KR101708173B1 - Novel lactic acid capable of controlling blood glucose level and use thereof - Google Patents

Abstract

The present invention provides a novel Lactobacillus sp. Strain isolated from kimchi or human feces. The specific strain of Bacillus sp. According to the present invention is isolated from kimchi or human feces and has high safety, excellent blood glucose control activity, anti-obesity activity, blood cholesterol lowering activity, blood neutral lipid lowering activity, arteriosclerosis inhibiting activity, And the like. Therefore, the specific Bacillus subtilis strain according to the present invention is useful as a medicament for preventing, ameliorating or treating metabolic syndrome including inflammation diseases such as diabetes, obesity, fatty liver, diabetes, hyperlipidemia, cardiovascular disease, hypertension, arteriosclerosis, It can be used as a material.

Description

Novel lactic acid capable of controlling blood glucose level and use thereof "

The present invention relates to a novel lactic acid bacterium, and more particularly to a novel lactic acid bacterium which is isolated from kimchi or a human feces and has a blood sugar controlling effect, an obesity inhibiting effect, a blood cholesterol lowering effect, a blood neutral lipid lowering effect, atherosclerosis inhibiting activity, And a novel lactic acid bacterium having various functionalities. The present invention also relates to various uses of novel lactic acid bacteria, and more particularly to the use of novel lactic acid bacteria for the prevention, improvement or treatment of diabetes, metabolic syndrome or lipid-related metabolic diseases.

As humanity develops into a prosperous society, the lifestyle is rapidly westernized, and the aspect of disease is also changing dramatically. In particular, abdominal obesity, hyperlipidemia, diabetes, hypertension, etc. are increasing in modern people. Such diseases are called lifestyle-related diseases in terms of diseases caused by lifestyle changes. Obesity, hyperlipidemia, diabetes, and hypertension among lifestyle-related diseases are known to be important risk factors for cerebrovascular disease, and metabolic syndrome is a case of simultaneous multiple cardiovascular risk factors. In other words, metabolic syndrome refers to a state of high insulin resistance, a high risk of diabetes and cardiovascular disease, a greater risk of developing cardiovascular disease in the presence of metabolic syndrome, and a 10-fold increase in the incidence of diabetes . In addition, the prevalence of various chronic diseases such as arthritis and cancer is increasing due to an increase in lifestyle-related diseases.

The increase in lifestyle diseases is largely due to westernized dietary habits and lack of exercise. Especially, changes in dietary habits lead to changes in human gut microbial counts, which causes endotoxins produced by gut microbial counts in the digestive tract. Increased endotoxin in the digestive tract induces digestive tract inflammation, increased uptake of endotoxin into the body, and promotes migration of macrophages to adipose tissue and the like, leading to obesity or hyperglycemia. Thus, controlling endotoxins produced by the gut microbial gasses can control blood sugar and ultimately improve or treat diabetes.

Diabetes mellitus is a typical chronic disease, and it is a chronic disease that causes microvascular complications such as retinas, kidneys and nerves, and macrovascular complications such as stroke, angina, myocardial infarction and peripheral vascular disease due to various metabolic disorders including glucose. Treatment of diabetes includes medication, exercise, and diet, and insulin medicines and various blood glucose control agents are used depending on the patient's symptoms. However, diabetes is a complex disease characterized by overgrowth of glucose in the liver, insulin resistance, and decreased glucose tolerance in muscle and adipocytes. Therefore, specific treatment alone can not prevent the occurrence of various side effects. Since the drug therapy uses insulin and chemicals, it has become a constant problem for side effects and tolerance of patients. Therefore, recently, diabetes treatment is possible and diabetes can be diarrhea. Prevention, improvement, or treatment.

In accordance with this trend, researches on the prevention and treatment of abdominal obesity, hyperlipidemia, diabetes, hypertension and the like are currently under way using lactic acid bacteria. For example, Korean Patent Registration No. 10-1061219 discloses a microorganism belonging to the genus Lactobacillus The present invention relates to a pharmaceutical composition for prevention and treatment of diabetes mellitus comprising a fermented extract of a porcine potato fermented by a fermentation method of the present invention as an active ingredient. Also, Korean Patent Registration No. 10-1010914 discloses a pharmaceutical composition comprising lactobacillus (hereinafter, referred to as " Lactobacillus ") for use in any one selected from the group consisting of body weight and fat reduction, plasma and liver lipid and carnitine improvement, blood leptin, Plantarum GLUS 42 (Accession No .: KCCM 10940P) is disclosed. In addition, Korean Patent Publication No. 10-1407980 discloses a method for reducing the insulin, ricin, glucose, C-peptide and triglyceride levels in blood. Lactobacillus < RTI ID = 0.0 > ( Lactobacillus < / RTI > Curvatus HY7601 (Accession No .: KCTC 11456BP) and Lactobacillus plantarum ) KY1032 (Accession No .: KCCM10430) as an active ingredient for the treatment of hyperinsulinemia, hyperglycemia and hypertriglyceridemia.

It is an object of the present invention to provide a novel lactic acid bacterium which is capable of inhibiting proliferation of intestinal microorganisms that secrete endotoxins or inhibiting endotoxin production of intestinal microorganisms and controlling blood glucose level .

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

The inventors of the present invention have found that, in order to develop an anti-diabetic material having higher safety than synthetic chemicals, the inventors of the present invention screen innumerable lactic acid bacteria from kimchi or human feces and inhibit proliferation of intestinal microorganisms in which specific Lactobacillus sp. Or endogenous endogenous microorganisms can be inhibited and blood glucose can be regulated. Thus, the present invention has been completed.

In order to achieve one object of the present invention, in still another aspect of the invention is Lactobacillus four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 to 16S rDNA sakei ), which has glucose-regulating activity. The Lactobacillus sakei is preferably a bacterium which is positive in Gram stain. The Lactobacillus sakei is preferably a bacterium having a carbon source such as Ribose, Galactose, Glucose, Fructose, Mannose, Mannitol, Sorbitol,? -Methyl-D mannoside, N-acetyl-glucosamine, Amygdalin, Arbutin, , Esculin, Salicin, Cellobiose, Maltose, Lactose, Melibiose, Sucrose, Trehalose, Melezitose, ), Gentiobiose (Gentiobiose) and Turanose. The Lactobacillus sakei preferably further has at least one activity selected from an anti-obesity activity, a blood cholesterol-lowering activity, a serum neutral lipid lowering activity, an arteriosclerosis-inhibiting activity or an inflammation-inhibiting activity in addition to a blood glucose controlling activity . In addition, the Lactobacillus sakei is preferably selected from the group consisting of Lactobacillus < RTI ID = 0.0 > sakei ) OK67 (accession number: KCCM 11670P). In addition, the Lactobacillus sakei preferably inhibits the proliferation of intestinal microorganisms that secrete endotoxins or inhibits endotoxin production of intestinal microorganisms, thereby lowering blood sugar.

In order to accomplish one object of the present invention, an example of the present invention relates to a method for producing Lactobacillus plantarum ) OK23, Lactobacillus paracasei OK9, and Lactobacillus ( Lactobacillus < RTI ID = 0.0 > ruminis ) OK17. The lactic acid bacterium is characterized in that it has a blood glucose controlling activity. The Lactobacillus < RTI ID = 0.0 > plantarum ) OK23, Lactobacillus paracasei OK9, and Lactobacillus ( Lactobacillus < RTI ID = 0.0 > rumen ) OK17 preferably inhibits the proliferation of intestinal microorganisms that secrete endotoxins, or inhibits endotoxin production of intestinal microorganisms, thereby lowering blood glucose.

In order to accomplish another object of the present invention, an example of the present invention is a method for producing Lactobacillus sakei comprising the nucleotide sequence of SEQ ID NO: 1 as 16S rDNA, a culture thereof, a lysate thereof or an extract thereof, By weight of the composition. Further, still another aspect of the invention is Lactobacillus four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 to 16S rDNA The present invention provides a pharmaceutical composition for preventing or treating metabolic syndrome including diabetes mellitus or diabetes mellitus comprising sakei , a culture thereof, a crushed product thereof or an extract thereof as an active ingredient. Further, still another aspect of the invention is Lactobacillus four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 to 16S rDNA The present invention provides a food composition for preventing or ameliorating a metabolic syndrome including diabetes or diabetes, which comprises a culture, a sakei thereof, a culture thereof, a crushed product thereof or an extract thereof as an active ingredient. At this time, the metabolic syndrome further includes at least one disease selected from obesity, fatty liver, hyperlipidemia, cardiovascular disease, hypertension or arteriosclerosis besides diabetes.

In order to accomplish another object of the present invention, an example of the present invention is a method for producing Lactobacillus plantarum ) OK23, Lactobacillus paracasei OK9, and Lactobacillus ( Lactobacillus < RTI ID = 0.0 > ruminis ) OK17, a culture thereof, a lysate thereof, or an extract thereof, as an active ingredient. The present invention also provides a composition for lowering blood glucose level, comprising at least one lactobacillus belonging to the genus Lactobacillus, In addition, one example of the present invention is a method for producing Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9 and Lactobacillus ruminis ) OK17, a culture thereof, a lysate thereof, or an extract thereof, as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating diabetes mellitus comprising at least one lactobacillus belonging to the genus Lactobacillus, In addition, one example of the present invention is a method for producing Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus the present invention provides a diabetic or improving food composition comprising as an active ingredient at least one lactobacillus of the genus Lactobacillus selected from the group consisting of Lactobacillus paracasei OK9 and Lactobacillus ruminis OK17, a culture thereof, a crushed product thereof or an extract thereof .

In order to achieve the object of the present invention, in still another aspect of the invention is Lactobacillus four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 to 16S rDNA sakei , a culture thereof, a crushed product thereof or an extract thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease. Further, still another aspect of the invention is Lactobacillus four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 to 16S rDNA sakei , a culture thereof, a disruption product thereof, or an extract thereof, as an active ingredient. At this time, the inflammatory disease is preferably selected from arthritis or colitis.

The specific strain of Bacillus sp. According to the present invention is isolated from kimchi or human feces and has high safety, and has excellent blood glucose control activity, anti-obesity activity, blood cholesterol lowering activity, blood neutral lipid lowering activity, arteriosclerosis inhibiting activity, And the like. Therefore, the specific Bacillus subtilis strain according to the present invention is useful as a medicament for preventing, ameliorating or treating metabolic syndrome including inflammation diseases such as diabetes, obesity, fatty liver, diabetes, hyperlipidemia, cardiovascular disease, hypertension, arteriosclerosis, It can be used as a material.

1 is a graph showing the effect of lactic acid bacteria isolated from cabbage kimchi, radish kimchi, pars kimchi, and human feces on the growth of E. coli.
Fig. 2 shows the results obtained by measuring the activity of lactic acid bacteria ( Lactobacillus < RTI ID = 0.0 > sakei) OK67, Lactobacillus Planta Room (Lactobacillus plantarum) OK23, Lactobacillus para Kasei (Lactobacillus paracasei ) OK9, Lactobacillus ( Lactobacillus ruminis ) OK17] on endotoxin production of intestinal flora.
FIG. 3 is a graph showing blood glucose levels of a model animal in which obesity is induced by a high fat diet; Lactobacillus < RTI ID = 0.0 > sakei ) OK67 by experimental group.
FIG. 4 is a graph comparing the plasma insulin levels of model animals induced by obesity with high-fat diets. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 by experimental group.
FIG. 5 is a graph showing the plasma intracellular toxin content of a model animal induced by obesity in a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei) and OK67 a graph showing the effect on each group, Figure 6 is Lactobacillus yarn K for the toxin content in the feces of obesity induced by high fat diet model animal (Lactobacillus sakei ) OK67 by experimental group.
FIG. 7 shows the results of oral glucose tolerance test of a model animal in which obesity was induced by a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei ) OK67, and FIG. 8 is a graph showing the results of the glucose load test of FIG. 7 as AUC (area under the glucose-time curve).
FIG. 9 is a graph showing changes in body weight of a model animal in which obesity was induced by a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei ) OK67. FIG. 10 is a graph showing the effect of OK67 on the weight gain of a model animal in which obesity was induced by high fat diet, and Lactobacillus sakei ) OK67 by experimental group.
FIG. 11 shows the change in the weight of epididymal adipose tissue of a model animal in which obesity was induced by a high fat diet, and Lactobacillus sakei ) OK67 by experimental group.
FIG. 12 shows the result of measurement of plasma total triglyceride content by experimental group, FIG. 13 shows the result of measurement of total cholesterol content by experimental group, FIG. 14 shows the result of measurement of HDL cholesterol content by experimental group, Hardening index measurement results.
As shown in FIG. 16, adipocyte differentiation factors and macrophage markers in the epididymal adipose tissues of experimental animals were significantly increased by high fat diet, but Lactobacillus sakei ) OK67, respectively.
LFD-67 "represents the low-fat dietary supplement and Lactobacillus sakei OK67-treated group with a dose of 1 x 10 ⁹ CFU / mouse, and" HFD "denotes a high-fat diet group benefit," HFD-OK67 "is high fat diet benefit and 1 × 10 Lactobacillus use of ⁹ CFU / mouse dose K (Lactobacillus sakei ) OK67 group.
FIG. 17 shows the results of a model animal experiment in which arthritis was induced by collagen. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strains on the severity of arthritis (arthritis severity).
FIG. 18 shows that in a model animal experiment in which arthritis was induced by collagen, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strains on the arthritis incidence.
In FIG. 17 and FIG. 18, "NOR" represents a group to which a vehicle is not induced by collagen but to which a vehicle is administered, "AC" represents a group to which arthritis is induced by collagen and a vehicle is administered, And arthritis is induced by the Lactobacillus casei ( Lactobacillus sakei ) OK67 strain.
FIG. 19 shows the results of a model animal experiment in which arthritis was induced by collagen. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strain on the increase in foot volume.
FIG. 20 shows the results of a model animal experiment in which arthritis was induced by collagen. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strain on the activity of myeloperoxidase (MPO) in the foot joint tissue.
FIG. 21 shows the results of a model animal experiment in which arthritis was induced by collagen. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strain on the microscopic appearance of paw joint tissue.
19 to 20, "represents collagen-induced arthritis, and" OK 67 " represents Lactobacillus sakei OK67 strain.

Hereinafter, terms used in the present invention will be described.

The term "culture product " used in the present invention means a product obtained by culturing a microorganism in a known liquid medium or solid medium, and includes a microorganism.

The terms " pharmaceutically acceptable "and" pharmaceutically acceptable "in the present invention are intended to mean not significantly irritating the organism and not interfering with the biological activity and properties of the administered active substance.

As used herein, the term "prophylactic " means any act that inhibits the symptoms of a particular disease or delays its progress by administration of the composition of the present invention.

The term "treatment" as used herein refers to any action that improves or alleviates the symptoms of a particular disease upon administration of the composition of the present invention.

The term "improvement" as used in the present invention means all actions that at least reduce the degree of symptom associated with the condition being treated.

The term "administering" as used herein is meant to provide any desired composition of the invention to an individual by any suitable method. The term " individual " means any animal such as a human, a monkey, a dog, a goat, a pig, or a mouse having a disease in which symptoms of a specific disease can be improved by administering the composition of the present invention.

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

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to a novel lactic acid bacterium having a blood glucose controlling activity (for example, hypoglycemic activity).

The novel lactic acid bacterium according to the present invention is 16S rDNA and is Lactobacillus sakei containing the nucleotide sequence as shown in SEQ ID NO: 1 and having a blood glucose controlling activity. The Lactobacillus sakei is preferably a bacterium which is positive in Gram stain. The Lactobacillus sakei is preferably a bacterium having a carbon source such as Ribose, Galactose, Glucose, Fructose, Mannose, Mannitol, Sorbitol,? -Methyl-D mannoside, N-acetyl-glucosamine, Amygdalin, Arbutin, , Esculin, Salicin, Cellobiose, Maltose, Lactose, Melibiose, Sucrose, Trehalose, Melezitose, ), Gentiobiose (Gentiobiose) and Turanose. The Lactobacillus sakei preferably further has at least one activity selected from an anti-obesity activity, a blood cholesterol-lowering activity, a serum neutral lipid lowering activity, an arteriosclerosis-inhibiting activity or an inflammation-inhibiting activity in addition to a blood glucose controlling activity . In addition, the above-mentioned Lactobacillus sakei ) is preferably selected from the group consisting of Lactobacillus < RTI ID = 0.0 > sakei ) OK67 (accession number: KCCM 11670P). In addition, the Lactobacillus sakei preferably inhibits the proliferation of intestinal microorganisms that secrete endotoxins or inhibits endotoxin production of intestinal microorganisms, thereby lowering blood sugar.

A novel lactic acid bacterium according to an example of the present invention is a lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9, and Lactobacillus ruminis OK17. The strain Lactobacillus sp. At this time, the Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9 and Lactobacillus ruminis OK17 have a blood glucose controlling activity, preferably a hypoglycemic activity. In addition, the above-mentioned Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9 and Lactobacillus ruminis OK17 preferably inhibit the proliferation of intestinal microorganisms that secrete endotoxins or inhibit endotoxin production of intestinal microorganisms and lower blood glucose levels.

The Lactobacillus sp. Strain of the present invention can be preferably isolated from kimchi or human feces. For example, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 was isolated from kimchi, and Lactobacillus ( Lactobacillus plantarum) OK23 will separate from Kimchi, Lactobacillus para Kasei (Lactobacillus paracasei ) OK9 and Lactobacillus OK17 is isolated from human feces.

Another aspect of the present invention relates to various uses of novel lactic acid bacteria. For example, the present invention Lactobacillus bacteria four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 by the above-mentioned 16S rDNA In one use of the novel lactic acid bacteria sakei , a culture thereof, a lysate thereof, or an extract thereof as an active ingredient. In addition, the present invention relates to a novel lactic acid bacterium for use as a Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9 and Lactobacillus ruminis ) OK17, a culture thereof, a lysate thereof, or an extract thereof, as an active ingredient. The present invention also provides a composition for lowering blood glucose level, comprising at least one lactobacillus belonging to the genus Lactobacillus, In addition, the present invention Lactobacillus bacteria four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 by the above-mentioned 16S rDNA In one use of the novel lactic acid bacteria The present invention provides a composition for preventing, ameliorating or treating metabolic syndrome including diabetes or diabetes, which comprises the sakei , a culture thereof, a crushed product thereof or an extract thereof as an active ingredient. The metabolic syndrome including diabetes further includes at least one disease selected from obesity, fatty liver, hyperlipidemia, cardiovascular disease, hypertension or arteriosclerosis. In the present invention, the metabolic syndrome refers to a disease in which various metabolic diseases such as diabetes and obesity occur simultaneously in one person, and narrowly refers to a lipid-related metabolic syndrome. In addition, the present invention Lactobacillus bacteria four K (Lactobacillus comprising the nucleotide sequence set forth in SEQ ID NO: 1 by the above-mentioned 16S rDNA In one use of the novel lactic acid bacteria The present invention also provides a composition for preventing, ameliorating or treating an inflammatory disease, which comprises a culture, a sakei thereof, a culture thereof, a lysate thereof or an extract thereof as an active ingredient. At this time, the inflammatory disease is not limited to the type of diseases caused by the inflammatory reaction, and is preferably selected from arthritis or colitis. In addition, the present invention relates to a novel lactic acid bacterium for use as a Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9 and Lactobacillus ruminis ) OK17, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient. The present invention also provides a composition for preventing, ameliorating or treating diabetes mellitus comprising at least one lactobacillus belonging to the genus Lactobacillus, The culture of lactic acid bacteria in the present invention is a product obtained by culturing a strain of Lactobacillus sp. In a culture medium. The culture medium may be selected from known liquid culture media or solid culture media, for example, MRS liquid medium, MRS agar medium, BL It may be an agar medium. Also, in the present invention, the composition may be formulated into pharmaceutical compositions, food additives, food compositions (particularly functional food compositions), or feed additives depending on the intended use or aspects thereof, and the specific Lactobacillus sp. Strain And the like can be adjusted in various ranges depending on the specific form of the composition, the purpose of use, and the manner of use.

In the pharmaceutical composition according to the present invention, the content of the novel lactic acid bacteria, the culture thereof, the lysate thereof or the extract thereof is not particularly limited and may be 0.01 to 99% by weight, 50% by weight, more preferably 1 to 30% by weight. In addition, the pharmaceutical composition according to the present invention may further contain, in addition to the active ingredient, an additive such as a pharmaceutically acceptable carrier, excipient or diluent. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition of the present invention may further comprise a known active ingredient having the effect of preventing or treating diabetes, obesity, fatty liver, hyperlipidemia, cardiovascular disease, hypertension, arteriosclerosis or metabolic syndrome in addition to the novel lactic acid bacteria, the culture thereof, May be further contained. The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration by a conventional method, and can be formulated into a pharmaceutical composition such as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention may be administered orally or parenterally to a mammal including a human according to a desired method. Examples of the parenteral administration method include external dermal application, intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, Intravenous injection or intra-thoracic injection. The dosage of the pharmaceutical composition of the present invention is not limited as long as it is a pharmacologically effective amount and is not limited as long as it depends on the body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, Varies. The typical daily dose of the pharmaceutical composition of the present invention is not particularly limited, but is preferably 0.1 to 3000 mg / kg, more preferably 1 to 2000 mg / kg, based on the active ingredient, once a day Or may be administered in divided doses.

The content of the novel lactic acid bacteria, the culture thereof, the lysate thereof or the extract thereof as an active ingredient in the food composition according to the present invention is preferably 0.01 to 50% by weight, more preferably 0.1 to 25% by weight, But is not limited thereto. The food composition of the present invention may be in the form of a pill, a powder, a granule, an infusion, a tablet, a capsule, or a liquid preparation. Examples of the food include meat, sausage, bread, chocolate, candy, snack, Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, functional water, drinks, alcoholic beverages and vitamin complexes. In addition to the active ingredient, the food composition of the present invention may contain a pharmaceutically acceptable carrier, various flavors or natural carbohydrates as an additional ingredient. In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Natural flavors such as tau Martin and stevia extract, and synthetic flavors such as saccharin and aspartame may be used as the flavor.

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

1. Isolation and Identification of Lactic Acid Bacteria

(1) Isolation of lactic acid bacteria from kimchi

Chinese cabbage kimchi, kimchi, and pork kimchi were each crushed and the crushed liquid was suspended in MRS broth (Difco, USA). The supernatant was then transferred to MRS agar medium (Difco, USA) and incubated at 37 ° C for 48 hours. Then, the colonies were isolated.

(2) Isolation of lactic acid bacteria from human feces

Human feces were suspended in GAM broth (Nissui Pharmaceutical, Japan). The supernatant was then transferred to a BL agar medium (Nissui Pharmaceutical, Japan), incubated at 37 ° C for 48 hours, and the colonies were isolated.

(3) Identification of selected lactic acid bacteria

The physiological characteristics and 16S rDNA sequence of strains isolated from kimchi or human feces were analyzed and the strains were identified and named. Table 1 below shows the control numbers and strain names of the lactic acid bacteria isolated from cabbage kimchi, radish kimchi, broccoli, and human feces. In Table 1, the lactic acid bacteria of Control Nos. 1, 3, 5, 7, 9, 11, 13, 17, 20, 22 and 25 were isolated from cabbage kimchi and control numbers 2, 4, 6, 8, , Lactic acid bacteria of 19 and 21 were isolated from kimchi, the lactic acid bacteria of control numbers 12, 15 and 18 were isolated from broccoli and the lactic acid bacteria of control numbers 24, 26, 27, 28, 29 and 30 were isolated from human feces It is separated.

Control Number Strain name Control Number Strain name One Leuconostoc mesenteroides OK1 16 Lactobacillus plantarum OK37 2 Leuconostoc mesenteroides OK15 17 Lactobacillus sakei OK26 3 Lactobacillus curvatus OK21 18 Lactobacillus sakei OK45 4 Lactobacillus curvatus OK25 19 Lactobacillus sakei OK67 5 Lactobacillus brevis OK11 20 Lactobacillus fermentum OK19 6 Lactobacillus brevis OK12 21 Lactobacillus fermented OK21 7 Lactobacillus acidophilus OK1 22 Lactobacillus gasseri OK1 8 Lactobacillus acidophilus OK7 23 Lactobacillus gasseri OK2 9 Lactobacillus lactis OK1 24 Lactobacillus johnsonii OK1 10 Lactobacillus lactis OK2 25 Lactobacillus johnsonii OK2 11 Lactobacillus helveticus OK1 26 Lactobacillus paracasei OK2 12 Lactobacillus helveticus OK2 27 Lactobacillus paracasei OK9 13 Lactobacillus plantarum OK23 28 Lactobacillus reuteri OK1 14 Lactobacillus plantarum OK32 29 Lactobacillus ruminis OK1 15 Lactobacillus plantarum OK36 30 Lactobacillus ruminis OK17

Among the strains described in Table 1, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 is an anaerobic bacterium that is positive in Gram stain. The carbon source availability of physiological characteristics is shown in Table 2 below. In Table 2, Lactobacillus < RTI ID = 0.0 > sakei ) The carbon source availability of OK67 was analyzed by sugar fermentation test using API Kit (API 50 CHL; manufacturer: BioMerieux's, USA). In the following table, "+" represents the case where the carbon source availability is positive, "-" represents the case where the carbon source availability is negative, "+" represents the case where the carbon source availability is ambiguous, and the blank represents the non-

Carbon source Strain name Lactobacillus sakei OK67 DSM20017 (ATCC15521) ^a Lactobacillus sakei K-7 ^b) glycerol - erythritol ± D-arabinose ± L-arabinose - + - D-ribose + + D-xylose - L-xylose - - adonitol - methyl-BD-xylopyranosicle - D-galactose + + - D-glucose + + D-fructose + + D-mannose + + L-sorbose - rhamnose - - dulcitol - inositol - mannitol + sorbitol + alpha -methyl-D-mannoside + alpha-methyl-D-glucoside - - N-acetyl-glucosamine + + amygdalin + arbutin + - esculin + + salicin + + cellobiose + - maltose + + ± lactose + - melibiose + + sucrose + + + trehalose + + inulin - melissose + raffinose - starch - glycogen - xylitol - ± gentiobiose + D-turanose + D-lyxose - D-tagatose - D-fucose - L-fucose - D-arabitol - L-arabitol - gluconate ± ± 2-keto-gluconate - 5-keto-gluconate -

^a) McLeod et al., Diversity of Lactobacillus strains investigated by phenotypic and genotypic methods. Systematic and applied microbiology 2008; 31: 393-403.

^b) Moon et al., Anticariogenic activities of Lactobacillus sakei K-7 isolated from kimchi. Korean society for biotechnology and bioengineering journal 2011; 26: 513-516.

In addition, Lactobacillus < RTI ID = 0.0 > sakei ) 16S rDNA was measured as a chemical taxonomic characteristic of OK67, and as a result, it was found to have the nucleotide sequence of SEQ ID NO: 1. Lactobacillus sakei ) The 16S rDNA sequence of OK67 was identified by BLAST search of Genebank (http://www.ncbi.nlm.nih.gov/). As a result, Lactobacillus ( Lactobacillus) strain having the same 16S rDNA base sequence sakei strain was not detected, and Lactobacillus sakei ) NBRC 15893 (NCBI ACCESSION: NR_113821).

2. Endotoxin ( endotoxin ) Selection of lactic acid bacteria inhibiting the production of Escherichia coli

(1) Experimental method

Escherichia coli pre-cultured in a TSB (tryptic soy broth) medium and lactic acid bacteria isolated from cabbage kimchi, radish kimchi, kimchi, and human feces were respectively added at a concentration of 1 × 10 ⁵ CFU and 1 × 10 ⁵ CFU TSB (tryptic soy broth) medium and incubated at 37 ° C for about 24 hours. Thereafter, a mixed culture medium of Escherichia coli and lactic acid bacteria was diluted 10 ⁶ times, 0.1 ml of the diluted culture was transplanted into DHL medium, and cultured at 37 캜 for about 24 hours. Thereafter, the number of E. coli in the final culture was measured. As a control, E. coli alone was transplanted into TSB (tryptic soy broth) medium in an amount of 1 × 10 ⁵ CFU instead of the mixed bacteria of E. coli and lactic acid bacteria, and the number of E. coli in the final culture was measured through the same procedure.

(2) Measurement results

1 is a graph showing the effect of lactic acid bacteria isolated from cabbage kimchi, radish kimchi, pars kimchi, and human feces on the growth of E. coli. As shown in Figure 1, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 exhibited the best ability to inhibit the growth of E. coli, followed by Lactobacillus plantarum ) OK23, Lactobacillus paracasei ( Lactobacillus paracasei ) OK9, Lactobacillus ( Lactobacillus ruminis ) OK17.

3. endotoxin of microbial gun of digestive tract ( endotoxin Selection of lactic acid bacteria to inhibit production

(1) Experimental method

Intestinal flora pre-cultured in GAM broth (GAM broth; Nissui Pharmaceutical, Japan) and lactic acid bacteria ( Lactobacillus ( Lactobacillus < (R) > sakei) OK67, Lactobacillus Planta Room (Lactobacillus plantarum) OK23, Lactobacillus para Kasei (Lactobacillus paracasei ) OK9, Lactobacillus ( Lactobacillus ruminis ) OK17] were transplanted into the anaerobic GAM broth (GAM broth; Nissui Pharmaceutical, Japan) at a concentration of 1 × 10 ⁸ CFU and 1 × 10 ⁵ CFU, respectively, and incubated at 37 ° C. for about 24 hours Lt; / RTI > Thereafter, the culture was treated with ultrasonic waves for about 1 hour to destroy the extracellular membrane of the microorganism, and centrifuged under the condition of 5000 x g to obtain a supernatant. The content of LPS (lipopolysaccharide), which is a typical endotoxin present in the supernatant, was measured by LAL (Limulus Amoebocyte Lysate) assay kit (manufactured by Cape Cod Inc., USA). In addition, as a control, the intestinal flora was replaced with GAM broth (GAM broth; Nissui Pharmaceutical, Japan) in an amount of 1 × 10 ⁸ CFU and cultured in the same manner as the mixed culture of intestinal flora and lactic acid bacteria. The content of LPS (lipopolysaccharide) present in the culture medium was measured.

(2) Measurement results

Fig. 2 shows the results obtained by measuring the activity of lactic acid bacteria ( Lactobacillus < RTI ID = 0.0 > sakei) OK67, Lactobacillus Planta Room (Lactobacillus plantarum) OK23, Lactobacillus para Kasei (Lactobacillus paracasei ) OK9, Lactobacillus ( Lactobacillus ruminis ) OK17] on endotoxin production of intestinal flora. In FIG. 2, the content of LPS (lipopolysaccharide) was shown as a relative multiple of the control. As shown in Fig. 2, the ability of the intestinal flora to inhibit the endotoxin production is also similar to the result of inhibiting the growth of Escherichia coli. Lactobacillus sakei ) OK67 was the best, followed by Lactobacillus ( Lactobacillus plantarum) OK23, Lactobacillus para Kasei (Lactobacillus paracasei) OK9, Lactobacillus Rumi Nice (Lactobacillus ruminis ) OK17.

4. Effect of Lactic Acid Bacteria on Blood Glucose Lowering Effect in - vivo  Experiment

(1) Experimental method

A total of 28 cows of 5-week-old male C57BL / 6J mice were divided into two groups and the low-fat diets (n = 14) were fed with low fat diets (product model: D12450B; (D12492; supplied by Research Diets, Inc., New Brunswick, NJ), 60% of the total calories were fed to a high fat diet (n = 14) Weekly. The LFD group was divided into two groups (LFD and LFD-67) in seven low-fat diet groups. LFD group was given low-fat diet and saline was administered orally for 4.5 weeks (31 days) When low-fat diets were administered, Lactobacillus ( Lactobacillus) sakei ) OK67 strain was orally administered at a dose of 1 × 10 ⁹ CFU for 4.5 weeks (31 days), and the experiment was terminated the next day. The HFD-67 group was divided into two groups (HFD, HFD-67) in the high fat diet group, followed by oral administration of saline to the HFD group for 4.5 weeks (31 days) At the same time as feeding, Lactobacillus sakei ) OK67 strain was orally administered at a dose of 1 × 10 ⁹ CFU for 4.5 weeks (31 days), and the experiment was terminated the next day. Lactobacillus The number of oral administration of OK67 strain was 6 days per week. During the experimental period, the mice were housed in a rabbit environment at a temperature of 20 ± 2 ° C, a humidity of 50 ± 5% and a light-dark cycle of 12 hours. Plasma glucose, plasma insulin, plasma endotoxin and fecal endotoxin levels were measured at the end of the experiment and oral glucose tolerance test was performed 5 days before the end of the experiment.

(2) Measurement of blood glucose, plasma insulin, plasma endotoxin and fecal endotoxin content

About 0.5 μl of blood was collected from the end of the mouse tail and blood glucose was measured using a kit for measuring glucose (manufactured by ASAN PHARM. CO. LTD., Korea). FIG. 3 is a graph showing blood glucose levels of a model animal in which obesity is induced by a high fat diet; Lactobacillus < RTI ID = 0.0 > sakei ) OK67 by experimental group. As shown in Figure 3, high fat diets supplemented with Lactobacillus saccharum sakei) glucose level in the group administered with OK67 strain was significantly decreased compared to the group fed high fat diet only.

Plasma insulin was measured using a mouse insulin ELISA kit (LINCO Research, St.Charles, MO). FIG. 4 is a graph comparing the plasma insulin levels of model animals induced by obesity with high-fat diets. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 by experimental group. As shown in FIG. 4, the high fat diet was supplemented with Lactobacillus saccharum sakei) plasma insulin content of the treated group, OK67 strain is significantly lower compared to the group fed high fat diet only.

FIG. 5 is a graph showing the plasma intracellular toxin content of a model animal induced by obesity in a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei) and OK67 a graph showing the effect on each group, Figure 6 is Lactobacillus yarn K for the toxin content in the feces of obesity induced by high fat diet model animal (Lactobacillus sakei ) OK67 by experimental group. As shown in FIG. 5 and FIG. 6, the high fat diet was supplemented with Lactobacillus < RTI ID = 0.0 > The intracellular toxin content and fecal endotoxin content of OK67 strain were significantly lower than those of high fat diet group.

(3) Oral glucose tolerance test (OGTT)

Five days before the end of the experiment, the animals were fasted for 6 hours, and glucose was orally administered at a dose of 2 g / kg body weight and blood was collected from the tail at 0, 15, 30, 60, 90 and 120 minutes Glucose concentration was measured. During the oral glucose load test, the animals were provided with a stable environment and water was allowed to ingest freely. The area under the glucose-time curve (AUC) was calculated using the following formula.

AUC = 0.5 占 0.5 C0 + C15 + C30 + C60 + C90 + 0.5 占 C120)

C0, C15, C30, C60, C90 and C120 are the glucose concentrations measured at 0 minute, 15 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes, respectively.

FIG. 7 shows the results of oral glucose tolerance test of a model animal in which obesity was induced by a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei ) OK67, and FIG. 8 is a graph showing the results of the glucose load test of FIG. 7 as AUC (area under the glucose-time curve). As shown in FIGS. 7 to 8, the high fat diet was supplemented with Lactobacillus < RTI ID = 0.0 > sakei ) AUC of the OK67 strain group was significantly lower than that of the high fat diet group.

5. Lactic acid bacteria Anti-obesity  Efficacy in - vivo  Experiment

(1) Experimental method

A total of 28 cows of 5-week-old male C57BL / 6J mice were divided into two groups and the low-fat diets (n = 14) were fed with low fat diets (product model: D12450B; (D12492; supplied by Research Diets, Inc., New Brunswick, NJ), 60% of the total calories were fed to a high fat diet (n = 14) Lt; / RTI > The LFD group was divided into two groups (LFD and LFD-67) in seven groups. Low-fat diets were administered to the LFD group and oral administration of saline was continued for 31 days. Low-fat diets were administered to the LFD-67 group At the same time, Lactobacillus sakei ) OK67 strain was orally administered at a dose of 1 × 10 ⁹ CFU for 31 days and the experiment was terminated the next day. The HFD group was divided into two groups (HFD and HFD-67) in seven groups of high fat diets, and the high fat diet was administered to the HFD group and the saline group was orally administered for 31 days. The HFD-67 group was fed a high fat diet At the same time, Lactobacillus sakei ) OK67 strain was orally administered at a dose of 1 × 10 ⁹ CFU for 31 days and the experiment was terminated the next day. Lactobacillus sakei ) The number of oral administration of OK67 strain was 6 days based on 7 days. During the experimental period, the mice were housed in a rabbit environment at a temperature of 20 ± 2 ° C, a humidity of 50 ± 5% and a light-dark cycle of 12 hours. After the experiment, the animals were sacrificed by cardiac puncture and epididymal fat (EF) was extracted and weighed.

(2) Weight change and weight change of adipose tissue

FIG. 9 is a graph showing changes in body weight of a model animal in which obesity was induced by a high fat diet. Lactobacillus < RTI ID = 0.0 > sakei ) OK67. FIG. 10 is a graph showing the effect of OK67 on the weight gain of a model animal in which obesity was induced by high fat diet, and Lactobacillus sakei ) OK67 by experimental group. The arrow in the downward direction in FIG. 9 represents the time point when oral administration of Lactobacillus sakei OK67 was administered. 11 is a graph showing the change in the weight of epididymal adipose tissue in a model animal in which obesity was induced by a high fat diet. The Lactobacillus case sakei ) OK67 by experimental group. As shown in Figs. 9 to 11, after the induction of obesity, the high fat diet was supplemented with Lactobacillus < RTI ID = 0.0 > In the group administered with OK67 strain, body weight was significantly decreased and weight of fat tissue was lower when compared to the group fed only high fat diet after induction of obesity.

(3) Measurement of plasma triglyceride, plasma total cholesterol and plasma HDL cholesterol content

Plasma triglyceride, plasma cholesterol and plasma HDL content of the experimental animals were measured as follows.

Plasma neutral lipids were measured using a kit for measuring neutral lipids (ASAN PHARM. CO. LTD., Korea). 1.5 ml of the enzyme solution and 10 쨉 l of plasma were added to the test tube, followed by stirring. The samples were then heated in a 37 ° C water bath for 10 minutes with standard solutions containing 0, 75, 150, 225 and 300 mg / dl, respectively, and the absorbance at 550 nm was measured. Respectively. Plasma total cholesterol was also measured using a kit for total cholesterol measurement (ASAN PHARM. CO. LTD., Korea). 1.5 ml of the enzyme solution and 10 쨉 l of plasma were added to the test tube, followed by stirring. Then, the samples were heated in a 37 ° C water bath for 10 minutes together with the standard solutions having total cholesterol contents of 0, 75, 150, 225, and 300 mg / dl, and the absorbance at 500 nm was measured, Respectively. Plasma HDL cholesterol was also measured using a kit for measuring HDL cholesterol (ASAN PHARM. CO. LTD., Korea). After adding 50 μl of needle-shaped reagent to 50 μl of plasma, the mixture was stirred, left at room temperature for 10 minutes, and centrifuged at 3000 rpm for 10 minutes to obtain 25 μl of the supernatant. Thereafter, 750 μl of the enzyme solution was added to the supernatant, followed by stirring. The samples were then heated for 5 minutes in a 37 ° C water bath with standard solutions containing HDL cholesterol levels of 0, 10, 20, 30, 40 and 50 mg / dl, respectively, and the absorbance at 500 nm was measured And colorimetrically determined.

In addition, the atherogenic index of plasma was calculated by the following equation.

Atherogenic index = (total cholesterol - HDL cholesterol) / HDL cholesterol

FIG. 12 shows the result of measurement of plasma total triglyceride content by experimental group, FIG. 13 shows the result of measurement of total cholesterol content by experimental group, FIG. 14 shows the result of measurement of HDL cholesterol content by experimental group, Hardening index measurement results. FIG Lactobacillus used together after the induction of obesity and the high fat diet benefit as seen in 12 to 15 K (Lactobacillus sakei ) OK67 was significantly decreased in plasma triglyceride, plasma total cholesterol and atherogenic index when compared to the group fed only with high fat diets after induction of obesity, and the HDL cholesterol content was increased Respectively.

(4) Analysis of lipid metabolism-related protein levels

High fat diets and Lactobacillus saccharides the sakei) for the administration of OK67 look at the impact on lipid metabolism-related protein level changes in experimental animals and epididymal adipose tissue of fat cell differentiation factor PPARγ, C / EBPα, FAS, level of macrophage markers in αFABP TNF-α , IL-1 [beta], F4 / 80, and CD68 levels were measured by Western blot analysis.

Specifically, the epididymal adipose tissue of the experimental animals was treated with RIPA buffer (50 mM Tris-HCl, pH 7.4, 1% NP-40, 0.25 mM) supplemented with protease inhibitor (Roche, USA), phosphatase inhibitor (Roche) and phenylmethanesulfonylfluoride Na-deoxycholate, 150 mM NaCl, 1 mM EDTA) and centrifuged at 14,000 rpm for 15 minutes to obtain supernatant. Proteins were separated from the supernatant by 10% SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). The separated protein samples were transferred to a PVDF membrane (Millipore, USA). After the sample was transferred, PVDF membrane was blocked with 5% skim milk (Difco, France) for 1 hour and 30 minutes in TBS-T buffer. The PPARγ (peroxisome proliferator-activated receptor-γ), C / EBPα (FAC), enhancer-binding protein-α, FAS (fatty acid synthase), αFABP, TNF-α, IL-1β, F4 / 80 and CD68, The reaction was allowed to proceed overnight. After washing with TBS-T buffer, goat anti-rabbit IgG (H + L) -HRP conjugate (BIORAD), a secondary antibody, was diluted at a ratio of 1: 5000 and reacted for 1 hour and 30 minutes . After washing with TBS-T buffer and reacting with ECL solution (Clarity western ECL substrate, BIORAD), proteins were detected by chemiluminescence (CLINX science instruments, USA). The density of each band was determined and the protein expression level of the other experimental groups was calculated on the basis of the amount of protein expression in the group fed only low fat diets. FIG. 16 is a graph showing changes in lipid metabolism-related protein levels in animal models of obesity induced by high-fat diet. Lactobacillus < RTI ID = 0.0 & sakei ) OK67 by experimental group. As shown in FIG. 16, adipocyte differentiation factors and macrophage markers in the epididymal adipose tissues of experimental animals were significantly increased by high fat diet, but Lactobacillus sakei ) OK67, respectively.

5. Effects of lactic acid bacteria on the anti-inflammatory effect in - vivo  Experiment

(1) Experimental method

To investigate the effect of Lactobacillus sakei OK67 strain on the arthritis-induced model animals, 21 male 8-week-old male DBA / 1J mice were divided into three groups (NOR, AC, AO) , Bovine type II collagen (bovine type Ⅱ collagen) was used as an immunogen to induce arthritis in the AC group and AO group except the normal NOR group. Specifically, 100 μg of collagen immunogen was injected intradermally into the proximal portion of the mouse tail, and primary immunization was performed. On the 21st day after primary immunization, the same amount of collagen immunogen was injected intradermally by the same method to secondary immunize. At this time, bovine type II collagen was dissolved in 0.05 M acetic acid as a collagen immunogen and emulsified with the same amount of Freund's complete adjuvant.

Secondary immunization From the next day, vehicle (50 mM sodium bicarbonate buffer containing 1% glucose) was orally administered daily for 20 days to NOR group and AC group, and AO group received Lactobacillus sakei ) 1 × 10 ⁹ CFU of OK67 strain was suspended in 0.1 ml of vehicle (50 mM sodium bicarbonate buffer containing 1% glucose) and orally administered daily for 20 days, and the experiment was terminated the next day.

(2) Analysis method

Arthritis severity and arthritis incidence of feet were measured from the stage of arthritis induction.

The arthritis severity of foot was visually observed around the foot and foot and assessed by a macroscopic score according to the criteria in Table 3 (Arii et al., 2008). The highest macroscopic appearance score that can be given to each mouse is 16.

step Appearance degree 0 normal One focal slight swelling and / or redness in one digit 2 moderate swelling and erythema 3 marked swelling and erythema of the limb 4 maximal swelling, erythema, deformity, and / or ankylosis

Myeloperoxidase (MPO) activity of the foot joint tissues, and microstructural microanalysis were performed after the end of the experiment.

At the end of the experiment, the animals were sacrificed and paw joint tissue was collected and immediately frozen at -70 ° C and used as a sample. The activity of myeloperoxidase (MPO) in the foot joint tissue was measured using the Mouse MPO assay ELISA kit (Hbt HK210, USA). In order to analyze the microstructure of the tissues, paw joint tissues were fixed with 4% paraformaldehyde, dried, embedded in paraffin, cut to a thickness of 20 쨉 m and then treated with hematoxylin- After staining with either hematoxylin-eosin, toluidine blue, or safranin O, the appearance of the tissue was microscopically evaluated.

(3) Experimental results

FIG. 17 shows the results of a model animal experiment in which arthritis was induced by collagen. Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strains on the severity of arthritis (arthritis severity). In addition, FIG. 18 shows that in a model animal experiment in which arthritis was induced by collagen, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strains on the arthritis incidence. In addition, FIG. 19 shows that in a model animal experiment in which arthritis was induced by collagen, Lactobacillus sakei ) OK67 strain on the increase in foot volume. In addition, FIG. 20 shows that in a model animal experiment in which arthritis was induced by collagen, Lactobacillus sakei ) OK67 strain on the activity of myeloperoxidase (MPO) in the foot joint tissue. Figure 21 also shows that in a model animal experiment in which arthritis was induced by collagen, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 strain on the microscopic appearance of paw joint tissue.

As shown in FIG. 17 to FIG. 21, Lactobacillus sakei OK67 strain showed high anti-inflammatory activity and showed excellent improvement or therapeutic effect on arthritis.

7. Preparation of pharmaceutical compositions containing lactic acid bacteria and the like

In the preparation of the following pharmaceutical compositions, Lactobacillus < RTI ID = 0.0 > sakei ) OK67 Cultures were cultured on Lactobacillus sakei ) OK67 It is possible to substitute the strain itself, its crushed product or its extract.

<7-1> Manufacture of powder

Lactobacillus sakei OK67 culture 20 mg

Lactose 100 mg

10 mg of talc

The above components were mixed and packed in airtight bags to prepare powders.

<7-2> Preparation of tablets

Lactobacillus sakei 10 mg of OK67 culture

Corn starch 100 mg

100 mg of milk

Magnesium stearate 2 mg

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

<7-3> Preparation of capsules

Lactobacillus sakei 10 mg of OK67 culture

3 mg of crystalline cellulose

15 mg of milk

Magnesium stearate 0.2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; 7-4 >

Lactobacillus sakei 10 mg of OK67 culture

Lactose 150 mg

100 mg of glycerin

Xylitol 50 mg

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<7-5> Preparation of granules

Lactobacillus sakei OK67 culture 15 mg

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

<7-6> Preparation of injection

Lactobacillus sakei 10 mg of OK67 culture

Sodium metabisulphite 3.0 mg

Methyl paraben 0.8 mg

0.1 mg of propylparaben

Sterile sterilized water for injection

After mixing the above ingredients, 2 ml of the mixture was filled in an ampoule and sterilized to prepare an injection.

8. Preparation of Food Composition Containing Lactic Acid Bacteria and the like

In the following food composition, Lactobacillus sakei OK67 culture can be replaced by Lactobacillus sakei OK67 strain itself, its lysate or its extract.

<8-1> Manufacture of Flour Food

To 100 parts by weight of wheat flour was added Lactobacillus sakei 0.5 part of OK67 culture was added to wheat flour and the mixture was used to prepare bread, cake, cookies, crackers and noodles.

<8-2> Manufacture of dairy products

100 parts by weight of milk was mixed with Lactobacillus sakei 0.5 part of OK67 culture was added to milk, and the milk was used to prepare various dairy products such as butter and ice cream.

<8-3> Manufacturing of Sunshine

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The cereals, seeds and Lactobacillus sakei OK67 cultures were prepared in the following proportions.

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

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

Lactobacillus sakei OK67 culture (1 part by weight),

(0.5 part by weight),

(0.5 parts by weight)

<8-4> Manufacture of health drinks

Lactobacillus (0.5 g), oligosaccharides (4 g), sugar (2 g), salt (0.5 g) and water (77 g) sakei 1 g of OK67 culture was homogeneously blended and sterilized in an instant, and packaged in glass bottles, plastic bottles, and plastic bottles.

<8-5> Manufacture of vegetable juice

A vegetable juice was prepared by adding 2 g of the fermentation product of Preparation Example 2 to 1,000 ml of tomato or carrot juice.

<8-6> Manufacture of fruit juice

Lactobacillus sakei One gram of the OK67 culture was added to 1,000 ml of apple or grape juice to prepare fruit juice.

9. Consignment information of lactic acid bacteria

The inventors of the present invention have found that Lactobacillus &lt; RTI ID = 0.0 &gt; sakei ) OK67 was deposited on February 23, 2015 with the Korea Microorganisms Conservation Center, a recognized depository institution, and granted the deposit number of KCCM 11670P.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Therefore, the scope of the present invention should be construed as including all embodiments falling within the scope of the appended claims.

Korea Microorganism Conservation Center (overseas) KCCM11670P 20150223

<110> University-Industry Cooperation Group of Kyung Hee University <120> Novel lactic acid capable of controlling blood glucose level and          use thereof <130> DP-15-186 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1376 <212> DNA <213> 16S rDNA of Lactobacillus sakei OK67 <400> 1 tgcagtcgaa cgcactctcg ttagattgaa gaagcttgct tctgattgat aacatttgag 60 tgagtggcgg acgggtgagt aacacgtggg taacctgccc taaagtgggg gataacattt 120 ggaaacagat gctaataccg cataaaacct agcaccgcat ggtgcaaggt tgaaagatgg 180 tttcggctat cactttagga tggacccgcg gtgcattagt tagttggtga ggtaaaggct 240 caccaagacc gtgatgcata gccgacctga gagggtaatc ggccacactg ggactgagac 300 acggcccaga ctcctacggg aggcagcagt agggaatctt ccacaatgga cgaaagtctg 360 atggagcaac gccgcgtgag tgaagaaggt tttcggatcg taaaactctg ttgttggaga 420 agaacgtatt tgatagtaac tgatcaggta gtgacggtat ccaaccagaa agccacggct 480 aactacgtgc cagcagccgc ggtaatacgt aggtggcaag cgttgtccgg atttattggg 540 cgtaaagcga gcgcaggcgg tttcttaagt ctgatgtgaa agccttcggc tcaaccgaag 600 aagtgcatcg gaaactggga aacttgagtg cagaagagga cagtggaact ccatgtgtag 660 cggtgaaatg cgtagatata tggaagaaca ccagtggcga aggcggctgt ctggtctgta 720 actgacgctg aggctcgaaa gcatgggtag caaacaggat tagataccct ggtagtccat 780 gccgtaaacg atgagtgcta ggtgttggag ggtttccgcc cttcagtgcc gcagctaacg 840 cattaagcac tccgcctggg gagtacgacc gcaaggttga aactcaaagg aattgacggg 900 ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 960 gtcttgacat cctttgacca ctctagagat agagctttcc cttcggggac aaagtgacag 1020 gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080 gcaaccctta ttactagttg ccagcattta gttgggcact ctagtgagac tgccggtgac 1140 aaaccggagg aaggtgggga cgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1200 acgtgctaca atggatggta caacgagtcg cgagaccgcg aggtttagct aatctcttaa 1260 aaccattctc agttcggatt gtaggctgca actcgcctac atgaagccgg aatcgctagt 1320 aatcgcggat cagcatgccg cggtgaatac gttcccgggc cttgtacaca ccgccc 1376

Claims (19)

Lactobacillus sakei OK67 (accession number: KCCM 11670P).
A composition for lowering blood glucose level comprising Lactobacillus sakei OK67 (Accession No .: KCCM 11670P) strain, a culture thereof, a lysate thereof or an extract thereof as an active ingredient.
( Lactobacillus sakei ) OK67 (accession number: KCCM 11670P), a culture thereof, a crushed product thereof or an extract thereof, as an active ingredient,
Pharmacy used for the prevention or treatment of metabolic syndrome in which at least two of at least two of diabetes, obesity, hypertriglycemia, hypercholesterolemia, hyperlipidemia, dyslipidemia, arteriosclerosis, fatty liver or the above diseases occur simultaneously Composition.
( Lactobacillus sakei ) OK67 (accession number: KCCM 11670P), a culture thereof, a crushed product thereof or an extract thereof, as an active ingredient,
A food used for the prevention or improvement of metabolic syndrome in which at least two of at least two of diabetes, obesity, hypertriglyceridemia, hypercholesterolemia, hyperlipidemia, dyslipidemia, arteriosclerosis, fatty liver or the above diseases occur simultaneously Composition.
Lactobacillus sakei OK67 (Accession No .: KCCM 11670P) A pharmaceutical composition for preventing or treating an inflammatory disease comprising a strain, a culture thereof, a crushed product thereof or an extract thereof as an active ingredient.
The pharmaceutical composition according to claim 5, wherein the inflammatory disease is arthritis or colitis.
Lactobacillus sakei OK67 (Accession No .: KCCM 11670P) A food composition for preventing or improving an inflammatory disease comprising a strain, a culture thereof, a crushed product thereof or an extract thereof as an active ingredient.
8. The food composition according to claim 7, wherein the inflammatory disease is arthritis or colitis. delete delete delete delete delete delete delete delete delete delete delete

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