KR101611830B1 - Use of bifidobacterium breve cbt br3 in the prevention or treatment of obesity and obesity-related metabolic syndrome and composition comprising the same - Google Patents

Abstract

The present invention relates to a Bifidobacterium breve CBT BR3 (KCTC 12201BP) strain for treating or preventing obesity and metabolic syndromes caused by obesity, and to a pharmaceutical composition and an anti-obesity composition for treating or preventing obesity and metabolic syndromes caused by obesity, comprising the strain.

Description

[0001] The present invention relates to a Bifidobacterium breve CBT BR3 strain for the prevention or treatment of metabolic diseases caused by obesity and obesity, and a composition comprising the same. BACKGROUND ART [0002] COMPRISING THE SAME}

The invention bipyridinium betting for the prevention or treatment of metabolic diseases caused by obesity and obesity Te Solarium Breve CBT BR3 (Bifidobacterium Breve CBT BR3 ) and a composition comprising the same, and more particularly to a composition comprising Bifidobacterium Breve CBT BR3 relates to a strain and a pharmaceutical or food composition comprising the same.

Obesity is a major health problem in most countries. The World Health Organization (WHO) estimates that over one billion adults worldwide are overweight, and 300 million of them are clinically obese (Hotamisligil, 2006). Obesity is a major risk factor for cardiovascular disease, stroke, insulin resistance, diabetes, liver disease, neurodegenerative diseases, respiratory diseases and other serious diseases, and is a risk factor for some cancers, including breast and colon cancer . In addition to the effects on physical health, obesity has a detrimental effect on quality of life and mental health, resulting in depression, dietary disturbances and low self-esteem.

The steady increase in food intake with high energy loads and the social changes associated with less exercise increase the incidence of metabolic diseases caused by obesity and obesity. Conventional treatments based on low-calorie diets and exercises have little effect on obesity control and only result in temporary weight loss. The development of safe and effective drugs to induce weight loss has been going on for decades. However, to date, efficacious drugs have had serious side effects or are not as effective in clinical trials. For example, amphetamine has been used effectively as an appetite suppressant, but has a strong risk of dependence with other side effects. The discovery of leptin has attracted attention as a breakthrough in the treatment of obesity, but leptin was not as effective as expected in clinical trials. Cannabinoid receptor antagonists have been developed as antiobesity drugs, but have resulted in serious psychiatric side effects. Therefore, a new approach is needed to improve metabolic diseases caused by obesity and obesity and to prevent such pathological conditions.

Probiotics are recognized as a novel factor associated with obesity and related diseases through their ability to control metabolic and immunological functions (Sanz et al., 2010. Proc. Nutr. Soc., 14: 1-8). In this regard, Korean Patent Publication No. 2010-0109661 discloses a novel Lactobacillus having the blood cholesterol lowering and obesity inhibitory effect Better Kerr switch and strains is disclosed, Korean Patent Publication No. 2012-0128260 discloses, and obesity and for the treatment or prevention of obesity-related disorders are disclosed compositions comprising a mixture of lactic acid bacteria of the Bifidobacterium, Korean Patent Publication No. 2010 - 0010015 discloses that lactobacillus having a blood cholesterol lowering and anti-obesity activity Johnsoni strain has been disclosed, but its effect has not reached a satisfactory level.

The present inventors have made intensive studies in order to discover an excellent anti-obesity effect probiotics, Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3) as an anti-obesity product. An object of the present invention is te bipyridinium gaming suitable for the prevention and treatment of metabolic diseases caused by obesity and obesity is excellent in anti-obesity effect Solarium Breve CBT BR3 (Bifidobacterium breve CBT BR3).

Another object of the present invention is to inhibit the differentiation into adipocytes in adipose precursor cells, reduce the secretion of IL-6, TNF-a and IL-1 beta in the central nervous system, reduce body weight and abdominal fat, decrease blood cholesterol Bifidobacterium The present invention relates to a new use of a strain of Bifidobacterium breve CBT BR3.

It is another object of the present invention to provide a method for the treatment of Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3). ≪ / RTI >

One aspect of the present invention in order to accomplish the above object is to provide a method for producing Bifidobacterium (Bifidobacterium tuberculosis) which is deposited with the Korean Collection for Type Culture (KCTC) under the deposit number KCTC 12201BP, Breve CBT BR3 (Bifidobacterium breve CBT BR3) relates to lactic acid.

Another aspect of the present invention for the prevention or treatment of metabolic diseases caused by obesity, or obesity, Bifidobacterium of the present invention Solarium The present invention relates to a pharmaceutical or food composition comprising a strain of Breve CBT BR3.

Another aspect of the present invention for achieving the above object is Bifidobacterium Rove L'CBT BR3 (KCTC 12201BP) strain and Lactobacillus Phil also know Ruth CBT CBT LA1 (KCTC 11906BP), Bifidobacterium Animal Malice Lactis CBT BL3 (KCTC 11904BP), Peddie Oh Caucus pen Tosa three mouse CBT SL4 (KCTC 10297BP)) and Lactobacillus strains Planta Room CBT LP3 (KCTC 10782BP) and Lactobacillus It relates to an antiobesity composition comprising ramno Versus CBT LR5 other prebiotic one selected from a group consisting of (KCTC 12202BP) or more (prebiotic).

The Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3 (KCTC 12201BP)) has excellent anti-obesity efficacy and improves metabolic diseases caused by obesity.

Also, the Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3 (KCTC 12201BP)) and a composition of the present invention comprising the same as an active ingredient are useful for inhibiting adipocyte differentiation, decreasing body fat, reducing the amount of visceral fat, decreasing total cholesterol concentration, plasma triglyceride and liver triglyceride Resulting in a prophylactic or therapeutic activity of metabolic diseases caused by obesity or obesity ultimately. In addition, the composition of the present invention also exhibits a significant reduction in blood glucose and cholesterol concentration, and also has the effect of improving insulin resistance.

The Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3 (KCTC 12201BP)) can be advantageously used as a pharmaceutical composition and an anti-obesity food composition for the treatment or prevention of metabolic diseases caused by obesity and obesity because of its excellent anti-obesity effect.

In another aspect of the invention, Bifidobacterium Breve CBT BR3 (KCTC 12201BP) strain and Lactobacillus Phil also know Ruth CBT LA1 (KCTC 11906BP), Bifidobacterium Annie Marlies lactis CBT BL3 (KCTC 11904BP), Peddie Oh Caucus Pen landslide three mouse CBT SL4 (KCTC 10297BP)) and Lactobacillus strains Planta Room CBT LP3 (KCTC 10782BP) and Lactobacillus Prevention of ramno Versus CBT LR5 (KCTC 12202BP) of metabolic disorders, anti-obesity composition comprising at least one other prebiotic (prebiotic) is selected from the group consisting of is caused by the high obesity and obesity because of its excellent anti-obesity efficacy in and It not only shows improvement effect but also has high safety and there is no fear of side effects.

≪ RTI ID = 0.0 > 1A < / RTI > Breve CBT BR3 (Bifidobacterium breve CBT BR3) 16S rRNA sequence of KCTC 12201BP strain.
Figures 1b-c show the Bifidobacterium This is a comparison of the homology and phylogenetic relationship of 16S rRNA sequences of breve CBT BR3 with related species.
FIG. 2 is a graph showing the activity of the Bifidobacterium BRAY a RAPD (Random Amplified Polymorphic DNA) analysis of the genome DNA of the chopping CBT BR3 strain.
Figure 3 is a graphical representation of the Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3) Pulsed Field Gel Electrophoresis (PFGE) analysis of the genomic DNA of KCTC 12201BP strain.
Figure 4 is a graphical representation of the Bifidobacterium Breve CBT BR3 (Bifidobacterium breve CBT BR3) hemolytic activity of KCTC 12201BP strain.
FIG. 5 is a graph showing the effect of Bifidobacterium of the present invention on fat accumulation in differentiated 3T3-L1 cells This is a photograph showing the effect of Brebe CBT BR3 strain.
FIG. 6 is a graph showing the effect of the Bifidobacterium of the present invention on proinflammatory cytokine secretion comprising IL-6, TNF-a, IL-1 beta in obesity-induced 3T3- A graph showing the effect of BR3 CBT breve strain and strain composition mixed (MIX).
Figure 7 shows that in co-cultured 3T3-L1 adipocytes-PBMCs in rats, Bifidobacterium This is a bar graph showing the inhibition of LPS-induced NO by Brave CBT BR3 strain.
Figure 8 is a graphical representation of the Bifidobacterium A photograph showing the result confirming the body fat accumulation inhibition by CBT BR3 breve strain and strain composition mixed (MIX).

As used herein, the terms "treat" and "treatment" refer to controlling effects caused as a result of a disease or pathological condition in a subject, Means prevention, reduction, alleviation or elimination of metabolic disease symptoms caused by obesity.

As used herein, the term "prevention" means preventing a disease or pathological condition from occurring when a subject is diagnosed with a metabolic disease caused by obesity and obesity.

Such obesity includes, but is not necessarily limited to, simple obesity, symptomatic obesity, childhood obesity, adult obesity, cell proliferative obesity, cell non-large obesity, upper body obesity, lower body obesity, visceral fat obesity and subcutaneous fat obesity.

One aspect of the present invention relates to a method of inhibiting the growth of Bifidobacterium (Bifidobacterium tuberculosis) which has been deposited with the Korean Collection for Type Culture (KCTC) A breve CBT BR3 (Bifidobacterium breve CBT BR3) lactic acid.

Obesity occurs as a result of a positive and long-term imbalance between consumption and energy consumption, causing an excessive increase in body fat. Insulin is the most important hormone in the control of the proper functioning of adipose tissue, the control of the accumulation of triglycerides therein, and the glucose uptake. Fat is a reactant to insulin and other hormones (leptin), which is stored in normal insulin-sensitive adipose tissue by stimulating lipoprotein lipase and inhibiting lipolysis. However, excessive accumulation of fatty acids in obesity-related adipose tissue reduces insulin sensitivity, which converts the accumulation of triglyceride-type free fatty acids into some organs and tissues, resulting in changes in leptin production or sensitivity, and synthesis of proinflammatory cytokines Which further increases the risk of developing related diseases such as metabolic syndrome and cardiovascular disease.

On the other hand, obesity is also recognized as a mild chronic inflammatory state and is characterized by high production in adipose tissue of cytokines, adipocytes and other proinflammatory proteins (Tilg and Moschen, 2006. Nat Rev Immunol., 6: 772 ~ 783). Inflammatory factors associated with obesity and metabolic disorders include in particular the proinflammatory cytokines TNF-a and IL6. In particular, TNF-a reduces expression of genes involved in the action of insulin, weakens insulin signaling, and inhibits the action of insulin-stimulated lipoprotein lipases.

The Bifidobacterium The Breve CBT BR3 strain restores the composition of intestinal microbial counts, normalizing the changes associated with metabolic diseases caused by obesity and obesity and the inflammatory effects that cause these changes, and also normalizing changes associated with other pathological conditions . The strain of the present invention significantly reduces the synthesis of TNF-a which inhibits body weight regulation and glucose metabolism regulation by increasing obesity in the central nervous system and contributing to the development of insulin and leptin resistance.

Obesity means that the body is over-accumulated in the body. Obesity is mainly caused by the accumulation and accumulation of fat in the food by degradation and absorption by lipase, which leads to the differentiation of lipid precursor cells into adipocytes, thereby increasing the volume and number of adipocytes. do.

As demonstrated in the following examples, the Bifidobacterium Breve CBT BR3 is to result in the inhibition of adipocyte differentiation, reduction in fat mass, decrease in visceral fat, decrease of total cholesterol, plasma triglycerides and reduction in liver tissue triglyceride exerts effects to significantly alleviate obesity. In addition, the Bifidobacterium Breve CBT BR3 is also demonstrated efficacy that results in a significant reduction by improving insulin resistance, thereby improving a metabolic disease which is caused by obesity thereto in blood glucose.

The Bifidobacterium Breve CBT BR3 strains demonstrate that to suppress the differentiation of adipocytes in preadipocytes, this fact is useful in the treatment of increased fat cell, which is of the present invention strains can lead to a metabolic disease caused by obesity and obesity. This is because large-sized adipocytes induce adipocyte formation through preadipocyte differentiation and secrete growth factors that produce the feedback process at higher concentrations. Hypertrophic adipocytes deepen abnormal high concentrations of inflammatory cytokines and chemokines (TNF-a, MCP-1, IL-6, resistin, etc.), which inhibit insulin signaling in hepatic cells, Other complications are caused. Thus, the Bifidobacterium CBT BR3 breve strain may be similar to those related to the prevention or improvement of metabolic diseases.

According to another aspect of the present invention, there is provided a pharmaceutical composition comprising Bifidobacterium The Breve CBT BR3 strain can be used as a probiotic lactic acid bacterium or can be used in a variety of dairy products and other fermented products, and can also be used as an animal feed additive, depending on the embodiment.

Bifidobacterium according to the present invention Breve strain because CBT BR3, as well as obesity inhibitory effect is very excellent and reduces the amount of cholesterol can be used as anti-obesity functional food and obesity treatment and prevention of a pharmaceutical composition, such as for. Another aspect of the present invention is a pharmaceutical composition useful for the prevention or treatment of metabolic diseases caused by obesity and obesity, and another aspect of the present invention relates to an anti-obesity food composition. The food composition is a food, a nutraceutical, a supplement, a probiotic or a symbiotic.

The term " food composition " in the context of the present invention refers to a food which, without regard to the nutrients provided to a subject ingesting it, serves to favor one or more functions of the organism to provide a better health condition. As a result, the food composition can be used for the prevention or treatment of disease or disease-inducing factors. Thus, the term "food composition" of the present invention may be used as a synonym for a functional food or a food or medicinal food for a specific nutritional purpose. The term "probiotic agent" as used herein refers to a live microorganism that is beneficial to the health of the host organism when supplied in an appropriate amount. As used herein, the term "symbiotic" refers to foods containing a mixture of prebiotic and probiotic.

The compositions of the present invention according to an embodiment Bifidobacterium In addition to Brevet CBT BR3 strain, lactobacillus Wu raised Bari's (Lactobacillus salivarius), Lactobacillus Brevis (Lactobacillus brevis), Lactobacillus Helveticus (Lactobacillus helveticus), Lactobacillus Fur lactofermentum (Lactobacillus fermentum), Lactobacillus The para-casein (Lactobacillus paracasei), Lactobacillus Kasei (Lactobacillus casei), Lactobacillus Del Brewer station (Lactobacillus delbrueckii), Lactobacillus Leu Terry (Lactobacillus reuteri), Lactobacillus Boots Tenerife (Lactobacillus buchneri), Lactobacillus In addition Lee (Lactobacillus gasseri), Lactobacillus Jones you (Lactobacillus johonsonii), Lactobacillus Kane pireu (Lactobacillus kefir), Nose Lactobacillus Syracuse Lactis (Lactococcus lactis), Bifidobacterium BP Doom (Bifidobacterium bifidum ), bifidobacterium Infante Tees (Bifidobacterium infantis), Bifidobacterium Also ronggum (Bifidobacterium pseudolongum), Bifidobacterium spent a brush Room (B ifidobacterium themophilus m, Bifidobacterium Adolfo sentiseu (Bifidobacterium adolescent < / RTI > bacteria), and at least one probiotic lactic acid bacteria selected from the group consisting of < RTI ID = 0.0 >

The Bifidobacterium The breve CBT BR3 strain can be recovered by growing it by culturing under the usual conditions using a medium usually used for culturing the lactic acid bacteria. The culture obtained after cultivation may be used as is, or further subjected to solid-liquid separation or sterilization by, for example, rough purification by centrifugation or the like and / or filtration. Preferably, centrifugation is carried out to recover only the microbial cells of the lactic acid bacteria. In addition, the lactic acid bacteria used in the present invention may be wet cells or dried cells. For example, it can be prepared into a biocide form by lyophilization and used.

The compositions of the present invention may be used to treat < RTI ID = In addition to Breve CBT BR3 strains, conventional pharmaceutical acceptable carriers or excipients may be further included. In addition, various additives commonly used in the pharmaceutical industry such as binders, disintegrating agents, coating agents, lubricants, .

The term "excipient" means a material that assists in the absorption of any component of the composition of the present invention and assists in the manufacture of the pharmaceutical composition in the sense of stabilizing the components. Thus, the excipient may be selected from the group consisting of a function of keeping the components bound to each other, a sweetening function, a coloring function, a protective function of the medicament, for example a function of separating the medicament from air or moisture, a function of a filler for tablets, capsules or any other form, And have the function of promoting dissolution and their intestinal absorption.

The "carrier" or vehicle is preferably an inert component. The function of the carrier is to promote the incorporation of other components so as to enable better dosing and administration. Thus, the carrier may be a substance used to dilute any of the components of the pharmaceutical composition of the present invention in a certain volume or weight in a medicament; Or a substance that does not dilute the components and that allows for better dosing and administration.

The composition of the present invention can be used to treat < RTI ID = Breve CBT BR3 can be strain and suitable carriers, excipients, by powder mixing of the secondary as an active ingredient, granules, tablets, formulations in the form of a capsule or liquefaction. In addition, the composition of the present invention can be commercialized using a known method after being passed through the stomach and reaching the small intestine so that the active ingredient, lactic acid bacteria, is quickly released into the intestines.

Excipients usable in the present invention include sugars such as sucrose, lactose, mannitol, glucose and the like and starches such as corn starch, potato starch, rice starch, and partially pregelatinized starch. The binder may be a natural-occurring macromolecular substance such as a polysaccharide such as dextrin, sodium alginate, carrageenan, guar gum, acacia or agar, tragacanth, gelatin or gluten, hydroxypropylcellulose, methylcellulose, Cellulose derivatives such as cellulose, ethyl cellulose, hydroxypropyl ethyl cellulose and carboxymethyl cellulose sodium, and cellulose derivatives such as polyvinyl pyrrolidone, polyvinyl alcohol, polyvinylacetate, polyethylene glycol, polyacrylic acid, polymethacrylic acid and vinyl acetate resin Polymer.

Examples of the decomposing agent include cellulose derivatives such as carboxymethylcellulose, carboxymethylcellulose calcium and low substituted hydroxypropylcellulose, and sodium carboxymethyl starch, hydroxypropyl starch, corn starch, potato starch, rice starch and partially pregelatinized starch Of starch can be used.

Examples of lubricants that can be used in the present invention include talc, stearic acid, calcium stearate, magnesium stearate, colloidal silica, hydrous silicon dioxide, various types of waxes and hydrogenated oils and the like.

Examples of the coating agent include dimethylaminoethyl methacrylate-methacrylic acid copolymer, polyvinyl acetal diethylaminoacetate, ethyl acrylate-methacrylic acid copolymer, ethyl acrylate-methyl methacrylate-chlorotrimethylammonium ethyl methacrylate A water-insoluble polymer such as a copolymer, a water-insoluble polymer such as ethylcellulose, a methacrylic acid-ethyl acrylate copolymer, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate and the like and a thickener such as methylcellulose, hydroxypropylmethylcellulose , Polyvinyl pyrrolidone, polyethylene glycol, and the like, but are not limited thereto.

In the composition for prevention or treatment of obesity of the present invention, the above-mentioned Bifidobacterium Breve dose of CBT BR3 strains considering the purpose of treatment or prevention, treatment, or the kind of patient to the prevention, symptoms, weight, age, gender, etc. can be appropriately determined. In one embodiment, the composition of the present invention comprises, as the active ingredient, Bifidobacterium Breve strain CBT BR3 comprises a therapeutically effective amount of a nutritional or an effective concentration, preferably in an amount of 10 ⁸ to 10 ¹² cfu / g, or comprises the culture with an equal number of live cells. Generally, in the case of adult patients, live cells of 1 x 10 ⁶ cfu / g or more, preferably 1 x 10 ⁸ to 1 x 10 ¹² cfu / g may be administered once or several times as needed .

In addition, the Bifidobacterium Breve anti-obesity composition containing as an active ingredient has the CBT BR3 Bifidobacterium Vitreous B1, B2, B5, B6, E and acetic acid ester, nicotinamide, oligosaccharide, etc. may be added as nutritional supplement components in addition to Brevet CBT BR3.

Another aspect of the present invention is a method of treating Bifidobacterium In addition to the breve CBT BR3 (Bifidobacterium breve CBT BR3) (KCTC 12201BP) strain, Lactobacillus Phil also know Ruth CBT LA1 (KCTC 11906BP), Bifidobacterium Animal Malice Lactis CBT BL3 (KCTC 11904BP), Peddie Oh Caucus pen Tosa three mouse CBT SL4 (Pediococcus pentosaceus CBT SL4) (KCTC 10297BP)) strains and Lactobacillus Basil Ruth Planta Room CBT LP3 (Lactobacillus plantarum CBT LP3) (KCTC 10782BP) and Lactobacillus relates to an antiobesity composition comprising a Bacillus ramno Versus CBT LR5 (KCTC 12202BP) add at least one other prebiotic (prebiotic) selected from a group consisting of. This mixed strain composition of the present invention inhibits differentiation into adipocytes in lipid precursor cells, reduces the secretion of IL-6, TNF-a and IL-1 beta in the central nervous system, reduces body weight and abdominal fat, , Or a culture thereof. Therefore, the present invention can provide a composition useful for improving obesity. Such a composition is useful as a medicine, a food, or a health functional food as a composition for improving or preventing obesity.

Such a composition may contain each strain in the same proportion or may contain lactobacillus lactic acid bacteria and Bifidobacterium bifidus bacteria in a ratio of 2: 1 to 10: 1. The composition may comprise a mixture of lactic acid bacteria strains as an active ingredient in an amount of 10 ⁸ to 10 ¹² CFU / g, or an equivalent number of live bacteria, relative to the total weight of the composition.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

Example

Example  One. Bifidobacterium Breve  CBT BR3 ( Bifidobacterium breve  CBT BR3 ) Isolation and Identification of Strain

1-1. Selection of strain

1 g of human feces was serially diluted in sterile anaerobic water and 1 ml of each dilution was poured into a Man-Rogosa-Sharpe (MRS, USA) solid medium and incubated for 3 days under anaerobic conditions. The resulting colonies were transferred to a lactic acid bacteria selective medium (BL solid medium) to which BCP (Bromocresol purple, 0.17 g / L) was added to MRS and then cultured for 3 days under the same conditions. The BCP indicator changes from purple to yellow when the lactic acid bacteria form lactic acid and the surrounding pH is lowered. The colony was transformed into a yellow color, and biochemical and molecular biochemical identification was carried out. One strain with the best functionality and stability was finally selected.

1-2. Identification of selected strains

1) API Kit  Biochemical identification using

The API 50 CHL Carbohydrate Test Kit (bioMerieux Co., France) was used to determine the sugar availability of selected strains. After culturing in 10 ml of MRS (Man-Rogosa-Sharpe) liquid medium at 37 ° C for 17 hours, 1 ml of the culture broth was collected and washed twice with CHL solution. The cells were then collected by centrifugation (MICRO-17, Hanil, KR) and resuspended in 9 mL of CHL solution. 150 [mu] L of strain suspension was placed in each well of the API 50 CHL kit, and autoclaved paraffin oil was dispensed onto the strain suspension in the wells. After 3 days of incubation at 37 ° C, the sugar availability was compared. 49 carbon sources were observed for the change of color due to microbial growth, and the use of each carbon source was observed. The results of the final identification were analyzed using the identification program API web, and the results are shown in Tables 1 and 2 .

No Carbohydrates Utilized No Carbohydrates Utilized 0  Control - 25  Esculine + One  Glycerol - 26  Salicine w 2  Erythritol - 27  Cellobiose - 3  D-Arabinose - 28  Maltose + 4  L-Arabinose - 29  Lactose + 5  Ribose + 30  Melibiose + 6  D-Xylose - 31  Saccharose + 7  L-Xylose - 32  Trehalose + 8  Adonitol - 33  Inuline - 9  β-Methyl-xyloside - 34  Melezitose - 10  Galactose w 35  D-Raffinose + 11  D-Glucose + 36  Amidon - 12  D-Fructose w 37  Glycogene - 13  D-Mannose w 38  Xylitol - 14  L-Sorbose - 39  β-Gentiobiose - 15  Rhamnose - 40  D-Turanose w 16  Dulcitol - 41  D-Lyxose - 17  Inositol - 42  D-Tagatose - 18  Mannitol - 43  D-Fucose - 19  Sorbitol - 44  L-Fucose w 20  alpha -Methyl-D-mannoside - 45  D-arabitol - 21  alpha -Methyl-D-glucoside w 46  L-Arabitol - 22  N-Acetyl glucosamine - 47  Gluconate - 23  Amygdaline - 48  2-Ceto-gluconate - 24  Arbutine w 49  5-Ceto-gluconate w W; weak change

B. breve B. breve * B. longum * B. infantis * (KCTC12201BP) (ATCC 15700) (ATCC 15707) (ATCC 27920) D-Ribose + + + - L-Arabinose - - + - Lactose + + + + Cellobiose - d - - Melezitose - d + - Raffinose + + + + Sorbitol - d - - Gluconate - - - - d; In the case of positive, it was gradually fermented.

2) 16s rDNA  Identification through gene sequencing

Genomic DNA was extracted from isolated strains and analyzed for 16s rRNA sequences. Genomic DNA was extracted from 1 ml of the pure culture of the strain isolated from the feces using Accuprep genome extraction kit (Bioneer, Korea). (MyCycler, BIO-RAD, USA) was performed using primers F (5'-AAGGAGGTGATCCAGCC-3 ') ³ and primer R (5'-AAGGAGGTGATCCAGCC-3') ³ in the 16s rRNA region using the extracted DNA as a template .

The PCR product was connected to the pGEM-Teasy vector (Promega, USA) E. coli The strain DH5α was transformed into LB / x-gal / ampplate and cultured overnight at 37 ° C. After the recombinant plasmid containing the insert was isolated from the transformant through screening, DNA sequencing was performed. DNA sequencing was carried out using the Cluster V method of the DNA star program using Bifidobacterium The homology with Breve ^T (ATCC 15700) was compared. 16s rRNA base sequence of the separated strain, as shown in Figure 1, Bifidobacterium Breve ^T showed a homology (ATCC 15700) and 99.3%.

3) RAPD  Analysis of DNA Fingerprint by Random Amplified Polymorphic DNA

RAPD analysis was performed by PCR-RAPD (MyCycler, BIO-RAD, USA) using primers (GTG) ₅ (5'-GTGGTGGTGGTGGTTG-3 ' Respectively. The final product, PCR product, was stained with EtBr (ethidium bromide) followed by G: BOX (SYNGENE, UK). A strain isolated from the RAPD result, as shown in Figure 2, the Bifidobacterium Breve ^T was confirmed that (ATCC 15700) and show a different band pattern. Therefore, the strains isolated from the feces from the above results were Bifidobacterium It was confirmed that this strain was different from Breve ^T (ATCC 15700). 2, lane 1 represents bifidobacterium Breve is the result of ^T (ATCC 15700), lane 2 represents the results of the Bifidobacterium breve CBT BR3 (KCTC 12201BP).

4) DNA fingerprint analysis by Pulsed Field Gel Electrophoresis ( PFGE )

In the MRS broth,Bifidobacterium Breve CBT BR3,Pedio Caucus Ashidakarta City(P. acidalactici)^T(DSM 20284) andBifidobacterium Breve ^T (ATCC 15700) was measured, and 2% Low Melting Agarose was used to measure the final OD₆₀₀= 4 plugs were made. The prepared plugs were placed in 1 ml lysozyme buffer (2 mg / ml Lysozyme (Sigma), 0.05% N-lauorylsarcosine (Sigma)) and 10 μl of 4 mg / ml Lysostaphin (Sigma) was added thereto and reacted overnight at 37 ° C. The plug was carefully removed and placed in 4 ml of NDS buffer (1 ml 1M Tris-HCl (pH = 8.0), 10 ml 100% SDS, 89 ml 0.5 M EDTA (pH = 8.5) and reacted overnight at 50 ° C. After washing the plug 6 times with 10 ml of 50 mM EDTA (pH 8.5), carefully transfer the plug to 400 μl of the enzyme buffer to be treated, and allow to stand for 30 minutes at room temperature. After transferring the plug to 400 μl of the new enzyme buffer, The electrophoresis was carried out using a CHEF system (BIO-RAD, USA) at 0.5X TBE at 5.3 cm / V, 1 s to 15 s pulse time , ≪ / RTI > for 20 hours.

After the electrophoresis was completed, the band pattern was observed with G: BOX (SYNGENE, UK) after staining with EtBr solution. As a result of PFGE using NotI, Fecal Bifidobacterium breve is CBT BR3 Peddie Oh Caucus Tea attic know when ^{(P. acidalactici) T (DSM 20284} ) and Bifidobacterium Was identified as breve ^T (ATCC 15700) and new strains exhibit different band patterns. In Fig. 3, lane 1 represents Bifidobacterium Breve is the result of ^T (ATCC 15700), lane 2 is Bifidobacterium The results of breve CBT BR3 (KCTC 12201BP).

5) Other Mycology  characteristic

Bifidobacterium according to the present invention Breve characteristics of CBT BR3 are as follows:

1) Types of bacteria Characteristics of bacteria when cultured in MRS agar plate medium at 37 ° C for 2 days
① Cell shape: Bacillus
② Mobility: None
③ Spore forming ability: None
④ Gram staining: positive 2) Morphology When cultured in MRS agar plate medium at 37 ° C for 2 days,
① Shape: Circular
② Bump: convex
③ Surface: Smooth 3) Physiological properties ① Growth temperature: Growth possible Growth temperature 15 ~ 40 ℃
Optimum growth temperature 37 ℃
② Growth pH: Growable growth pH 5.0 ~ 7.5
Optimum pH 6.0 to 6.5
③ Effect on oxygen: anaerobic 4) Catalase - 5) Whether gas formation - 6) Growth at 15 캜 - 7) Growth at 45 캜 + 8) Indole production - 9) Production of lactic acid +

Based on the above results, the strain was designated " Bifidobacterium Breve deposited in the CBT BR3 (Bifidobacterium breve CBT BR3) " named the strain and the strain deposited with the Republic of Korea patent on May 7, 2012, Microbiological institute Resource Center (KCTC) and was given the accession number KCTC 12201BP.

1-3. Functionality and stability

1) Antibiotic resistance test

Isolated Bifidobacterium To verify the safety of breve CBT BR3 (KCTC 12201BP) antibiotic resistance were analyzed for. Antibiotic resistance tests were carried out using the micro-dilution method recommended by the European Food Safety Authority (EFSA). Ten antibiotics were used: ampicillin (AMP), vancomycin (VAN), gentamicin (GEN), kanamycin (KAN), streptomycin (STM), erythromycin (ERM), cinnamic acid (Q / D), clindamycin (CLM), tetracycline (TET) and chloramphenicol (CP).

For antibiotics except for clindamycin, the concentrations of 256, 128, 64, 32, 16, 8, 4, 2, 1 and 0.5 ㎍ / ㎖ were added to broth mixed with ISO-sensitized broth 10% and MRS broth 90% Since clindamycin has an EFSA breakpoint value of less than 0.25 μg / ml in the Lactobacillus group, antibiotics are added at concentrations of 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 and 0.03125 μg / Respectively. In addition, the thinner was carried out using an E-test strip of BioMeriux.

The microplate was incubated at 37 캜 under anaerobic conditions for 48 hours, and then the MIC was measured at the lowest antibiotic concentration at which no visible growth was observed. Based on the EFSA breakpoints, Bifidobacterium breve CBT BR3 (Bifidobacterium breve CBT BR3 ) were tested for resistance to each antibiotic and are shown in Table 4 below.

Strain Antibiotics (μg / ml) AMP VAN GEN KAN STM ERM CLM QU + DA TET CP B. breve CBT BR3 <0.5 <0.5 <8 <64 <32 <0.5 <0.06 <0.38 <4 <2 EFSA break point 2 2 64 nr 128 0.5 0.25 One 8 4

Isolated Bifidobacterium Breve CBT BR3 was confirmed due to the resistance to any antibiotic used in the experiment appeared to be lower than EFSA antibiotic resistance criteria to be suitable for the stability criteria for the EFSA antibiotics. Since Bifidobacterium has been reported to be resistant to aminoglycosides such as kanamycin due to the lack of a cytochrome-mediated drug delivery system, EFSA does not require the MIC value of a drug for Bifidobacterium.

2) Long-term fixability  Experiment

Bifidobacterium The measurement of intestinal fixation of BREVE CBT BR3 was carried out on Bifidobacterium Breve in the ^T (ATCC 15700) as a control was carried out in the HT-29 cell line derived from human colonic epithelial cells. HT-29 cell lines treated with each strain for 1 hour, and then the number of viable cells and the number of viable cells were measured. The results are shown in Table 5 below.

Bifidobacterium breve CBT BR3 Bifidobacterium breve ^T Long-term settlement rate (%) 91.25 77.24

As a result of the measurement of the fixation property of the bifidobacterium The Breve CBT BR3 strain is Bifidobacterium It was confirmed that breve ^T (ATCC 15700) chapter looks superior to strain strain fixation. These results indicate that the strain of the present invention can improve the intestinal environment by adhering to the epithelial cells.

3) Hemolytic test

The hemolysis test was conducted to confirm the absence of hemolytic toxicity of lactic acid bacteria in the human body. By the method of Baumgartner et al., The test strain was grown in MRS medium supplemented with 5% horse blood and incubated at 37 ° C for 48 hours under anaerobic conditions. The hemolysis was judged by whether a transparent ring was formed around the cells. As a result of examining the hemolytic property of the strain of the present invention, as shown in FIG. 4,Bifidobacterium Breve The CBT BR3 strain was found to be harmless to the human body because it was not hemolytic to the blood of the horse.

4) Acute toxicity test

The Bifidobacterium In order to verify the safety of Breve CBT BR3 strain, acute toxicity test was performed on experimental animals. A lyophilized strain of the present invention was orally administered to a 6-week-old male Sprague-Dawley (SD) rats at 1.0 × 10 ¹¹ cfu / kg. The control group received 0.85% saline in the stomach. Clinical symptoms for all experimental animals were observed for 1 day from day 1 to day of autopsy for 14 days. Observation results are shown in Table 6 below.

Bifidobacterium After administering the CBT BR3 breve strain, it was unable to observe the mortality in all treated groups and the control group, also could not find an object that represents a specific clinical symptoms. In addition, no statistically significant difference was observed between the administration and control groups after 14 days of body weight change, food and water intakes, organ weights, and body temperature. The autopsy conducted at the end of the experiment showed no gross changes in appearance and internal organs in rats of all individuals.

gender Strain Days Since Administration Mortality (%) LD ₅₀ One 2 3 4 5 6 7 8 9 10 11 12 13 14 cock CBT BR3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 > 10 ¹¹
cfu / kg Control group 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 female CBT BR3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 > 10 ¹¹
cfu / kg Control group 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 * Numbers in the above table indicate the number of dead

Example  2. Obesity Bifidobacterium Breve  Effect of administration of CBT BR3 strain

The following examples demonstrate the characteristics and anti-obesity efficacy of the strains of the present invention. The results of all the experiments were expressed as mean ± SD. Statistical analysis of the experimental results was performed using GraphPad Prism ^™ 6.0. The mean difference between the experimental groups was determined by one-way ANOVA and then Tukey's multiple range test.

2.1. The culture of the strain of the invention and the preparation of a composition comprising it

For in vitro experiments, Bifidobacterium Breve CBT BR3 strain (KCTC 12201BP) to MRS agar was cultured at 24 hours 37 ℃ in (BD Diagnstics, Sparks, MD) , a phosphate buffer solution of 10 to (PBS, 10 mM sodium phosphate, 130 mM sodium chloride, pH 7.4) ¹¹ After the supernatant was diluted with CFU / ml, the supernatant was centrifuged, filtered through a 0.45 μm pore size filter, lyophilized and stored at -20 ° C. until in vivo experiment.

- Anti-obesity  Composition manufacturing

The fermentation medium for the cultivation of the lactic acid bacteria strain contained in the mixed strain composition of the present invention is prepared by preparing a 1% -10% (w / v) aqueous solution of skimmed milk powder and soy protein isolate, respectively, (Delvolase; DSM, Netherlands) solution in the range of 0.01% -1% (w / w).

The mixed strain composition was prepared as follows. After the enzyme treatment, the yeast extract in the range of 0.1% to 5% (w / v), the yeast extract in the range of 0.1% to 5% (w / v), the starch such as sucrose, Addition of ionic constituents of soymilk citrate, sodium acetate, dipotassium phosphate, magnesium sulfate, manganese sulfate, sodium chloride in the range of 5% (w / v) soy peptone, 0.01% -0.1% (w / v) And dissolved to prepare a lactic acid bacteria culture medium. The prepared culture medium for lactic acid bacteria was sterilized using a heat exchanger. The lactic acid bacteria that had been pre-cultured in the prepared lactic acid fermentation medium were inoculated at a level of 0.1% -1% (v / v) and fermented for 8 hours to 10 hours so as to maintain the pH at 5.0-6.0 at 37 ° C. After the cultivation of the lactic acid bacteria was completed, the lactic acid bacteria were separated and concentrated using a continuous centrifuge. The isolated lactic acid bacteria were lyophilized by adding a cryoprotectant, etc., followed by pulverizing the lactic acid bacteria using a pulverizer to have a uniform particle size and then producing lactic acid bacteria. The prepared lactic acid bacteria starter was Bifidobacterium Brevet CBT BR3 bifidobacteria (1.25 x ^{10 8} CFU / g or more), Pediococcus Pen soil three mouse CBT SL4 lactic acid (more than 1.25X10 ⁸ CFU / g), Lactobacillus Planta Room CBT LP3 Lactic acid bacteria (1.25 × ^{10 8} CFU / g or more), lactobacillus Phil also know Ruth CBT LA1 lactic acid bacteria (1.25X10 ⁸ CFU / g or more), Bifidobacterium lactis Annie Marlies CBT BL3 bifidobacteria (1.25X10 ⁸ CFU / g or more) and lactobacillus Ramno Seuss CBT LR5 lactic acid bacteria (1.25X10 ⁸ CFU / g or more). Each lactic acid bacterium was mixed with excipient and mixed to 500 mg per capsule. Hydroxypropylmethylcellulose capsule was prepared so that the total viable cell count was 7.5 x 10 ⁹ CFU / g.

2.2. Obese animal models and sampling

Animal experiments were conducted in accordance with the Animal Use and Care Protocol of the Institutional Animal Care and Use Committee (IACUC). Experimental animals were purchased for 6 weeks for male SD rats from Uiwang, Korea. After 6 weeks of adaptation period, they were fed for 8 weeks. Breeding conditions were 24 ℃, 55% The light cycle was maintained for 12 hours. To induce obesity, high fat diets (60 kcal% fat; Research Diets Inc, NJ, USA) were consumed for 8 weeks.

Animals were divided into random groups of 4:

 Experimental group I (NC): normal diet, normal control group,

Experimental group II (PC): high fat diets fed obesity group

Experimental group III (CBT BR3) High fat diet fed obesity + Bifidobacterium Brevet CBT BR3 strain treated group.

Experimental group IV (MIX) High fat diet feeds Obesity induction + Bifidobacterium Breve strain CBT BR3 + PD five caucuses Three pens Tosa funny CBT SL4 + Lactobacillus Planta Room CBT LP3 + Lactobacillus even know Phil Ruth CBT LA1 + Bifidobacterium Animal Malice Lactis CBT BL3 + Lactobacillus Lambsos CBT LR5 group.

In the case of experimental groups III and IV, after induction of obesity by high-fat diet, the freeze-dried Bifidobacterium Breve and Bifidobacterium strains CBT BR3 Breve mix the mixed culture in drinking water comprising a BR3 CBT through the gastrointestinal tract was injected with 10 ⁷ daily dose of cfu / tablets / day for 8 weeks. Normal control (NC) and obesity induction group (PC) were administered PBS in the same manner as placebo.

2.3 The 3T3-L1 of the strain of the present invention In fat precursor cells  Inhibitory effect on the differentiation of adipocytes

(Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% calf serum (Gibco, Grand Island, NY, USA) was inoculated with 1% penicillin - streptomycin (PS, Lonza, Allendale, NJ, USA). To induce differentiation of adipocytes, 0.5 mM 3-isobutyl-1-methylxanthine, 100 μM indomethacin, 0.25 μM dexamethasone, and 167 nM insulin were added to the culture medium for two days. After that, the medium was replaced with DMEM supplemented with 167 nM insulin and 10% fetal bovine serum (Gibco) at intervals of two days to induce differentiation for 10 days. Bifidobacterium The Breve CBT BR3 strain supernatant (100 [mu] g / ml) was treated with the medium for 10 days from the start of differentiation and treated with replacement every two days.

For 24 hours in 3T3-L1 preadipocytes lipid polysaccharide (LPS) (10 ㎍ / ㎖ ; Sigma, St. Lpuis, MO, USA) and Bifidobacterium breve CBT BR3 stimulation with the strain or supernatant alone stimulated only LPS Respectively. The Bifidobacterium In order to confirm whether Breve CBT BR3 strain acts on the differentiation of 3T3-L1 cells from adipocyte precursor cells into adipocytes and inhibits the formation of lipid spheres, it was stained with oil red O (Sigma Chemicals Co.) Respectively. The lipids produced in the stained 3T3-L1 cells were observed under a microscope. The lipid content was then desalted with isopropanol for 30 minutes to recover the dye, and the absorbance of the cell monolayer was measured at 520 nm and calculated as follows. The results are shown in FIG.

Fat accumulation rate (%) = 100- (A-B) / A x 100

A: A _{520 nm} [control group]

B: A _{520 nm} [CBT BR3 sample]

Adipogenesis is a process in which preadipocytes are transformed into mature adipocytes through proliferation and differentiation, and obesity occurs mainly when the volume or number of adipocytes is increased.

Figure 5A-E to see if, treated with a differentiation medium (differentiation media, DM) for 6 days Bifidobacterium breve CBT BR3 strain supernatant of the present invention as compared to the treatment group (100 ㎍ / ㎖) in differentiation medium The number of lipids in the group was decreased to 26.5%. These results suggest that the Bifidobacterium It can be confirmed that Breve CBT BR3 strain is excellent in inhibiting differentiation from adipocytes into adipocytes. Also, the amount of fat accumulated in the oil-red-O stained cells was quantified by a spectroscopic analyzer, and the OD value was also greatly reduced (FIG. 5E).

2.4 Bifidobacterium Breve  CBT BR3 strain IL-6, TNF -α, and IL- 1β  Secretion inhibitory effect

- in mice 3T3-L1 cells TNF -α, IL-6 and IL- 1β  Inhibition

3T3-L1 cells were treated with LPS (10 g / ml) and Bifidobacterium Breve the CBT BR3 strain supernatant proinflammatory cytokine secretion of IL-6, TNF-α and IL-1β after stimulation with (100 ㎍ / ㎖) or alone stimulated only LPS was measured by ELISA. In addition, the same manner with a mixed culture composition (MIX) comprising a Bifidobacterium breve CBT BR3 strains were compared to the results.

When LPS alone stimulated 3T3-L1 cells, the secretion of IL-6, TNF-α and IL-1β was increased (p <0.001, FIG. 6A-C). Whereas LPS for Bifidobacterium breve CBT BR3 strain supernatant or was the secretion of IL-6, TNF-α and IL-1β compared to LPS alone stimulation decreases when stimulated with the supernatant of the mixed culture in (p <0.001) .

Recently, the term 'metaflammation' has emerged for the inflammatory response caused by overfeeding of nutrients or metabolites, and obesity is interpreted as 'chronic and low-level inflammation' Research on the correlation with the system is actively underway. For example, toll-like receptor 4 (TLR4), which is involved in the innate immune response, plays an important role in the inflammatory response and insulin resistance pathway by using dietary fat (especially saturated fatty acid) as a ligand . In this study, we investigated the effect of NF-κB on the proliferation of NF-κB and pro-inflammatory cytokine (TNF) -α, IL-6, and the like. In addition, TNF-α and IL-6 phosphorylate the serine residues of the insulin receptor substrate (IRS) by activating SOCS3 (cytokine signaling 3) and JNK, inhibiting glucose transport, It is known to induce insulin resistance in peripheral tissues. In particular, the synthesis of TNF-α increases obesity and contributes to the development of insulin and leptin resistance, thereby inhibiting weight control and glucose metabolism (De Souza et al., 2005. Endocrinology., 146: 4192-9).

As can be seen from the results of FIG. 6, the Bifidobacterium CBT BR3 breve strain, as well as Bifidobacterium The mixed strain composition (MIX) comprising Breve CBT BR3 strain is effective in reducing TNF-a, IL-1 beta and IL-6 and thus significantly improves the chronic inflammatory activation of visceral fat tissue induced by obesity, It can be confirmed that there is an effect of controlling glucose metabolism and preventing or treating insulin resistance.

- Co-cultured rat 3T3-L1 adipocytes and In peripheral blood mononuclear cells (PBMC) LPS  Inhibition of induced NO

Rats of 3T3-L1 fat cells and peripheral blood mononuclear cells (PBMC) a co-culture on plates of 96 wells, according to the present invention, Bifidobacterium breve Use the CBT BR3 cultured for 1 hour LPS (10 ㎍ / mL) for 48 hours. The supernatant was analyzed for NO by an ELISA kit (R & D Biosystems) and the results are shown in FIG. As can be seen in Figure 7, the Bifidobacterium Breve administration of CBT BR3 was inhibited LPS experiments since nitric oxide (NO) production in these cells.

2.5 Biochemical Analysis of Serum Associated with Metabolic Syndrome

Obesity is closely related to metabolic syndrome such as insulin resistance, glucose tolerance, hyperlipidemia, and cardiovascular disease. Thus, the Bifidobacterium Breve strains was CBT BR3 within a biochemical analysis of serum to the end of the experiment in relation to the effect on the blood levels of glucose, triglycerides, total protein and cholesterol that are associated with metabolic syndrome.

Experimental rats were fasted for about 15 hours and blood was collected from the heart after anesthesia with Zoletil (30 mg / kg body weight). Cholesterol, HDL-cholesterol, LDL-cholesterol, total protein, triglyceride, and the like were measured using an automatic blood chemical analyzer 7020 (Hitachi, Japan) after centrifuging the collected blood at 2,000 × for 20 minutes. Seride (TG) concentrations were measured by ELISA (BDBioscience, San Diego, CA, USA).

Bifidobacterium Breve CBT BR3 was a result of intake measured in blood chemistry inspector to investigate the effects on lipid metabolism of the strain, total protein, cholesterol, HDL, LDL, glucose, statistical significance in triglycerides portion (p <0.001).

It's normal. Obesity induction group CBT BR3 MIX B. breve ^T Glucose 142.08 ± 12.48 205.63 ± 48.43 133.40 ± 11.87 ^# 130 ± 12.97 ^# 140.40 ± 11.98 Total protein 6.12 ± 0.26 6.98 + 0.24 5.72 ± 0.11 ^### 5.54 + - 0.22 ^### 5.92 + 0.15 cholesterol 49.73 ± 12.19 85.64 8.38 52.14 + - 4.55 ^### 47.83 ± 4.39 ^### 60.37 ± 11.91 Triglyceride 37.97 ± 18.71 68.36 + - 11.52 38.63 + - 25.24 32.16 ± 18.78 50.67 ± 26.17 HDL 37.97 ± 18.47 16.16 + - 2.52 20.74 ± 2.16 ^## 24.25 ± 2.91 ^## 17.14 + - 2.87 LDL  5.91 + 1.54 14.08 ± 2.18 9.37 ± 2.00 ^## 8.34 ± 1.26 ^### 11.34 + 1.89 * The number (##) next to the number indicates a significant difference from other dietary groups at P <0.001 using one-way ANOVA followed by Turkey's multiple range test.

As can be seen from the results of Table 7, the concentration of cholesterol, total protein, HDL, LDL, glucose and triglyceride in the serum was higher in the high fat diet group than in the normal control group (NC) And it was confirmed that obesity caused by LPS was induced. On the other hand, the Bifidobacterium It was confirmed that cholesterol, total protein, LDL, and glucose were significantly decreased by the administration of breve CBT BR3 strain as compared with obesity induction group (PC). In particular, it was confirmed that the CBT BR3-treated group had an effect of significantly reducing the concentrations of glucose, cholesterol, and triglyceride as compared with the obesity induction group (PC).

In obesity, when the amount of cytoplasmic triglyceride per cell increases, adipose tissue mass increases due to the production of new adipocytes, resulting in the development of adipocytes. And reduced triglyceride by 43% or more. In addition, CBT BR3-treated group showed an increase in HDL, which is capable of eliminating excess cholesterol present in tissue cells, compared with obese induction group (PC), and a decrease of about 33% in LDL that can cause atherosclerosis . Moreover, BP gaming of the present invention, Te Solarium breve CBT BR3 strain and Rhodococcus Phedi O Three pens Tosa funny CBT SL4 and Lactobacillus It was confirmed that when the mixed strain (MIX) of Planta room CBT LP3 was administered, the blood glucose lowering effect was further increased, the triglyceride was further decreased, and the HDL increase and LDL reduction effects were further increased.

2.6 Bifidobacterium Breve  CBT BR3 strain Induction of obesity  In the experimental animals, Liver tissue  Effect on weight

Bifidobacterium In order to observe the inhibitory effect of Breve CBT BR3 on obesity, obesity was induced by taking a high fat diet for 8 weeks in experimental rats. Before and after the experiment, the body weight of the experimental rats of each group was measured. After completion of the experiment, the mice were anesthetized with Zolethyl, and then the liver was harvested and weighed. The results are shown in Table 7 below .

Weight change, liver and adipose tissue weight were measured. The body weights of the experimental animals were increased by obesity induction group (NC: 515.67vs PC; 650.71) compared to the normal control group Was induced. The body weight change was significantly lower in the CBT BR3 group than in the obese group (p <0.001). Bifidobacterium It was found that the group receiving Brevet CBT BR3 significantly inhibited weight gain compared with the group fed only high fat diet (PC). As shown in the following Table 8, when the weight increase amount for 8 weeks was observed, Bifidobacterium The Brebeber CBT BR3 group was 555.62, the high-fat diet group was 650.71, and Bifidobacterium The breve weight gain by the CBT BR3 was confirmed that approximately 70% inhibition.

Bifidobacterium ( Bifidobacterium ) was also found in the liver Breve CBT was not significantly different from one group (CBT BR3) intake of BR3 (p <0.05). In the case of obesity, it is known that body fat metabolism and glucose metabolism occur abnormally and lipid components accumulate in the liver to increase the weight of the liver. In the case of Bifidobacterium In the BRTB BRT strain, the liver weight was reduced to a nearly normal level. Therefore, Bifidobacterium breve CBT BR3 strain of the present invention, it was confirmed that an excellent body weight and visceral fat reducing effect. Also, the Bifidobacterium Breve and Bifidobacterium strains CBT BR3 A synergistic effect was observed with respect to the excellent effect of reducing the body weight and visceral fat when mixed with Brev CBT BR3 strain.

It's normal. Obesity induction group CBT BR3 MIX B. breve ^T Initial weight 231.81 + - 10.66 231.26 ± 12.24 230.29 + - 10.41 233.86 8.38 231.97 + - 11.65 Final weight 515.67 ± 23.61 650.71 + - 24.38 555.62 ± 13.66 ^### 504.27 ± 22.29 ^### 610.16 + 21.35 Liver weight (g) 14.2 ± 1.3 19.6 ± 2.7 14.0 ± 0.7 ^## 13.8 ± 0.7 ^## 18.5 ± 2.8 The symbol next to the number (##) is used for one-way ANOVA followed by Turkey's multiple range test
And significantly different from other dietary groups at P <0.001.

2.7 for abdominal fat Bifidobacterium Breve  Effect of CBT BR3 strain administration

The Bifidobacterium CBT BR3 breve strain (CBT BR3) and Bifidobacterium Breve CBT BR3 and Peddie Oh Caucus Three pens Tosa funny CBT SL4 and Lactobacillus The lumbar spine (L2-L5) region was photographed using Magnetic Resonance Imaging (MRI) through the Korea Science and Technology Research Institute (KITA) to evaluate the abdominal fat according to the intake of the mixed strain (MIX) of Planta room CBT LP3. The volume of abdominal visceral fat and abdominal subcutaneous fat was observed in the obtained MRI images, and the results are shown in Fig.

As a result of measurement of abdominal fat using in vivo MRI, as shown in FIG. 8, it was found that Bifidobacterium Total abdominal fat and abdominal subcutaneous fat content decreased in the group that received Brevet CBT BR3 . These results suggest that Bifidobacterium Breve CBT BR3 This has the effect of decreasing body fat, particularly visceral fat reduction proves that it can help to suppress obesity.

Also, Bifidobacterium It was also confirmed that Brebeber CBT BR3 and the mixed strain (MIX) showed excellent effects of reducing body fat and abdominal fat.

As it viewed on the basis of the above results with the embodiments of the invention, the Bifidobacterium breve of the present invention Solarium CBT BR3 strain (KCTC 12201BP ) and the mixed strain (MIX) have excellent anti-obesity activity.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Accordingly, the true scope of protection of the present invention is not limited to the above-described embodiments, but should be interpreted within the scope of the following claims and equivalents thereof.

Korea Biotechnology Research Institute KCTC12201BP 20120507 Korea Biotechnology Research Institute KCTC10297BP 20020629 Korea Biotechnology Research Institute KCTC12202BP 20120507 Korea Biotechnology Research Institute KCTC10782BP 20050321 Korea Biotechnology Research Institute KCTC11906BP 20110428 Korea Biotechnology Research Institute KCTC11904BP 20110428

Claims (8)

Chapter fixability is excellent, IL-6, TNF-α and Bifidobacterium breve CBT BR3 (Bifidobacterium breve CBT BR3) to inhibit the secretion of IL-1β having anti-inflammatory activity (accession number: KCTC 12201BP) strains A drug for treating or preventing obesity, overweight or hyperlipemia.
The method according to claim 1, wherein the strain inhibits differentiation into adipocytes in adipose precursor cells, reduces body weight and abdominal fat, and reduces blood cholesterol, in order to treat or prevent obesity, overweight or hyperlipemia medicine.
Superior section carrier and a pharmaceutically acceptable fixability and, IL-6, TNF-α and IL-1β Bifidobacterium breve CBT BR3 (Bifidobacterium breve CBT BR3) having to suppress anti-inflammatory activity secretion ( Accession No .: KCTC 12201BP), which is used for treating or preventing obesity, overweight or hyperlipidemia.
The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition comprises 10 ⁸ to 10 ¹² cfu / g of the Bifidobacterium breve CBT BR3 (Accession No: KCTC 12201BP) strain relative to the total weight of the composition. A pharmaceutical composition for treating or preventing hyperlipidemia.
Chapter fixability is excellent, IL-6, TNF-α and Bifidobacterium breve CBT BR3 (Bifidobacterium breve CBT BR3) to inhibit the secretion of IL-1β having anti-inflammatory activity (accession number: KCTC 12201BP) strains &Lt; / RTI &gt;
6. The food composition according to claim 5, wherein the food composition comprises 10 ⁸ to 10 ¹² cfu / g of a live or dried bacterium of Bifidobacterium breve CBT BR3 (Accession No: KCTC 12201BP) An anti-obesity functional food composition.
The composition of claim 5, wherein the food composition is selected from the group consisting of Lactobacillus acidophilus CBT LA1 (KCTC 11906BP), Bifidobacterium animalis lactis CBT BL3 (KCTC 11904BP), Pediococcus pentosaceus CBT SL4 10297BP)) and lactobacillus plantarum CBT LP3 (KCTC 10782BP) and Lactobacillus lambusus CBT LR5 (KCTC 12202BP), which is characterized in that it further comprises at least one other prebiotic selected from the group consisting of: Wherein said composition is an anti-obesity functional food composition.
8. An anti-obesity functional food product according to claim 7, wherein the food composition comprises a lactic acid bacterium mixture in an amount of 10 ⁸ to 10 ¹² CFU / g, or an equivalent number of live bacteria, Composition.

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