KR20210080182A - Composition for preventing and treating of obesity comprising powder of lactic acid cell lysate - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, and treating metabolic diseases, comprising atomized Lactobacillus plantarum K8 dead cells. The atomized Lactobacillus plantarum K8 dead cells according to the present invention have effects of inhibiting the differentiation and adipogenesis of adipocytes, thereby being able to be useful as a pharmaceutical composition, a health functional food composition, and a health food composition for preventing, alleviating or treating metabolic diseases such as obesity, and as a cosmetic composition for reducing or preventing cellulite.

Description

Composition for preventing and treating obesity comprising powder of lactic acid cell lysate, comprising atomized dead cells of lactic acid bacteria

The present invention relates to a composition for the prevention, improvement and treatment of metabolic diseases comprising atomized dead cells of lactic acid bacteria, and more specifically, the prevention of obesity including atomized Lactobacillus plantarum K8 dead cells, It relates to a pharmaceutical composition for improvement or treatment, a health functional food composition, and a health food composition.

Currently, most of the research on functional enhancement and disease prevention using lactic acid bacteria has mainly used live lactic acid bacteria as the concept of probiotics to induce changes in the intestinal flora. In Korea, consumer awareness of lactic acid bacteria dead cells is still insufficient, and the reality is that lactic acid bacteria manufacturers lack information about dead cells. In particular, since there are many consumers, manufacturers, and researchers who think that the lactic acid bacteria dead cells will not have any effect on the body, studies applied in the form of dead bacteria or lysate of lactic acid bacteria are relatively insignificant.

The lactic acid bacteria dead cells have the advantage of live bacteria, but the distribution of lactic acid bacteria sterilized through heat treatment is much more advantageous than the live bacteria, and it is not affected by the gastointestinal environment such as stomach acid, so it has the advantage of being able to safely move to the intestinal tract. have. According to a paper published in Nutrition Reviews in 2009, it was suggested that “for the safe use of probiotics, if possible, slain bacteria, that is, microbial carcass, should be used.” The idea of being active is being withdrawn.

It is known that live lactic acid bacteria settle in the intestine and increase their number to compete with harmful bacteria to normalize the intestinal flora, whereas dead bacteria are known to exhibit functionality through the role of prebiotics as food for beneficial bacteria in the intestine. In fact, it has been reported that ingestion of dead cells normalizes the intestinal flora by inducing rapid production and proliferation of beneficial bacteria such as Bifidobacteria and Lactobacilli, as well as delayed and growth inhibition of harmful bacteria such as Enterobacteriae. Due to this complex action, lactic acid bacteria can be expected to function as synbiotics alone.

In 2012, it was reported that lactic acid bacteria products (including dead bacteria) other than live bacteria accounted for more than 25% of the total lactic acid bacteria market in the Japanese lactobacillus market in 2012. There is a need to create Judging from these results, it is expected that the R&D of various physiologically active materials using lactic acid bacteria dead cells will have a very important industrial value.

According to a recent survey, the adult obesity rate is 19.5% of the OECD average, which has been continuously increasing since 1990. Obesity has been shown to increase the risk of cardiovascular disease, diabetes, kidney disease, etc., and the number of years of potential lifespan loss, and the number of deaths due to obesity in 2015 is estimated to be about 4 million. As the social cost of obesity is rapidly increasing, each country is preparing various national obesity management measures, such as providing financial incentives to combat obesity, and strengthening taxes on obesity-causing food and beverages. According to the National Health Insurance Corporation, it has been reported that the socioeconomic cost of obesity exceeds 11 trillion won per year as of 2018.

The diet industry was worth 7.6 trillion won a year as of 2016, and the size of the diet food and other supplies market is estimated to be about 42% or 3.2 trillion won a year, and the diet food industry is the largest in the diet market. occupies size. According to the '2017 Health Functional Food Market Status and Consumer Actual Survey' report published by the Korea Health Functional Food Association, the health functional food most preferred by women in their 20s and 30s was identified as a diet product. It is expected to increase steadily with the increase in prevalence. Accordingly, the development and industrialization of new types of health functional foods and pharmaceutical materials for the prevention and treatment of obesity are in great demand.

The present inventors have completed the present invention by confirming that the atomized lactic acid bacteria dead cells (lactic acid bacteria disrupted body, hereinafter LAB-C) has an anti-obesity effect that inhibits adipogenesis and fat differentiation.

Korean Patent Publication No. 10-2012-0128260 Korean Patent Publication No. 10-2011-0113026

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating metabolic diseases.

Another object of the present invention is to provide a health functional food and health food for the prevention or improvement of metabolic diseases.

Another object of the present invention is to provide a cosmetic composition for reducing or preventing cellulite.

In order to achieve the above object,

The present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) for the prevention or treatment of metabolic diseases comprising at least one selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts as an active ingredient A pharmaceutical composition is provided.

In addition, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) Live cells, dead cells, culture, culture solution, lysate, and any one or more selected from the group consisting of extracts as an active ingredient for the prevention of metabolic diseases or Provides health functional food and health food for improvement.

Furthermore, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) of live cells, dead cells, culture, culture solution, lysate and cellulite containing any one or more selected from the group consisting of extracts as an active ingredient reduction or prevention A cosmetic composition is provided.

Furthermore, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) live cells, dead cells, culture, culture solution, lysate and lipoteichoic acid isolated from any one or more selected from the group consisting of extracts (lipoteichoic acid) ) provides a pharmaceutical composition, a health functional food composition and a health food composition for the prevention, treatment or improvement of metabolic diseases comprising as an active ingredient.

Furthermore, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) live cells, dead cells, culture, culture solution, lysate and lipoteichoic acid isolated from any one or more selected from the group consisting of extracts (lipoteichoic acid) ) provides a cosmetic composition for reducing or preventing cellulite comprising as an active ingredient.

Since the atomized Lactobacillus plantarum K8 dead cells according to the present invention have an effect of inhibiting adipocyte differentiation and adipogenesis, a pharmaceutical composition for prevention, improvement or treatment of metabolic diseases such as obesity, health It may be useful as a nutraceutical composition, a health food composition, and a cosmetic composition for reducing or preventing cellulite.

1 is a 3T3-L1 according to the treatment concentration of the atomized Lactobacillus plantarum K8 dead cells (Example 2, cell lysate) and Lactobacillus plantarum K8 live cells (Comparative Example 1, live cell) according to the present invention; It is a graph showing the survival rate (%) of preadipocytes.
Figure 2 shows the dead cells (Example 2, cell lysate) of the atomized Lactobacillus plantarum K8 strain according to the present invention and the fat according to the treatment concentration of the Lactobacillus plantarum K8 live cells (Comparative Example 1, live cell) It is a graph showing the degree of generation (%).
3 is a dead cell (Example 2, L. plantarum K8) of an atomized Lactobacillus plantarum K8 strain according to the present invention and a dead cell of another type of atomized Lactobacillus plantarum strain (Comparative Example 2, L .plantarum) is a graph showing the degree of fat production (%) according to the treatment concentration.
Figure 4 shows the protein expression level (%) of the lipodifferentiation-related genes (PPARγ, C/EBPa and FABP4) according to the treatment concentration of the dead cells (Example 2, LAB-C) of the atomized Lactobacillus plantarum K8 strain according to the present invention; ) is the result.
Figure 5 is a dead cell (Example 2, LAB-C) of the atomized Lactobacillus plantarum K8 strain according to the present invention adipogenesis-related genes (C / EBPβ, DLK1, KLF4, KLF5, KROX20 and C / EBPδ) is a result showing the protein expression level (%).
6 shows the survival rate (%) and adipogenesis degree (%) of 3T3-L1 preadipocytes according to the treatment concentration of lipoteichoic acid (LTA), a cell wall component of the Lactobacillus plantarum K8 strain of the present invention. ) is a graph showing

Hereinafter, the present invention will be described in detail.

Pharmaceutical composition for the prevention or treatment of metabolic diseases

The present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) for the prevention or treatment of metabolic diseases comprising at least one selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts as an active ingredient A pharmaceutical composition is provided.

The Lactobacillus plantarum K8 of the present invention may be a strain isolated from kimchi, and is preferably deposited at the Korea Institute of Biotechnology and Biotechnology Biological Resources Center (KCTC: Korean Collection for Type Cultures) and given an accession number of KTCT 10887BP. It may be the received Lactobacillus plantarum K8 strain.

In the pharmaceutical composition according to the present invention, the Lactobacillus plantarum K8 may be an atomized Lactobacillus plantarum K8 dead cell.

The 'micronized Lactobacillus plantarum K8 dead cells' may mean a shredded body of the Lactobacillus plantarum K8 strain, and the live cells are atomized into dead cells through the crushing process, or the dead cells themselves are atomized. may be used.

In the pharmaceutical composition according to the present invention, the atomized Lactobacillus plantarum K8 dead cells is a Lactobacillus plantarum K8 (Lactobacillus plantarum K8) strain (accession number: KTCT 10887BP) live or dead cells of microflora It may be a powder prepared by a method comprising; crushing with an ider and lyophilizing to obtain a crushed body (step 1).

In the pharmaceutical composition according to the present invention, the number of live or dead cells of step 1 ^{may be adjusted to be 1×10 8} to 1×10 ¹⁴ CFU, preferably 1×10 ¹¹ to 1×10 ^It may be adjusted to be 13 CFU, more preferably 1×10 ¹² to 1×10 ¹³ CFU, but is not limited thereto.

In the pharmaceutical composition according to the present invention, the live cells of the Lactobacillus plantarum K8 strain of step 1 may be prepared by culturing before crushing to obtain the cells. In this case, the cells may be recovered through centrifugation, and the method may further include washing the obtained cells with sterile distilled water.

In the pharmaceutical composition according to the present invention, the live or dead cells of the Lactobacillus plantarum K8 strain may include lipoteichoic acid (LTA).

In the pharmaceutical composition according to the present invention, the metabolic disease may be any one or more selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver. Preferably, it may be obese.

According to one embodiment of the present invention, it was confirmed that the atomized Lactobacillus plantarum K8 dead cells according to the present invention can prevent or treat metabolic diseases such as obesity by inhibiting adipocyte differentiation and adipogenesis (experimental). see Examples 1-4).

The atomized Lactobacillus plantarum K8 dead cells according to the present invention may be administered in various oral and parenteral formulations during clinical administration, preferably oral administration, transdermal administration, topical administration, inhalation administration, intranasal administration , intravenous administration, intradermal administration, intraperitoneal administration, injection, and may be administered in any one or more formulations selected from the group consisting of subcutaneous injection formulations. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, and these solid preparations include at least one excipient in one or more atomized Lactobacillus plantarum K8 dead cells of the present invention. For example, it is prepared by mixing starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, or syrups. 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. can

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, creams, gels, sprays, drops, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, etc. may be used.

The effective dose for the human body of the atomized Lactobacillus plantarum K8 dead cells of the present invention may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and is generally about 0.01-1000 mg /kg/day, preferably 0.1-500 mg/kg/, may be administered in divided doses once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist.

The pharmaceutical composition of the present invention may additionally include a known therapeutic agent for metabolic diseases such as obesity, in addition to the atomized Lactobacillus plantarum K8 dead cells as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases. have.

Health functional food and health food composition for the prevention or improvement of metabolic diseases

In addition, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) Live cells, dead cells, culture, culture solution, lysate, and any one or more selected from the group consisting of extracts as an active ingredient for the prevention of metabolic diseases or It provides a health functional food composition for improvement.

The term "health functional food" as used in the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful in the human body. Here, the term 'functionality' refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the ordinary technical field, and at the time of the preparation, it can be prepared by adding raw materials and components commonly added in the conventional technical field. In addition, the dosage form of the health functional food may also be manufactured without limitation as long as the dosage form is recognized as a health functional food. The health functional food composition of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long period of time using food as a raw material, and has excellent portability, The health functional food of the present invention can be ingested as an adjuvant to enhance the effect of a therapeutic agent for metabolic diseases.

In addition, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) Live cells, dead cells, culture, culture solution, lysate, and any one or more selected from the group consisting of extracts as an active ingredient for the prevention of metabolic diseases or A health food composition for improvement is provided.

In the health functional food composition or health food composition according to the present invention, the Lactobacillus plantarum K8 may be an atomized Lactobacillus plantarum K8 dead cell.

The 'micronized Lactobacillus plantarum K8 dead cells' may mean a shredded body of the Lactobacillus plantarum K8 strain, and the live cells are atomized into dead cells through the crushing process, or the dead cells themselves are atomized. may be used.

In the health functional food composition or health food composition according to the present invention, the atomized Lactobacillus plantarum K8 dead cells is a live cell of Lactobacillus plantarum K8 (Lactobacillus plantarum K8) strain (accession number: KTCT 10887BP) or It may be a powder prepared by a method comprising; crushing the dead cells with a microfluidizer and lyophilizing to obtain a crushed body (step 1).

In the health functional food composition or health food composition according to the present invention, the number of live or dead cells of step 1 ^{may be adjusted to be 1×10 8} to 1×10 ¹⁴ CFU, preferably 1×10 ¹¹ to 1×10 ¹³ CFU, more preferably 1×10 ¹² to 1×10 ¹³ CFU may be adjusted to be, but is not limited thereto.

In the health functional food composition or health food composition according to the present invention, the live cells of the Lactobacillus plantarum K8 strain of step 1 may be prepared by further comprising the step of culturing before crushing to obtain the cells. In this case, the cells may be recovered through centrifugation, and the method may further include washing the obtained cells with sterile distilled water.

In the health functional food composition or health food composition according to the present invention, the live or dead cells of the Lactobacillus plantarum K8 strain may include lipoteichoic acid (LTA).

In the health functional food composition or health food composition according to the present invention, the metabolic disease may be any one or more selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver. Preferably, it may be obese.

According to one embodiment of the present invention, it was confirmed that the atomized Lactobacillus plantarum K8 dead cells according to the present invention can prevent or treat metabolic diseases such as obesity by inhibiting adipocyte differentiation and adipogenesis (experimental). see Examples 1-4).

When the composition of the present invention is used as a health functional food composition or health food composition, the atomized Lactobacillus plantarum K8 dead cells can be added as it is or used with other foods or food ingredients, and appropriate according to a conventional method. can be used In addition to the active ingredient, the composition may contain a food additive that is pharmaceutically acceptable, and the mixing amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).

As used in the present invention, the term "food supplement additive" refers to a component that can be supplementally added to food, and is added to manufacture a health functional food or health food of each formulation, and those skilled in the art can appropriately select and use it. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.

In addition, there is no limitation on the type of health functional food or health food in which the composition of the present invention can be used. In addition, the composition comprising the atomized Lactobacillus plantarum K8 dead cells of the present invention as an active ingredient can be prepared by mixing known additives with other suitable auxiliary ingredients that may be contained in health functional foods according to the selection of those skilled in the art. . Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes and the like, and it can be prepared by adding to juice, tea, jelly, juice, etc. prepared by using the atomized Lactobacillus plantarum K8 dead cells according to the present invention as a main component.

Cosmetic composition for reducing or preventing cellulite

The present invention provides a cosmetic for reducing or preventing cellulite comprising at least one selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts of Lactobacillus plantarum K8 as an active ingredient A composition is provided.

In the cosmetic composition according to the present invention, the Lactobacillus plantarum K8 may be an atomized Lactobacillus plantarum K8 dead cell.

The 'micronized Lactobacillus plantarum K8 dead cells' may mean a shredded body of the Lactobacillus plantarum K8 strain, and the live cells are atomized into dead cells through the crushing process, or the dead cells themselves are atomized. may be used.

In the cosmetic composition according to the present invention, the atomized Lactobacillus plantarum K8 dead cells are Lactobacillus plantarum K8 (Lactobacillus plantarum K8) strains (accession number: KTCT 10887BP) live or dead cells of microflora. It may be a powder prepared by a method comprising; crushing with low and lyophilizing to obtain a crushed body (step 1).

In the cosmetic composition according to the present invention, the number of live or dead cells of step 1 ^{may be adjusted to be 1×10 8} to 1×10 ¹⁴ CFU, preferably 1×10 ¹¹ to 1×10 ¹³ CFU, more preferably 1×10 ¹² to 1×10 ¹³ CFU may be adjusted to be, but is not limited thereto.

In the cosmetic composition according to the present invention, the live cells of the Lactobacillus plantarum K8 strain of step 1 may be prepared by culturing before crushing to obtain the cells. In this case, the cells may be recovered through centrifugation, and the method may further include washing the obtained cells with sterile distilled water.

In the cosmetic composition according to the present invention, the live or dead cells of the Lactobacillus plantarum K8 strain may include lipoteichoic acid (LTA).

The cosmetic composition according to the present invention is, for example, a solution, a gel, a solid or kneaded anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic (liposome), nonionic It may be provided in the form of a vesicular dispersion in the form of a cream, skin, lotion, oil, powder, ointment, spray or in the form of a cone stick. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition contains a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent in addition to the atomized Lactobacillus plantarum K8 dead cells of the present invention. agent), fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, It may contain adjuvants commonly used in the cosmetic field, such as lipid vesicles or any other ingredients commonly used in cosmetics.

In the cosmetic composition of the present invention, the atomized Lactobacillus plantarum K8 dead cells of the present invention may be added in an amount of 0.01 to 50% by weight, preferably 0.1 to 10% by weight, in the cosmetic composition usually contained therein. .

When the atomized Lactobacillus plantarum K8 dead cells according to the present invention is used as an external preparation for skin, additionally, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, and foaming agents (foaming agent), fragrance, surfactant, water, ionic or nonionic emulsifier, filler, sequestering and chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic or lipophilic It may contain adjuvants commonly used in the field of dermatology, such as active agents, lipid vesicles or any other ingredients commonly used in external preparations for skin. In addition, the ingredients may be introduced in an amount generally used in the field of dermatology.

Composition for the prevention, treatment or improvement of metabolic diseases comprising lipoteichoic acid

The present invention is effective for Lactobacillus plantarum K8 (Lactobacillus plantarum K8) isolated from any one or more selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts. It provides a pharmaceutical composition for the prevention or treatment of metabolic diseases comprising as a component.

In addition, the present invention is Lactobacillus plantarum K8 (Lactobacillus plantarum K8) lipoteichoic acid isolated from any one or more selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts. It provides a health functional food composition for the prevention or improvement of metabolic diseases comprising as an active ingredient.

In the pharmaceutical composition, health functional food composition or health food composition according to the present invention, the Lactobacillus plantarum K8 may be an atomized Lactobacillus plantarum K8 dead cell.

The 'micronized Lactobacillus plantarum K8 dead cells' may mean a shredded body of the Lactobacillus plantarum K8 strain, and the live cells are atomized into dead cells through the crushing process, or the dead cells themselves are atomized. may be used.

In the pharmaceutical composition, health functional food composition or health food composition according to the present invention, the atomized Lactobacillus plantarum K8 dead cells are Lactobacillus plantarum K8 (Lactobacillus plantarum K8) strain (accession number: KTCT 10887BP) It may be a powder prepared by a method comprising; crushing live or dead cells of the microfluidizer and freeze-drying to obtain a disrupted body (step 1).

In the pharmaceutical composition, health functional food composition or health food composition according to the present invention, an organic solvent extraction method using butanol is used from the live or dead cells of the lipoteichoic acid strain. It may be separated or purified by performing a hydrophobic column using Octyl-Sepharose CL-4B or ion-exchange column chromatography using DEAE-Sephadex. , but is not limited thereto, and may be using a method for isolating or purifying lipoteichoic acid from a generally known strain.

In the pharmaceutical composition, health functional food composition or health food composition according to the present invention, the metabolic disease may be any one or more selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver. Preferably, it may be obese.

According to an embodiment of the present invention, it was confirmed that lipoteichoic acid according to the present invention can inhibit the differentiation and adipogenesis of adipocytes, thereby confirming that it can be used to prevent, improve or treat metabolic diseases such as obesity. (see Experimental Example 5).

Cosmetic composition for reducing or preventing cellulite comprising lipoteichoic acid

The present invention is effective for Lactobacillus plantarum K8 (Lactobacillus plantarum K8) isolated from any one or more selected from the group consisting of live cells, dead cells, cultures, cultures, lysates and extracts. It provides a cosmetic composition for reducing or preventing cellulite comprising as an ingredient.

In the cosmetic composition according to the present invention, the Lactobacillus plantarum K8 may be an atomized Lactobacillus plantarum K8 dead cell.

The 'micronized Lactobacillus plantarum K8 dead cells' may mean a shredded body of the Lactobacillus plantarum K8 strain, and the live cells are atomized into dead cells through the crushing process, or the dead cells themselves are atomized. may be used.

In the cosmetic composition according to the present invention, the atomized Lactobacillus plantarum K8 dead cells are Lactobacillus plantarum K8 (Lactobacillus plantarum K8) strains (accession number: KTCT 10887BP) live or dead cells of microflora. It may be a powder prepared by a method comprising; crushing with low and lyophilizing to obtain a crushed body (step 1).

The present invention will be described in more detail through the following examples. However, the following examples are only for specifying the contents of the present invention, and the present invention is not limited thereto.

<Example 1> Lactobacillus plantarum K8 strain isolation and identification

The lactic acid bacteria used in Examples and Experimental Examples of the present invention were provided with Lactobacillus plantarum K8 strain isolated from kimchi, a traditional Korean fermented food, and used.

Cells were isolated from kimchi, a traditional Korean fermented food, and the isolated cells were identified by 16S rRNA sequencing and biochemical tests. As a result, they were named Lactobacillus plantarum K8, and as a new strain, Korea The number of KTCT 10887BP has been given to it by depositing it with the Biotechnology Research Institute (KCTC: Korean Collection for Type Cultures).

The strain was cultured using Lactobacillus MRS medium and stored frozen at -70°C by adding 15% of glycerol, and rapidly thawed in a 37°C thermostat to stabilize the cells during raw material production.

<Example 2> Preparation of atomized Lactobacillus plantarum K8 dead cells

The kimchi lactic acid bacteria Lactobacillus plantarum K8 (accession number: KTCT 10887BP) of Example 1 was crushed and lyophilized to prepare an atomized lactic acid bacteria dead cell (lactic acid bacterial cell lysate: LAB-C).

Specifically, Lactobacillus plantarum K8 stock was suspended in MRS medium and activated at 37° C. for 9 hours, then only the cells were recovered through 12 hours of main culture and centrifugation (8000 rpm, 8 minutes). Cells were washed 3 times with sterile distilled water, and the recovered cells were adjusted to ^{about 4×10 12 CFU through cell count.}

Then, after adding 500 mL of sterile distilled water, the cells are crushed through a microfluidizer and freeze-dried to obtain finely divided lactic acid bacterial cell lysate (LAB-C: Lactic acid bacterial cell lysate). prepared.

<Comparative Example 1> Preparation of lyophilized Lactobacillus plantarum K8 live cells

Lactobacillus plantarum K8 stock was cultured in the same manner as in Example 2 using the same strain as in Example 1, and the cells were adjusted to ^{about 4×10 12 CFU.}

Then, lyophilization only proceeded without crushing, to prepare a lactic acid bacterial live cell lyophilisate.

The freeze-dried product of live lactic acid bacteria prepared only by freeze-drying is a freeze-dried product with the same number of bacteria as the lactic acid bacteria dead cells (shredded body) atomized by crushing and freeze-drying of Example 2.

<Comparative Example 2> Preparation of atomized Lactobacillus plantarum dead cells

Instead of the Lactobacillus plantarum K8 strain of Example 1, the Lactobacillus plantarum strain (accession number: CT 5045) purchased from the Korean Collection for Type Cultures was used in the same procedure as in Example 2 above. by culturing the strain, crushing and freeze-drying, to prepare an atomized lactic acid bacteria dead cell (shrapnel) of L. plantarum.

The atomized lactic acid bacteria dead cells (shrapnel) of the L. plantarum is a freeze-dried product of the same number of cells as the atomized lactic acid bacteria dead cells (shrapnel) of L. plantarum K8 of Example 2.

<Experimental Example 1> Cytotoxicity evaluation of atomized Lactobacillus plantarum K8 dead cells

The toxicity of the atomized Lactobacillus plantarum K8 dead cells (LAB-C, cell lysate) prepared in Example 2 to 3T3-L1 pre-adipocytes was evaluated. In addition, the cell viability was compared with the case of treating the live cell freeze-dried product (Comparative Example 1, live cell) of the same strain as the dead cell.

Specifically, 3T3-L1 cells used in the experiment were purchased from the American type culture collection (ATCC, Rockville, MD, USA), and 10% bovine serum (BS, Gibco, NY, USA), 100 U/mL of penicillin, Using DMEM medium containing 100 μg/mL of streptomycin, subculture was performed every 3 days at _{37° C. and 5% CO 2 conditions.} To evaluate cytotoxicity, 3T3-L1 preadipocytes were seeded at 1×10 ⁴ cells/well in a 96-well tissue culture plate and stabilized for 24 hours. Then, all of the culture medium was removed, and 200 μL of the culture medium containing each sample (Example 2 and Comparative Example 1) suspended at an appropriate concentration was added and cultured for 24 hours. The toxicity of 3T3-L1 cells by the sample was measured by a colorimetric method using a 0.1 mg/mL MTT (Invitrogen) solution after removing all of the culture supernatant, and the cell viability (%) compared to the untreated control (ND). indicated.

1 is a 3T3-L1 according to the treatment concentration of the atomized Lactobacillus plantarum K8 dead cells (Example 2, cell lysate) and Lactobacillus plantarum K8 live cells (Comparative Example 1, live cell) according to the present invention; It is a graph showing the survival rate (%) of preadipocytes.

1 is a cytotoxicity test for (pre) adipocytes before measuring the effect of Lactobacillus live and dead cells on fat accumulation. As shown in FIG. 1, the micronized Lactobacillus plantarum K8 dead cells (LAB-C) according to Example 2 did not have toxicity to cells up to a concentration of 1000 μg/mL, but rather 16-31 μg/ At concentrations above mL, the cell viability was further increased. In addition, it was found that the viability of 3T3-L1 cells at the same concentration was significantly higher when treated with micronized dead cells than with live cells.

<Experimental Example 2> Evaluation of adipogenesis inhibitory activity of atomized Lactobacillus plantarum K8 dead cells and live cells

Oil red O staining method for the atomized Lactobacillus plantarum K8 dead cells (LAB-C, cell lysate) prepared in Example 2 and the lyophilized product of the same strain (Comparative Example 1, live cell) was used to compare the adipogenesis inhibitory activity.

Specifically, to check the ability to inhibit adipogenesis, 3T3-L1 preadipocytes were seeded at 5×10 ⁴ cells/well in a 24-well tissue culture plate, and during the cell growth phase, medium with DMEM containing 10% BS until confluent. was cultured while exchanging. After 2 days from confluent, 10% FBS and 5 μg/mL of insulin (INS, Sigma), 517 nM of 3-isobutyl-1-methylxanthine (IBMX, Sigma), DMEM containing 1 μM dexamethasone (DEX, Sigma) was exchanged, and 3 days after differentiation induction, DMEM medium containing only 10% FBS and insulin was exchanged. Thereafter, the adipogenic state of the cells was confirmed while exchanging the medium with DMEM containing only 10% FBS at intervals of 2 days, and the samples (Example 2 and Comparative Example 1) were treated twice on day 0 and day 3, respectively, and the control medium was treated instead of the sample. Seven days after induction of differentiation (day 7), the culture medium was removed to measure the amount of generated triglycerides, washed twice with PBS, and the recovered cells were fixed with PBS containing 10% formalin for 20 minutes. . After removing formalin, cells washed 4 times with distilled water were stained with intracellular triglycerides for 20 minutes by adding 0.5% (w/v) Oil red O working solution/60% isopropanol solution. Stained intracellular triglycerides were extracted with 100% isopropanol and absorbance was measured at 520 nm, and the degree of differentiation of fat was expressed as relative activity (%) to the undifferentiated control, and the inhibitory activity for the differentiation control instead of the sample in the differentiation induction medium. (%).

Figure 2 is fat according to the treatment concentration of the dead cells (Example 2, cell lysate) and Lactobacillus plantarum K8 live cells (Comparative Example 1, live cell) of the atomized Lactobacillus plantarum K8 strain according to the present invention It is a graph showing the degree of generation (%).

As a result of measuring the effect on fat accumulation during the differentiation period of adipocytes using Oil-red O staining, Lactobacillus live cells (LP-live cells) and dead cells (LAB-C, cell lysates) were concentration-dependently accumulated in fat. was observed to inhibit It was confirmed that the adipogenesis inhibitory activity of 3T3-L1 preadipocytes was significantly increased when compared to live cells at the same concentration in the concentration range of 25 to 200 μg/mL. In particular, at a concentration of 250 mg/mL, the live cells showed a 40% inhibitory effect on fat accumulation and the dead cells showed a 60% inhibitory effect, indicating that the fat accumulation inhibitory effect of dead cells (micronized dead cells) was superior to that of live cells.

<Experimental Example 3> Evaluation of adipogenesis inhibitory activity of atomized Lactobacillus plantarum K8 dead cells and Lactobacillus plantarum dead cells

Fat of the atomized Lactobacillus plantarum K8 dead cells (LAB-C, L. plantarum K8) prepared in Example 2 and the atomized Lactobacillus plantarum dead cells (L. plantarum) prepared in Comparative Example 2 The production inhibitory efficacy was comparatively evaluated. The experimental method is shown in FIG. 3 by calculating the lipid accumulation (%) through Oil red O staining using the same method as in Experimental Example 2.

3 is a dead cell of an atomized Lactobacillus plantarum K8 strain according to the present invention (Example 2, L. plantarum K8) and a dead cell of another type of atomized Lactobacillus plantarum strain (Comparative Example 2, L .plantarum) is a graph showing the degree of fat production (%) according to the treatment concentration.

As a result of comparing the fat accumulation inhibitory effect of L. plantarum (CT 5045) and L. plantarum K8 (KTCT 10887BP) through Oil-red O staining, the same concentration of atomized L. plantraum in the concentration range of 6.25~500μg/mL Compared with dead cells, it was shown that the degree of 3T3-L1 fat accumulation was significantly reduced by the treatment of the atomized L. plantarum K8 dead cells according to the present invention. In particular, at a concentration of 500 μg/mL, L. plantarum had almost no effect. Although not shown, L. plantarum K8 showed more than 60% inhibitory effect. Accordingly, it was confirmed that the adipogenesis inhibitory activity of the K8 strain of the present invention was remarkably excellent among L. plantarum strains.

<Experimental Example 4> Efficacy of inhibiting expression of genes related to adipogenic differentiation of micronized Lactobacillus plantarum K8 dead cells

In order to confirm the effect of the atomized Lactobacillus plantarum K8 dead cells (LAB-C) prepared in Example 2 on adipogenesis and adipogenesis, the related gene expression inhibition efficacy was evaluated.

During or after differentiation of adipocytes, the cells were lysed using a lysis buffer containing proteolytic enzyme and phosphatase inhibitor to obtain a protein extract. The cell lysate was centrifuged to recover only the supernatant, and after quantifying the protein, 50 μg of the protein was electrophoresed in 10% or 12% acrylamide and then transferred to a PVDF membrane. After blocking the membrane with 5% skim milk for 30 minutes, the primary antibody and secondary antibody were reacted, and the detection of the target protein was confirmed by chemiluminescence using LAS imaging software. The amount of protein in each group was quantified using the Image J program.

Figure 4 shows the protein expression level (%) of the lipodifferentiation-related genes (PPARγ, C/EBPa and FABP4) according to the treatment concentration of the dead cells (Example 2, LAB-C) of the atomized Lactobacillus plantarum K8 strain according to the present invention; ) is the result.

The effect of the atomized L. plantarum K8 dead cells (Example 2, LAB-C) on the expression patterns of the fat differentiation factors PPARγ and C/EBPa and their target protein FABP4 was analyzed by Western blot. As a result, it was observed that the atomized dead K8 cells (K8 lysate) significantly reduced the protein expression of adipogenic factors during adipogenesis. At a concentration of 200 μg/mL of micronized K8 dead cells, protein expression of fat differentiation factor and FABP4 was reduced by more than 80%.

Figure 5 is a dead cell (Example 2, LAB-C) of the atomized Lactobacillus plantarum K8 strain according to the present invention adipogenesis-related genes (C / EBPβ, DLK1, KLF4, KLF5, KROX20 and C / EBPδ) is a result showing the protein expression level (%).

As a result of analyzing the expression patterns of factors involved in the initial stage of adipogenesis through Western blot and real-time PCR, the atomized dead cells of L. plantarum K8 (Example 2, LAB-C) were C It significantly reduced the amount of /EBPb protein and suppressed the mRNA expression of KLF4/5 and KROX20 by more than 50%. On the other hand, DLK1, a factor that decreases during fat differentiation, showed a significant increase when the atomized K8 dead cells were treated. From these results, the micronized K8 dead cells of the present invention mainly act at the initial stage when 3T3-L1 preadipocytes are converted to adipogenic cells, thereby reducing factors that promote early adipogenesis and factors inhibiting early adipogenesis. was confirmed to increase.

<Experimental Example 5> Adipogenesis inhibitory activity evaluation of lipoteichoic acid isolated from Lactobacillus plantarum K8

In general, as one of the major cell wall components of Gram-positive bacteria including Lactobacillus plantarum, lipoteichoic acid (LTA) is known, and the lipoteichoic acid has been reported to have immune-enhancing and anti-inflammatory activity. have. It was confirmed that the lipoteichoic acid was also contained in the micronized Lactobacillus plantarum K8 dead cells and the Lactobacillus plantarum K8 live cells according to the present invention. Adipogenesis inhibitory activity was evaluated in the same manner as in Experimental Example 2. The fat accumulation inhibitory activity of lipoteichoic acid in the (pre)adipocyte cytotoxicity test and in the concentration range in which toxicity is not shown is shown in FIG.

6 shows the survival rate (%) and adipogenesis degree (%) of 3T3-L1 precursor cells according to the treatment concentration of lipoteichoic acid (LTA), a cell wall component of the Lactobacillus plantarum K8 strain of the present invention. ) is a graph showing

As a result of the analysis, lipoteichoic acid was not observed to be toxic to cells up to a concentration of 150-300 μg/mL, but rather the survival of the cells was further increased. As a result of measuring the effect on fat accumulation in the non-toxic concentration range using the Oil-red O staining method, lipoteichoic acid inhibits adipogenesis in 3T3-L1 preadipocytes in the concentration range of 31.25 to 250 μg/mL. was found to increase in a concentration-dependent manner. Through this, it was confirmed that lipoteichoic acid can play a key role in the expression of adipogenesis inhibitory activity in live or dead Lactobacillus plantarum cells.

<Formulation Example 1> Preparation of pharmaceutical formulations

The atomized Lactobacillus plantarum K8 dead cells (hereinafter, LAB-C), which are the active substances according to the present invention, can be formulated in various forms depending on the purpose. The following exemplifies several formulation methods containing LAB-C as an active ingredient according to the present invention, but the present invention is not limited thereto.

<1-1> Preparation of powder

LAB-C 2 g

lactose 1 g

After mixing the above ingredients, the powder was prepared by filling in an airtight cloth.

<1-2> Preparation of tablets

LAB-C 100 mg

corn starch 100 mg

lactose 100 mg

magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional method for manufacturing tablets.

<1-3> Preparation of capsules

LAB-C 100 mg

corn starch 100 mg

lactose 100 mg

magnesium stearate 2 mg

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

<1-4> Preparation of injection

LAB-C 10 μg/ml

dilute hydrochloric acid BP until pH 3.5

Sodium Chloride BP for Injection up to 1 ml

Dissolve the LAB-C according to the invention in an appropriate volume of sodium chloride BP for injection, adjust the pH of the resulting solution to pH 3.5 with dilute hydrochloric acid BP, adjust the volume with sodium chloride BP for injection and mix thoroughly did. The solution was filled in a 5 ml Type I ampoule made of clear glass, sealed under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120° C. for 15 minutes or more to prepare an injection solution.

<1-5> Preparation of liquid preparation

LAB-C 100 mg

Lee Seonghwadang 10 g

mannitol 5 g

Purified water appropriate amount

According to a conventional method for preparing a solution, add each component to purified water to dissolve it, add lemon flavor, mix the above components, add purified water to adjust the total to 100 mL, fill a brown bottle, and sterilize to prepare a solution did.

<Formulation Example 2> Manufacture of health functional food and health food

LAB-C according to the present invention can be manufactured into various types of health functional food or health food depending on the purpose. The following exemplifies several health functional foods and methods for producing health foods containing LAB-C as an active ingredient according to the present invention, but the present invention is not limited thereto.

<2-1> Manufacture of health functional food

LAB-C 100 mg

vitamin mixture appropriate amount

vitamin A acetate 70 μg

vitamin E 1.0 mg

vitamin B1 0.13 mg

vitamin B2 0.15 mg

vitamin B6 0.5 mg

vitamin B12 0.2 μg

vitamin C 10 mg

biotin 10 μg

Nicotinamide 1.7 mg

folic acid 50 μg

Calcium Pantothenate 0.5 mg

mineral mixture appropriate amount

ferrous sulfate 1.75 mg

zinc oxide 0.82 mg

magnesium carbonate 25.3 mg

monobasic potassium phosphate 15 mg

Dibasic Calcium Phosphate 55 mg

Potassium Citrate 90 mg

calcium carbonate 100 mg

magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food, but the composition is mixed in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules can be prepared and used in the preparation of a health functional food composition according to a conventional method.

<2-2> Manufacture of health functional beverages

LAB-C 100 mg

citric acid 100 mg

oligosaccharide 100 mg

Plum Concentrate 2 mg

Taurine 100 mg

Purified water is added to 500 mL

After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in one sterilized container, sealed and sterilized, and then refrigerated. used in the manufacture of health beverage compositions. Although the composition ratio is a composition that is relatively suitable for a beverage of preference in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, demand country, and use purpose.

<2-3> Production of dairy products

0.01-1 parts by weight of LAB-C of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-4> Manufacture of wire type

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. It was prepared by mixing the dry powder of grains, seeds and LAB-C prepared above in the following ratio.

Grains (34 parts by weight of brown rice, 19 parts by weight of barley, 20 parts by weight of barley),

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame),

LAB-C (2 parts by weight),

Reishi (1.5 parts by weight), and

Rehmannia (1.5 parts by weight).

<Formulation Example 3> Preparation of cosmetics

The LAB-C according to the present invention can be prepared into various types of cosmetics depending on the purpose. The following exemplifies the manufacturing method of several cosmetics containing LAB-C as an active ingredient according to the present invention, but the present invention is not limited thereto.

<3-1> Manufacture of softened lotion

LAB-C 10.0 parts by weight

1,3-butylene glycol 1.00 parts by weight

Disodium EDIT 0.05 parts by weight

allantoin 0.10 parts by weight

Dipotassium glycyrrhizate 0.05 parts by weight

citric acid 0.01 parts by weight

sodium citrate 0.02 parts by weight

Glyceres-26 1.00 parts by weight

arbutin 2.00 parts by weight

Hydrogenated Castor Oil 1.00 parts by weight

ethanol 30.0 parts by weight

preservative a very small amount

coloring agent a very small amount

flavoring agent a very small amount

Purified water remaining amount

<3-2> Preparation of nourishing cream

LAB-C 10.0 parts by weight

1,3-butylene glycol 7.00 parts by weight

glycerin 1.00 parts by weight

D-Panthenol 0.10 parts by weight

plant extract 3.20 parts by weight

Magnesium Aluminum Silicate 0.30 parts by weight

PEG-40 Stearate 1.20 parts by weight

stearic acid 2.00 parts by weight

Polysorbate 60 1.50 parts by weight

Lipophilic Glyceryl Stearate 2.00 parts by weight

Sorbitan sesquioleate 1.50 parts by weight

cetearyl alcohol 3.00 parts by weight

mineral oil 4.00 parts by weight

squalane 3.80 parts by weight

Carlylic/Capric Triglycerides 2.80 parts by weight

vegetable oil 1.80 parts by weight

dimethicone 0.40 parts by weight

Dipotassium glycyrrhizate a very small amount

allantoin a very small amount

Sodium Hyaluronate a very small amount

Tocopheryl Acetate appropriate amount

triethanolamine appropriate amount

preservative appropriate amount

flavoring agent appropriate amount

Purified water remaining amount

So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than in the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Korea Institute of Biotechnology and Biotechnology Biological Resource Center KCTC10887BP 20060120

Claims (16)

Lactobacillus plantarum K8 (Lactobacillus plantarum K8) of live cells, dead cells, culture, culture medium, lysate, and a pharmaceutical composition for the prevention or treatment of metabolic diseases comprising any one or more selected from the group consisting of extracts and extracts as an active ingredient . According to claim 1,
The Lactobacillus plantarum K8 is a pharmaceutical composition, characterized in that the atomized Lactobacillus plantarum K8 dead cells. 3. The method of claim 2,
The atomized Lactobacillus plantarum K8 dead cells,
A method comprising: crushing live or dead cells of Lactobacillus plantarum K8 strain (accession number: KTCT 10887BP) with a microfluidizer and freeze-drying to obtain a shredded body (step 1); A pharmaceutical composition, characterized in that prepared with. 4. The method of claim 3,
The number of live or dead cells of step 1 ^{is adjusted to be 1×10 8} to 1×10 ¹⁴ CFU, the pharmaceutical composition. According to claim 1,
The metabolic disease is characterized in that any one or more selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver, a pharmaceutical composition. 6. The method of claim 5,
The metabolic disease is characterized in that obesity, the pharmaceutical composition. According to claim 1,
The Lactobacillus plantarum K8 is a pharmaceutical composition, characterized in that it comprises lipoteichoic acid (lipoteichoic acid). Lactobacillus plantarum K8 (Lactobacillus plantarum K8) Live cells, dead cells, culture, culture medium, lysate and any one or more selected from the group consisting of extracts and extracts as an active ingredient Health functional food for preventing or improving metabolic diseases composition. 9. The method of claim 8,
The Lactobacillus plantarum K8 is a health functional food composition, characterized in that the atomized Lactobacillus plantarum K8 dead cells. Lactobacillus plantarum K8 (Lactobacillus plantarum K8) Live cells, dead cells, culture, culture solution, lysate, and any one or more selected from the group consisting of extracts and extracts as an active ingredient Health food composition for the prevention or improvement of metabolic diseases . 11. The method of claim 10,
The Lactobacillus plantarum K8 is a health food composition, characterized in that the atomized Lactobacillus plantarum K8 dead cells. A cosmetic composition for reducing or preventing cellulite comprising, as an active ingredient, any one or more selected from the group consisting of live cells, dead cells, cultures, cultures, lysates, and extracts of Lactobacillus plantarum K8 (Lactobacillus plantarum K8). 13. The method of claim 12,
The Lactobacillus plantarum K8 is a cosmetic composition, characterized in that the atomized Lactobacillus plantarum K8 dead cells. Lactobacillus plantarum K8 (Lactobacillus plantarum K8) of live cells, dead cells, culture, culture medium, lysate, and lipoteichoic acid isolated from any one or more selected from the group consisting of extracts as an active ingredient A pharmaceutical composition for the prevention or treatment of metabolic diseases. Lactobacillus plantarum K8 (Lactobacillus plantarum K8) of live cells, dead cells, culture, culture medium, lysate, and lipoteichoic acid isolated from any one or more selected from the group consisting of extracts as an active ingredient A health functional food composition for the prevention or improvement of metabolic diseases. Lactobacillus plantarum K8 (Lactobacillus plantarum K8) of live cells, dead cells, culture, culture medium, lysate, and lipoteichoic acid isolated from any one or more selected from the group consisting of extracts as an active ingredient A cosmetic composition for reducing or preventing cellulite.

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