WO2015122717A1 - Novel lactic acid bacteria having obesity inhibitory effect and use thereof - Google Patents

Abstract

The present invention provides a novel Lactobacillus strain isolated from kimchi. A particular Bacillus strain according to the present invention is isolated from kimchi, and has various functionalities, such as an excellent anti-obesity activity, blood cholesterol lowering activity, blood triglyceride lowering activity, or antioxidant activity. Therefore, the particular Bacillus strain according to the present invention can be used as a food and drug material useful to prevent, alleviate, or treat diseases, such as obesity, fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, atherosclerosis, and lipid related metabolic syndrome. Further, a product obtained by fermenting soybeans, Codonopsis lanceolata, ginseng, and Cornus officinalis seeds with the particular Bacillus strain according to the present invention, or an extract thereof, can also be used as a food and drug material useful to prevent, alleviate, or treat obesity or obesity-related diseases.

Description

Novel lactic acid bacteria having obesity inhibitory effect and use thereof

The present invention relates to a novel lactic acid bacterium having an obesity inhibitory effect and its use, and more particularly, it is isolated from kimchi, and has various novel functions such as obesity inhibitory effect, blood cholesterol lowering effect, blood triglyceride lowering effect, antioxidant effect, and the like. It relates to the use of the prophylaxis, amelioration or treatment of lactic acid bacteria and obesity or obesity-related diseases.

As mankind has developed into a prosperous society, obesity has emerged as one of the serious diseases, and the World Health Organization (WHO) has declared it the object of the disease to be treated. Obesity is a metabolic disease caused by an imbalance between calorie intake and consumption and is morphologically caused by hypertrophy or hyperplasia of fat cells in the body. Obesity is not only the most common malnutrition in Western society, but the importance of treatment and prevention has been highlighted in recent years in Korea, as the frequency of obesity is rapidly increasing due to the improvement of dietary life and the westernization of lifestyle. have. Obesity is an important factor that not only psychologically diminishes individuals but also increases the risk of developing various adult diseases. Obesity is known to be directly related to the increased prevalence of various adult diseases, such as type 2 diabetes, hypertension, hyperlipidemia, and heart disease (Cell 87: 377, 1999), and the combination of obesity-related diseases together with metabolic syndrome or insulin resistance It is called syndrome (insulin resistance syndrome), and these have been found to be the cause of atherosclerosis and cardiovascular disease. It can be inferred that fat-rich fat cells mediate this phenomenon by the fact that obesity increases the incidence of various metabolic diseases and actual weight loss significantly reduces the incidence of these diseases.

In the past, adipose tissue was only thought of as an energy storage organ that stores excess energy in the form of triglyceryl (triacylglycerol) and releases it when needed, but recently, adiponectin, leptin, and resistin Various adipokines are accepted as important endocrine organs that regulate the homeostasis of energy (Trends Endocrinol Metab 13:18, 2002). Therefore, understanding of the proliferation of fat cells and the substances secreted from fat cells and their in vivo regulation mechanisms are expected to be the foundation for understanding obesity and various diseases and developing effective treatments. Studies on the regulation of adipocyte differentiation are being actively conducted, and the main mechanism is that differentiation from preadipocytes in the body is associated with increased adipocyte derivation in obese patients. The process of differentiation of pro-adipocytes into adipocytes has been studied using cells such as 3T3-L1, and several transcription factors, especially transcription factors known to be involved in localization, C / EBPs (CAAT enhancers). It is known that binding proteins, PPARs (Peroxisome Proliferator Activated receptor) and ADD / SREBPs (Adipocyte determination and differentiation dependent factor 1 / sterol response element binding proteins) are expressed over time and regulate the process (Bart A Jessen). et al., Gene, 299, pp95-100, 2002; Darlington et al., J. Biol. Chem., 273, pp30057-30060, 1998; Brun RP et al., Curr. Opin. Cell. Biol., 8 , pp826-832, 1996). Given the stimulation of hormones such as isobutylmethylxanthin, dexamethasone and insulin (MDI), C / EBP β and δ are the first, transiently expressed, to initiate differentiation into adipocytes (Reusch J. E et al., Mol Cell Biol., 20, pp 1008-1020, 2000). This continues to lead to increased expression of C / EBPa and PPARγ (James M. N. et al., J. Nutr., 130, pp 3122S-3126S, 2000). PPARγ is a particularly important transcription factor for adipocyte differentiation, and forms dimers with the retinoic acid X receptor protein (RXR), which is present in the promoters of various adipocyte genes. Binds to peroxisome proliferator response elements (Tontonoz PE et al., Genes Dev., 8, pp1224-1234, 1994; Hwang, C. S et al., Cell Dev. Biol., 13, pp873-877) . The interaction of PPARγ with C / EBP α is critical for differentiation into mature adipocytes. These transcription factors and adipocyte regulatory factors promote differentiation into adipocytes, and ap2 (adipocyte fatty acid-binding protein 2) The expression levels of fat cell specific proteins such as and fat metabolizing enzymes such as fat acid synthase (Fas) are increased. In addition, ADD1 / SREBPs play an important role in fat metabolism, but are also known to be involved in the differentiation process. Expression of ADD1 / SREBP1c in immature adipocytes is believed to contribute to the activation of PPARγ (Rosen ED et al., Annu. Rev. Cell Dev. Biol., 16, pp 145-171, 2000; Osborn TF, J. Biol Chem., 275, pp 32379-32382, 2000). Only the differentiated fat cells synthesize fatty acids and store triglycerides. Therefore, current research trends are focused on the discovery of substances that can inhibit metabolic processes related to adipocyte differentiation as a method for preventing or treating obesity and lipid-related metabolic diseases. In other words, attempts have been made to treat obesity through the regulation of fat cells based on the mechanism of the development of obesity. To reduce cell numbers, transcription factors, proteins and adipokines, known to mediate or regulate these processes, have emerged as targets for the development of new anti-obesity drugs. In fact, the adipose cell differentiation transcription factor PPAR (Peroxisome proliferator-activated receptor) family, the adipocyte secreting substances leptin and adiponectin have been the target of many new drug development.

Known obesity treatments such as Xenical (Roche Pharmaceuticals, Switzerland), Reductil (Eboth, USA), Exolise (Atopama, France), etc. Classified as inhibitors, most anti-obesity agents are appetite suppressants that suppress appetite by regulating neurotransmitters associated with the hypothalamus. However, conventional treatments have low side effects such as heart disease, respiratory disease, and neurological disease, and thus have low sustainability. Therefore, development of an improved obesity treatment agent is needed, and currently developed products have satisfactory treatment effects without side effects. Since there are few therapeutic agents, development of new obesity agents is required.

On the other hand, many efforts have been made to reduce blood cholesterol levels using lactic acid bacteria, which have been considered safe microorganisms. Lactobacillus has been reported to show the effects of maintaining normal intestinal flora, improving intestinal flora, antidiabetic and antihyperlipidemic effects, inhibiting carcinogenesis, inhibiting colitis, and nonspecific activity of the host's immune system. Among them, the strain Lactobacillus is a major member of the normal microbial community in the human intestine, which has long been known to be important for maintaining a healthy digestive system and the vaginal environment, and the US Public Health Service (US Public Health Service). guidelines) are currently classified as 'Bio-safty Level 1', where none of the Lactobacillus strains currently deposited with the American Strain Deposit Organization (ATCC) is known about the potential risk of causing disease in humans or animals. have.

On the other hand, kimchi lactic acid bacteria are reported to have the effect of immuno-enhancing, anti-microbial, antioxidant, anti-cancer, anti-obesity, hypertension or constipation as a lactic acid bacteria involved in kimchi fermentation [Hivak P, Odrska J, Ferencik M, Ebringer L, Jahnova E, Mikes Z .: One-year application of Probiotic strain Enterococcus facium M-74 decreases Serum cholesterol levels. Bratisl lek Listy 2005; 106 (2); 67-72; Agerholm-Larsen L. Bell ML. Grunwald GK. Astrup A.: The effect of a probiotic milk product on plasma cholesterol: a metaanalysis of short-term intervention studies; Eur J Clin Nutr. 2000; 54 (11) 856-860; Renato Sousa, Jaroslava Helper, Jian Zhang, Strephen J Lewis and Wani O Li; Effect of Lactobacillus acidophilus supernants on body weight and leptin expression in rats; BMC complementary and alternative medicine. 2008; 8 (5) 1-8]. For example, Korean Laid-Open Patent Publication No. 10-2012-0034482 discloses Lactobacillus plantarum KY1032 (Accession No .: KCCM-10430) having an inhibitory effect on 3T3-L1 adipocyte differentiation, and Korea Korean Patent Laid-Open Publication No. 10-0996577 discloses a novel Lactobacillus curvatus HY7601 (Accession No .: KCTC 11456BP) having a blood cholesterol lowering effect and an anti-obesity effect, and Korean Patent Publication No. 10-2013 -0046896 discloses a composition for the prevention and treatment of obesity, including Lactobacillus plantarum DSR920 (Accession Number: KCCM 11210P) as an active ingredient, Korean Patent Publication No. 10-2010-0010015 A Lactobacillus johnsonii HF1 108 strain (KCTC 11356BP) having blood cholesterol lowering and anti-obesity activity is disclosed. However, there is no technology related to lactic acid bacteria having an excellent anti-obesity effect enough to be commercially successful. Therefore, it is necessary to screen new strains having excellent anti-obesity effects and to identify various functions thereof.

The present invention is derived under such a conventional background, and an object of the present invention is to provide a novel lactic acid bacteria having various functionalities such as anti-obesity activity.

Another object of the present invention is to provide various uses of the novel lactic acid bacteria.

In order to develop an anti-obesity material that is safer than synthetic chemicals, the inventors of the present invention screen countless lactic acid bacteria from kimchi or human feces, and among them, certain Lactobacillus strains are excellent for preventing or treating obesity-induced model animals. It was found that it had an effect, and the present invention was completed.

In order to achieve the object of the present invention, the present invention is Lactobacillus brevis ( Lactobacillus brevis ) comprising the nucleotide sequence of SEQ ID NO: 1 as 16S rDNA, Lactobacillus company comprising the nucleotide sequence of SEQ ID NO: 2 as 16S rDNA Lactobacillus plantarum (K) ( Lactobacillus sakei ) and Lactobacillus plantarum ( Lactobacillus plantarum ) comprising the nucleotide sequence of SEQ ID NO: 4 in 16S rDNA, Lactic acid bacterium selected from the group consisting of lactic acid bacteria, anti-obesity activity, blood cholesterol lowering activity To provide lactic acid bacteria having a neutral lipid lowering activity or antioxidant activity in the blood. The Lactobacillus brevis ( Lactobacillus brevis ) is preferably Lactobacillus brevis ( Lactobacillus brevis ) OK56 (Accession Number: KCCM 11516P). In addition, the Lactobacillus sakei ( Lactobacillus sakei ) is preferably Lactobacillus sakei ( Lactobacillus sakei ) OK101 (Accession Number: KCCM 11664P). In addition, the Lactobacillus plantarum ( Lactobacillus plantarum ) is preferably Lactobacillus plantarum OK169 (Accession Number: KCCM 11663P).

In order to achieve another object of the present invention, one example of the present invention is Lactobacillus brevis comprising the nucleotide sequence of SEQ ID NO: 1 as 16S rDNA, comprising the nucleotide sequence of SEQ ID NO: 2 as 16S rDNA At least one Lactobacillus lactic acid bacterium selected from the group consisting of Lactobacillus sakei and Lactobacillus plantarum comprising the nucleotide sequence shown in SEQ ID NO: 4 as 16S rDNA, cultures thereof, and lysates thereof Or it provides a composition for the prevention, improvement or treatment of obesity or obesity-related diseases comprising an extract thereof as an active ingredient. The composition for preventing, ameliorating or treating the obesity or obesity-related disease is preferably a pharmaceutical composition or a food composition. In addition, the Lactobacillus brevis ( Lactobacillus brevis ) is preferably Lactobacillus brevis ( Lactobacillus brevis ) OK56 (Accession Number: KCCM 11516P). In addition, the Lactobacillus sakei ( Lactobacillus sakei ) is preferably Lactobacillus sakei ( Lactobacillus sakei ) OK101 (Accession Number: KCCM 11664P). In addition, the Lactobacillus plantarum ( Lactobacillus plantarum ) is preferably Lactobacillus plantarum OK169 (Accession Number: KCCM 11663P).

In order to achieve another object of the present invention, another example of the present invention is Lactobacillus brevis comprising at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with the base sequence of SEQ ID NO: 1 as 16S rDNA ( Lactobacillus brevis ), Lactobacillus sakei comprising the nucleotide sequence of SEQ ID NO: 2 as 16S rDNA and Lactobacillus plantarum comprising the nucleotide sequence of SEQ ID NO: 4 as 16S rDNA It provides a composition for the prevention, improvement or treatment of obesity or obesity-related diseases comprising a fermentation product obtained by fermenting with one or more Lactobacillus lactic acid bacteria selected from the group consisting of or extract of the fermentation as an active ingredient. The composition for preventing, ameliorating or treating the obesity or obesity-related disease is preferably a pharmaceutical composition or a food composition. In addition, the Lactobacillus brevis ( Lactobacillus brevis ) is preferably Lactobacillus brevis ( Lactobacillus brevis ) OK56 (Accession Number: KCCM 11516P). In addition, the Lactobacillus sakei ( Lactobacillus sakei ) is preferably Lactobacillus sakei ( Lactobacillus sakei ) OK101 (Accession Number: KCCM 11664P). In addition, the Lactobacillus plantarum ( Lactobacillus plantarum ) is preferably Lactobacillus plantarum OK169 (Accession Number: KCCM 11663P).

Specific Bacillus strain according to the present invention is isolated from kimchi has a high safety, and has various functions such as excellent anti-obesity activity, blood cholesterol lowering activity, blood triglyceride lowering activity or antioxidant activity. Therefore, certain Bacillus strains according to the present invention can be used as a food and pharmaceutical material useful for preventing, ameliorating or treating diseases such as obesity, fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis and lipid-related metabolic syndrome have. In addition, the product or extract thereof fermented soybean, deodeok, ginseng and cornus seed as a specific Bacillus strain according to the present invention can also be used as a food and pharmaceutical material useful for preventing, improving or treating obesity or obesity-related diseases.

1 is a graph showing the effect of lactic acid bacteria on the differentiation inhibition of 3T3-L1 precursor adipocytes into adipocytes.

Figure 2 is a graph showing the antioxidant efficacy of the lactic acid bacteria at 50% inhibitory concentration (50% Scavenging Concentration).

Hereinafter, the term used by this invention is demonstrated.

As used herein, the term "culture" means a product obtained by culturing a microorganism in a known liquid medium or a solid medium, and is a concept in which a microorganism is included.

As used herein, "pharmaceutically acceptable" and "food acceptable" means that they do not significantly stimulate the organism and do not inhibit the biological activity and properties of the administered active substance.

As used herein, the term "prevention" refers to any action that inhibits the symptoms or delays the progression of a particular disease (eg colitis) by administration of a composition of the invention.

As used herein, the term "treatment" refers to any action that improves or beneficially alters the symptoms of a particular disease (eg, colitis) by administration of a composition of the present invention.

The term "improvement" as used herein refers to any action that at least reduces the parameters associated with the condition being treated, for example, the extent of symptoms.

As used herein, the term "administration" means providing a subject with a composition of the present invention in any suitable manner. At this time, the subject refers to all animals, such as humans, monkeys, dogs, goats, pigs or mice having a disease that can improve the symptoms of a particular disease by administering the composition of the present invention.

As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, which refers to the type of disease, the severity, the activity of the drug, the drug, and the like. Sensitivity, time of administration, route of administration and rate of excretion, duration of treatment, factors including drug used concurrently, and other factors well known in the medical arts.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

One aspect of the present invention relates to a novel lactic acid bacterium having various functionalities such as anti-obesity activity, blood cholesterol lowering activity, blood triglyceride lowering activity or antioxidant activity. The novel lactic acid bacterium according to the present invention is Lactobacillus brevis comprising the nucleotide sequence of SEQ ID NO: 1 as 16S rDNA, Lactobacillus sakei comprising the nucleotide sequence of SEQ ID NO: 2 as 16S rDNA, and At least one Lactobacillus genus strain selected from the group consisting of Lactobacillus plantarum comprising the nucleotide sequence of SEQ ID NO: 4 as 16S rDNA. At this time, the Lactobacillus brevis ( Lactobacillus brevis ) is a bacillus that shows positive upon gram staining, glucose (Glucose), Amigdalin (Amygdalin), Arabinos (Arabinose), Esculin, Fructose, Galactose, Lactose, Mannose, Mannose, Raffinose, Rhamnose, Sorbitol, Sobitol, Sucrose, and Xylose are used. Lactobacillus brevis OK56 with accession number KCCM 11516P. In addition, the Lactobacillus sakei ( Lactobacillus sakei ) is a bacillus that shows positive upon gram staining, glucose (Glucose), Amigdalin (Amygdalin), Arabinos (Arabinose), Esculin, Fructose as a carbon source , Using galactose, glycerol, lactose, lactose, mannose, melibiose, raffinos, sorbitol, sucrose and xylose It is characterized in that, preferably Lactobacillus sakei OK101 with accession number KCCM 11664P. In addition, the Lactobacillus plantarum ( Lactobacillus plantarum ) is a bacilli that show positive gram staining, and as a carbon source glucose (Glucose), Amigdalin (Amygdalin), Arabinos (Arabinose), Esculin (Fructose) ), Galactose, Glycerol, Lactose, Mannose, Melezitose, Meliviose, Raffinose, Rhamnose, Sorbitol It is characterized by using sucrose (Sucrose) and xylose (Xylose), preferably the accession number KCCM 11663P is Lactobacillus plantarum OK169.

Further, the novel lactic acid bacterium according to the present invention is Lactobacillus sakei OK156 comprising the nucleotide sequence of SEQ ID NO: 3 as 16S rDNA or Lactobacillus plantarum comprising the nucleotide sequence of SEQ ID NO: 5 as 16S rDNA. ( Lactobacillus plantarum ) OK215 may further include. The Lactobacillus sakei OK156 is a bacillus that shows positive gram staining, and as a carbon source, glucose (Glucose), amigdalin (Amygdalin), Arabinos (Arabinose), Esculin, Fructose, Galactose (Galactose), Glycerol (Glycerol), Lactose, (Lactose), Mannose (Mannose), Melibiose (Melibiose), Raffinose (Sorbitol) and sucrose (Sucrose) is characterized by using. In addition, the Lactobacillus plantarum OK215 is a bacilli that show positive gram staining, and as a carbon source, glucose (Glucose), amygdalin (Amygdalin), arabinose (Esculin), fructose ( Fructose, Galactose, Lactose, Lactose, Mannose, Melezitose, Melibiose, Raffinose, Rhamnose, Sorbitol, and Sucrose It is characterized by using).

The Lactobacillus strain of the present invention is preferably isolated from kimchi, such as cabbage kimchi or gad kimchi.

Another aspect of the invention relates to various uses of the novel lactic acid bacteria. For example, the present invention prevents or improves obesity or obesity-related diseases comprising lactic acid bacteria, cultures thereof, lysates or extracts thereof, as one of the above-described Lactobacillus strains, as an active ingredient. Or a therapeutic composition. The culture of the lactic acid bacteria is a product obtained by culturing in Lactobacillus medium, the medium may be selected from known liquid medium or solid medium, for example, may be MRS liquid medium, MRS agar medium.

In another aspect, the present invention is a fermentation product obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed as a lactic acid bacterium consisting of at least one strain of Lactobacillus sp. It provides a composition for the prevention, improvement or treatment of obesity or obesity-related diseases comprising the extract of the fermentation as an active ingredient. Extracts of the fermentation may be prepared by various methods. For example, a method for obtaining an extract from a product obtained by fermenting a material selected from soybean, deodeok, ginseng and cornus seed with the novel lactic acid bacteria of the present invention, or an extract of a material selected from soybean, deodeok, ginseng and cornus seed. Fermentation with novel lactic acid bacteria. On the other hand, in order to obtain an extract of the fermented product, an extraction process is required, and as the extraction method, a conventional extraction method known in the art, for example, a solvent extraction method may be used. Extractive solvents that may be used when preparing extracts of fermentation using solvent extraction are water, lower alcohols having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol and butanol) or hydrous lower alcohols which are mixtures thereof, Propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, and may be selected from the group consisting of double water, alcohol or It is preferred to be selected from mixtures of. When water is used as the extraction solvent, the water is preferably hot water. In addition, when alcohol is used as the extraction solvent, the alcohol is preferably a lower alcohol having 1 to 4 carbon atoms, and the lower alcohol is more preferably selected from methanol or ethanol. In addition, when using a hydrous alcohol as the extraction solvent, the alcohol content is preferably 50 ~ 90%. On the other hand, it will be apparent to those skilled in the art that the extract of the fermentation product can obtain an extract having substantially the same effect using not only the extraction solvent but also other extraction solvents. For example, extraction by supercritical fluid extraction using carbon dioxide, extraction by ultrasound, separation using ultrafiltration membranes with constant molecular weight cut-off values, or various chromatography (size, charge, Active fractions obtained through various purification and extraction methods additionally carried out, such as separation by hydrophobicity or affinity), are also included in the extract of the present invention. In general, supercritical fluids have the properties of liquids and gases when the gas reaches a critical point at high temperature and high pressure, and have a chemically similar polarity to nonpolar solvents. J. Chromatogr. A. 1998; 479: 200-205. Carbon dioxide becomes a supercritical fluid with both liquid and gaseous properties as the pressure and temperature reach a critical point through the operation of a supercritical fluid device, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from the supercritical carbon dioxide. After extracting the fat-soluble substance, the supercritical carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction container to extract components that were not extracted with pure supercritical carbon dioxide alone. The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract the active ingredient by using supercritical carbon dioxide or a mixed fluid in which a co-solvent is added to carbon dioxide. As such cosolvent, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether can be used. Most of the extracted samples contain carbon dioxide, which is volatilized into the air at room temperature, and the cosolvent can be removed by a reduced pressure evaporator. In addition, the ultrasonic extraction method is an extraction method using the energy generated by the ultrasonic vibration, the ultrasonic wave can destroy the insoluble solvent contained in the sample in the water-soluble solvent, due to the high local temperature is generated Since the kinetic energy of the reactant particles is increased, sufficient energy required for the reaction is obtained, and the extraction efficiency is increased by inducing high pressure by the impact effect of ultrasonic energy to increase the mixing effect of the substance and the solvent contained in the sample. The extraction solvent that can be used in the ultrasonic extraction method may be used one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether. The extracted sample is vacuum filtered to recover the filtrate, and then removed by a vacuum evaporator, the extract can be obtained through a conventional extract preparation method of freeze drying.

The fermented product fermented with a specific Lactobacillus strain or a specific Lactobacillus strain of the present invention has various functionalities such as anti-obesity activity, blood cholesterol lowering activity, blood triglyceride lowering activity, etc., thus preventing or improving obesity or obesity-related diseases. Or as an active ingredient for treatment. The obesity-related diseases are not limited in kind as long as they are caused by obesity or have a high correlation with obesity, for example, fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. It can be any one. In addition, the lipid-related metabolic syndrome refers to a disease in which various metabolic diseases, such as diabetes and obesity, appear simultaneously in one person. In addition, the composition for preventing, ameliorating or treating obesity or obesity-related diseases of the present invention may be embodied as a pharmaceutical composition, a food additive, a food composition (particularly a functional food composition), a feed additive, or the like, depending on the purpose or aspect of use. The content of the fermentation product fermented with a specific Lactobacillus strain or a specific Lactobacillus strain, which is an active ingredient in the composition, may also be adjusted in various ranges according to the specific form of the composition, purpose of use, and aspects.

New lactic acid bacteria, cultures thereof, lysates thereof or extracts thereof as an active ingredient in the pharmaceutical composition for the prevention or treatment of obesity or obesity-related diseases according to the present invention; Or the content of fermentation products or extracts thereof obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with new lactic acid bacteria is not particularly limited, for example 0.01 ~ based on the total weight of the composition 99% by weight, preferably 0.5-50% by weight, more preferably 1-30% by weight. In addition, the pharmaceutical composition according to the present invention may further include an additive such as a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. Carriers, excipients and diluents that may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, the pharmaceutical composition for the prevention or treatment of obesity or obesity-related diseases of the present invention is a novel lactic acid bacteria, its culture, its lysate or extract thereof; Or a known active ingredient having a prophylactic or therapeutic effect of obesity or obesity-related diseases in addition to fermentation products or extracts thereof obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with new lactic acid bacteria. It may contain more than one species. The pharmaceutical composition of the present invention may be formulated into a formulation for oral administration or a parenteral administration by a conventional method, and when formulated, such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. Diluents or excipients may be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose in active ingredients. ), Lactose (Lactose) or gelatin can be prepared by mixing. In addition to simple excipients, lubricants such as magnesium styrate talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents, water and liquid paraffin. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used. Furthermore, it may be preferably formulated according to each disease or component by an appropriate method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention can be administered orally or parenterally to mammals including humans according to a desired method, and parenteral administration methods include external skin, intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, muscle Intra-injection or intrathoracic injection; The dosage of the pharmaceutical composition of the present invention is not particularly limited as long as it is a pharmaceutically effective amount, and the range thereof depends on the weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate and severity of the disease. Varies. Typical daily dosages of the pharmaceutical compositions of the present invention are not particularly limited but are preferably 0.1 to 3000 mg / kg, more preferably 1 to 2000 mg / kg, once daily based on the active ingredient. Or divided into several doses.

In addition, the novel lactic acid bacteria, its culture, its lysate or extract thereof as an active ingredient in a food composition for preventing or improving obesity or obesity-related diseases of the present invention; Or a fermentation product or extract thereof obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with new lactic acid bacteria, based on the total weight of the composition, preferably 0.01 to 50% by weight, preferably 0.1 25 wt%, more preferably 0.5-10 wt%, but is not limited thereto. The food composition of the present invention includes the form of pills, powders, granules, acupuncture, tablets, capsules, or liquids, and examples of specific foods include meat, sausage, bread, chocolate, candy, snacks, confectionary, pizza, ramen, Other noodles, gums, dairy products, including ice cream, various soups, beverages, tea, functional water, drinks, alcoholic beverages and vitamin complexes, and includes all of the health food in the usual sense. In addition to the active ingredient, the food composition of the present invention may contain various flavors or natural carbohydrates as additional ingredients. In addition, the food composition of the present invention is a variety of nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols And carbonation agents used in carbonated beverages. In addition, the food composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, natural flavoring agents such as taumartin, stevia extract, synthetic flavoring agents such as saccharin, aspartame, etc. may be used.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are only intended to clearly illustrate the technical features of the present invention, and do not limit the protection scope of the present invention.

1. Isolation and Identification of Lactic Acid Bacteria

(1) Isolation of Lactic Acid Bacteria from Kimchi

Cabbage kimchi and fresh kimchi were suspended in MRS liquid medium (MRS Broth; Difco, USA). Then, the supernatant was taken and transplanted into MRS agar medium (DRS, Aifco, USA), and cultured anaerobicly at 37 ° C. for about 24 hours, and then colonies were formed.

(2) Isolation of Lactic Acid Bacteria from Human Feces

Human feces were suspended in GAM liquid medium (GAM broth; Nissui Pharmaceutical, Japan). Then, the supernatant was taken, transplanted into BL agar medium (Nissui Pharmaceutical, Japan), and anaerobicly cultured at 37 ° C. for about 48 hours, and then colonies were formed.

(3) Identification of selected lactic acid bacteria

Physiological characteristics and 16S rDNA sequences of strains isolated from kimchi or human feces were analyzed to determine the species of the strains and to give a strain name. Table 1 shows the control numbers and strain names of lactic acid bacteria isolated from Chinese cabbage kimchi and gad kimchi, and Table 2 shows the control numbers and strain names of lactic acid bacteria isolated from human feces.

Table 1 Control Number Strain name Control Number Strain name One Leuconostoc mesenteroides C1 17 Lactobacillus pentosus C15 2 Leuconostoc mesenteroides C2 18 Lactobacillus pentosus C27 3 Lactobacillus mesenteroides OK2 19 Lactobacillus sakei C14 4 Lactobacillus curvatus C3 20 Lactobacillus sakei C21 5 Lactobacillus brevis C4 21 Lactobacillus sakei C22 6 Lactobacillus brevis C5 22 Lactobacillus sakei C23 7 Lactobacillus brevis OK55 23 Lactobacillus sakei OK101 8 Lactobacillus brevis OK56 24 Lactobacillus sakei OK156 9 Lactobacillus acidophilus C9 25 Lactobacillus plantarum C12 10 Lactobacillus acidophilus C20 26 Lactobacillus plantarum C24 11 Lactobacillus acidophilus OK75 27 Lactobacillus plantarum C25 12 Lactobacillus lactis C10 28 Lactobacillus plantarum C26 13 Lactobacillus lactis OK78 29 Lactobacillus plantarum C27 14 Lactobacillus helveticus C11 30 Lactobacillus plantarum OK169 15 Lactobacillus helveticus OK95 31 Lactobacillus plantarum OK215 16 Lactobacillus casei C13 32 Lactobacillus plantarum OK216

TABLE 2 Control Number Strain name Control Number Strain name 33 Bifidobacterium adolescentis OK1 35 Bifidobacterium breve OK6 34 Bifidobacterium longum OK11 36 Bifidobacterium bifidum OK22

Among the strains described in Table 1 and Table 2, Lactobacillus brevis OK56, Lactobacillus sakei OK101, Lactobacillus sakei OK156, Lactobacillus plantarum OK169 And Lactobacillus plantarum OK215 physiological characteristics and chemical classification characteristics are shown in Table 3.

TABLE 3 Test Items Strain name Lactobacillus brevis OK56 Lactobacillus sakei OK101 Lactobacillus sakei OK156 Lactobacillus plantarum OK169 Lactobacillus plantarum OK215 Morphology Bacillus Bacillus Bacillus Bacillus Bacillus Gram dyeing + + + + + Catalase activity - - - - - Growth temperature 15 ℃ + + + + + 35 ℃ + + + + + Growth pH 4 + + + + + 7 + + + + + Heat resistance (50 ℃) + + + + + Fermentation test per API 50CH Kit Glucose + + + + + Amygdalin + + + + + Arabinose + + + + + Esulin + + + + + Fructose + + + + + Galactose + + + + + Glycerol - + + + - Lactose + + + + + Mannose + + + + + Melezitose - - - + + Melibiose - + + + + Raffinose + + + + + Rhamnose + - - + + Sorbitol + + + + + Sucrose + + + + + Xylose + + - + - 16S rDNA SEQ ID NO: 1 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5

16S rDNA nucleotide sequence of each strain was identified by BLAST search in Genebank (http://www.ncbi.nlm.nih.gov/). As a result, Lactobacillus brevis OK56 16S rDNA (see SEQ ID NO: 1) was identified. Lactobacillus brevis ATCC 367 (NCBI ACCESSION: NR_075024) showed 99% homology with Lactobacillus sakei OK101 16S rDNA (see SEQ ID NO: 2). Lactobacillus sakei NBRC 15893 (NCBI ACCESSION: NR_113821) showed 98% homology with the 16S rDNA subsequence, and Lactobacillus sakei OK156 16S rDNA (see SEQ ID NO: 3) was ( Lactobacillus sakei ) 100% homology with 16S rDNA subsequence of NBRC 15893 (NCBI ACCESSION: NR_113821), and 16S rDNA of Lactobacillus plantarum OK169 (see SEQ ID NO: 4) Room ( Lactobacillus plantarum ) 97% homology with the 16S rDNA subsequence of CIP 103151 (NCBI ACCESSION: NR_104573), and the 16S rDNA of Lactobacillus plantarum OK215 (see SEQ ID NO: 5) is shown in Lactobacillus plantarum (see SEQ ID NO: 5). Lactobacillus plantarum ) showed 99% homology with the 16S rDNA subsequence of JCM 1149 ((NCBI ACCESSION: NR_115605)).

2. In-vitro experiment on anti-obesity effect of lactic acid bacteria

Mouse preadipocytes 3T3-L1 were dispensed into 12 well plates at a concentration of 2 × 10 ⁴ / mL and cultured at 37 ° C. and 5% CO ₂ . At this time, use 3T3-L1 complete media [DMEM / high glucose (Thermo scientific, USA), newborn calf serum (Thermo scientific), 1% penicillin-streptomycin solution (Thermo scientific)] until the 100% confluency point as a medium. It was. At the 100% confluency time point, it was maintained for two more days. Progenitor cells were then incubated with MDI media (DMEM / high glucose, 10% fetal bovine serum, 1% penicillin-streptomycin solution, 0.5 mM 3-isobutyl-1-methylxanthine, 1 uM dexamethazone, 5 ㎍ / ml insulin) for 2 days. Differentiation into adipocytes was induced for 2 days, and differentiation media (DMEM / high glucose, 10% fetal bovine serum, 1% penicillin-streptomycin solution, 5 ㎍ / ml insulin) with lactic acid bacteria 2 days after induction of differentiation. ) Was added and incubated for 2 days. At this time, the lactic acid bacteria were added by suspension in phosphate buffered saline (PBS), the amount was 1 × 10 ⁴ CFU / well. After that, the cells were replaced with growth media (DMEM / high glucose, 10% fetal bovine serum, 1% penicillin-streptomycin solution) every 4 days to complete the differentiation of adipocytes.

After inducing the differentiation of profatal cells into adipocytes for 8 days, the proliferative cell cultures were washed three times with PBS to remove the cultures and treated with 10% formalin for 1 hour to fix the cells. Subsequently, the cells were treated with oil red O solution in the dark for 30 minutes and washed with distilled water to stain the fat globules. Thereafter, isopropanol was added to dissolve the aliphatic sample stained with an oil red O solution, and the absorbance was measured at 520 nm using a Spectrophotometer. The inhibition rate of adipocyte differentiation of each lactic acid bacterium was calculated by the following equation.

In the above formula, Control O.D. is the absorbance of the test solution to which ethanol was added instead of the sample, and Sample O.D. is the absorbance of the test solution to which the sample was added.

1 is a graph showing the effect of lactic acid bacteria on the differentiation inhibition of 3T3-L1 precursor adipocytes into adipocytes. Lactobacillus brevis (Lactobacillus brevis), as also seen in the first eruption was inhibitory ability is best for OK56, followed by Lactobacillus Planta Room (Lactobacillus plantarum) OK169, Lactobacillus four K (Lactobacillus sakei) OK101, Lactobacillus Planta Lactobacillus plantarum OK215 and Lactobacillus sakei OK156.

3. In-vitro experiment on the antioxidant efficacy of lactic acid bacteria

The lactic acid bacteria were suspended in 10 mM phosphate buffered saline (PBS) to prepare samples having lactic acid bacteria concentrations of 10, 1, 0.1 and 0.01 mg / ml, respectively, and the same amount of 0.1 mM DPPH solution (2,2). 39.4 mg of -diphenyl-1-picrylhydrazyl was dissolved in ethanol to prepare 1000 ml), and then stirred for 10 seconds and reacted at 37 ° C. for 30 minutes. The reaction was then measured at 515 nm using a Spectrophotometer. On the other hand, a test solution in which ethanol was added instead of the sample was used as a control. In addition, Vitamin C was used as a reference for comparing the antioxidant efficacy. Inhibition rate of each lactic acid bacteria was calculated by the following equation.

In the above formula, Control O.D. is the absorbance of the control in which ethanol was added instead of the sample, and Sample O.D. is the absorbance of the test solution in which the sample was added.

Figure 2 is a graph showing the antioxidant efficacy of the lactic acid bacteria at 50% inhibitory concentration (50% Scavenging Concentration). 50% Scavenging Concentration refers to the concentration of lactic acid bacteria in the sample required to reduce the absorbance of the control to 1/2, that is, the concentration of lactic acid bacteria in the sample with 50% inhibition rate. This means that the antioxidant effect is great. As shown in FIG. 2, the anti-oxidant effect was the best Lactobacillus brevis ( Lactobacillus brevis ) OK56 as well as the anti-obesity effect, followed by the Lactobacillus plantarum OK169, Lactobacillus sakei ( Lactobacillus sakei ) OK101 , Lactobacillus plantarum OK215 and Lactobacillus sakei OK156.

4. In-vivo experiment on anti-obesity effect of lactic acid bacteria

(1) Experiment Method

Four 42-week-old male C57BL / 6J mice were fed for a week on a chow diet (CD; Purina, Korea) under a 20-hour temperature room at 20 ± 2 ° C, 50 ± 5% humidity, and a 12-hour light-dark cycle. Adapted. Thereafter, the experimental animals were divided into two groups, and the normal diet group (CON, n = 6) was fed a general diet of 4.5% of total calories for 5 weeks and the high fat diet group (n = 36) was 45 Obesity was induced by feeding a high fat diet (D12451, Research Diets, New Brunswick, NJ), where% was fat for 5 weeks. Subsequently, the obese high fat diet group was divided into six groups of 6 animals (HF, HF + OK56, HF + OK169, HF + OK101, HF + OK215, HF + OK156), and then the HF group was fed a high fat diet. At the same time, phosphate buffered saline (PBS) was orally administered for 5 weeks, and HF + OK56, HF + OK169, HF + OK101, HF + OK215, and HF + OK156 groups were fed a high-fat diet. Other lactic acid bacteria were suspended in phosphate buffered saline (PBS) and orally administered for 5 weeks. In addition, the normal diet group was fed orally administered the same amount of phosphate buffered saline (PBS) for 5 weeks at the same time as the normal diet.

(2) weight change and dietary intake

The body weight and dietary intake of the experimental animals were measured at a constant time every day during the entire experimental period, and the results are shown in Table 4 below.

Table 4 Measurement item Experimental group CON HF HF + OK56 HF + OK169 HF + OK101 HF + OK215 HF + OK156 Initial weight (g) 24.2 26.8 27.0 26.9 26.8 26.7 26.6 Final weight (g) 27.2 38.5 33.5 34.3 34.5 35.1 35.2 Weight gain (g) 4.0 11.5 6.5 7.4 7.7 8.4 8.6 % Weight gain 16.5 42.9 24.0 27.5 28.7 31.4 32.8 Dietary Intake (g) 4.1 2.9 3.1 2.9 2.9 2.9 2.9

CON: General Dietary Salary

* HF: High Fat Dietary Salary Group

* HF + OK56: Lfatobacillus brevis ( Lactobacillus brevis ) OK56 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK169: high fat diet and 1 × 10 ⁹ CFU / mouse dose of Lactobacillus plantarum OK169 administration group

* HF + OK101: Lfatobacillus sakei OK101 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK215: Lactobacillus plantarum OK215 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK156: Lactobacillus sakei OK156 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

As shown in Table 4, the group of oral administration of lactic acid bacteria with high-fat diet after induction of obesity had no significant difference in dietary intake compared to the group receiving only high-fat diet after induction of obesity, but significantly reduced weight. Showed results. It can be seen that the specific lactic acid bacteria isolated from kimchi has excellent anti-obesity effect.

(3) Measurement of weight change of liver and adipose tissue

After inducing obesity and sacrificing experimental animals for 5 weeks with cardiac puncture, subcutaneous fat (SF), epididymal fat (EF), and peripheral kidney tissue ( Perirenal fat (PF), mesenteric fat (mesenteric fat, MF), brown adipose tissue (BAT) and liver were extracted and weighed, and the results are shown in Table 5 below.

Table 5 Measurement item Experimental group CON HF HF + OK56 HF + OK169 HF + OK101 HF + OK215 HF + OK156 Liver (g) 1.11 1.12 1.11 1.03 1.12 1.15 1.09 Epididymal Adipose Tissue (g) 0.38 2.11 1.50 1.55 1.65 1.75 1.85 Fat tissue around the kidneys (g) 0.11 0.91 0.71 0.76 0.76 0.81 0.87 Subcutaneous adipose tissue (g) 0.41 2.01 1.67 1.73 1.73 1.75 1.85 Mesenteric adipose tissue (g) 0.20 0.72 0.57 0.58 0.68 0.61 0.62 Brown adipose tissue (g) 0.07 0.17 0.16 0.16 0.16 0.17 0.17

CON: General Dietary Salary

* HF: High Fat Dietary Salary Group

* HF + OK56: Lfatobacillus brevis ( Lactobacillus brevis ) OK56 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK169: high fat diet and 1 × 10 ⁹ CFU / mouse dose of Lactobacillus plantarum OK169 administration group

* HF + OK101: Lfatobacillus sakei OK101 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK215: Lactobacillus plantarum OK215 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK156: Lactobacillus sakei OK156 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

As shown in Table 5, the group of oral administration of lactic acid bacteria with high fat diet after induction of obesity showed a lower weight of adipose tissue when compared to the group receiving only high fat diet after induction of obesity.

(4) Determination of plasma triglycerides, total plasma cholesterol and plasma HDL cholesterol content

Plasma trigiyceride, plasma cholesterol, and plasma HDL cholesterol levels of the experimental animals that induced obesity and received lactic acid bacteria for 5 weeks were measured as follows.

Plasma neutral lipids were measured using a kit for measuring neutral lipids (ASAN PHARM. CO. LTD, South Korea). 1.5 ml of enzyme solution and 10 µl of plasma were added to the test tube, followed by stirring. Thereafter, the sample was warmed for 10 minutes in a water bath at 37 ° C. with a standard solution having a neutral lipid content of 0, 75, 150, 225, 300 mg / dL, and the absorbance at 550 nm was measured for colorimetric determination. It was. In addition, plasma total cholesterol was measured using a total cholesterol measurement kit (ASAN PHARM. CO. LTD, South Korea). 1.5 ml of enzyme solution and 10 µl of plasma were added to the test tube, followed by stirring. Thereafter, the sample was warmed for 10 minutes in a water bath at 37 ° C. with a standard solution having a total cholesterol content of 0, 75, 150, 225, 300 mg / dL, and the absorbance at 500 nm was measured and colorimetrically determined. Quantification In addition, plasma HDL cholesterol was measured using a kit for measuring HDL cholesterol (ASAN PHARM. CO. LTD, South Korea). 50 μl of the needle was added to 50 μl of plasma, stirred, and allowed to stand at room temperature for 10 minutes, followed by centrifugation at 3000 rpm for 10 minutes to obtain 25 μl of supernatant. Thereafter, 750 µl of the enzyme solution was added to the supernatant and stirred. The sample was then warmed for 5 minutes in a water bath at 37 ° C. with a standard solution having an HDL cholesterol content of 0, 10, 20, 30, 40 and 50 mg / dl, respectively, and the absorbance at 500 nm was measured. Colorimetric quantification was carried out.

Table 6 shows the results of the measurement of plasma total triglyceride, total cholesterol and HDL cholesterol content for each experimental group.

Table 6 Measurement item Experimental group CON HF HF + OK56 HF + OK169 HF + OK101 HF + OK215 HF + OK156 Plasma TG (mg / dl) 65.5 87.7 70.5 71.5 73.2 73.2 73.2 Plsama cholesterol (mg / ㎗) 93.3 184.5 149.5 156.5 158.9 161.2 166.5 Plasma HDL (mg / dl) 55.2 95.2 89.5 95.4 93.2 97.3 92.6

CON: General Dietary Salary

* HF: High Fat Dietary Salary Group

* HF + OK56: Lfatobacillus brevis ( Lactobacillus brevis ) OK56 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK169: high fat diet and 1 × 10 ⁹ CFU / mouse dose of Lactobacillus plantarum OK169 administration group

* HF + OK101: Lfatobacillus sakei OK101 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK215: Lactobacillus plantarum OK215 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

* HF + OK156: Lactobacillus sakei OK156 administration group of high fat diet and 1 × 10 ⁹ CFU / mouse dose

As shown in Table 6, the group of oral administration of lactic acid bacteria with high fat diet after induction of obesity significantly reduced plasma triglyceride and total cholesterol content when compared with the group receiving high fat diet after induction of obesity. It was.

5. In-vivo experiments on anti-obesity effects of fermented lactic acid bacteria

(1) Preparation of lactic acid bacteria fermented product or extract thereof

* Preparation of lactic acid bacteria fermented soybean

Lactobacillus brevis OK56 was inoculated in 10 ml of MRS broth (Difco, USA) and incubated for about 24 hours. Thereafter, the Lactobacillus brevis OK56 culture was centrifuged to remove the medium and cells were obtained. Thereafter, 10 g of skim soybean powder was suspended in 90 ml of water and sterilized, and the cells were inoculated to a concentration of 1 × 10 ⁹ / ml and incubated for about 3 days. Thereafter, the mixed culture of soybean and lactic acid bacteria was lyophilized to obtain lactic acid bacteria fermented soybean powder.

* Lactic acid bacteria fermentation deodeok and its extract preparation

Lactobacillus brevis OK56 was inoculated in 10 ml of MRS broth (Difco, USA) and incubated for about 24 hours. Thereafter, the Lactobacillus brevis OK56 culture was centrifuged to remove the medium and cells were obtained. Thereafter, 10 g of Deodeok powder was suspended in 90 ml of water and sterilized, and the cells were inoculated to a concentration of 1 × 10 ⁹ / ml and incubated for about 3 days. Thereafter, the mixed culture solution of Deodeok and lactic acid bacteria was lyophilized to obtain lactic acid bacteria fermentation deodeok powder. In addition, 2 times the volume of ethanol was added to the mixed culture of Deodeok and lactic acid bacteria, followed by extraction. The extract was concentrated under reduced pressure and lyophilized to obtain an extract of Lactobacillus fermented Deodeok.

* Preparation of lactic acid bacteria fermented ginseng and extracts thereof

Lactobacillus brevis OK56 was inoculated in 10 ml of MRS broth (Difco, USA) and incubated for about 24 hours. Thereafter, the Lactobacillus brevis OK56 culture was centrifuged to remove the medium and cells were obtained. Thereafter, 10 g of ginseng powder was suspended in 90 ml of water and sterilized, and the cells were inoculated to a concentration of 1 × 10 ⁹ / ml and incubated for about 3 days. Thereafter, the mixed culture of ginseng and lactic acid bacteria was lyophilized to obtain lactic acid bacteria fermented ginseng powder. In addition, two times the volume of ethanol was added to the mixed culture of ginseng and lactic acid bacteria to extract the extract, the extract was concentrated under reduced pressure and lyophilized to obtain an extract of lactic acid bacteria fermented ginseng.

* Lactic acid bacteria fermented cornus seeds and extracts thereof

Lactobacillus brevis OK56 was inoculated in 10 ml of MRS broth (Difco, USA) and incubated for about 24 hours. Thereafter, the Lactobacillus brevis OK56 culture was centrifuged to remove the medium and cells were obtained. Thereafter, 10 g of degreasing cornus seed powder was suspended and sterilized in 90 ml of water, and the cells were inoculated to a concentration of 1 × 10 ⁹ / ml and incubated for about 3 days. Thereafter, the mixed culture solution of cornus seeds and lactic acid bacteria was lyophilized to obtain lactic acid bacteria fermented cornus seeds powder. In addition, two times the volume of ethanol was added to the mixed culture solution of cornus seeds and lactic acid bacteria, and the extract was concentrated under reduced pressure and lyophilized to obtain an extract of lactic acid bacteria fermented cornus oil.

(2) Animal test method

A total of 36 four-week-old male C57BL / 6J mice were fed with a chow diet (CD; Purina, Korea) for 20 weeks at a temperature of 20 ± 2 ° C, 50 ± 5% humidity, and a 12-hour light-dark cycle. Adapted. Thereafter, the experimental animals were divided into two groups, and the normal diet group (CON, n = 6) was fed a general diet of 4.5% of the total calories for 5 weeks and the high fat diet group (n = 30) was 45% of the total calories. Obesity was induced by feeding a high fat diet (D12451, Research Diets, New Brunswick, NJ), where% was fat for 5 weeks. The obese high fat diet group was divided into five groups (HF, HF + SOK56, HF + COK56, HF + POK56, and HF + OOK56), each of which was fed a high-fat diet and phosphate buffered saline. (phosphate buffered saline, PBS) was orally administered for 5 weeks, and HF + SOK56, HF + COK56, HF + POK56, and HF + OOK56 groups were fed high-fat diets, and at the same time, different fermenters or extracts thereof were phosphate buffered. It was suspended in saline (phosphate buffered saline (PBS) and administered orally for 5 weeks. In addition, the normal diet group was fed orally administered the same amount of phosphate buffered saline (PBS) for 5 weeks at the same time as the normal diet.

(3) weight change and dietary intake

The body weight and dietary intake of the test animals were measured at a constant time every day during the entire test period, and the results are shown in Table 7 below.

TABLE 7 Measurement item Experimental group CON HF HF + SOK56 HF + COK56 HF + POK56 HF + OOK56 Initial weight (g) 25.2 27.0 26.8 26.9 26.8 27.8 Final weight (g) 28.2 38.1 33.1 33.1 33.3 34.5 Weight gain (g) 4.0 11.1 6.3 6.2 6.5 5.7 % Weight gain 16.6 42.7 27.0 26.5 26.5 20.5 Dietary Intake (g) 4.1 2.9 3.1 2.9 2.9 3.1

CON: General Dietary Salary

* HF: High Fat Dietary Salary Group

* HF + SOK56: high fat diet and lactic acid bacteria fermented soybean powder dose group of 100mg / kg body weight

* HF + COK56: high fat dietary supplement and 50mg / kg body weight dose of lactic acid bacteria fermentation deodeok extract administration group

* HF + POK56: high fat diet and 50mg / kg body weight dose of lactic acid bacteria fermented ginseng extract administration group

* HF + OOK56: high fat dietary supplement and 50mg / kg body weight dose lactobacillus fermented cornus seed extract group

As shown in Table 7, in the case of the group fed oral with lactic acid bacteria fermented product or its extract after the induction of obesity, the dietary intake was not significantly different compared to the group fed the high fat diet after induction of obesity. This significantly decreased. From this, it can be seen that soybean, deodeok, ginseng, wild corn seed or its extract fermented with a specific lactic acid bacterium has excellent anti-obesity effect.

(3) Measurement of weight change of liver and adipose tissue

After inducing obesity and sacrificing the fermented product or its extract for 5 weeks by cardiac puncture, subcutaneous fat (SF), epididymal fat (EF), Peripheral fat (PF), mesenteric fat (MF), brown adipose tissue (BAT) and liver were measured and weighed. The results are shown in Table 8 below. .

Table 8 Measurement item Experimental group CON HF HF + SOK56 HF + COK56 HF + POK56 HF + OOK56 Liver (g) 1.07 1.12 1.08 1.05 1.08 1.10 Epididymal Adipose Tissue (g) 0.38 2.10 1.20 1.35 1.40 1.54 Fat tissue around the kidneys (g) 0.11 0.92 0.61 0.66 0.68 0.77 Subcutaneous adipose tissue (g) 0.41 2.11 1.65 1.63 1.65 1.63 Mesenteric adipose tissue (g) 0.21 0.71 0.58 0.58 0.57 0.56 Brown adipose tissue (g) 0.08 0.17 0.15 0.15 0.15 0.17

CON: General Dietary Salary

* HF: High Fat Dietary Salary Group

* HF + SOK56: high fat diet and lactic acid bacteria fermented soybean administration group of 100mg / kg body weight dose

* HF + COK56: high fat dietary supplement and 50mg / kg body weight dose of lactic acid bacteria fermentation deodeok extract administration group

* HF + POK56: high fat diet and 50mg / kg body weight dose of lactic acid bacteria fermented ginseng extract administration group

* HF + OOK56: high fat dietary supplement and 50mg / kg body weight dose lactobacillus fermented cornus seed extract group

As shown in Table 8, the group of oral administration of lactic acid bacteria fermented product or extract thereof with high fat diet after induction of obesity showed a lower weight of adipose tissue when compared to the group receiving only high fat diet after induction of obesity.

6. Preparation of pharmaceutical composition comprising lactic acid bacteria

In the following pharmaceutical compositions, Lactobacillus brevis OK56 cultures were Lactobacillus sakei OK101, Lactobacillus sakei OK156, Lactobacillus plantarum OK169 and Lactobacillus plantarum OK169. Cultures of lactic acid bacteria such as Planta room ( Lactobacillus plantarum OK215) or the like can be replaced with fermented products prepared by fermenting soybean, deodeok, ginseng, cornus seed and the like or extracts of the fermented products.

<6-1> Preparation of Powder

Lactobacillus brevis OK56 Culture 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

<6-2> Preparation of Tablet

Fermented Deodeok Extract of Preparation Example 2 10 mg

Corn starch 100 mg

Lactose 100 mg

Stearic Acid Magnesium 2 mg

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

<6-3> Preparation of Capsule

Fermented Deodeok Extract of Preparation Example 2 10 mg

3 mg of crystalline cellulose

Lactose 15 mg

Magnesium Stearate 0.2mg

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

<6-4> Preparation of the ring

Fermented Deodeok Extract of Preparation Example 2 10 mg

Lactose 150 mg

Glycerin 100 mg

Xylitol 50 mg

After mixing the above components, it was prepared to be 4 g per ring in a conventional manner.

<6-5> Preparation of Granules

Fermented Deodeok Extract of Preparation Example 2 15 mg

Soy extract 50 mg

Glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added, dried at 60 ° C. to form granules, and then filled into fabrics.

<6-6> Preparation of Injection

Fermented Deodeok Extract of Preparation Example 2 10 mg

Sodium Metabisulfite 3.0 mg

Methylparaben 0.8 mg

Propylparaben 0.1 mg

Appropriate sterile distilled water for injection

After the above ingredients were mixed, 2 ml of this was filled into ampoules and sterilized to prepare an injection.

7. Preparation of food composition containing lactic acid bacteria

In the following food composition, Lactobacillus brevis OK56 cultures were Lactobacillus sakei OK101, Lactobacillus sakei OK156, Lactobacillus plantarum OK169 and Lactobacillus plantarum OK169. Cultures of lactic acid bacteria such as Planta room ( Lactobacillus plantarum OK215) or the like can be replaced with fermented products prepared by fermenting soybean, deodeok, ginseng, cornus seed and the like or extracts of the fermented products.

<7-1> Preparation of Flour Food

0.5 part by weight of the fermented deodeok extract of Preparation Example 2 was added to the flour, and bread, cake, cookies, crackers, and noodles were prepared using the mixture.

<7-2> Production of Dairy Products

0.5 parts by weight of the fermented deodeok extract of Preparation Example 2 was added to 100 parts by weight of milk, and various dairy products such as butter and ice cream were prepared using the milk.

<7-3> manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh.

Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

The grains, seeds and the fermented deodeok extract of Preparation Example 2 prepared above were formulated in the following ratio.

Cereals (30 parts by weight brown rice, 17 parts by weight of barley, 20 parts by weight of barley),

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

Fermented deodeok extract of Preparation Example 2 (1 part by weight),

Ganoderma lucidum (0.5 parts by weight),

Foxglove (0.5 part by weight)

<7-4> Preparation of health drink

A homogeneous blend of subsidiary materials such as liquid fructose (0.5 g), oligosaccharide (4 g), sugar (2 g), salt (0.5 g) and water (77 g) and 1 g of fermented deodeok extract of Preparation Example 2 After sterilization, it was prepared by packing it in a small packaging container such as a glass bottle or a plastic bottle.

<7-5> Preparation of vegetable juice

2 g of the fermented deodex extract of Preparation Example 2 was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.

<7-6> Preparation of Fruit Juice

1 g of the fermented deodeok extract of Preparation Example 2 was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

8. Accession Information of Lactic Acid Bacteria

The inventors of the present invention patented the Lactobacillus brevis OK56 on February 5, 2014 to the Korea Microorganism Conservation Center, an official depository, and received the accession number of KCCM 11516P. In addition, the inventors of the present invention secured the Lactobacillus sakei OK101 to the Korea Microorganism Conservation Center, an official depository institution on February 5, 2014, and received the accession number of KCCM 80070, and on January 28, 2015. Changed to patent deposit and received the accession number of KCCM 11664P. In addition, the inventors of the present invention secured the Lactobacillus plantarum OK169 to the Korea Microorganism Conservation Center, an authorized depository institution on February 5, 2014, and received the accession number of KCCM 80069, January 28, 2015. Changed to a patent deposit on the date was given an accession number of KCCM 11663P.

Although the present invention has been described through the above embodiments as described above, the present invention is not necessarily limited thereto, and various modifications can be made without departing from the scope and spirit of the present invention. Therefore, the protection scope of the present invention should be construed as including all embodiments falling within the scope of the claims appended to the present invention.

* Lactobacillus brevis ( Lactobacillus brevis ) OK56 Patent Deposit Certificate

[Revision under Rule 26 08.04.2015]

* Lactobacillus sakei OK101 safety deposit certificate and patent deposit certificate

[Revision under Rule 26 08.04.2015]

[Revision under Rule 26 08.04.2015]

* Lactobacillus plantarum OK169 safety deposit certificate and patent deposit certificate

[Revision under Rule 26 08.04.2015]

Claims (17)

Lactobacillus brevis comprising the nucleotide sequence of SEQ ID NO: 1 as 16S rDNA, Lactobacillus sakei comprising the nucleotide sequence as shown in SEQ ID NO: 2 as 16S rDNA, and SEQ ID NO: 4 as 16S rDNA. As one or more Lactobacillus genus lactic acid bacteria selected from the group consisting of Lactobacillus plantarum containing the base sequence described, Lactic acid bacteria having anti-obesity activity, blood cholesterol lowering activity, blood triglyceride lowering activity or antioxidant activity. According to claim 1, The Lactobacillus brevis ( Lactobacillus brevis ) is a bacilli that are positive when Gram staining, Glucose, Amygdalin, Arabinose, Esculin, Fructose as a carbon source (Fructose), Galactose, Galactose, Lactose, Mannose, Mannose, Raffinose, Rhamnose, Sorbitol, Sobitol, Sucrose and Xylose Lactic acid bacteria. According to claim 1, The Lactobacillus brevis ( Lactobacillus brevis ) is Lactobacillus brevis ( Lactobacillus brevis ) OK56 (Accession Number: KCCM 11516P), characterized in that the lactic acid bacteria. According to claim 1, The Lactobacillus sakei ( Lactobacillus sakei ) is a bacillus that is positive when Gram staining, Glucose, Amygdalin, Arabinos (Arabinose), Esculin, Fruc as a carbon source Fructose, Galactose, Glycerol, Lactose, Mannose, Melibiose, Raffinose, Sobitol, Sucrose and Xylose Lactobacillus characterized in using. According to claim 1, Lactobacillus sakei ( Lactobacillus sakei ) is Lactobacillus sakei ( Lactobacillus sakei ) OK101 (Accession Number: KCCM 11664P) characterized in that the. According to claim 1, The Lactobacillus plantarum ( Lactobacillus plantarum ) is a bacillus that is positive when Gram staining, glucose (Glucose), Amygdalin (Amygdalin), Arabinos (Arabinose), Esculin (Esculin), Fructose, Galactose, Glycerol, Lactose, Mannose, Melezitose, Meliviose, Raffinose, Rhamnose, Lactobacillus characterized in using sorbitol (Sorbitol), sucrose (Sucrose) and xylose (Xylose). The lactobacillus of claim 1, wherein the Lactobacillus plantarum is Lactobacillus plantarum OK169 (Accession Number: KCCM 11663P). A pharmaceutical composition for preventing or treating obesity or obesity-related diseases comprising the lactic acid bacterium of any one of claims 1 to 7, a culture thereof, a lysate thereof or an extract thereof as an active ingredient. The method of claim 8, wherein the obesity-related disease is any one selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. Or therapeutic pharmaceutical compositions. A food composition for the prevention or improvement of obesity or obesity-related diseases comprising the lactic acid bacterium of any one of claims 1 to 7, its culture, its lysate or extract thereof as an active ingredient. 11. The method for preventing or improving obesity or obesity-related diseases according to claim 10, wherein the obesity-related diseases are selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. Food composition. Obesity comprising a fermentation product obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with any one of claims 1 to 7 or the extract of the fermentation as an active ingredient or Pharmaceutical compositions for the prevention or treatment of obesity-related diseases. The pharmaceutical composition for preventing or treating obesity or obesity-related diseases according to claim 12, wherein the extraction solvent for obtaining the extract is water, alcohol or a mixture thereof. 13. The method of claim 12, wherein the obesity-related disease is any one selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. Or therapeutic pharmaceutical compositions. Obesity comprising a fermentation product obtained by fermenting at least one selected from the group consisting of soybean, deodeok, ginseng and cornus seed with any one of claims 1 to 7 or the extract of the fermentation as an active ingredient or Food composition for the prevention or improvement of obesity-related diseases. The method of claim 15, wherein the extraction solvent for obtaining the extract is a food composition for the prevention or improvement of obesity or obesity-related diseases, characterized in that water, alcohol or a mixture thereof. 16. The method of claim 15, wherein the obesity-related disease is selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis, and lipid-related metabolic syndrome. Food composition.

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