cultured several times in TSA for isolation. After the culture was completed, the colonies were suspended in 20% glycerol and stored in a -80°C freezer. To identify the isolated strains, the nucleotide sequence of the 16S ribosomal RNA gene (16S rDNA) was analyzed to confirm their phylogenetic position. The 16S rDNA of the isolated strains was amplified by PCR using a universal 16S rDNA primer consisting of a forward primer represented by SEQ ID NO: 2 (5'-AGAGTTTGATCCTGGCTCAG-3') and a reverse primer represented by SEQ ID NO: 3 (5'-AAGGAGGTGATCCAGCC-3'). For more detailed identification, a specific gene of Lactococcus lactis strains was amplified and analyzed by PCR using a forward primer represented by SEQ ID NO: 4 (5'-GTTGTATTAGCTAGTTGGTGAGGTAAA-3') and a reverse primer represented by SEQ ID NO: 5 (5'-GTTGAGCCACTGCCTTTTAC-3'). PCR was carried out under the following conditions: 1 minute at 94°C for DNA denaturation, 1 minute at 50°C for DNA binding, and 1 minute 50 seconds at 72°C for DNA elongation, repeated 30 times to amplify the DNA.

[0024] PCR analysis revealed that the 16S rRNA gene sequence of the isolated novel strain was highly similar to that of the standard strain Lactococcus lactis, and analysis of the specific genes of Lactococcus lactis confirmed that the novel strain was a Lactococcus lactis subsp. Morphological analysis of the novel strain also confirmed that it was a Gram-positive coccus, similar to the lactic acid bacterium Lactococcus lactis subsp. Therefore, the novel strain was named Lactococcus lactis subsp. lactisLB1022 and deposited at the Korea Institute of Bioscience and Technology (KCTC) on February 14, 2023 (Accession Number: KCTC 15324BP).

[0025] Example 2 Evaluation of bile and acid tolerance of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain, Lactococcus lactis subsp. lactis LB1022, is a functional lactic acid bacterium, we analyzed the bile and acid tolerance of the novel strain and evaluated its intestinal growth efficiency.

[0026] To evaluate the bile tolerance of the new strains, we used TSA (Tryptic Soy Agar) medium containing 0.4% bile or Alsan medium for Lactococcus selection. The new strain, Lactococcus lactis subsp. lactis LB1022, and the control strain, Lactococcus lactis subsp. lactis KCTC 3769, were cultured on the same medium. T (Lactococcus lactis subsp. lactis KCTC 3769 T The strains were inoculated at a concentration of 1% v / v and incubated at 30°C for 6 hours. The culture medium was obtained either immediately after inoculation or after 6 hours of incubation, and then plated onto TSA (Tryptic Soy Agar) medium or Lactococcus selective medium (Alsan media) containing 0.4% bile. To evaluate the acid tolerance of the novel strains, TSA (Tryptic Soy Agar) medium or Lactococcus selective medium (Alsan medium) adjusted to pH 2.0 was used. The novel strain, Lactococcus lactis subsp. lactis LB1022, or the control strain, Lactococcus lactis subsp. lactis KCTC 3769, was cultured in TSA (Tryptic Soy Agar) medium or Lactococcus selective medium (Alsan medium) adjusted to pH 2.0. T (Lactococcus lactis subsp. lactis KCTC 3769 T The strains were inoculated at a concentration of 1% (v / v) and cultured at 30°C for 2 hours. The culture broth was obtained immediately after inoculation or after 2 hours of culture, and plated on TSA (Tryptic Soy Agar) medium or Lactococcus selective medium (Alsan media) adjusted to pH 2.0. The plated strains were cultured at 30°C for 48 hours, and after the culture was completed, the number of colonies was counted to analyze bile resistance and acid resistance.

[0027] As shown in Figure 1, after 6 hours of incubation in bile-containing medium, the control strain Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 T The survival rate of the Lactococcus lactis subsp. lactis LB1022 strain, a novel strain, was shown to be maintained without any decrease in survival rate. Furthermore, after 2 hours of incubation under acidic pH conditions, the survival rate of the control strain, Lactococcus lactis subsp. lactis KCTC 3769, was significantly higher than that of the control strain, Lactococcus lactis subsp. lactis LB1022. T (Lactococcus lactis subsp. lactis KCTC 3769 TThe survival rate of the 100-kJ / ml strain decreased slightly from the initial stage, but the survival rate of the new strain, Lactococcus lactis subsp. lactis LB1022, was maintained without any decrease. These results indicate that the bile and acid tolerance of the new strain, Lactococcus lactis subsp. lactis LB1022, is higher than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain, suggesting that it can grow in the intestinal environment and can be used as a useful lactic acid bacterium.

[0028] Example 3 Evaluation of the intestinal adhesion ability of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain, Lactococcus lactis subsp. lactis LB1022, is a functional lactic acid bacterium, we evaluated its intestinal adhesion ability by culturing it with Caco-2 cells, a human intestinal epithelial cell line.

[0029] Minimum essential medium (MEM) containing 10% fetal bovine serum (FBS) and antibiotics (100 U / mL penicillin and 100 mg / mL streptomycin) was dispensed into T-75 flasks, and Caco-2 cells were inoculated and cultured at 37°C under 5% CO2 conditions until a cell monolayer was formed. After the culture was completed, Caco-2 cells were plated in a 24-well plate at 1 x 10 5The new strain, Lactococcus lactis subsp. lactis LB1022, or the control strain, Lactococcus lactis subsp. lactis KCTC 3769, was inoculated at a concentration of 1000 cells / ml and cultured until a cell monolayer was formed. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain in PBS (phosphate buffered saline) at 1 x 10 8 The strains were diluted at various concentrations and applied to Caco-2 cell monolayers formed in 24-well plates. The cells were incubated at 37°C under 5% CO2 for 2 hours, and unattached bacteria were removed by washing twice with PBS. Adherent cells were obtained by treatment with 0.25% trypsin-EDTA (ethylene-diamine-tetraacetic acid). The cells were serially diluted in PBS and plated onto Tryptic Soy Agar (TSA) or Lactococcus selective medium (Alsan media), followed by incubation at 30°C for 48 hours. After incubation, colonies were counted and compared with the initial inoculum concentration to assess the ability of each strain to adhere to intestinal epithelial cells.

[0030] As shown in Figure 2, when co-cultured with Caco-2 cells, an intestinal epithelial cell line, for 48 hours, the control group, Lactococcus lactis subsp. lactis KCTC 3769, showed a significant increase in the number of serotypes. T (Lactococcus lactis subsp. lactis KCTC 3769 T The rate at which the 100-kJ strain adhered to Caco-2 cells was significantly reduced to 89±1.29%, whereas the rate at which the novel strain Lactococcus lactis subsp. lactis LB1022 adhered to Caco-2 cells was 96±0.85%, showing almost no reduction. These results indicate that the adhesion ability of the novel strain, Lactococcus lactis subsp. lactis LB1022, to the intestinal epithelium is significantly greater than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) strain, suggesting that it can grow in the intestinal environment and can be used as a useful lactic acid bacterium.

[0031] Example 4 Anti-inflammatory evaluation of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain, Lactococcus lactis subsp. lactis LB1022, is a functional lactic acid bacterium with anti-inflammatory properties, the strain was cultured with inflammation-induced macrophages and analyzed for nitric oxide (NO) production levels, pro-inflammatory cytokine secretion levels, and expression of the NF-kB pathway, which is an inflammatory immune response, to evaluate the strain's anti-inflammatory effects.

[0032] RAW264.7 cells, a mouse macrophage cell line, were used as macrophages in which inflammation was induced. DMEM (Dulbecco's Modified Eagle's Medium) containing 10% fetal bovine serum (FBS) and antibiotics (20 μg / mL gentamicin) was dispensed into T-75 flasks, and RAW264.7 cells were inoculated and cultured at 37°C under 5% CO2 conditions. After the culture was completed, RAW264.7 cells were plated at 1 x 10 in a 24-well plate. 5The cells were inoculated at a concentration of 1000 cells / ml and cultured until a cell monolayer was formed. To induce an inflammatory response, the cells were treated with 1 μg / ml of lipopolysaccharide (LPS). The novel strain, Lactococcus lactis subsp. lactis LB1022, or the control strain, Lactococcus lactis subsp. lactis KCTC 3769, were used. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain in PBS (phosphate buffered saline) at 1 x 10 8 The solutions were diluted to a concentration of cells / ml and then applied to a monolayer of RAW264.7 cells formed in a 24-well plate.

[0033] 4-1. Evaluation of NO generation level Nitric oxide (NO), a pro-inflammatory substance induced by LPS in cells, is an important mediator of inflammatory diseases. To evaluate the anti-inflammatory effects of the strains, we analyzed the nitric oxide (NO) production levels in the supernatant of RAW264.7 cells in which an inflammatory response was induced. RAW264.7 cells were centrifuged at 1,000 rpm for 5 minutes to obtain the supernatant. The supernatant was mixed with Griess reagent at a 1:1 ratio, and the absorbance at 540 nm was measured using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland).

[0034] As shown in Figure 3a, when the NO production level in the LPS-induced inflammatory response was set at 100%, the NO production level in the control group, Lactococcus lactis subsp. lactis KCTC 3769, was significantly higher than that in the control group. T (Lactococcus lactis subsp. lactis KCTC 3769 TTreatment with the novel Lactococcus lactis subsp. lactis LB1022 strain reduced NO production by 52.5±0.95%. On the other hand, treatment with the novel Lactococcus lactis subsp. lactis LB1022 strain reduced NO production by 7.28±0.98%. These results indicate that the inhibitory effect of the novel Lactococcus lactis subsp. lactis LB1022 strain on NO production in inflammatory responses is significantly greater than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain, and that the new strain, Lactococcus lactis subsp. lactis LB1022, has superior anti-inflammatory effects.

[0035] 4-2. Cytokine secretion levels The secretion levels of pro-inflammatory cytokines in induced RAW264.7 cells were analyzed using a Quantikine immunoassay kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer's instructions. The secretion levels of the pro-inflammatory cytokines TNF-α, IFN-γ, and IL-1β secreted by RAW264.7 cells were analyzed by measuring absorbance at 540 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland).

[0036] As shown in Figure 3b, the secretion levels of TNF-α, IFN-γ, and IL-1β under LPS-induced inflammatory responses were 7.31 ± 0.06 ng / ml, 13.12 ± 0.41 pg / ml, and 35.74 ± 0.49 pg / ml, respectively. T (Lactococcus lactis subsp. lactis KCTC 3769 T When treated with the Lactococcus lactis subsp. lactis LB1022 strain, the secretion levels of TNF-α, IFN-γ, and IL-1β were 3.6±0.06 ng / ml, 7.27±0.17 pg / ml, and 17.2±0.97 pg / ml, respectively. When treated with the novel Lactococcus lactis subsp. lactis LB1022 strain, the secretion levels of TNF-α, IFN-γ, and IL-1β were 0.71±0.09 ng / ml, 2.06±0.46 pg / ml, and 4.88±0.31 pg / ml, respectively, significantly lower than those of the control strain. These results indicate that the inhibitory effect of the novel strain, Lactococcus lactis subsp. lactis LB1022, on inflammatory responses to cytokine secretion is greater than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain, and that the new strain, Lactococcus lactis subsp. lactis LB1022, has superior anti-inflammatory effects.

[0037] 4-3. Evaluation of the expression of the NF-kB pathway Activation of the NF-kB (Nuclear Factor Kappa B) pathway, an inflammatory immune response, leads to the expression of various inflammatory cytokines, which may lead to the onset of various inflammatory diseases. To evaluate the anti-inflammatory effects of the strains, we quantitatively analyzed the gene expression levels of the NF-kB subunits p65 and p50 in inflammatory-induced RAW264.7 cells using real-time PCR.

[0038] RNA was extracted from RAW264.7 cells using Trizol (Sigmaldrich, Korea), and cDNA synthesis kit (PrimeScript TM cDNA was synthesized using a 1st strand cDNA Synthesis Kit (Takara, Japan). 2 μl of the synthesized cDNA was mixed with 0.5 μl of each primer set (10 μM) and RNase-free water in SYBR Green PCR master mix (Applied Biosystems, USA). Real-time PCR was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) at 50°C for 2 minutes and 95°C for 10 minutes. Expression levels were evaluated relative to those of the housekeeping gene GAPDH. The primer information used for PCR is listed in Table 1 below.

[0039] [Table 1]

[0040] As shown in Figure 3c, the expression levels of p65 (100 ± 1.44%) and p50 (100 ± 3.38%) in L. lactis subsp. lactis KCTC 3769 cells were significantly lower than those in L. lactis subsp. lactis KCTC 3769 cells, which had been treated with LPS for inflammatory responses. T (Lactococcus lactis subsp. lactis KCTC 3769 TIn cells treated with the novel Lactococcus lactis subsp. lactis LB1022 strain, the p65 expression level was 51.83 ± 9.73%, and the p50 expression level was 56.13 ± 3.14%. In cells treated with the novel Lactococcus lactis subsp. lactis LB1022 strain, the p65 expression level was 4.86 ± 0.41%, and the p50 expression level was 17.07 ± 3.04%. In normal cells, the p65 expression level was 1.88 ± 0.31%, and the p50 expression level was 1.45 ± 0.27%. These results indicate that the NF-kB pathway inhibitory effect of the novel strain, Lactococcus lactis subsp. lactis LB1022, on inflammatory responses is greater than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) strain, and that the new strain, Lactococcus lactis subsp. lactis LB1022, has superior anti-inflammatory effects.

[0041] Example 5 Evaluation of the hangover efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional lactic acid bacterium for hangovers, its alcohol and acetaldehyde decomposition efficacy was evaluated by measuring alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities.

[0042] ADH activity was measured by comparing the relative activity with that of the control group through absorbance data. ADH enzyme was used as the control group. The enzyme reaction mixture (3 ml) was prepared in 1.0 M Tris / HCl buffer (pH 8.8), 20 mM NAD + Lactococcus lactis subsp. lactis KCTC 3769, composed of 0.2 M ethanol and distilled water. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) or a novel strain, Lactococcus lactis subsp. lactis LB1022, was treated and incubated in a thermostatic water bath at 30°C for 5 minutes, after which the absorbance of the supernatant was measured at 340 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland). ALDH activity was measured by comparing the relative activity with that of the control group through absorbance data. ALDH enzyme was used as the control group. The reaction mixture (3 ml) was prepared in 1.0 M Tris / HCl buffer (pH 8.0), 20 mM NAD + The buffer solution consisted of 3.0 M KCl, 0.33 M 2-mercaptoehanol, 1.0 M acetaldehyde, and distilled water. Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 T ) or a novel strain, Lactococcus lactis subsp. lactis LB1022, was treated and incubated in a thermostatic water bath at 30°C for 5 minutes. The absorbance of the supernatant was measured at 340 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland).

[0043] As shown in Figure 4, when compared with the ADH enzyme activity of Lactococcus lactis subsp. lactis KCTC 3769T (Lactococcus lactis subsp. lactis KCTC 3769 T The ADH activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 95 ± 1.77%, whereas the ADH activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 166.77 ± 1.08%. In addition, when compared with the activity of ALDH enzyme, Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 T Treatment with Lactococcus lactis subsp. lactis LB1022 (a novel strain) resulted in an ADH activity of 296.95 ± 6.91%. The results showed that the ADH and ALDH activity of the novel strain Lactococcus lactis subsp. lactis LB1022 on hangover was significantly higher than that of the existing standard strain Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ), and that the new strain, Lactococcus lactis subsp. lactis LB1022, has excellent hangover prevention and improvement effects.

[0044] Example 6 Evaluation of the liver function protecting efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for protecting liver function, its efficacy was evaluated through the analysis of indicators for alcoholic liver damage using Sprague-Dawley (SD) rats.

[0045] 6-1. Check blood AST and ALT indicators Lactococcus lactis subsp. lactis KCTC 3769 was introduced into Sprague-Dawley (SD) rats that had been fasted for 16 hours. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) or a novel strain, Lactococcus lactis subsp. lactis LB1022. Thirty minutes later, 25% alcohol (10 ml / kg) was orally administered. 24 hours after alcohol ingestion, Sprague-Dawley (SD) rats were sacrificed and blood was obtained. AST (Aspartate Aminotransferase) and ALT (Alanine Aminotransferase) values ​​were obtained by requesting data from Green Cross Labocel Co., Ltd. The blood AST and ALT values ​​were then compared with those of rats orally administered alcohol only.

[0046] As shown in Figure 5a, blood AST levels were significantly higher in Lactococcus lactis subsp. lactis KCTC 3769 rats than in rats administered alcohol alone (359 ± 41.96 U / L). T (Lactococcus lactis subsp. lactis KCTC 3769 TThe AST level of rats administered the novel strain Lactococcus lactis subsp. lactis LB1022 was confirmed to be 327 ± 15.01 U / L. On the other hand, the AST level of rats administered the novel strain Lactococcus lactis subsp. lactis LB1022 was confirmed to be 189 ± 2.31 U / L. The AST level of normal rats was confirmed to be 205 ± 2.89 U / L. Furthermore, the blood ALT level of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of rats administered alcohol only (82.5 ± 4.70 U / L). T (Lactococcus lactis subsp. lactis KCTC 3769 T The ALT level in rats administered the novel strain Lactococcus lactis subsp. lactis LB1022 was 72.5 ± 3.77 U / L. The ALT level in rats administered the novel strain Lactococcus lactis subsp. lactis LB1022 was 39.5 ± 7.55 U / L. The ALT level in normal rats was 49.5 ± 0.64 U / L.

[0047] 6-2. Confirmation of mRNA expression levels of factors involved in hepatoprotection Liver tissue was collected from Sprague-Dawley (SD) rats, and tissue RNA was extracted using an RNeasy mini kit (Qiagen, Germany). cDNA was synthesized from the extracted tissue RNA using a cDNA synthesis kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to confirm the expression levels of hepatoprotective factors (AMPK, SIRT1). The primer information used for PCR is shown in Table 2 below.

[0048] [Table 2]

[0049] As shown in Figure 5b, the expression levels of AMPK (100 ± 3.19%) and SIRT1 (100 ± 1.87%), which are hepatoprotective factors, in rats administered only alcohol, were significantly lower in Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T In rats treated with Lactococcus lactis subsp. lactis LB1022, the AMPK expression level was 101.94 ± 3.16% and the SIRT1 expression level was 99.37 ± 1.65%. In rats treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the AMPK expression level was 143.48 ± 2.81% and the SIRT1 expression level was 177.09 ± 11.56%. In normal rats, the AMPK expression level was 193.22 ± 4.15% and the SIRT1 expression level was 198.55 ± 0.93%. These results indicate that the hepatoprotective effect of the novel strain, Lactococcus lactis subsp. lactis LB1022, on activating the AMPK / SIRT1 pathway is significantly greater than that of the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T ), and that the new strain, Lactococcus lactis subsp. lactis LB1022, has excellent hepatoprotective effects.

[0050] Example 7: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against allergic diseases To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for allergic diseases, its effects on allergic dermatitis and asthma were evaluated using female BALB / c mice.

[0051] 7-1. Confirmation of serum IgE and mRNA expression levels of factors involved in allergic dermatitis Female BALB / c mice were sensitized intraperitoneally with a mixture of 50 μg / ml ovalbumin and 50 μg / ml alum (Sigma-Aldrich, USA), an immunopotentiator, at weeks 1, 3, 5, and 7. Allergic disease was then induced by periodically exposing the mice to 2% ovalbumin (w / v) in PBS via aerosols. Subsequently, the mice were exposed to Lactococcus lactis subsp. lactis KCTC 3769 for 8 weeks. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) or a novel strain, Lactococcus lactis subsp. lactis LB1022, was orally administered. After the study was completed, blood was collected from sacrificed BALB / c mice, stored at 4°C for 1 hour, and then centrifuged at 5,000 x g for 1 hour to separate serum. Serum IgE analysis was performed according to the manufacturer's instructions for the ELISA kit (R&D systems, USA), and absorbance was measured at 450 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland). RNA was extracted from the harvested skin tissue using an RNeasy mini kit (Qiagen, Germany). cDNA was synthesized from the extracted skin tissue RNA using a cDNA synthesis kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to confirm the expression levels of factors involved in allergic dermatitis (TARC, eotaxin). The primer information used for PCR is shown in Table 3 below.

[0052] [Table 3]

[0053] As shown in Figure 6a, the serum IgE level of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of Lactococcus lactis subsp. lactis KCTC 3769 compared with that of Lactococcus lactis subsp. lactis KCTC 3769 in mice with induced allergic disease (19.34 ± 1.39 ng / ml). T (Lactococcus lactis subsp. lactis KCTC 3769 T The serum IgE level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 18.96 ± 0.59 ng / ml. In contrast, the serum IgE level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 8.19 ± 0.54 ng / ml. The serum IgE level in normal mice was 7.38 ± 0.63 ng / ml. In addition, when compared with the expression levels of TARC (100%) and Eotaxin (100%), which are factors involved in dermatitis in mice with induced allergic diseases, Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactisKCTC 3769 TIn mice administered the novel strain Lactococcus lactis subsp. lactis LB1022, the TARC expression level was 92.22 ± 2.19% and the Eotaxin expression level was 91.16 ± 2.06%. In mice administered the novel strain Lactococcus lactis subsp. lactis LB1022, the TARC expression level was 52.26 ± 2.51% and the Eotaxin expression level was 26.94 ± 1.12%. In normal mice, the TARC expression level was 32.97 ± 3.81% and the Eotaxin expression level was 18.06 ± 0.34%.

[0054] 7-2. Confirmation of serum IgG1, IgG2a, and cytokine expression levels in bronchoalveolar lavage fluid (BALF) After the study, blood was obtained from sacrificed BALB / c mice, stored at 4°C for 1 hour, and centrifuged at 5,000 x g for 1 hour to separate serum. Serum IgG1 and IgG2a analysis was performed according to the manufacturer's guidelines for the ELISA kit (Abcam, UK), and absorbance was measured at 450 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland). To obtain alveolar fluid, the skin on the neck and chest of each mouse was incised to expose the bronchi and lungs. The alveoli were then lavaged with PBS three times, 0.5 ml per aliquot. The collected alveolar fluid was centrifuged at 1,000 rpm for 10 minutes at 4°C, and the supernatant was analyzed for cytokines using an ELISA kit (R&D Systems, USA) according to the manufacturer's instructions.

[0055] As shown in Figure 6b, the serum IgG1 level of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of Lactococcus lactis subsp. lactis KCTC 3769 when compared with that of Lactococcus lactis subsp. lactis KCTC 3769 in mice with induced allergic asthma (58.85 ± 0.47 ng / ml). T (Lactococcus lactis subsp. lactis KCTC 3769 TThe serum IgG1 level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 57.77 ± 2.52 ng / ml. In contrast, the serum IgG1 level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 15.35 ± 0.03 ng / ml. The serum IgG1 level in normal mice was 8.15 ± 0.05 ng / ml. Furthermore, serum IgG2a levels in mice with induced allergic asthma (35.88 ± 1.37 ng / ml) were significantly higher in Lactococcus lactis subsp. lactis KCTC 3769 than in mice with induced allergic asthma (35.88 ± 1.37 ng / ml). T (Lactococcus lactis subsp. lactis KCTC 3769 T The serum IgG2a level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 33.26 ± 1.28 ng / ml. In contrast, the serum IgG2a level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 20.06 ± 1.29 ng / ml. The serum IgG2a level in normal mice was 5.41 ± 0.17 ng / ml. Compared with the intraalveolar IL-17 level (58.36 ± 2.82 ng / ml) in mice with induced allergic asthma, Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactisKCTC 3769 T In rats treated with Lactococcus lactis subsp. lactis LB1022, the alveolar IL-17 level was 53.69 ± 3.81 ng / ml. In mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the alveolar IL-17 level was 29.47 ± 2.43 ng / ml. In normal mice, the alveolar IL-17 level was 13.21 ± 1.78 ng / ml. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769, in treating allergic diseases. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior effects in preventing and improving allergic diseases.

[0056] Example 8: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain on skin lesions To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional material for skin damage, its skin regeneration efficacy was evaluated using human skin epithelial cells.

[0057] HaCaT cells, a human skin epithelial cell line, were used to induce skin wounds. HaCaT cells were inoculated into T-75 flasks containing Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) and antibiotics (100 units / ml penicillin, 100 μg / ml streptomycin) and cultured at 37°C and 5% CO2. After the culture was completed, HaCaT cells were seeded into 6-well plates and cultured until a cell monolayer was formed. When the cell monolayer approached 100%, a scratch was made on the cells with a tip to induce skin wounds, and Lactococcus lactis subsp. lactis KCTC 3769 was added. T (Lactococcus lactis subsp. lactis KCTC 3769 TThe cells were treated with either Lactococcus lactis subsp. lactis LB1022 or a novel strain, Lactococcus lactis subsp. lactis LB1022. The changes in the cells over time were photographed under a microscope, and the scratch size was measured using ImageJ software (http: / / imagej.nih.gov / ij / index.html) to quantify the degree of re-ephithelialization.

[0058] As shown in Figure 7, the skin regeneration rate of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of HaCaT cells (14.62 ± 3.08%) 8 hours after scratch injury. T (Lactococcus lactis subsp. lactis KCTC 3769 T The skin regeneration rate for cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 36.57 ± 0.03%, while the skin regeneration rate for cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 45.58 ± 3.21%. Compared with the skin regeneration rate of HaCaT cells (25.05 ± 2.19%) 16 hours after skin injury, Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 T The skin regeneration rate for cells treated with Lactococcus lactis subsp. lactis LB1022 was 39.94 ± 1.35%, while the skin regeneration rate for cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 64.96 ± 2.99%. Compared with the skin regeneration rate of HaCaT cells (29.27 ± 2.27%) 24 hours after skin injury, Lactococcus lactis subsp. lactis KCTC 3769 T(Lactococcus lactis subsp. lactis KCTC 3769 T The degree of skin regeneration in cells treated with Lactococcus lactis subsp. lactis LB1022 was 48.58 ± 2.38%, while the degree of skin regeneration in cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 81.49 ± 0.87%. These results indicate that the novel strain, Lactococcus lactis subsp. lactis LB1022, is more effective against skin wounds than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior skin regeneration (wound healing) effects.

[0059] Example 9: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against obesity To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for obesity, its efficacy in reducing body fat was evaluated using male C57BL / 6 mice.

[0060] Male C57BL / 6 mice were fed a 60% kcal / kg fat diet (Research Diets, USA) for 10 weeks to induce high-fat diet-mediated obesity. At the same time, Lactococcus lactis subsp. lactis KCTC 3769 was introduced. T (Lactococcus lactis subsp. lactis KCTC 3769 T) or a novel strain, Lactococcus lactis subsp. lactis LB1022, was orally administered to the mice. After the study, the body composition of the C57BL / 6 mice was analyzed using dual-energy X-ray absorptiometry with an Inalyzer (Medikors Inc, Korea). The mice were then subjected to whole-body scans, and the scan images and body fat mass values ​​were measured using Inalyzer software.

[0061] As shown in Figure 8, the body weight-to-fat ratio of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of mice in which obesity was induced by a 12-week high-fat diet (54.59 ± 0.32%). T (Lactococcus lactis subsp. lactis KCTC 3769 T The fat-to-body weight ratio of mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 52.96 ± 0.97%. Meanwhile, the fat-to-body weight ratio of mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 33.59 ± 1.43%. The fat-to-body weight ratio of normal rats was 26.21 ± 0.41%. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769, in preventing obesity. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior obesity prevention and improvement effects compared to other foods.

[0062] Example 10: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against diabetes To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for diabetes, its efficacy was evaluated by analyzing its glycolytic enzyme activity inhibitory effect and blood glucose lowering indicators using male C57BL / 6 mice.

[0063] 10-1. Confirmation of glycolytic enzyme activity inhibition To confirm the inhibitory effect on α-amylase activity, 11 U / ml of α-amylase enzyme was added to Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T The culture medium was mixed with either Lactococcus lactis subsp. lactis LB1022 or a novel strain, Lactococcus lactis subsp. lactis LB1022, and incubated at room temperature (25°C) for 10 minutes. After incubation, 1% starch was added as a substrate, and the mixture was incubated again at room temperature (25°C) for 10 minutes. A DNS color reagent was prepared by adding 45 mM 3,5-dinitrosalicylic acid to 30% sodium potassium tartrate. The mixture was then added to the culture medium and heated in boiling water at 100°C for 5 minutes to develop the color. The reaction mixture was cooled, and the absorbance was measured at a wavelength of 540 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland). To confirm the inhibitory effect on α-glucosidase enzyme activity, 1 U / ml of α-glucosidase enzyme was added to Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 TThe mixture was mixed with either Lactococcus lactis subsp. lactis LB1022 or a novel strain, Lactococcus lactis subsp. lactis LB1022, and incubated at 37°C for 10 minutes. After incubation, 3 mM pNPG (p-nitrophenyl α-D-glucopyranoside) was added, and the mixture was incubated at 37°C for 20 minutes. The reaction was stopped with 1 M sodium carbonate, and the absorbance was measured at 405 nm.

[0064] As shown in Figure 9a, when compared with the α-amylase enzyme activity level (100 ± 1.78%) of Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 T The α-amylase activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 82.96 ± 1.61 ng / ml, while the α-amylase activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 51.05 ± 1.32 ng / ml. The activity level of α-glucosidase enzyme (100 ± 0.73%) was compared with that of Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T The α-glucosidase activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 85.57 ± 1.63 ng / ml, while the α-glucosidase activity of the lactic acid bacteria treated with Lactococcus lactis subsp. lactis LB1022 was 339.64 ± 2.17 ng / ml.

[0065] 10-2. Confirmation of blood glucose lowering effect in diabetic mice Male C57BL / 6 mice were fed a 60% kcal / 100% fat diet (Research Diets, USA) for 10 weeks to induce type 2 diabetes through a high-fat diet. At the same time, Lactococcus lactis subsp. lactis KCTC 3769 was introduced. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) or a novel strain, Lactococcus lactis subsp. lactis LB1022, was orally administered to the subjects. After the study, fasting insulin (uU / L) and fasting glucose (mg / dL) data were obtained through blood analysis, and the Homeostasis Model Assessment for Insulin Resistance (HOMA-IR) was calculated based on the results using the following formula: HOMA-IR = [fasting insulin concentration (uU / L)] × [fasting glucose concentration (mg / dL)] / 405

[0066] As shown in Figure 9b, the HOMA-IR value of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of mice with 12-week type 2 diabetes (8.21 ± 0.23). T (Lactococcus lactis subsp. lactis KCTC 3769 T The HOMA-IR value of mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 7.63 ± 0.21. In contrast, the HOMA-IR value of mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 4.16 ± 0.08%. The body weight-to-fat ratio of normal mice was 2.96 ± 0.14. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective against diabetes than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior diabetes prevention and improvement effects compared to other foods.

[0067] Example 11: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against muscle loss To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for muscle loss, its efficacy was evaluated through the analysis of indicators related to muscle differentiation using muscle cells.

[0068] C2C12 cells, a mouse skeletal muscle cell line, were used for muscle differentiation. Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) and antibiotics (100 U / mL penicillin and 100 mg / mL streptomycin) was dispensed into T-75 flasks, C2C12 cells were inoculated, and cultured at 37°C in 5% CO2 until the cells reached 70-80% growth. After the culture was completed, C2C12 cells were plated in 12-well plates at 5 x 10 5C2C12 cells were seeded at a concentration of 1000 cells / ml and cultured until a cell monolayer was formed. To induce differentiation, the cells were cultured in DMEM medium (Lonza, USA) supplemented with 2% horse serum (Gibco, USA). To induce muscle loss, C2C12 cells were treated with 1 μM dexamethasone (Sigma-Aldrich, USA). Fresh medium of the same composition was replaced every two days. On day 6 of C2C12 cell differentiation, Lactococcus lactis subsp. lactis KCTC 3769 was cultured. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) or a new strain, Lactococcus lactis subsp. lactis LB1022, and reacted for 24 hours. RNA was extracted from C2C12 cells using Trizol (Sigmaldrich, Korea), and cDNA was synthesized using a PrimeScript cDNA synthesis kit ( TM cDNA was synthesized using a 1st strand cDNA Synthesis Kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to determine the expression levels of mitochondrial biogenesis-related factors (TFAM, NRF1) involved in muscle differentiation. The primers used for PCR are listed in Table 4 below.

[0069] [Table 4]

[0070] As shown in Figure 10, the expression levels of TFAM (79.19 ± 4.06%) and NRF1 (71.09 ± 7.88%), which are involved in mitochondrial biogenesis, were significantly lower in C2C12 cells in which dexamethasone-induced muscle loss was observed during differentiation. T (Lactococcus lactis subsp. lactis KCTC 3769 T In cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the TFAM expression level was 86.21 ± 1.85% and the NRF1 expression level was 68.41 ± 3.01%. In cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the TFAM expression level was 116.33 ± 0.63% and the NRF1 expression level was 91.96 ± 3.78%. In normal cells, the TFAM and NRF1 expression levels were 100%. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769, in reducing muscle loss. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior muscle loss prevention and improvement effects compared to other

[0071] Example 12: Evaluation of the gastrointestinal protective efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for gastrointestinal protection, its efficacy was evaluated through the analysis of indicators related to gastrointestinal protection using gastrointestinal epithelial cells.

[0072] AGS cells, a human gastric adenocarcinoma cell line primarily used in gastrointestinal protection studies, were used. RPMI 1640 (Gibco, USA) medium containing 10% fetal bovine serum (FBS) and antibiotics (100 U / mL penicillin and 100 mg / mL streptomycin) was dispensed into T-75 flasks, and AGS cells were inoculated and cultured at 37°C under 5% CO2 conditions. After the culture was completed, 5 x 10 AGS cells were plated in a 12-well plate. 4 The cells were seeded at a concentration of 1 × 10 cells / ml and cultured until a cell monolayer was formed. 48 Helicobacter pylori was treated with Lactococcus lactis subsp. lactis KCTC 3769 equivalent to CFU / ml. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) or a new strain, Lactococcus lactis subsp. lactis LB1022, were treated together and cultured for 2 hours. RNA was extracted from AGS cells using Trizol (Sigmaldrich, Korea), and cDNA was synthesized using a PrimeScript cDNA synthesis kit ( TM cDNA was synthesized using a 1st strand cDNA Synthesis Kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to confirm the expression levels of pathogen interaction-related factors (integrin α5, integrin β1). The primers used for PCR are listed in Table 5 below.

[0073] [Table 5]

[0074] As shown in Figure 11, the expression levels of integrin α5 (100 ± 0.53%) and integrin β1 (100 ± 3.35%), which are involved in pathogen interaction, were significantly lower in AGS cells in which cell damage was induced by Helicobacter pylori. T (Lactococcus lactis subsp. lactis KCTC 3769 T In cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the integrin α5 expression level was 66.39 ± 3.18% and the integrin β1 expression level was 86.39 ± 1.14%. In cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the integrin α5 expression level was 6.95 ± 0.22% and the integrin β1 expression level was 20.72 ± 1.81%. In normal cells, the integrin α5 expression level was 12.65 ± 0.71% and the integrin β1 expression level was 15.78 ± 2.35%. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769, against gastrointestinal damage caused by pathogens, including Helicobacter pylori strains. T (Lactococcus lactis subsp. lactis KCTC 3769 T ) has been shown to have superior gastrointestinal protection compared to

[0075] Example 13: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against gout To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for gout, the efficacy was evaluated by analyzing indicators related to gouty inflammation using human-derived macrophages.

[0076] The cells used in gout research were THP-1 cells, a monocytic cell line. RPMI 1640 (Gibco, USA) medium containing 10% fetal bovine serum (FBS) and antibiotics (100 U / mL penicillin and 100 mg / mL streptomycin) was dispensed into T-75 flasks, and THP-1 cells were inoculated and cultured at 37°C in 5% CO2 until the cells reached approximately 70-80% growth. After the culture was completed, THP-1 cells were plated in 12-well plates at 5 x 10 5 Lactococcus lactis subsp. lactis KCTC 3769 was seeded at a concentration of 100 cells / ml and treated with 250 ng / ml of phorbol 12-myristate 13-acetate (PMA) for 3 hours, followed by replacement with fresh medium to differentiate the cells. To induce gouty cell inflammation, the cells were treated with 100 μg / ml of MSU (Invivogen, USA). T (Lactococcus lactis subsp. lactis KCTC 3769 T ) or a new strain, Lactococcus lactis subsp. lactis LB1022, and reacted for 24 hours. RNA was extracted from THP-1 cells using Trizol (Sigmaldrich, Korea), and cDNA was synthesized using a PrimeScript cDNA synthesis kit ( TMcDNA was synthesized using a 1st strand cDNA Synthesis Kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to confirm the expression levels of inflammatory inflammasome factors (NLRP3, IL-1β). The primers used for PCR are listed in Table 6 below.

[0077] [Table 6]

[0078] As shown in Figure 12, the expression levels of NLRP3 (100 ± 6.50%) and IL-1β (100 ± 12.07%), which are inflammatory inflammasome factors, in THP-1 cells in which gouty inflammation was induced, were significantly lower in Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactis KCTC 3769 T In cells treated with Lactococcus lactis subsp. lactis LB1022, the NLRP3 expression level was 92.32 ± 1.54% and the IL-1β expression level was 81.64 ± 2.18%. In cells treated with the novel strain Lactococcus lactis subsp. lactis LB1022, the NLRP3 expression level was 33.28 ± 3.91% and the IL-1β expression level was 24.79 ± 2.77%. In normal cells, the NLRP3 expression level was 77.29 ± 7.98% and the IL-1β expression level was 47.91 ± 6.95%. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective against gout than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T(Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior gout prevention and improvement effects compared to other

[0079] Example 14: Evaluation of the efficacy of the novel Lactococcus lactis subsp. lactis LB1022 strain against stress-related diseases To prove that the novel strain Lactococcus lactis subsp. lactis LB1022 is a functional ingredient for stress-related diseases, we evaluated its effects on reducing corticosterone and the expression of stress-related factors in adipose tissue using male C57BL / 6 mice.

[0080] Male C57BL / 6 mice were subjected to cold stress by being placed on ice for 1 hour each day without food or water for 12 weeks. After the cold stress period, the mice were placed on a 60% kcal / % fat diet (Research Diets, USA) to induce stress-induced diabetes. During the induction of stress-induced diabetes, Lactococcus lactis subsp. lactis KCTC 3769 was used. T (Lactococcus lactis subsp. lactis KCTC 3769 T Mice were orally administered either Lactococcus lactis subsp. lactis LB1022 or a novel strain, Lactococcus lactis subsp. lactis LB1022. After the study, blood was obtained from the mice and serum was separated. Serum corticosterone analysis was performed according to the manufacturer's instructions using an ELISA kit (R&D systems, USA), and absorbance was measured at 405 nm using a NanoQuant spectrophotometer (Infinite M200, Tecan, Switzerland). Adipose tissue was collected from C57BL / 6 mice and tissue RNA was extracted using an RNeasy mini kit (Qiagen, Germany). cDNA was synthesized from the extracted tissue RNA using a cDNA synthesis kit (Takara, Japan). Real-time PCR quantitative analysis was performed using an ABI 7500 real-time thermocycler (Biosystems, Foster City, CA, USA) in the same manner as in Example 4-3 to determine the expression level of NPY (neuropeptide Y), a stress-related factor in adipose tissue. The primer information used for PCR is shown in Table 7 below.

[0081] [Table 7]

[0082] As shown in Figure 13, the serum corticosterone level of Lactococcus lactis subsp. lactis KCTC 3769 was significantly higher than that of stress-induced mice (221.61 ± 4.27 ng / ml). T (Lactococcus lactis subsp. lactis KCTC 3769 T The serum corticosterone level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 216.89 ± 7.13 ng / ml. Meanwhile, the serum corticosterone level in mice treated with the novel strain Lactococcus lactis subsp. lactis LB1022 was 112.09 ± 4.08 ng / ml. The serum IgE level in normal mice was 63.82 ± 3.47 ng / ml. In addition, when compared with the expression of NPY (100 ± 3.92%), a stress-related factor in the fat of mice with stress-induced diabetes, Lactococcus lactis subsp. lactis KCTC 3769 T (Lactococcus lactis subsp. lactis KCTC 3769 TThe NPY expression level in mice administered with the novel strain Lactococcus lactis subsp. lactis LB1022 was 99.34 ± 1.55%. In contrast, the NPY expression level in mice administered with the novel strain Lactococcus lactis subsp. lactis LB1022 was 31.22 ± 1.54%. The NPY expression level in normal mice was 15.31 ± 2.92%. The results showed that the novel strain, Lactococcus lactis subsp. lactis LB1022, was more effective against stress-related diseases than the existing standard strain, Lactococcus lactis subsp. lactis KCTC 3769. T (Lactococcus lactis subsp. lactisKCTC 3769 T ) has been proven to have superior effects in preventing and improving stress-related diseases.

[0083] Although certain parts of the present invention have been described in detail above, it is obvious to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the present invention, and the true scope of the present invention is defined by the appended claims and their equivalents.

[0084] Numerical ranges are inclusive of the numerical values ​​defined within the range. Every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitation were expressly written. Every minimum numerical limitation given throughout this specification includes every higher numerical limitation, as if such higher numerical limitation were expressly written. Every numerical limitation given throughout this specification includes every finer numerical range within the broader numerical range, as if such narrower numerical limitation were expressly written. [Accession number]

[0085] Depository institution: Korea Center for Biological Resources (KCTC) Accession number: KCTC15324BP Date of acceptance: 20230214 JPEG2026508199000009.jpg221170

Claims

1. Lactococcus lactis subsp. lactis strain LB1022, deposited under accession number KCTC 15324BP.

2. The strain according to claim 1, characterized in that the strain is acid- and bile-tolerant.

3. The strain according to claim 1, characterized in that it has the ability to adhere to intestinal epithelial cells.

4. 2. The bacterial strain according to claim 1, wherein the bacterial strain inhibits production of any one inflammatory factor selected from the group consisting of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β).

5. The bacterial strain according to claim 1, characterized in that it suppresses the expression of p65 or p50 genes in the NF-kB pathway (Nuclear Factor Kappa B pathway).

6. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

7. A health food composition for preventing or improving inflammatory diseases, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

8. A health functional food composition for preventing or alleviating hangovers, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

9. A health food composition for preventing or improving liver function, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

10. A pharmaceutical composition for preventing or treating allergic diseases, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

11. The pharmaceutical composition for preventing or treating an allergic disease according to claim 10, wherein the allergic disease is at least one disease selected from the group consisting of allergic dermatitis, atopic dermatitis, asthma, and allergic rhinitis.

12. A health food composition for preventing or ameliorating allergic diseases, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

13. The health functional food composition for preventing or improving allergic diseases according to claim 12, characterized in that the allergic disease is any one or more diseases selected from the group consisting of allergic dermatitis, atopic dermatitis, asthma, and allergic rhinitis.

14. A pharmaceutical composition for preventing or treating skin damage, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

15. A cosmetic composition for preventing or improving skin damage, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

16. A pharmaceutical composition for preventing or treating obesity, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

17. A health food composition for preventing or improving obesity, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

18. A pharmaceutical composition for preventing or treating diabetes, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

19. A health food composition for preventing or improving diabetes, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

20. A pharmaceutical composition for preventing or treating muscle loss, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

21. A health food composition for preventing or improving muscle loss, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

22. A health food composition for gastrointestinal protection, comprising live or killed bacteria of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

23. A pharmaceutical composition for preventing or treating gout, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

24. A health food composition for preventing or ameliorating gout, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

25. A pharmaceutical composition for preventing or treating stress-related diseases, comprising live or killed cells of the strain according to any one of claims 1 to 5, or a culture solution thereof, as an active ingredient.

26. 26. The pharmaceutical composition for preventing or treating a stress-related disease according to claim 25, wherein the stress-related disease is one or more diseases selected from the group consisting of stress-related gastritis, stress-related diabetes, stress-related headache, stress-related tension, stress-related anxiety, and stress-related depression.

27. The pharmaceutical composition for preventing or treating a stress-related disease according to claim 25, wherein the pharmaceutical composition has a regulating effect of alleviating or reducing stress.

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