Composition for alleviating male menopause symptoms containing probiotics

Abstract

The present invention relates to a composition for alleviating male menopause symptoms, the composition containing probiotics selected from among eight novel strains of probiotics, that is, two strains of Lactis paracasei, four strains of Lactis plantarum, one strain of Lymosilactobacillus fermentum, and one strain of Streptococcus thermophilus. The probiotics are characterized by enhancing the secretion of the male hormone testosterone, and enhancing the expression of a gene, selected from among 3β-hydroxysteroid dehydrogenase I (3β-HSD-I), 3β-hydroxysteroid dehydrogenase VI (3β-HSD-VI), androgen receptor (AR), and steroidogenic acute regulatory protein (StAR), involved in testosterone secretion.

Description

Composition containing probiotics for improving male menopausal symptoms

[0001] The present invention relates to a composition for improving male menopausal symptoms, comprising probiotics selected from eight novel probiotics, namely two strains of Lacticaceibacillus paracasei, four strains of Lactiplantibacillus plantarum, one strain of Rimosylactobacillus fermentum, and one strain of Streptococcus thermophilus.

[0002]

[0003] The term "menopause" was once limited to women, but the concept has recently been developed in men as well, as endocrine changes caused by a gradual decline in androgen production with aging occur. In men in their 40s and 50s, androgen secretion gradually declines, and by the age of 70, androgen secretion has decreased to half the level of that in their 30s, and target cells become less sensitive to testosterone. This leads to the appearance of various menopausal symptoms similar to those in women, and this is called andropause (Partial Androgen Deficiency of Aging Male, PADAM; androgen deficiency for aging male, ADAM; Male climateric, andropause).

[0004] The main symptoms of male menopause include easy fatigue, memory loss, and frequent depression. Physically, muscle strength declines, body fat increases, and bones become weak. Sexual function also declines, leading to problems such as erectile dysfunction and low libido. The main causes include a decrease in testosterone due to the aging of the brain and testicles, environmental factors such as excessive drinking, smoking, and stress, and physical factors such as high blood pressure, diabetes, and liver disease. In addition, as the activity of Leydig cells, male germ cells, decreases, the death rate of Leydig cells increases, and the expression of PDE (phosphodiesterase) proteins, which are associated with erectile dysfunction, increases.

[0005] Meanwhile, for the treatment of male menopausal syndrome, improvement effects have been confirmed using androgen replacement therapy such as testosterone undecanoate, but side effects of hormone therapy may include liver, lipid status, cardiovascular and prostate diseases, etc., so recently, attempts are being made to develop androgenetic amelioration agents using natural or fermented products as the main ingredients.

[0006] Accordingly, the inventors of the present invention confirmed that probiotics selected from among eight new probiotics, namely, two strains of Lacticaceibacillus paracasei, four strains of Lactiplantibacillus plantarum, one strain of Rimosylactobacillus fermentum, and one strain of Streptococcus thermophilus, increase the expression of testosterone, and confirmed that they can be used as a natural composition for improving male menopause, thereby completing the present invention.

[0007]

[0008] [Prior art literature]

[0009] [Patent Document]

[0010] (Patent Document 1) Republic of Korea Patent No. 10-2151372 (Title of Invention: Food composition for preventing and improving male menopausal syndrome containing fermented noni and its manufacturing method, Applicant: Eunyoung Park, Registration Date: August 27, 2020)

[0011] (Patent Document 2) Republic of Korea Patent No. 10-2362968 (Title of invention: Composition for preventing, alleviating, or treating menopausal symptoms containing lactic acid bacteria and prebiotics, Applicant: Cell Biotech Co., Ltd., Registration date: February 10, 2022)

[0012] (Patent Document 3) Republic of Korea Patent Publication No. 10-2022-0126621 (Title of the invention: Composition for preventing, improving, or treating male menopausal syndrome containing fermented green tea as an effective ingredient, Applicant: Biocare Co., Ltd., Publication date: September 16, 2022)

[0013] (Patent Document 4) Republic of Korea Patent Publication No. 10-2015-0106478 (Title of invention: Lactic acid bacteria-containing complex extract for enhancing immunity and improving menopausal symptoms, applicant: Ulsan University Industry-Academic Cooperation Foundation and 2 others, publication date: September 22, 2015)

[0014]

[0015] The purpose of the present invention is to provide a composition for improving male menopausal symptoms, containing eight novel probiotics. More preferably, the composition comprises Lacticaseibacillus paracasei ATG-V7 deposited under KCTC15545BP of [First Invention], and Lacticaseibacillus paracasei ATG-K22 deposited under KCTC15540BP. [Second invention] Lactiplantibacillus plantarum ATG-V1 deposited under KCTC15547BP, Lactiplantibacillus plantarum ATG-V2 deposited under KCTC15546BP, Lactiplantibacillus plantarum ATG-V8 deposited under KCTC15544BP, and Lactiplantibacillus plantarum ATG-K20 deposited under KCTC15541BP; A composition for improving male menopausal symptoms can be provided, containing probiotics selected from among Limosilactobacillus fermentum ATG-F11 deposited under KCTC15542BP of [the third invention]; Streptococcus thermophilus ATG-V11 deposited under KCTC15543BP of [the fourth invention];

[0016]

[0017] [Technical solution of the first invention]

[0018] The present invention relates to a composition for improving male menopausal symptoms, comprising at least one strain of Lacticaseibacillus paracasei ATG-V7 (Lacticaseibacillus paracaseiATG-V7, KCTC15545BP) and Lacticaseibacillus paracasei ATG-K22 (Lacticaseibacillus paracaseiATG-K22, KCTC15540BP).

[0019] The strain may include live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, or a metabolite isolated from cells or a culture solution.

[0020] The strain is characterized by having an effect of enhancing the production of testosterone. The strain can also increase the expression of one or more genes selected from the group consisting of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), AR (Androgen Receptor), and StAR (Steroidogenic Acute Regulatory Protein), which are involved in the production of testosterone.

[0021] The present invention relates to a pharmaceutical composition for treating or improving male menopausal symptoms, comprising the composition.

[0022] The present invention provides a health functional food or general food composition for preventing or improving male menopausal symptoms, comprising the composition.

[0023] The present invention also relates to a novel Lacticaseibacillus paracasei ATG-V7 (Lacticaseibacillus paracaseiATG-V7, KCTC15545BP) or Lacticaseibacillus paracasei ATG-K22 (Lacticaseibacillus paracaseiATG-K22, KCTC15540BP) strain.

[0024] Hereinafter, the present invention will be described in detail.

[0025] The above Lacticaceibacillus paracasei ATG-V7 or Lacticaceibacillus paracasei ATG-K22 strain is characterized by not producing biogenic amines of histamine, tyramine, putrecine, and cadaverine.

[0026] The above Lacticaceibacillus paracasei ATG-V7 or Lacticaceibacillus paracasei ATG-K22 strain is acid-resistant or bile-resistant.

[0027] A composition comprising the Lacticaceibacillus paracasei ATG-V7 or Lacticaceibacillus paracasei ATG-K22 strain of the present invention may include at least one selected from the group consisting of live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, and a metabolite isolated from cells or a culture solution.

[0028] The strain of the present invention can be cultured in liquid or solid medium (broth or agar) of MRS, and can be cultured at a density of about 1 × 10 10 It can be cultured up to a concentration of CFU / mL.

[0029] The above Lacticaceibacillus paracasei ATG-V7 or ATG-K22 strain is preferably cultured at 35 to 40°C for 8 to 20 hours. The optimal temperature for culture is 37°C, the minimum temperature is 15°C, the maximum temperature is 45°C, the optimal pH is 6.0, the minimum pH for culture is 4.0, and the maximum pH is 8.0. The optimal culture time is 16 hours, the minimum culture time is 8 hours, and the maximum culture time is 24 hours.

[0030] When the above Lacticaceibacillus paracasei ATG-V7 strain is treated, testosterone secretion is characterized by an increase of 115 to 180% compared to the control group. In addition, 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) is characterized by an increase of 1.3 to 1.8 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) is characterized by an increase of 3.8 to 6.0 times, AR (Androgen Receptor) is characterized by an increase of 1.5 to 2.0 times, and StAR (Steroidogenic Acute Regulatory Protein) is characterized by an increase of 12.0 to 18.0 times.

[0031] In addition, the Lacticaceibacillus paracasei ATG-K22 strain increases testosterone secretion by approximately 115-150% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) is increased by 4-27 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) is increased by 3-17 times, AR (Androgen Receptor) is increased by 1.7-6.2 times, and StAR (Steroidogenic Acute Regulatory Protein) is increased by 2.5-5.0 times.

[0032]

[0033] [Technical solution of the second invention]

[0034] The present invention relates to a composition for improving male menopausal symptoms, comprising at least one strain of Lactiplantibacillus plantarum ATG-V1 (Lactiplantibacillus plantarumATG-V1, KCTC15547BP), Lactiplantibacillus plantarum ATG-V2 (Lactiplantibacillus plantarumATG-V2, KCTC15546BP), Lactiplantibacillus plantarum ATG-V8 (Lactiplantibacillus plantarumATG-V8, KCTC15544BP), or Lactiplantibacillus plantarum ATG-K20 (Lactiplantibacillus plantarumATG-K20, KCTC15541BP).

[0035] The strain may include live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, or a metabolite isolated from cells or a culture solution.

[0036] The strain is characterized by having an effect of enhancing the production of testosterone. The strain can also increase the expression of one or more genes selected from the group consisting of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), AR (Androgen Receptor), and StAR (Steroidogenic Acute Regulatory Protein), which are involved in the production of testosterone.

[0037] The present invention relates to a pharmaceutical composition for treating or improving male menopausal symptoms, comprising the composition.

[0038] The present invention provides a health functional food or general food composition for preventing or improving male menopausal symptoms, comprising the composition.

[0039] The present invention also relates to a novel Lactiplantibacillus plantarum ATG-V1 (Lactiplantibacillus plantarumATG-V1, KCTC15547BP), Lactiplantibacillus plantarum ATG-V2 (Lactiplantibacillus plantarumATG-V2, KCTC15546BP), Lactiplantibacillus plantarum ATG-V8 (Lactiplantibacillus plantarumATG-V8, KCTC15544BP) or Lactiplantibacillus plantarum ATG-K20 (Lactiplantibacillus plantarumATG-K20, KCTC15541BP) ​​strain.

[0040] Hereinafter, the present invention will be described in detail.

[0041] The above Lactibacillus plantarum ATG-V1, Lactibacillus plantarum ATG-V2, Lactibacillus plantarum ATG-V8 or Lactibacillus plantarum ATG-K20 strains are characterized by not producing biogenic amines of histamine, tyramine, putrecine and cadaverine.

[0042] The above Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8 or Lactobacillus plantarum ATG-K20 strains are acid-resistant or bile-resistant.

[0043] The composition comprising Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8 or Lactobacillus plantarum ATG-K20 of the present invention may include at least one selected from the group consisting of live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, and a metabolite isolated from cells or a culture solution.

[0044] The strain of the present invention can be cultured in liquid or solid medium (broth or agar) of MRS, and can be cultured at a density of about 1 × 10 10 It can be cultured up to a concentration of CFU / mL.

[0045] The above Lactibacillus plantarum ATG-V1, ATG-V2, ATG-V8 or ATG-K20 strains are preferably cultured at 35 to 40°C for 8 to 20 hours. The optimal temperature for culture is 37°C, the minimum temperature is 15°C, the maximum temperature is 45°C, the optimal pH is 6.0, the minimum pH for culture is 4.0 and the maximum pH is 7.8. The optimal culture time is 16 hours, the minimum culture time is 8 hours and the maximum culture time is 24 hours.

[0046] When treated with the above Lactibacillus plantarum ATG-V1 strain, testosterone secretion is characterized by an increase of 120 to 180% compared to the control group. In addition, 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) is characterized by an increase of 2 to 5 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) is characterized by an increase of 4 to 10 times, AR (Androgen Receptor) is characterized by an increase of 3.5 to 9.0 times, and StAR (Steroidogenic Acute Regulatory Protein) is characterized by an increase of 1.2 to 2.0 times.

[0047] In addition, the Lactibacillus plantarum ATG-V2 strain increases testosterone secretion by 130-200% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) by 1.5-3.0 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) by 4-7 times, AR (Androgen Receptor) by 2-4 times, and StAR (Steroidogenic Acute Regulatory Protein) by 1.5-3.0 times is increased.

[0048] In addition, the Lactibacillus plantarum ATG-V8 strain increases testosterone secretion by 120-200% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) by 1.5-3.0 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) by 3.8-6.0 times, AR (Androgen Receptor) by 2.5-4.0 times, and StAR (Steroidogenic Acute Regulatory Protein) by 15-30 times is increased.

[0049] In addition, the Lactibacillus plantarum ATG-K20 strain increases testosterone secretion by 120-170% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) by 3.0-4.5 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) by 2.7-3.5 times, and StAR (Steroidogenic Acute Regulatory Protein) by 1.5-3.0 times is increased.

[0050]

[0051] [Technical solution of the third invention]

[0052] The present invention relates to a composition for improving male menopausal symptoms containing Limosilactobacillus fermentum ATG-F11 (KCTC15542BP) strain.

[0053] The strain may include live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, or a metabolite isolated from cells or a culture solution.

[0054] The strain is characterized by having the ability to enhance the production of testosterone. The strain can also increase the expression of one or more genes selected from the group consisting of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), and AR (Androgen Receptor), which are involved in the production of testosterone.

[0055] The present invention relates to a pharmaceutical composition for treating or improving male menopausal symptoms, comprising the composition.

[0056] The present invention provides a health functional food or general food composition for preventing or improving male menopausal symptoms, comprising the composition.

[0057] The present invention also relates to a novel strain of Limosilactobacillus fermentum ATG-F11 (Limosilactobacillus fermentumATG-F11, KCTC15542BP).

[0058] Hereinafter, the present invention will be described in detail.

[0059] The above Limosilactobacillus fermentum ATG-F11 (Limosilactobacillus fermentumATG-F11, KCTC15542BP) strain is characterized by not producing biogenic amines such as histamine, tyramine, putrecine, and cadaverine.

[0060] The above Limosilactobacillus fermentum ATG-F11 (Limosilactobacillus fermentumATG-F11, KCTC15542BP) strain is acid-resistant or bile-resistant.

[0061] A composition comprising the Limosilactobacillus fermentum ATG-F11 (Limosilactobacillus fermentumATG-F11, KCTC15542BP) strain of the present invention may include at least one selected from the group consisting of live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, and a metabolite isolated from cells or a culture solution.

[0062] The strain of the present invention can be cultured in liquid or solid medium (broth or agar) of MRS, and can be cultured at a density of about 1 × 10 10 It can be cultured up to a concentration of CFU / mL.

[0063] The above-mentioned Limosyl Lactobacillus fermentum ATG-F11 strain is preferably cultured at 35-40°C for 8-20 hours. The optimal temperature for culture is 37°C, the minimum temperature is 15°C, the maximum temperature is 45°C, the optimal pH is 6.0, the minimum pH for culture is 4.0, and the maximum pH is 7.8. The optimal culture time is 16 hours, the minimum culture time is 8 hours, and the maximum culture time is 24 hours.

[0064] In addition, the Limosyl Lactobacillus fermentum ATG-F11 strain increases testosterone secretion by approximately 110-130% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) is increased by 2-3 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) is increased by 3-5 times, and AR (Androgen Receptor) is increased by 2-3 times.

[0065]

[0066] [Technical Solution of the Fourth Invention]

[0067] The present invention relates to a composition for improving male menopause symptoms containing a Streptococcus thermophilus ATG-V11 (Streptococcus thermophilusATG-V11, KCTC15543BP) strain.

[0068] The strain may include live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, or a metabolite isolated from cells or a culture solution.

[0069] The strain is characterized by having the ability to enhance the production of testosterone. The strain can also increase the expression of one or more genes selected from the group consisting of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), and AR (Androgen Receptor), which are involved in the production of testosterone.

[0070] The present invention relates to a pharmaceutical composition for treating or improving male menopausal symptoms, comprising the composition.

[0071] The present invention provides a health functional food or general food composition for preventing or improving male menopausal symptoms, comprising the composition.

[0072] The present invention also relates to a novel Streptococcus thermophilus ATG-V11 (Streptococcus thermophilusATG-V11, KCTC15543BP) strain.

[0073] Hereinafter, the present invention will be described in detail.

[0074] The above Streptococcus thermophilus ATG-V11 (Streptococcus thermophilusATG-V11, KCTC15543BP) strain is characterized by not producing biogenic amines such as histamine, tyramine, putrecine, and cadaverine.

[0075] A composition comprising the Streptococcus thermophilus ATG-V11 (Streptococcus thermophilusATG-V11, KCTC15543BP) strain of the present invention may include at least one selected from the group consisting of live cells, dead cells, a culture, a culture solution containing cells, a culture solution from which cells have been removed, a concentrate of a culture solution, and a metabolite isolated from cells or a culture solution.

[0076] The strain of the present invention can be cultured in a liquid or solid medium (broth or agar) of BL, and can be cultured at a density of about 1 × 10 8 It can be cultured up to a concentration of CFU / mL.

[0077] The above Streptococcus thermophilus ATG-V11 strain is preferably cultured at 35-40°C for 18-48 hours. The optimal temperature for culture is 37°C, the minimum temperature is 20°C, the maximum temperature is 45°C, the optimal pH is 6.0, the minimum pH for culture is 4.0, and the maximum pH is 8.0. The optimal culture time is 24 hours, the minimum culture time is 18 hours, and the maximum culture time is 48 hours.

[0078] In addition, the Streptococcus thermophilus ATG-V11 strain increases testosterone secretion by approximately 130-150% compared to the control group, and gene expression of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) is increased by 2-4 times, 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) is increased by 4-6 times, and AR (Androgen Receptor) is increased by 2-4 times.

[0079]

[0080] [Common features of the technical solutions of the first to fourth inventions]

[0081] The above male menopause symptoms are characterized by one or more symptoms selected from among hypogonadism, benign prostatic hyperplasia, lower urinary tract symptoms, decreased urogenital function, joint or muscle pain, osteoporosis, dizziness, hot flashes, flushing, hyperhidrosis, decreased physical strength, decreased exercise capacity, decreased muscle strength, decreased body hair, decreased testosterone, decreased bone density, increased arteriosclerosis, increased carotid artery thickness, increased insulin resistance, decreased libido, erectile dysfunction, premature ejaculation, decreased energy, decreased sperm count and decreased sperm motility, nervousness, emotional instability, depression, facial flushing, sleep disorders, decreased energy, lethargy, fatigue, decreased work ability, decreased concentration, decreased cognitive ability, decreased spatial perception, and decreased memory.

[0082] The above probiotics, strains, or compositions derived therefrom can be extracted by solvent extraction, and the extract can be obtained by extracting each strain using water, C1 to C4 alcohol, or a mixed solution thereof as a solvent, and the C1 to C4 alcohol can be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, and isobutanol. The extract can also be a water or 30 to 90% (v / v) alcohol aqueous solution extract, preferably a water or 50 to 80% (v / v) alcohol aqueous solution extract. The water, C1 to C4 alcohol, or a mixed solution thereof used in the preparation of the above extract can be used in an amount of 1 to 40 times the volume (1 to 40 ℓ per 1 kg) or 1 to 40 times the weight based on the weight of the strain used, and preferably 5 to 40 times the volume or 5 to 40 times the weight. The extraction conditions of the above extract may be 20 to 100°C, preferably 20 to 70°C, for 1 minute to 48 hours. The above process may be repeated 1 to 4 times. The above strain or its extract may be fractionated or purified using a single or appropriate combination of known methods used for separating and extracting plant components, such as extraction using an organic solvent (alcohol, ether, acetone, etc.), partitioning of hexane and water, and column chromatography, according to a commercial method.

[0083] In addition, the present invention provides a pharmaceutical composition for preventing or treating male menopausal symptoms, comprising the strain-containing composition. The strain-containing composition may be added to the pharmaceutical composition of the present invention in an amount of 0.001 to 30 wt%.

[0084] The above pharmaceutical composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. Carriers, excipients, and diluents that can be included in the above pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, it is prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the extract of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cocoa butter, laurin, and glycerogelatin.

[0085] The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and body weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the prescriber's judgment. Determining the dosage based on these factors is within the skill of those skilled in the art, and the dosage typically ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 1 mg / kg / day to 500 mg / kg / day. Administration may be administered once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0086] The pharmaceutical composition of the present invention can be administered to mammals, including rats, livestock, and humans, via various routes. All modes of administration are conceivable, including oral, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebrovascular injection. The composition of the present invention exhibits minimal toxicity and side effects, making it a safe drug for long-term use for preventive purposes.

[0087] In addition, a health functional food for improving male menopausal symptoms, which comprises a composition containing the strain of the present invention and a food-wise acceptable food additive, is provided. The strain-containing composition can be added to the health functional food of the present invention in an amount of 0.001 to 100 wt%. The health functional food of the present invention includes forms such as tablets, capsules, pills, or liquids, and foods to which the extract of the present invention can be added include, for example, various drinks, meat, sausage, bread, candy, snacks, noodles, ice cream, dairy products, soups, sports drinks, beverages, alcoholic beverages, gum, tea, and vitamin complexes.

[0088]

[0089] The present invention relates to a composition for improving male menopause symptoms, comprising probiotics selected from eight novel probiotics, namely two strains of Lacticaceibacillus paracasei, four strains of Lactiplantibacillus plantarum, one strain of Rimosylactobacillus fermentum, and one strain of Streptococcus thermophilus.

[0090] The above probiotics are characterized by enhancing the secretion of testosterone, a male hormone, and enhancing the expression of genes selected from 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), AR (Androgen Receptor), and StAR (Steroidogenic Acute Regulatory Protein) related to the secretion thereof.

[0091]

[0092] [Description of the drawing of the first invention]

[0093] Figures 1a and 1b are graphs showing the results of confirming the cytotoxicity of Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains.

[0094] Figures 2a and 2b are graphs showing the results of comparing the testosterone secretion ability of the Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains with that of the Lactobacillus paracasei ATG-E1 (KCTC 14245BP) strain.

[0095] Figures 3a and 3b are graphs showing the results of comparing the expression of the 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) gene of Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains with that of Lactobacillus paracasei ATG-E1 (KCTC 14245BP) strain.

[0096] Figures 4a and 4b are graphs showing the results of comparing the expression of the 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) gene of the Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains with that of the Lactobacillus paracasei ATG-E1 (KCTC 14245BP) strain.

[0097] Figures 5a and 5b are graphs showing the results of comparing the AR (Androgen Receptor) gene expression of Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains with that of Lactobacillus paracasei ATG-E1 (KCTC 14245BP) strain.

[0098] Figures 6a and 6b are graphs showing the results of comparing the expression of the StAR (Steroidogenic Acute Regulatory Protein) gene of the Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains with that of the Lactobacillus paracasei ATG-E1 (KCTC 14245BP) strain.

[0099]

[0100] [Description of the drawing of the second invention]

[0101] Figures 7a to 7d are graphs showing the results of confirming the cytotoxicity of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains.

[0102] Figures 8a to 8d are graphs showing the testosterone secretion ability of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains compared to Lactobacillus plantarum HAC01 (KCTC 12647BP) strain.

[0103] Figures 9a to 9d are graphs showing the results of comparing the expression of the 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) gene of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains with that of Lactobacillus plantarum HAC01 (KCTC 12647BP) strain.

[0104] Figures 10a to 10d are graphs showing the results of comparing the expression of the 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) gene of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains with that of Lactobacillus plantarum HAC01 (KCTC 12647BP) strain.

[0105] Figures 11a to 11c are graphs showing the results of comparing AR (Androgen Receptor) gene expression of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains with Lactobacillus plantarum HAC01 (KCTC 12647BP) strains.

[0106] Figures 12a to 12d are graphs showing the results of comparing the expression of the StAR (Steroidogenic Acute Regulatory Protein) gene of the Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strains with that of the Lactobacillus plantarum HAC01 (KCTC 12647BP) strain.

[0107]

[0108] [Description of the drawing of the third invention]

[0109] Figure 13 is a graph showing the results of confirming the cytotoxicity of the Limosyl Lactobacillus fermentum ATG-F11 strain.

[0110] Figure 14 is a graph showing the testosterone secretion ability of the Limosyl Lactobacillus fermentum ATG-F11 strain compared to that of the same strain, ATG-V5.

[0111] Figure 15 is a graph showing the results of comparing the expression of the 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) gene of the Limosyl Lactobacillus fermentum ATG-F11 strain with that of the ATG-V5 strain, which is the same species.

[0112] Figure 16 is a graph showing the results of comparing the expression of the 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) gene of the strain Rimosyl Lactobacillus fermentum ATG-F11 with that of the strain of the same species, ATG-V5.

[0113] Figure 17 is a graph showing the results of comparing the AR (Androgen Receptor) gene expression of the Limosyl Lactobacillus fermentum ATG-F11 strain with that of the same strain, ATG-V5.

[0114]

[0115] [Description of the drawing of the fourth invention]

[0116] Figure 18 is a graph showing the results of confirming the cytotoxicity of the Streptococcus thermophilus ATG-V11 strain.

[0117] Figure 19 is a graph showing the results of confirming the testosterone secretion ability of Streptococcus thermophilus ATG-V11 strain.

[0118] Figure 20 is a graph showing the results of comparing the expression of the 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) gene of the Streptococcus thermophilus ATG-V11 strain with that of the Streptococcus thermophilus TA40 strain.

[0119] Figure 21 is a graph showing the results of comparing the expression of the 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) gene of the Streptococcus thermophilus ATG-V11 strain with that of the Streptococcus thermophilus TA40 strain.

[0120] Figure 22 is a graph showing the results of comparing the AR (Androgen Receptor) gene expression of the Streptococcus thermophilus ATG-V11 strain with that of the Streptococcus thermophilus TA40 strain.

[0121]

[0122] Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the contents introduced herein are provided to ensure thoroughness and completeness, and to sufficiently convey the spirit of the present invention to those skilled in the art.

[0123]

[0124] [Form for implementing the first invention]

[0125] <Example 1 of the first invention. Isolation and identification of novel lactic acid bacteria - ATG-V7 and ATG-K22>

[0126] Put 90 ml of saline solution (0.85% NaCl, g / L) into a sterile plastic bag, add 10 g of each mushroom or kimchi, and mix well. Then, 1 ml of each homogenate was serially diluted 10-fold in 9 ml of saline solution and spread on MRS solid medium in 3 replicates. After culturing in an incubator at 37°C for more than 48 hours, the grown bacteria were analyzed by catalase test using 0.3% hydrogen peroxide and Gram staining. After that, catalase-negative, Gram-positive, and rod-shaped bacteria were selected. The 16S rRNA gene sequences of the two isolated strains were analyzed by requesting Macrogen, and the sequences obtained through 16S rRNA sequencing were compared with the NCBI BLAST database. As a result, the 16S rRNA sequences of both strains were 99.9% identical to those of Lacticaseibacillus paracasei, indicating that they belonged to Lacticaseibacillus paracasei in terms of taxonomic position. They were identified as novel strains with the gene sequences shown in Tables 1 and 2. The strain isolated from matsutake mushrooms was named Lacticaseibacillus paracaseiATG-V7, and the strain isolated from kimchi was named Lacticaseibacillus paracaseiATG-K22, and they were deposited with the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology as KCTC15545BP (ATG-V7) and KCTC15540BP (ATG-K22), respectively.

[0127] 서열번호 1 :Lcb. paracaseiATG-V7tgcagtcgaa cgagttctcg ttgatgatcg gtgcttgcac cgagattcaa catggaacga 60gtggcggacg ggtgagtaac acgtgggtaa cctgccctta agtgggggat aacatttgga 120aacagatgct aatacccat accact acct agggct t aagatggcgt 180aagctatcgc ttttggatgg acccgcggcg tattagctag ttggtgaggt aatggctcac 240caaggcgatg atacgtagcc gaactgagag gttgatcggc cacattggga ctgagacacg 300gcccaaactc ctagcgcggc aagtctgatg 360gagcaacgcc gcgtgagtga agaaggcttt cgggtcgtaa aactctgttg ttggagaaga 420atggtcggca gagtaactgt tgtcggcgtg acggtatcca accagaaagc cacggctaac 480tacgtgccag cagccgcggtcgtcgtc tattgggcgt 540aaagcgagcg caggcggttt tttaagtctg atgtgaaagc cctcggctta accgaggaag 600cgcatcggaa actgggaaac ttgagtgcag aagaggacag tggaactcca tgtgtagcgg 660tgaaatgcgt agatatatcactcgc gtctgtaact 720gacgctgagg ctcgaaagca tgggtagcga acaggattag ataccctggt agtccatgcc 780gtaaacgatg aatgctaggt gttggagggt ttccgccctt cagtgccgca gctaacgcat840taagcattcc gcctggggag tacgaccgca aggttgaaac tcaaaggaat tgacgggggc 900ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 960ttgacatctt ttgatcacct gagagatcag gtttcccctt cgggggcaaa atgacaggtg 1020gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080acccttatga ctagttgcca gcatttagtt gggcactcta gtaagactgc cggtgacaaa 1140ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1200tgctacaatg gatggtacaa cgagttgcga gaccgcgagg tcaagctaat ctcttaaagc 1260cattctcagt tcggactgta ggctgcaact cgcctacacg aagtcggaat cgctagtaat 1320cgcggatcag cacgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac 1380catgagagtt tgtaacaccc gaagccggtg gcgtaaccct tttagggagc gagccgtcta 1440a 1441

[0128]

[0129] 서열번호 2 :Lcb. paracaseiATG-K22tatttatg agagtttgat cctggctcag gatgaacgct ggcggcgtgc ctaatacatg 60caagtcgaac gagttctcgt tgatgatcgg tgctgcact gagattcaac atggaccgag 120tggcggatacataac cgtgagggct gtgggggata acatttggaa 180acagatgcta ataccgcata gatccagaa ccgcatggtt cttggctga agatggcgta 240agctacgct tttggatgga cccgcggcgt attagctagt tggtgaggta atggctcacc 30acagatgcgaggtacc acattgggac tgagacacgg 360cccaaactcc tacgggaggc agcagtaggg aatctccac aatggacgca agtctgatgg 420agcaacgccg cgtgagtgaa gaaggcttc gggtcgttaa actctgttgt tggagagagtagt gggcattga 480 cggtatccaa ccagaaagcc acggctaact 540acgtgccagc agccgggta atacgtaggt ggcaagcgtt atccggattt attgggcgta 600aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ctgacggagcacttaa cc60cggaggcacttaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 720gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 780acgtgaggc tcgaaagcat caggggattagaccacct840taaacgatga atgctaggtg ttggagggtt tccgcccttc agtgccgcag ctaacgcatt 900aagcattccg cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc 960cgcacaagcg gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggtct 1020tgacatcttt tgatcacctg agagatcagg tttccccttc gggggcaaaa tgacaggtgg 1080tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1140cccttatgac tagttgccag catttagttg ggcactctag taagactgcc ggtgacaaac 1200cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1260gctacaatgg atggtacaac gagttgcgag accgcgaggt caagctaatc tcttaaagcc 1320attctcagtt cggactgtag gctgcaactc gcctacacga agtcggaatc gctagtaatc 1380gcggatcagc acgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1440atgagagttt gtaacacccg aagccggtgg cgtaaccctt ttagggagcg agccgtctaa 1500ggtgggacaa atgattaggg tgaagtcgta acaaggtagc cgtaggagaa cctgcggctg 1560gatcacctcc tttcta 1576

[0130]

[0131] <제1발명의 실시예 2. 신규 유산균의 특성 확인>

[0132] 제1발명의 실시예 2-1. 당 이용 특성

[0133] The sugar utilization characteristics of the Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains isolated in the present invention were investigated using the API 50 CHL test (Biomerieux) according to the method in the manual. As a result, each strain of the present invention was observed to have sugar utilization characteristics as shown in Table 3 below.

[0134] CarbohydratesATG-V7ATG-K221Glycerol--2Erythritol--3D-Arabinose--4L-Arabinose--5Ribose++6D-Xylose--7L-Xylose--8Adonitol--9β-Methyl-D-Xyloside--10Galactose++11D-Gluc ose++12D-Fructose++13D-Mannose++14L-Sorbose--15Rhamnose--16Dulcitol++17Inositol--18Mannitol++19Sorbitol++20α-Methyl-D-Mannoside--21α-Methyl-D-Glucoside++22N-Acetyl glucosamine++23Amygdalin++24Arbutin++25Esculin++26Salicin++27Cellobiose++28Maltose++29Lactose--30Melibios e--31Saccharose++32Trehalose++33Inulin++34Melezitose++35D-Raffinose--36Starch--37Glycogen--38Xylitol--39β Gentiobiose++40D-Turanose++41D-Lyxose--42D-Tagatose++43D-Fucose--44L-Fucose--45D-Arabitol--46L-Arabitol--47Gluconate++482-Keto-Gluconate--495-Keto-Gluconate--

[0135]

[0136] The above results are from Biomerieux's analysis site (APIWEB TM ) and both strains were identified as Lcb. paracasei99.7%.

[0137]

[0138] Example 2-2 of the First Invention. Acid and Bile Resistance Tests

[0139] Acid resistance and bile tolerance of the strains were tested using the SSDP (simulated stomach duodenum passage) method, which was set up under conditions similar to the human (stomach and duodenum) environment. The isolated Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains were cultured in MRS liquid medium for 18 hours, respectively. 1 ml of the strain solution was placed in a tube, centrifuged (3000 x g, 5 min, 4°C), and the supernatant was discarded. The cell pellet was washed with saline solution. This process was repeated twice. The saline solution was then removed, and the precipitated bacteria were mixed in 10 ml of MRS liquid medium adjusted to pH 3. 1 ml of the suspension was serially diluted 10-fold and spread on MRS solid medium. After incubation at 37℃ for 24-48 hours, the number of bacteria produced was counted to determine the initial CFU / ml of bacteria. The remaining 9 ml was incubated at 37℃ for 1 hour, and 1 ml was taken to analyze the number of viable bacteria and determine the survival rate from the initial bacteria. 1 ml of the acid resistance test culture was spread on MRS plate medium containing bile acids (0.3%, w / v), and after incubation for 48 hours, the number of surviving bacteria on the MRS plate was compared. Or, 1 ml of the acid resistance test culture medium was continuously mixed with 4 ml of bile acid solution (10 g oxgall mixed with 100 ml of distilled water, autoclaved) and 17 ml of duodenal juice (6.4 g / L NaHC03, 0.239 g / L KCl, and 1.28 g / L NaCl mixed well in distilled water, adjusted to pH 7.4, autoclaved) and incubated at 37℃ for 2 hours, after which the number of viable cells was counted. The survival rate was expressed by comparing the CFU / ml of the initial counted bacteria with the CFU / ml of the surviving bacteria.

[0140] Initial sample survival rate (%) Acid resistance test survival rate (%) Bile resistance test survival rate (%) ATG-V710088.9107.4 ATG-K22100122.697.1

[0141]

[0142] As confirmed through Table 4, the survival rates of Lactobacillus paracasei ATG-V7 and Lactobacillus paracasei ATG-K22 strains under gastric acid conditions and through bile to the small intestine can be confirmed.

[0143]

[0144] Example 2-3 of the First Invention. Antibiotic Resistance Test

[0145] The minimum inhibitory concentration (MIC) values ​​of nine antibiotics, including ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, clindamycin, erythromycin, tetracycline, and chloramphenicol, were confirmed using E-test strips (Liofilchem, USA). Briefly, the lactic acid bacteria to be tested were measured by absorbance (OD 600 ) were each suspended at about 0.8 and spread on MRS solid medium using a sterilized cotton swab. The solid medium on which lactic acid bacteria were spread was dried for about 3 minutes, and then the E-test strip was placed on it and cultured at 37℃ for about 48 hours. However, since lactic acid bacteria may develop natural intrinsic resistance to aminoglycosides such as gentamicin, kanamycin, and streptomycin, plate count agar (PCA, Difco Laboratories, USA) or Mueller-Hinton agar (MHA, Difco Laboratories, USA) was used as the test medium for these antibiotics. The types of antibiotics and the standards for the lowest inhibitory concentration that can be judged to be safe were referenced from the guidelines published by the European food safety authority (EFSA).

[0146] SampleAMPVANGENKANSTRERYCDTETCMATG-V70.5N.R.16323.20.190.380.193ATG-K220.5N.R.1232240.640.190.753EFSA(European food safety authority)2NR1632641184AMP: ampicillin; VAN: vancomycin, GEN: Gentamicin, KAN: kanamycin; STR: streptomycin;ERY: erythromycin; CD: clindamycin; TET: tetracycline; CM: chloramphenicol;NR: not required (each unit: ㎍ / ㎖)

[0147] The results of each analysis are presented in Table 5 as the results of the minimum inhibitory concentration (MIC) measurement experiment for the Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains against the major antibiotics.

[0148] Accordingly, it was confirmed that the strain of the present invention has antibiotic susceptibility significantly lower than the limit of the EFSA guideline, and accordingly, it was found that there is no risk of exchange of antibiotic resistance genes between the Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains.

[0149]

[0150] <Example 3 of the first invention. Confirmation of cytotoxicity>

[0151] TM-3 cells (mouse Leydig cells) for the experiments of the present invention were cultured in a medium containing 90% DMEM (Dulbecco's modified Eagle's medium) and 10% FBS (heat-inactivated fetal bovine serum).

[0152] TM-3 cells were cultured at 5×10 3 / well were cultured in 96-well plates, and the number of bacteria was calculated by the ratio of MOI (Multiplicity of Infection) to treat each Lacticaceae Bacillus paracasei ATG-V7 and Lacticaceae Bacillus paracasei ATG-K22 strain at a certain ratio to a certain number of cells. For this, TM-3 cells: Lactic acid bacteria strains were mixed at a ratio of 1:1 (5 × 10 3 CFU / well) to 1:100 (5×10 5 The cells were treated with a strain concentration of 10 CFU / well and cultured for 24 hours, and the proliferation ability of TM-3 cells was measured at an absorbance of 450 nm using a CCK substrate (D-Plus™), and the results are shown in Figures 1a and 1b.

[0153] As shown in Figures 1a and 1b, it can be confirmed that both the Lacticaceaebacillus paracasei ATG-V7 and Lacticaceaebacillus paracasei ATG-K22 strains of the present invention are not cytotoxic. (CTL and Con in Figure 1a below indicate the control group, i.e., the untreated group.)

[0154]

[0155] <Example 4 of the first invention. Confirmation of testosterone secretion>

[0156] For comparison of testosterone secretion, a strain of the same species, Lacticaceibacillus paracasei ATG-E1 (KCTC 14245BP), was prepared.

[0157] TM-3 cells were cultured at 1×10 5 / well, and cells and each lactic acid bacteria were treated at a multiplicity of infection (MOI) of 1:1 to 1:10 and cultured for 24 hours. Afterwards, the culture medium was collected and centrifuged at 4°C, 1500 rpm, for 5 minutes to obtain the supernatant, and the expression level of testosterone was measured using an ELISA kit (ADI-901-065). The results are shown in Fig. 2 and Tables 6 and 7.

[0158] Lcb. paracaseiATG-V7 Testosterone expression level (%) Control group 100 1:1 treatment group - MOI (Cell:bacteria ratio) 122 1:5 treatment group - MOI (Cell:bacteria ratio) 127 1:10 treatment group - MOI (Cell:bacteria ratio) 142

[0159] Lcb. paracaseiATG-K22 Testosterone expression level (%) Untreated group 100 1:1 treatment group - MOI (Cell:bacteria ratio) 119 1:10 treatment group - MOI (Cell:bacteria ratio) 134

[0160] As shown in Figures 2a and 2b and Tables 6 and 7, both Lacticaceibacillus paracasei ATG-V7 and Lacticaceibacillus paracasei ATG-K22 strains were confirmed to increase testosterone expression in a concentration-dependent manner. In contrast, the Lacticaceibacillus paracasei ATG-E1 (KCTC 14245BP) strain, although it is a strain of the same species, did not show such an effect.

[0161]

[0162] <Example 5 of the first invention. Confirmation of expression of genes involved in testosterone production>

[0163] Using TM-3 cells prepared under the same conditions as Example 3, mRNA was extracted from the cells using the RNeasy mini Kit (Qiagen, 74104), and cDNA was synthesized using the ReverTraAce-α- TM (TOYOBO) kit. Afterwards, the gene expression levels of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), AR (Androgen Receptor), and StAR (Steroidogenic Acute Regulatory Protein), enzymes involved in testosterone production, were confirmed through real-time PCR.

[0164] The results are shown in Figures 3 to 6. Both Lacticaceaebacillus paracasei ATG-V7 and Lacticaceaebacillus paracasei ATG-K22 strains significantly increased the gene expression of 3β-HSD-I, 3β-HSD-VI, AR, and StAR, but the Lacticaceaebacillus paracasei ATG-E1 (KCTC 14245BP) strain had little effect.

[0165]

[0166] [Form for implementing the second invention]

[0167] <Example 1 of the second invention. Isolation and identification of novel lactic acid bacteria - ATG-V1, ATG-V2, ATG-V8, and ATG-K20>

[0168] Put 90 ml of saline solution (0.85% NaCl / L) into an autoclaved bag, add 10 g of kimchi or vegetable samples obtained from the Daejeon area, mix well, dispense 1 ml, serially dilute 10-fold in 9 ml of saline solution, and spread on MRS agar plate three times. After growing in an incubator at 37℃ for more than 48 hours, the grown bacteria were analyzed using a catalase test using 0.3% hydrogen peroxide and Gram staining. After that, catalase-negative, Gram-positive, rod-shaped bacteria were selected, and strains with high testosterone secretion ability were isolated. The 16S rRNA gene sequences of the two isolated strains were analyzed by requesting Macrogen, and the sequences obtained through 16S rRNA sequencing were compared with the NCBI BLAST database. As a result, the 16S rRNA sequences of both strains were 99.9% identical to those of Lactiplantibacillus plantarum, indicating that they belong to Lactiplantibacillus plantarum in terms of taxonomy. They were identified as novel strains having the gene sequences shown in Tables 8 to 11. Accordingly, these strains were deposited at the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology as Lactiplantibacillus plantarum ATG-V1 (Lactiplantibacillus plantarumATG-V1, KCTC15547BP), Lactiplantibacillus plantarum ATG-V2 (Lactiplantibacillus plantarumATG-V2, KCTC15546BP), Lactiplantibacillus plantarum ATG-V8 (Lactiplantibacillus plantarumATG-V8, KCTC15544BP), and Lactiplantibacillus plantarum ATG-K20 (Lactiplantibacillus plantarumATG-K20, KCTC15541BP).

[0169] 서열번호 3 :Lactiplantibacillus plantarumATG-V1ttaatttgag agtttgatcc tggctcagga cgaacgctgg cggcgtgcct aatacatgca 60agtcgaacga actctggtat tgattggtgc ttgcatcatg atttacatt gagtgactgact2 gggaaacctg cccagaagcg gggataaca cctggaaaca 180gatgctaata ccgcataaca acttggaccg catggtccga gtttgaaaga tggcttcggc 240tatcactttt ggatggtccc gcggcgtatt agctagatgg tggggtaacg gcacgc tgagagggta atcggccaca ttgggactga gacacggccc 360aaactcctac gggaggcagc agtagggaat cttccacaat ggacgaaagt ctgatggagc 420aacgccgcgt gagtgaagaa gggtttcggc tcgtaaaact ctggaacttacatta80agtgaagt8 aactgttcag gtattgacgg tatttaacca gaaagccacg gctaactacg 540tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc cggatttatt gggcgtaaag 600cgagcgcagg cggtttttta agtctgatgt gaagactttgacgc 660tcggaaactg ggaaacttga gtgcagaaga ggacagtgga actccatgtg tagcggtgaa 720atgcgtagat atatggaaga acaccagtgg cgaaggcggc tgtctggtct gtaactgacg 780ctgaggctcg aaagtatggg tagcaa taggattacct ccataccgtaa 840acgatgaatg ctaagtgttg gagggtttcc gcccttcagt gctgcagcta acgcattaag 900cattccgcct ggggagtacg gccgcaaggc tgaaactcaa aggaattgac gggggcccgc 960acaagcggtg gagcatgtgg tttaattcga agctacgcga agaaccttac caggtcttga 1020catactatgc aaatctaaga gattagacgt tcccttcggg gacatggata caggtggtgc 1080atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc 1140ttattatcag ttgccagcat taagttgggc actctggtga gactgccggt gacaaaccgg 1200aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta cacacgtgct 1260acaatggatg gtacaacgag ttgcgaactc gcgagagtaa gctaatctct taaagccatt 1320ctcagttcgg attgtaggct gcaactcgcc tacatgaagt cggaatcgct agtaatcgcg 1380gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg 1440agagtttgta acacccaaag tcggtggggt aaccttttag gaaccagccg cctaaggtgg 1500gacagatgat tagggtgaag tcgtaacaag gtagccgtag gagaacctgc ggctggatca 1560cctcctttct a 1571

[0170] 서열번호 4 :Lactiplantibacillus plantarumATG-V2tcattaattt gagagtttga tcctggctca ggacgaacgc tggcggcgtg cctaatacat 60gcaagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca ttgagtgaggg 120gctgc gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa 180acagatgcta ataccgcata acaacttgga ccgcatggtc cgagcttgaa agatggcttc 240ggctatcact tttggatggt cccgcggcgt attagctta3 acggcggctcagggatcaggatc tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 360cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 420agcaacgccg cgtgagtgaa gaagggtttc ggctcgttaaa actagactagta80tacatgacg agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact 540acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 600aagcgagcgc aggcggtttt ttaaggcgta tgtgaaagctccc tcggcagc 660gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 720gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 780acgctgaggc tcgaaagtat gggtact caggaca taggtagtccataccg 840taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 900aagcattccg cctggggagat acggccgcaa ggctgaaact caaaggaatt gacggggcc 960cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 1020tgacatacta tgcaaatcta agagattaga cgttcccttg ggggacatgg atacaggtgg 1080tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1140cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc ggtgacaaac 1200cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1260gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc tcttaaagcc 1320attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc gctagtaatc 1380gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1440atgagagttt gtaacaccca aagtcggtgg ggtaaccttt taggaaccag ccgcctaagg 1500tgggacagat gattagggtg aagtcgtaac aaggtagccg taggagaacc tgcggctgga 1560tcacctcctt tcta 1574

[0171] 서열번호 5 :Lactiplantibacillus plantarumATG-V8tcattaattt gagagtttga tcctggctca ggacgaacgc tggcggcgtg cctaatacat 60gcaagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca ttgagtgagggc 120gctgc gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa 180acagatgcta ataccgcata acaacttgga ccgcatggtc cgagtttgaa agatggcttc 240ggctatcact tttggatggt cccgcggcgt attagggtacact acgggct300cggcagcacga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 360cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 420agcaacgccg cgtgagtgaa gaagggtttc ggctcgttaaa actagactagta80tacatgacg agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact 540acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 600aagcgagcgc aggcggtttt ttaaggcgta tgtgaaagctccc tcggcagc 660gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 720gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 780acgctgaggc tcgaaagtat gggtact caggaca taggtagtccataccg 840taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 900aagcattccg cctggggagat acggccgcaa ggctgaaact caaaagaatt gacggggcc 960cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 1020tgacatacta tgcaaatcta agagattaga cgttcccttg ggggacatgg atacaggtgg 1080tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1140cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc ggtgacaaac 1200cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1260gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc tcttaaagcc 1320attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc gctagtaatc 1380gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1440atgagagttt gtaacaccca aagtcggtgg ggtaaccttt taggaaccag ccgcctaagg 1500tgggacagat gattagggtg aagtcgtaac aaggtagccg taggagaacc tgcggctgga 1560tcacctcctt tcta 1574

[0172] 서열번호 6 :Lactiplantibacillus plantarumATG-K20tcattaattt gagagtttga tcctggctca ggacgaacgc tggcggcgtg cctaatacat 60gcaagtcgaa cgaactctgg tattgattga tgcttgcatc atgatttaca ttgactgaggtc 120ggtc gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa 180acagatgcta ataccgcata acaacttgga ccgcatggtc cgagtttgaa agatggcttc 240ggctatcact tttggatggt cccgcggcgt attagggtacact acgggct300cggcagcacga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 360cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 420agcaacgccg cgtgagtgaa gaagggtttc ggctcgttaaa actagactagta80tacatgacg agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact 540acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 600aagcgagcgc aggcggtttt ttaaggcgta tgtgaaagctccc tcggcagc 660gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 720gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 780acgctgaggc tcgaaagtat gggtact caggaca taggtagtccataccg 840taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 900aagcattccg cctggggagt acggccgcaa ggctgaaact caaaggaatt gacgggggcc 960cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 1020tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg atacaggtgg 1080tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1140cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc ggtgacaagc 1200cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1260gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc tcttaaagcc 1320attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc gctagtaatc 1380gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1440atgagagttt gtaacaccca aagtcggtgg ggtaaccttt taggaaccag ccgcctaagg 1500tgggacagat gattagggtg aagtcgtaac aaggtagccg taggagaacc tgcggctgga 1560tcacctcctt tcta 1574

[0173]

[0174] <제2발명의 실시예 2. 신규 유산균의 특성 확인>

[0175] 제2발명의 실시예 2-1. 당 이용 특성

[0176] The sugar utilization characteristics of the strains Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, and Lactobacillus plantarum ATG-K20 isolated in the present invention were investigated using the API 50 CH test (Biomerieux) according to the method in the manual. As a result, each strain of the present invention was observed to have sugar utilization characteristics as shown in Table 12 below.

[0177] CarbohydratesATG-V1ATG-V2ATG-V8ATG-K201Glycerol+++-2Erythritol----3D-Arabinose----4L-Arabinose+ ++-5Ribose++++6D-Xylose----7L-Xylose----8Adonitol----9β-Methyl-D-Xyloside----10Galactose++++11D -Glucose++++12D-Fructose++++13D-Mannose++++14L-Sorbose--+-15Rhamnose--+-16Dulcitol----17Inosito l----18Mannitol++++19Sorbitol++++20α-Methyl-D-Mannoside++-+21α-Methyl-D-Glucoside---+22N-Acetyl glucosamine++++23Amygdalin++++24Arbutin++++25Esculin++++26Salicin++++27Cellobiose++++28Maltose++++29Lactose++++30Melibiose ++++31Saccharose++++32Trehalose++++33Inulin--+-34Melezitose++++35D-Raffinose-++-36Starch----37Glycogen----38Xylitol----39β Gentiobiose++++40D-Turanose+++-41D-Lyxose----42D-Tagatose----43D-Fucose----44L-Fucose----4 5D-Arabitol++-+46L-Arabitol----47Gluconate++++482-Keto-Gluconate----495-Keto-Gluconate----

[0178] Table 12 above is from Biomeriux's analysis site (APIWEB TM ) As a result of identification, ATG-V1, ATG-V2, and ATG-K20 were confirmed to be Lpb. plantarum 99.9% and ATG-V8 was confirmed to be Lpb. plantarum 91.8%.

[0179]

[0180] Example 2-2 of the Second Invention. Acid and Bile Resistance Tests

[0181] Acid resistance and bile tolerance tests of the strains were performed using the SSDP (simulated stomach duodenum passage) method set under conditions similar to the human (stomach and duodenum) environment. The isolated Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 were cultured in MRS liquid medium for 18 hours, respectively. 1 ml of the strain solution was placed in a tube, centrifuged (3000 × g, 5 min, 4°C), and the supernatant was discarded. The cell pellet was washed with saline solution. This process was repeated twice. The saline solution was then removed, and the precipitated bacteria were mixed in 10 ml of MRS liquid medium adjusted to pH 3. 1 ml of the suspension was serially diluted 10-fold and spread on MRS agar. After incubation at 37°C for 24 to 48 hours, the number of bacteria produced was counted to determine the CFU / ml of the initial bacteria. The remaining 9 ml was incubated at 37°C for 1 hour, and 1 ml was taken to analyze the number of viable bacteria and determine the survival rate from the initial bacteria. 1 ml of the acid resistance test culture was spread on MRS plate medium containing bile acids (0.3%, w / v), and after incubation for 48 hours, the number of surviving bacteria on the MRS plate was compared. Alternatively, 1 ml of the acid-resistant experimental culture was continuously mixed with 4 ml of bile acid solution (10 g oxgall mixed with 100 ml of distilled water, autoclaved) and 17 ml of duodenal juice (6.4 g / L NaHC03, 0.239 g / L KCl, and 1.28 g / L NaCl mixed well in distilled water, adjusted to pH 7.4, autoclaved) and incubated at 37°C for 2 hours, after which the number of viable cells was counted. The survival rate was expressed by comparing the CFU / ml of the initial counted bacteria with the CFU / ml of the surviving bacteria.

[0182] Initial survival rate of the experimental strain (%) Survival rate after one hour of incubation at pH 3.0 (%) 0.3% Survival rate after two hours of incubation in Oxgal (%) ATG-V110066.782.9 ATG-V2100131.581.3 ATG-V8100107.380.5 ATG-K20100111.1117.0

[0183] As confirmed through Table 13, the strains Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, and Lactobacillus plantarum ATG-K20 each have very high survival rates under gastric acid conditions and through bile to the small intestine.

[0184]

[0185] Example 2-3 of the Second Invention: Antibiotic Resistance Test

[0186] The minimum inhibitory concentration (MIC) values ​​were confirmed using E-test strips (Liofilchem, USA) for nine antibiotics, including ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, clindamycin, erythromycin, tetracycline, and chloramphenicol. Briefly, the lactic acid bacteria to be tested were measured by absorbance (OD 600) were each suspended at about 0.8 and spread on MRS solid medium using a sterilized cotton swab. The solid medium on which lactic acid bacteria were spread was dried for about 3 minutes, and then the E-test strip was placed on it and cultured at 37℃ for about 48 hours. However, since lactic acid bacteria may develop natural intrinsic resistance to aminoglycosides such as gentamicin, kanamycin, and streptomycin, plate count agar (PCA, Difco Laboratories, USA) or Mueller-Hinton agar (MHA, Difco Laboratories, USA) was used as the test medium for these antibiotics. The types of antibiotics and the standards for the lowest inhibitory concentration that can be judged safe were referenced from the guidelines published by the European food safety authority (EFSA).

[0187] test subject StrainAMPVANGENKANSTRERYCDTETCMATG-V11.5N.R.1248N.R.0.250.016126ATG-V21N.R.312N.R.0.50 .02388ATG-V80.047N.R.624N.R.0.750.522ATG-K200.023N.R.212N.R.0.190.1921EFSA(European food safety authority)2N.R.1664N.R.12328AMP: ampicillin; VAN: vancomycin, GEN: Gentamicin, KAN: kanamycin; STR: streptomycin; ERY: erythromycin; CD: clindamycin; TET: tetracycline; CM: chloramphenicol; NR: not required. (Each unit: ㎍ / ㎖)

[0188] The results of each analysis are presented in Table 14 as the results of the minimum inhibitory concentration (MIC) measurement experiment for the Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, and Lactobacillus plantarum ATG-K20 strains against the major antibiotics.

[0189] Accordingly, it was confirmed that the strain of the present invention had antibiotic susceptibility significantly lower than the limit of the EFSA guideline, and thus it was found that the Lactibacillus plantarum ATG-V1, Lactibacillus plantarum ATG-V2, Lactibacillus plantarum ATG-V8, and Lactibacillus plantarum ATG-K20 strains had no risk of exchanging antibiotic resistance genes.

[0190]

[0191] <Example 3 of the Second Invention. Confirmation of Cytotoxicity>

[0192] TM-3 cells for the experiments of the present invention were cultured in a medium containing 90% DMEM (Dulbecco's modified Eagle's medium) and 10% FBS (heat-inactivated fetal bovine serum).

[0193] TM-3 cells were cultured at 5×10 3 / well were cultured in 96-well plates, and each Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, or Lactobacillus plantarum ATG-K20 strain was treated at a certain ratio to a certain number of cells, and the number of bacteria was calculated by multiplicity of infection (MOI) ratio. For this, TM-3 cells: Lactobacillus strains were treated at a ratio of 1:1 (5 × 10 3 CFU / well) to 1:100 (5×10 5The cells were treated with a strain concentration of 10 CFU / well and cultured for 24 hours, and the proliferation ability of TM-3 cells was measured at an absorbance of 450 nm using a CCK substrate (D-Plus™), and the results are shown in Figures 7a to 7d.

[0194] When checking FIGS. 7a to 7d, it can be confirmed that all of the Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, and Lactobacillus plantarum ATG-K20 strains of the present invention are free of cytotoxicity.

[0195]

[0196] <Example 4 of the Second Invention: Confirmation of Testosterone Secretion>

[0197] For comparison of testosterone secretion, a strain of the same species, Lactiplantibacillus plantarum ATG-HAC01 (KCTC 12647BP, registered patent 10-1500974), was prepared.

[0198] TM-3 cells were cultured at 1×10 5 / well, and cells and each lactic acid bacteria were treated at a multiplicity of infection (MOI) of 1:1 to 1:10, and cultured for 24 hours. Afterwards, the culture medium was collected and centrifuged at 4°C, 1500 rpm, for 5 minutes to obtain the supernatant, and the testosterone expression level was measured using an ELISA kit (Enzo Life Science, ADI-901-065). The results are shown in Figure 8.

[0199] As shown in FIGS. 8A to 8D, it was confirmed that all strains of Lactobacillus plantarum ATG-V1, Lactobacillus plantarum ATG-V2, Lactobacillus plantarum ATG-V8, and Lactobacillus plantarum ATG-K20 concentration-dependently enhanced testosterone expression. In contrast, Lactobacillus plantarum HAC01 (KCTC 12647BP, registered patent 10-1500974) did not show such an effect, even though it was a strain of the same species.

[0200]

[0201] <Example 5 of the Second Invention: Confirmation of Expression of Genes Involved in Testosterone Production>

[0202] Using TM-3 cells prepared under the same conditions as Example 3, mRNA was extracted from the cells using the RNeasy mini Kit (Qiagen, 74104), and cDNA was synthesized using the ReverTraAce-α- TM (TOYOBO) kit. Afterwards, the gene expression levels of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), AR (Androgen Receptor), and StAR (Steroidogenic Acute Regulatory Protein), enzymes involved in testosterone production, were confirmed through real-time PCR.

[0203] The results are shown in Figs. 9 to 12, and all of the Lactibacillus plantarum ATG-V1, Lactibacillus plantarum ATG-V2, Lactibacillus plantarum ATG-V8, and Lactibacillus plantarum ATG-K20 strains significantly increased the gene expression of 3β-HSD-I, 3β-HSD-VI, AR, and StAR, but the Lactibacillus plantarum HAC01 (KCTC 12647BP) strain had little effect.

[0204]

[0205] [Form for implementing the third invention]

[0206] <Example 1 of the Third Invention. Isolation and Identification of Novel Lactic Acid Bacteria - ATG-V11>

[0207] Put 90 ml of saline solution (0.85% NaCl / L) into an autoclaved bag, add 10 g of a fecal sample of a newborn obtained from a postpartum care center in the Jeonju area, mix well, and dispense 1 ml, serially dilute 10-fold in 9 ml of saline solution, and spread on MRS agar plate three times. After growing in an incubator at 37℃ for more than 48 hours, the grown bacteria were analyzed by catalase test using 0.3% hydrogen peroxide and Gram staining. After that, catalase-negative, Gram-positive, rod-shaped bacteria were selected, and strains with high testosterone secretion ability were isolated. The 16S rRNA gene sequences of the two isolated strains were analyzed by requesting Macrogen, and the sequences obtained through 16S rRNA sequencing were compared with the NCBI BLAST database. As a result, the 16S rRNA sequence of this strain was 99.9% identical to that of Limosilactobacillus fermentum, indicating that it belonged to Limosilactobacillus fermentum in terms of taxonomy. It was determined to be a novel strain with the gene sequence shown in Table 15. Accordingly, this strain was named Limosilactobacillus fermentumATG-F11 and deposited with the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology under the accession number KCTC15542BP.

[0208] 서열번호 7 :Limosilactobacillus fermentumATG-F11gcaagtcgaa cgcgttggcc caattgattg atggtgcttg cacctgattg attttggtcg 60ccaacgagtg gcggacgggt gagtaacacg taggtaacct gggcac 120atttggaaac agatgctaat accgcataac agcgttgttc gcatgaacaa cgcttaaaag 180atggcttctc gctatcactt ctggatggac ctgcggtgca ttagcttgtt ggtggggtaa 240cggcctacca aggcgatgat gcatagcc 300gagacacggc ccatactcct acgggaggca gcagtaggga atcttccaca atgggcgcaa 360gcctgatgga gcaacaccgc gtgagtgaag aagggttcg gctcgtaaag ctctgttgtt 420aaagaac acgtatgaga gtacttc cagaaagtca 480cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgtta tccggattta 540ttgggcgtaa agagagtgca ggcggttttc taagtctgat gtgaaagcct tcggcttaac 600cggagaagtg caggacggatc gagggtagtg gaactccatg 660tgtagcggtg gaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctacctggt 720ctgcaactga cgctgagact cgaaagcatg ggtagcgaac aggattagat accctggtag 780tccatgtagtc ccgcccttcagtgccggagc 840taacgcatta agcactccgc ctggggagta cgaccgcaag gttgaaactc aaaggaattg 900acgggggccc gcacaagcgg tggagcatgt ggtttaattc gaagctacgc gaagaacctt 960accaggtctt gacatcttgc gccaacccta gagatagggc gtttccttcg ggaacgcaat 1020gacaggtggt gcatggtcgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa 1080cgagcgcaac ccttgttact agttgccagc attaagttgg gcactctagt gagactgccg 1140gtgacaaacc ggaggaaggt ggggacgacg tcagatcatc atgcccctta tgacctgggc 1200tacacacgtg ctacaatgga cggtacaacg agtcgcgaac tcgcgagggc aagcaaatct 1260cttaaaaccg ttctcagttc ggactgcagg ctgcaactcg cctgcacgaa gtcggaatcg 1320ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc 1380cgtcacacca tgagagtttg taacacccaa agtcggtggg gtaaccttta ggagccagcc 1440gcctaa 1446

[0209] <제3발명의 실시예 2. 신규 유산균의 특성 확인>

[0210] 제3발명의 실시예 2-1. 당 이용 특성

[0211] The sugar utilization characteristics of the strain Limosyl Lactobacillus fermentum ATG-F11 isolated in the present invention were investigated using the API 50 CHL test (Biomerieux) according to the method in the manual. As a result, each strain of the present invention was observed to have sugar utilization characteristics as shown in Table 16 below.

[0212] CarbohydratesLimosilactobacillus fermentumATG-F111Glycerol-2Erythritol-3D-Arabinose-4L-Arabinose-5Ribose+6D-Xylose-7L-Xylose-8Adonitol-9β-Methyl-D-Xyloside-10Galactose+11D-Glucose+ 12D-Fructose+13D-Mannose+14L-Sorbose-15Rhamnose-16Dulcitol-17Inositol-18Mannitol-19Sorbitol-20α-Methyl-D-Mannoside-21α-Methyl-D-Glucoside-22N-Acetyl glucosamine-23Amygdalin-24Arbutin-25Esculin+26Salicin-27Cellobiose-28Maltose+29Lactose+30Melibiose+31Saccharose+32Trehalose-33Inulin-34Melezitose-35D-Raffinose+36Starch-37Glycogen-38Xylitol-39β Gentiobiose-40D-Turanose-41D-Lyxose-42D-Tagatose-43D-Fucose-44L-Fucose-45D-Arabitol-46L-Arabitol-47Gluconate+482-Keto-Gluconate-495-Keto-Gluconate-

[0213] The above results are from Biomeriux's analysis site (APIWEB TM ) was identified as Lim. fermentum99.6%.

[0214]

[0215] Example 2-2 of the Third Invention. Acid and Bile Resistance Tests

[0216] Acid resistance and bile tolerance of the strains were tested using the SSDP (simulated stomach duodenum passage) method, which was set up under conditions similar to the human (stomach and duodenum) environment. The isolated Rimosyl Lactobacillus fermentum ATG-F11 strain was cultured in MRS liquid medium for 18 hours, and then 1 ml of the strain solution was placed in a tube, centrifuged (3000 x g, 5 minutes, 4℃), and the supernatant was discarded, and the cell pellet was washed with physiological saline. This process was repeated twice. After that, the physiological saline solution was removed, and the precipitated bacteria were mixed with 10 ml of MRS liquid medium adjusted to pH 3. 1 ml of the suspension was taken, serially diluted 10-fold, plated on MRS solid medium, and incubated at 37℃ for 24 to 48 hours. The number of bacteria produced was counted to determine the CFU / ml of the initial bacteria. The remaining 9 ml was cultured at 37℃ for 1 hour, and 1 ml was taken to analyze the viable cell count and confirm the survival rate from the initial bacteria. 1 ml of the acid-resistant test culture was spread on MRS plate medium containing bile acids (0.3%, w / v), cultured for 48 hours, and compared with the number of surviving bacteria on the MRS plate. Alternatively, 1 ml of the acid-resistant test culture was continuously mixed with 4 ml of bile acid solution (10 g oxgall mixed with 100 ml of distilled water, autoclaved, used) and 17 ml of duodenal juice (6.4 g / L NaHC03, 0.239 g / L KCl, and 1.28 g / L NaCl mixed well in distilled water, adjusted to pH 7.4, autoclaved, used), cultured at 37℃ for 2 hours, and then the viable cell count was counted. And the survival rate was expressed by comparing the CFU / ml of the initial counted bacteria with the CFU / ml of the surviving bacteria.

[0217] Initial viability of the sample (%) Viability after one hour of incubation at pH 3.0 (%) 0.3% Viability after two hours of incubation in Oxgal (%) ATG-F1110094.792.1

[0218] As confirmed through Table 17, the survival rate of the Limosyl Lactobacillus fermentum ATG-F11 strain was confirmed under gastric acid conditions and through bile to the small intestine.

[0219]

[0220] Example 2-3 of the Third Invention: Antibiotic Resistance Test

[0221] The minimum inhibitory concentration (MIC) values ​​were confirmed using E-test strips (Liofilchem, USA) for nine antibiotics, including ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, clindamycin, erythromycin, tetracycline, and chloramphenicol. Briefly, the lactic acid bacteria to be tested were measured by absorbance (OD 600 ) were each suspended at about 0.8 and spread on MRS solid medium using a sterilized cotton swab. The solid medium on which lactic acid bacteria were spread was dried for about 3 minutes, and then the E-test strip was placed on it and cultured at 37℃ for about 48 hours. However, since lactic acid bacteria may develop natural intrinsic resistance to aminoglycosides such as gentamicin, kanamycin, and streptomycin, plate count agar (PCA, Difco Laboratories, USA) or Mueller-Hinton agar (MHA, Difco Laboratories, USA) was used as the test medium for these antibiotics. The types of antibiotics and the standards for the lowest inhibitory concentration that can be judged safe were referenced from the guidelines published by the European food safety authority (EFSA).

[0222] SampleAMPVANGENKANSTRERYCDTETCMLimosilactobacillus fermentumATG-F110.047N.R.1.516120.50.0321.01.5EFSA(European food safety authority)2N.R.1632641184AMP: ampicillin; VAN: vancomycin, GEN: Gentamicin, KAN: kanamycin; STR: streptomycin; ERY: erythromycin; CD: clindamycin; TET: tetracycline; CM: chloramphenicol; NR: not required. (Each unit: ㎍ / ㎖)

[0223] The results of each analysis are presented in Table 18 as the results of the minimum inhibitory concentration (MIC) measurement experiment for the Rimosyl Lactobacillus fermentum ATG-F11 strain against major antibiotics.

[0224] Accordingly, it was confirmed that the strain of the present invention had antibiotic susceptibility that was significantly lower than the limit of the EFSA guideline, and thus it was found that the Rimosyl Lactobacillus fermentum ATG-F11 strain had no risk of exchanging antibiotic resistance genes.

[0225]

[0226] <Example 3 of the Third Invention. Confirmation of Cytotoxicity>

[0227] TM-3 cells for the experiments of the present invention were cultured in a medium containing 90% DMEM (Dulbecco's modified Eagle's medium) and 10% FBS (heat-inactivated fetal bovine serum).

[0228] TM-3 cells were cultured at 5×10 3 / well were cultured in 96-well plates, and the number of bacteria was calculated by the ratio of MOI (Multiplicity of Infection) to treat the Limosyl Lactobacillus fermentum ATG-F11 strain at a certain ratio to a certain number of cells. For this, TM-3 cells: Lactobacillus strain were mixed at a ratio of 1:1 (5 × 10 3 CFU / well) to 1:100 (5×10 5 The cells were treated with a strain concentration of 10 CFU / well and cultured for 24 hours, and the proliferation ability of TM-3 cells was measured at an absorbance of 450 nm using a CCK substrate (D-Plus™), and the results are shown in Figure 13.

[0229] As shown in Fig. 13, it can be confirmed that the Limosyl Lactobacillus fermentum ATG-F11 strain of the present invention is not cytotoxic.

[0230]

[0231] <Example 4 of the Third Invention: Confirmation of Testosterone Secretion>

[0232] Limosyl Lactobacillus fermentum ATG-V5, a strain of the same species * (KCTC14481BP) strain was used for comparative experiments. * ATG-V5 strain: Lactobacillus fermentum ATG-V5 (see Korean Patent Publication No. 10-2022-0147328)

[0233] For comparison of testosterone secretion, TM-3 cells were cultured at 1×10 5 / well, and cells and each Streptococcus thermophilus strain were treated at a multiplicity of infection (MOI) of 1:1 to 1:10, and cultured for 24 hours. Afterwards, the culture medium was collected and centrifuged at 4°C, 1500 rpm, for 5 minutes to obtain the supernatant, and the expression level of testosterone was measured using an ELISA kit (Enzo Life Science, ADI-901-065). The results are shown in Figure 14.

[0234] As shown in Figure 14, the strain Rimosyl Lactobacillus fermentum ATG-F11 was found to increase testosterone expression in a concentration-dependent manner. In contrast, the strain Rimosyl Lactobacillus fermentum ATG-V5, although of the same species, did not exhibit this effect.

[0235]

[0236] <Example 5 of the Third Invention: Confirmation of Expression of Genes Involved in Testosterone Production>

[0237] Using TM-3 cells prepared under the same conditions as Example 3, mRNA was extracted from the cells using the RNeasy mini Kit (Qiagen, 74104), and cDNA was synthesized using the ReverTraAce-α- TM (TOYOBO) kit. Afterwards, the gene expression levels of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), and AR (Androgen Receptor), enzymes involved in testosterone production, were confirmed through real-time PCR.

[0238] The results are shown in Figures 15 to 17. The strain Rimosyl Lactobacillus fermentum ATG-F11 significantly increased the gene expression of 3β-HSD-I, 3β-HSD-VI, and AR, but the strain Rimosyl Lactobacillus fermentum ATG-V5 had little effect.

[0239]

[0240] [Form for implementing the fourth invention]

[0241] <Example 1 of the Fourth Invention. Isolation and Identification of Novel Lactic Acid Bacteria - ATG-V11>

[0242] Put 90 ml of saline solution (0.85% NaCl / L) into an autoclaved bag, add 10 g of homemade yogurt collected in the Daejeon area, mix well, dispense 1 ml, serially dilute 10-fold in 9 ml of saline solution, and spread on BL solid medium three times. After growing in an incubator at 37℃ for more than 48 hours, the grown bacteria were analyzed using a catalase test using 0.3% hydrogen peroxide and Gram staining. After that, catalase-negative, Gram-positive, rod-shaped bacteria were selected, and strains with high testosterone secretion ability were isolated. The 16S rRNA gene sequences of the two isolated strains were analyzed by requesting Macrogen, and the sequences obtained through 16S rRNA sequencing were compared with the NCBI BLAST database. As a result, the 16S rRNA sequence of this strain was 99.9% identical to that of Streptococcus thermophilus, indicating that it belonged to Streptococcus thermophilus in terms of taxonomy. It was determined to be a novel strain with the gene sequence shown in Table 19. Accordingly, this strain was named Streptococcus thermophilus ATG-V11 and deposited with the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology under the accession number KCTC15543BP.

[0243] 서열번호 8 :Streptococcus thermophilusATG-V11tgcagtagaa cgctgagaga ggagcttgct cttcttggat gagttgcgaa cgggtgagta 60acgcgtaggt aacctgcctt gtagcggggg ataactaccgataggcta cgctgagcta 120ataacaatgg atgacacatg tcatttattt gaaaggggca attgctccac tacaagatgg 180acctgcgttg tattagctag taggtgaggt aatggctcac ctaggcgacg atacatagcc 240gacctgagag ggtgatcggc cacactggga ctcgacacac 300cagcagtagg caatgggggc aaccctgacc gagcaacgcc gcgtgagtga 360agaaggtttt cggatcgtaa agctctgttg taagtcaaga acgggtgtga gagtggaaag 420ttcacactgt gacggtagct taccagaaag cggcgcgc 480taatacgtag gtcccgagcg ttgtccggat ttattgggcg taagcgagc gcaggcggtt 540tgataagtct gaagttaaag gctgtggctc aaccatagtt cgctttggaa actgtcaaac 600ttgagtgcag aaggggagag tgggaattcgatc 660aggaacaccg gtggcgaaag cggctctctg gtctgtaact gacgctgagg ctcgaaagcg 720tggggagcga acaggattag ataccctggt agtccacgcc gtaaacgatg agtgctaggt 780gttggatcct ttccgggatca gcgcctcgcctggggag 840tacgaccgca aggttgaaac tcaaaggaat tgacgggggc ccgcacaagc ggtggagcat 900gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc ttgacatccc gatgctattt 960ctagagatag aaagttactt cggtacatcg gtgacaggtg gtgcatggtt gtcgtcagct 1020cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccctattg ttagttgcca 1080tcattcagtt gggcactcta gcgagactgc cggtaataaa ccggaggaag gtggggatga 1140cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg gttggtacaa 1200cgagttgcga gtcggtgacg gcgagctaat ctcttaaagc caatctcagt tcggattgta 1260ggctgcaact cgcctacatg aagtcggaat cgctagtaat cgcggatcag cacgccgcgg 1320tgaatacgtt cccgggcctt gtacacaccg cccgtcacac cacgagagtt tgtaacaccc 1380gaagtcggtg aggtaacctt ttggagccag ccgccta 1417

[0244]

[0245] <Example 2 of the Fourth Invention. Confirmation of the Characteristics of Novel Lactic Acid Bacteria>

[0246] Example 2-1 of the Fourth Invention. Characteristics of Sugar Utilization

[0247] The sugar utilization characteristics of the Streptococcus thermophilus ATG-V11 strain isolated in the present invention were investigated using the API 50 CHL test (Biomerieux) according to the method in the manual. As a result, each strain of the present invention was observed to have sugar utilization characteristics as shown in Table 20 below.

[0248] CarbohydratesATG-V71Glycerol-2Erythritol-3D-Arabinose-4L-Arabinose-5Ribose-6D-Xylose-7L-Xylose-8Adonitol-9β-Methyl-D-Xyloside-10Galactose-11D-Glucose +12D-Fructose-13D-Mannose-14L-Sorbose-15Rhamnose-16Dulcitol-17Inositol-18Mannitol-19Sorbitol-20α-Methyl-D-Mannoside-21α-Methyl-D-Glucoside-22N-Acetyl glucosamine-23Amygdalin-24Arbutin-25Esculin-26Salicin-27Cellobiose-28Maltose-29Lactose+30Melibiose-31Saccharose+32Trehalose-33Inulin-34Melezitose-35D-Raffinose-36Starch-37Glycogen-38Xylitol-39β Gentiobiose-40D-Turanose-41D-Lyxose-42D-Tagatose-43D-Fucose-44L-Fucose-45D-Arabitol-46L-Arabitol-47Gluconate-482-Keto-Gluconate-495-Keto-Gluconate-

[0249] The above results are from Biomeriux's analysis site (APIWEB TM ) was identified as Str. thermophilus99.2%.

[0250]

[0251] Example 2-2 of the Fourth Invention: Antibiotic Resistance Test

[0252] The minimum inhibitory concentration (MIC) values ​​of nine antibiotics, including ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, clindamycin, erythromycin, tetracycline, and chloramphenicol, were confirmed using E-test strips (Liofilchem, USA). Briefly, the lactic acid bacteria to be tested were measured by absorbance (OD 600 ) were each suspended at about 0.8 and spread on MRS solid medium using a sterilized cotton swab. The solid medium on which lactic acid bacteria were spread was dried for about 3 minutes, and then the E-test strip was placed on it and cultured at 37℃ for about 48 hours. However, since lactic acid bacteria may develop natural intrinsic resistance to aminoglycoside drugs such as gentamicin, kanamycin, and streptomycin, plate count agar (PCA, Difco Laboratories, USA) or Mueller-Hinton agar (MHA, Difco Laboratories, USA) was used as the test medium for these antibiotics. The types of antibiotics and the standards for the lowest inhibitory concentration that can be judged to be safe were referenced from the guidelines published by the European food safety authority (EFSA).

[0253] SampleAMPVANGENKANSTRERYCDTETCMSreptococcus thermophilusATG-V1100.0942420.1250.0160.0161.5EFSA(European food safety authority)2432326464224AMP: ampicillin; VAN: vancomycin, GEN: Gentamicin, KAN: kanamycin; STR: streptomycin; ERY: erythromycin; CD: clindamycin; TET: tetracycline; CM: chloramphenicol; NR: not required. (Each unit: ㎍ / ㎖)

[0254] The results of each analysis are presented in Table 21 as the results of the minimum inhibitory concentration (MIC) measurement experiment for the Streptococcus thermophilus ATG-V11 strain against major antibiotics.

[0255] Accordingly, it was confirmed that the strain of the present invention had antibiotic susceptibility that was significantly lower than the limit of the EFSA guideline, and thus it was found that there was no risk of the Streptococcus thermophilus ATG-V11 strain exchanging antibiotic resistance genes.

[0256]

[0257] <Example 3 of the Fourth Invention. Confirmation of Cytotoxicity>

[0258] TM-3 cells for the experiments of the present invention were cultured in a medium containing 90% DMEM (Dulbecco's modified Eagle's medium) and 10% FBS (heat-inactivated fetal bovine serum).

[0259] TM-3 cells were cultured at 5×10 3 / well were cultured in 96-well plates, and the number of bacteria was calculated by multiplicity of infection (MOI) ratio to treat Streptococcus thermophilus ATG-V11 strain to a certain amount of cells at a certain ratio. For this, TM-3 cells: Lactic acid bacteria strain were mixed at a ratio of 1:1 (5 × 10 3 CFU / well) to 1:100 (5×10 5 The cells were treated with a strain concentration of 10 CFU / well and cultured for 24 hours, and the proliferation ability of TM-3 cells was measured at an absorbance of 450 nm using a CCK substrate (D-Plus™), and the results are shown in Figure 18.

[0260] As shown in Fig. 18, it can be confirmed that the Streptococcus thermophilus ATG-V11 strain of the present invention is not cytotoxic.

[0261]

[0262] <Example 4 of the Fourth Invention. Confirmation of Testosterone Secretion>

[0263] Streptococcus thermophilus TA40 (TA 40 LYO 250 DCU, dairyconnection.com), a strain of the same species, was purchased from Danisco and used for comparative experiments.

[0264] For comparison of testosterone secretion, TM-3 cells were cultured at 1×10 5 The cells and each Streptococcus thermophilus strain were treated at a multiplicity of infection (MOI) of 1:1 to 1:10 in a 24-well plate and cultured for 24 hours. The culture medium was collected and centrifuged at 4°C, 1500 rpm, for 5 minutes to obtain the supernatant, and the testosterone expression level was measured using an ELISA kit (Enzo Life Science, ADI-901-065). The results are shown in Fig. 19.

[0265] As shown in Figure 19, the Streptococcus thermophilus ATG-V11 strain was found to increase testosterone expression in a concentration-dependent manner. In contrast, the Streptococcus thermophilus TA40 strain, although of the same species, did not exhibit this effect.

[0266]

[0267] <Example 5 of the Fourth Invention: Confirmation of Expression of Genes Involved in Testosterone Production>

[0268] Using TM-3 cells prepared under the same conditions as Example 3, mRNA was extracted from the cells using the RNeasy mini Kit (Qiagen, 74104), and cDNA was synthesized using the ReverTraAce-α- TM (TOYOBO) kit. Afterwards, the gene expression levels of 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I), 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI), and AR (Androgen Receptor), enzymes involved in testosterone production, were confirmed through real-time PCR.

[0269] The results are shown in Figures 20 to 22. The Streptococcus thermophilus ATG-V11 strain significantly increased the gene expression of 3β-HSD-I, 3β-HSD-VI, and AR, but the Streptococcus thermophilus TA40 strain had little effect.

[0270]

[0271] [Accession number]

[0272] [First invention strain]

[0273] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0274] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0275] Accession number: KCTC15545BP

[0276] Deposit date: 20230809

[0277]

[0278] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0279] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0280] Accession number: KCTC15540BP

[0281] Deposit date: 20230809

[0282]

[0283] [Second invention strain]

[0284] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0285] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0286] Accession number: KCTC15547BP

[0287] Deposit date: 20230809

[0288]

[0289] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0290] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0291] Accession number: KCTC15546BP

[0292] Deposit date: 20230809

[0293]

[0294] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0295] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0296] Accession number: KCTC15544BP

[0297] Deposit date: 20230809

[0298]

[0299] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0300] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0301] Accession number: KCTC15541BP

[0302] Deposit date: 20230809

[0303]

[0304] [Third invention strain]

[0305] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0306] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0307] Accession number: KCTC15542BP

[0308] Deposit date: 20230809

[0309]

[0310] [Fourth Invention Strain]

[0311] Name of depositor: Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology

[0312] Address of the depository: 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do (56212)

[0313] Accession number: KCTC15543BP

[0314] Deposit date: 20230809

[0315]

[0316]

[0317]

[0318]

Claims

1. A composition for improving male menopause symptoms, characterized by containing the Lacticaseibacillus paracasei ATG-V7 strain deposited as KCTC15545BP.

2. In paragraph 1, A composition for improving male menopause symptoms, characterized in that the above strain has the effect of enhancing the production of testosterone.

3. In paragraph 1, A composition for improving male menopausal symptoms, characterized in that the strain has the effect of increasing the expression of the 3β-HSD-I (3β Hydroxysteroid Dehydrogenase I) gene.

4. In paragraph 1, The above strain is a composition for improving male menopausal symptoms, characterized in that it has the effect of increasing the expression of 3β-HSD-VI (3β Hydroxysteroid Dehydrogenase VI) and AR (Androgen Receptor) genes.

5. In paragraph 1, The above strain is a composition for improving male menopause symptoms, characterized in that it has the effect of increasing the expression of the AR (Androgen Receptor) gene.

6. In paragraph 1, A composition for improving male menopause symptoms, characterized in that the strain has the effect of increasing the expression of the StAR (Steroidogenic Acute Regulatory Protein) gene.

7. A health functional food for preventing or improving male menopausal symptoms, characterized by containing the composition of paragraph 1.

8. A food composition for preventing or improving male menopause symptoms, characterized by containing the composition of paragraph 1.

9. A pharmaceutical composition for preventing or treating male menopause symptoms, characterized by containing the composition of paragraph 1.

10. Lacticaseibacillus paracasei ATG-V7 strain deposited as KCTC15545BP, which has the effect of enhancing testosterone production.

11. A method for preventing, improving or treating male menopause symptoms using the Lacticaseibacillus paracasei ATG-V7 strain deposited as KCTC15545BP.

12. A composition for improving male menopausal symptoms, characterized by containing the Lacticaseibacillus paracasei ATG-K22 strain deposited under the designation KCTC15540BP.

13. A composition for improving male menopausal symptoms, characterized by containing the Lactiplantibacillus plantarum ATG-V1 strain deposited as KCTC15547BP.

14. A composition for improving male menopause symptoms, characterized by containing the Lactiplantibacillus plantarum ATG-V2 strain deposited under the designation KCTC15546BP.

15. A composition for improving male menopausal symptoms, characterized by containing the Lactiplantibacillus plantarum ATG-V8 strain deposited under KCTC15544BP.

16. A composition for improving male menopausal symptoms, characterized by containing the Lactiplantibacillus plantarum ATG-K20 strain deposited under the designation KCTC15541BP.

17. A composition for improving male menopause symptoms, characterized in that it contains Limosilactobacillus fermentum ATG-F11 strain deposited as KCTC15542BP.

18. A composition for improving male menopause symptoms, characterized by containing the Streptococcus thermophilus ATG-V11 strain deposited as KCTC15543BP.

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