KR20180118363A - Lactobacillus plantarum having anti-inflammation and metabolic disease improvement effect and uses thereof - Google Patents

Abstract

The present invention relates to a Lactobacillus plantarum strain having anti-inflammatory effects and metabolic disease-improving effects, and a use thereof. More specifically, the present invention relates to a Lactobacillus plantarum strain having anti-inflammatory effects and metabolic disease-improving effects, a probiotic preparation containing the same, a health functional food composition, and a pharmaceutical composition. The Lactobacillus plantarum HAC07 strain of the present invention is excellent in acid resistance and bile resistance, and has no resistance to various kinds of antibiotics and no production ability for bioamine. In addition, the strain also promotes production of short-chain fatty acid in the intestines, and thus is used as functional starters for producing various lactic acid fermented products, functional food additives, functional foods capable of alleviating chronic inflammation, and probiotics preparations. In addition, the strain is expected to be industrially useful by being used as health functional foods or pharmaceutical compositions for anti-inflammation and prevention and amelioration of metabolic diseases.

Description

Lactobacillus plantarum strains having an anti-inflammatory and metabolic disease improving effect and their use {

The present invention relates to a Lactobacillus plantarum strain HAC07 having an anti-inflammatory and metabolic disease improving effect, and a method for producing the Lactobacillus plantarum strain HAC07. A health functional food composition, and a pharmaceutical composition.

Lactobacillus has been associated with humans and its history, and its usefulness as a microorganism that has a very beneficial effect on human health is increasing day by day. In recent years, studies on lactic acid bacteria have been actively carried out, and the application range thereof has been expanded not only as general foods but also as health foods and drugs. These lactic acid bacteria play an important role in the maintenance of intestinal health by decomposing nutrients and fibrin which are consumed by animals in the intestines of animals and using them as an energy source and producing lactic acid and antibiotics to suppress the development of harmful bacteria in the intestines. Lactic acid bacteria have also been used to promote animal growth and feed utilization, to increase resistance to diseases, to inhibit the growth of harmful bacteria, to reduce mortality, to inhibit the production of toxic substances, and to produce various vitamins. However, in order to exert the above-mentioned effect, the lactic acid bacteria which flows from the outside must reach the intestines without any obstruction and exhibit its function. In order to do this, oral administration should be less destructive to gastric acid and should be resistant to bile acids.

Although obesity is a social issue in appearance, it is the most serious problem of obesity that it can cause serious health risks such as complications of metabolic diseases such as diabetes and hypertension. Symptoms related to the pathological condition of obesity are systemic chronic inflammation in obese individuals. Inflammation is one of the immune mechanisms that occur in the body, and when it occurs locally, it is an important reaction to defend the body against invasion of germs or viruses from the outside. However, such an inflammatory reaction causes systemic and chronic hyperactivity due to the balance collapse of the body 's immune response, which leads to a disorder in the metabolism in the body.

Chronic inflammation caused by obesity is known to be a cause of various metabolic diseases such as diabetes, cardiovascular disease and arteriosclerosis, and it is also the most important factor that defines obesity as a disease. Obesity is simply a cosmetic problem without the onset of secondary metabolic diseases caused by chronic inflammatory reactions. Recently, the World Health Organization has been criticized for having a chronic inflammatory reaction that can lead to secondary metabolic diseases such as diabetes, .

Korean Patent Laid-Open Publication No. 2011-0000871 discloses Lactobacillus plantarum L. wortii 7711 having antioxidant and anti-inflammatory properties and a product containing the same as an active ingredient. However, the anti-inflammatory and metabolic disease improvement Lactobacillus plantarum strains that have all of their efficacy have not been disclosed.

The present invention has been made in view of the above-mentioned needs. The present inventors have succeeded in isolating Lactobacillus plantarum HAC07 strains which are resistant to antibiotics and which have strong acid-fast bacilli, The present inventors have completed the present invention by confirming that when the strain is inoculated into the cecum of rats, the production of short chain fatty acids is remarkably higher than that of the control group.

The present invention provides a Lactobacillus plantarum strain HAC07 (accession number: KCTC13243BP) which has the effect of preventing or ameliorating anti-inflammatory and metabolic diseases.

The present invention also provides a probiotic preparation comprising at least one selected from the group consisting of the strain, the culture of the strain, the concentrate of the culture and the dried product as an active ingredient.

The present invention also provides a food and beverage comprising at least one selected from the group consisting of the strain, the culture of the strain, the concentrate of the culture and the dried product as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating an anti-inflammatory and metabolic disease comprising at least one member selected from the group consisting of the strain, the culture of the strain, the concentrate of the culture and the dried product do.

The present invention also provides a pharmaceutical composition for the prophylaxis or treatment of anti-inflammatory and metabolic diseases comprising, as an active ingredient, at least one selected from the group consisting of the strain, the culture of the strain, the concentrate of the culture and the dried product .

The Lactobacillus plantarum HAC07 strain of the present invention is excellent in acid resistance and biliary excretion, has no resistance to various kinds of antibiotics, has no bioamine production ability, and has an effect of inhibiting the expression of various inflammation-inducing cytokines , A probiotic preparation, a functional food capable of alleviating chronic inflammation, a functional food additive, and the like, and is expected to be used as a fermented food functional starter for producing various fermented milk products and fermented products. In addition, the Lactobacillus plantarum HAC07 strain of the present invention promotes the production of short-chain fatty acids in the intestines, thereby alleviating the levels of glucose, cholesterol and triglycerides in the blood, thereby providing a health functional food for preventing or improving anti- inflammatory and metabolic diseases, It is expected to be useful industrially.

Fig. 1 shows the result of analysis of the amount of short chain fatty acids after inoculation of lactic acid bacteria in the mouse cecum. (Control: HAC02: Lactobacillus plantraum HAC02 inoculation, HAC03: Lactobacillus plantraum HAC03 inoculation, HAC07: Lactobacillus plantraum HAC07, Lactobacillus plantraum HAC02 inoculation, Inoculation, 299v (comparative): Lactobacillus plantraum 299v inoculation). Results were expressed as mean and standard deviation, and compared with control (control) and ANOVA, p <0.05 when *, p <0.01 and ** when p <0.001.

In order to achieve the object of the present invention, the present invention provides Lactobacillus plantarum HAC07 strain having an anti-inflammatory and metabolic disease preventing or improving effect.

The present inventors isolated lactic acid bacteria Lactobacillus plantarum from the white kimchi and analyzed the acid resistance, bile resistance, antibiotic resistance and biogenic amine production ability of the isolated lactic acid bacteria. As a result, the isolated lactic acid bacteria showed excellent acid resistance and biliary properties , It was confirmed that there was no antibiotic resistance and no bioamine production ability. The isolated strain was named Lactobacillus plantarum HAC07 and deposited on Apr. 10, 2017 (KCTC 13243BP) at Korea Biotechnology Research Institute.

In the strain according to an embodiment of the present invention, the metabolic disease may be, but not limited to, obesity, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, arteriosclerosis, fatty liver, cardiovascular disease and the like.

The strain according to an embodiment of the present invention is not limited to the above-mentioned antibiotics, but is resistant to erythromycin, gentamicin, ampicillin, tetracycline, chloramphenicol, streptomycin, ciprofloxacin or penicillin.

The strain according to an embodiment of the present invention is characterized in that it does not produce a biogenic amine such as tyramine, histamine, putrescine or cadaverine from a tyrosine, histidine, ornithine or lysine amino acid precursor. However, .

The strain according to an embodiment of the present invention is characterized by having the effect of preventing or improving anti-inflammatory and metabolic diseases by producing short chain fatty acids. The short chain fatty acid may be acetic acid, propionic acid or butric acid, but is not limited thereto.

The present invention also provides a probiotic preparation comprising as an active ingredient at least one selected from the group consisting of the Lactobacillus plantarum HAC07 strain, the culture of the strain, the concentrate of the culture, and the dried product.

The Lactobacillus plantarum HAC07 strain of the present invention has excellent acid resistance and biliary cholesterol, has no bio-amine producing ability capable of inducing an irritable immune response, and has no erythromycin, gentamicin, ampicillin, tetracycline, chloramphenicol, streptomycin , Ciprofloxacin, penicillin, and the like, and thus can be usefully used as probiotics.

The probiotic agent can be prepared and administered in various formulations and methods according to methods known in the art. For example, the Lactobacillus plantarum HAC07 strain of the present invention, a culture thereof, a concentrate of the culture broth or a dried product thereof may be mixed with a carrier commonly used in the pharmaceutical field to prepare powders, liquids and solutions, May be formulated and administered in the form of tablets, capsules, syrups, suspensions or granules, and the like. Examples of the carrier include, but are not limited to, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, coloring matters and fragrances. The administration dose can be appropriately selected depending on the degree of absorption of the active ingredient in the body, the inactivation rate, the excretion rate, the age, sex, strain, condition and severity of the disease.

Further, the present invention relates to a pharmaceutical composition for preventing or ameliorating an anti-inflammatory and metabolic disease comprising at least one member selected from the group consisting of the lactobacillus plantarum HAC07 strain, the culture of the strain, the concentrate of the culture and the dried product, Functional food composition.

The health functional food composition of the present invention may be prepared in any one form selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings. The health functional food composition according to the present invention may be formulated into a form such as a powder, a liquid, a tablet, a soft capsule, a granule, a tea bag, an instant tea or a drink by incorporating the Lactobacillus plantarum HAC07 strain or the like as an active ingredient . The content of the Lactobacillus plantarum HAC07 strain as an active ingredient can be appropriately determined depending on the intended use (for prevention or improvement). Generally, the amount of the Lactobacillus plantarum HAC07 strain contained in the food composition may be 0.1 to 90% by weight based on the total weight of the food. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range. Further, the food composition according to the present invention may contain, in addition to the Lactobacillus plantarum HAC07 strain, other components that can give rise to a synergistic effect on the main effect, within a range not impairing the intended main effect of the present invention It is acceptable.

The food composition formulated in this form can be added directly to the food or used together with other food or food ingredients, and can be suitably used according to conventional methods. Examples of food products include dairy products including drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, , Dairy products and dairy products, and all health functional foods in the ordinary sense are included.

When the food composition of the present invention is a beverage, Lactobacillus plantarum HAC07 strain is contained as an essential ingredient in the indicated ratio, and there are no particular restrictions on other ingredients used for the purpose of producing other beverages. Flavoring agents, natural carbohydrates and the like as additional components. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors and synthetic flavors and the like can be used as the flavors other than those described above. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), synthetic flavors and flavors such as natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the inventive food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not so important, it is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the Lactobacillus plantarum HAC07 strain of the present invention.

The present invention also relates to a pharmaceutical composition for preventing or treating anti-inflammatory and metabolic diseases, which contains at least one selected from the group consisting of the lactobacillus plantarum HAC07 strain, the culture of the strain, the concentrate of the culture, Gt;

The amount of the pharmaceutical composition for preventing or treating anti-inflammatory and metabolic diseases used in the present invention should be a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and the effective dose level will vary depending on the species and severity, The type of virus being infected, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and rate of release, the duration of the treatment, factors including co-administered drugs and other well known factors in the medical field. Effective amounts may vary depending on the route of treatment, the use of excipients, and the likelihood of use with other agents, as will be appreciated by those skilled in the art.

The pharmaceutical compositions for the prevention or treatment of anti-inflammatory and metabolic diseases of the present invention may be formulated into pharmaceutical formulations using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal . In the preparation of the formulations, it is preferred that the active ingredient is mixed with or diluted with the carrier, or enclosed in a carrier in the form of a container.

Accordingly, the pharmaceutical composition for the prevention or treatment of anti-inflammatory and metabolic diseases of the present invention may be formulated into oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations and patches And may further comprise suitable carriers, excipients or diluents conventionally used in the preparation of the compositions.

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 Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example  1. Isolation and Identification of Strain from White Kimchi

After fermentation, fermented white kimchi was purchased for 10 ~ 15 days and pH was measured and lactic acid bacteria were isolated.

90 mL of physiological saline (0.85% NaCl / L) was added to a high-pressure sterilized vinyl bag, 10 g of white kimchi sample was added, and the mixture was uniformly mixed at 200 rpm for 5 minutes. (1.5% Lactobacillus MRS broth, BD Difco, USA and 1.5% Bacteriological Agar, Affymetrix, USA) by stairwise dilution in 9 ml of physiological saline (0.85% NaCl / L) . It was then grown in a 37 ° C incubator for 24 to 48 hours, and then isolated and identified.

First, colony morphology was compared by microscopic observation to estimate the type of strains on the plate. Lactobacillus brevis , a rod-like microorganism with rounded cell shape, The rod shape of medium length was estimated to be Lactobacillus plantarum and the rod shape of the rod was estimated to be Lactobacillus sakei .

 Next, catalase activity and Gram stain analysis were performed while culturing the next grown bacteria at 37 ° C for 24 to 48 hours using 0.3% hydrogen peroxide. The catalase negative strains identified taxonomic species by analyzing the bi - directional 16S rDNA gene sequence. The nucleotide sequence of the strain was identified by SolGent (Korea).

The strain of the present invention isolated from white kimchi was a catalase-negative, gram-positive, rod-like strain identified as Lactobacillus plantarum and designated as Lactobacillus plantarum HAC07.

The sugar-using characteristics of the Lactobacillus plantarum HAC07 strain isolated in the present invention were tested using the API 50 CH kit (Biomerieux) according to the method of the corresponding manual. The sugar-using characteristics of each strain were as shown in Table 1 below.

Per type Availability Per type Availability CONTROL - Esculin ferric citrate ++ Glycerol + Salicin ++ Erythritol - D-celobiose ++ D-arabinose - D-maltose ++ L-arabinose - D-lactose ++ D-rivose ++ D-melibiose ++ d-xylose - D-saccharose (sucrose) ++ 1-xylose - D-trehalose ++ d-adonitol - Inulin - methyl-β-D-xylopyranoside - D-melezitose ++ d-galactose ++ d-raffinose + d-glucose ++ Amdon (starch) - D-fructose ++ Glycogen - D-manose ++ Xylitol - L-sorbose - Gentiobiose ++ L-rhamnose - D-turanose - dulcitol - D-lyxose - inositol - D-tagatose - D-mannitol ++ D-fucose - D-sorbitol ++ L-fucose - Methyl-aD-Mannopyranoside - D-arabitol + Methyl-aD-glucopyranoside - L-arabitol - N-acetylglucosamine ++ Potassium GlucoNaTe - Amygdalin ++ Potassium 2-KetoGluconate - Arbutin ++ Potassium 5-KetoGluconate -

Example  2. Lactobacillus Flora Room HAC07  The organism of the strain Amine  Generate Whether

Biogenic amines are produced by the fermentation of foodstuffs and may vary depending on the type of microorganism or chemical and physical conditions. Biomolecules produced in fermented foods are an important criterion for choosing a food-engineered strain because it can cause food poisoning or allergic reactions.

In order to confirm whether or not the strain of the present invention was formed with a biomolecule, the MRS liquid medium was transferred to a special medium according to Bover-Cid and Holzapfel (1999) for 16 hours at 37 ° C. and incubated at 37 ° C. for 48 hours Respectively.

MRS liquid medium supplemented with amino acid precursors of each of tyrosine, histidine, ornithine and lysine was prepared and the biomolecules (tyramine, histamine (histamine), putrescine and cadaverine) were produced. Specifically, 1% of the isolated Lactobacillus plantarum strain was inoculated into the MRS liquid medium supplemented with 0.1% of the amino acid precursor, and then cultured for 5-10 times to activate the decarboxylase. The concentration of dextrinase enzyme medium [tryptone 0.5%, yeast extract 0.5%, cocoon extract 0.5%, sodium chloride 0.5%, glucose 0.25%, tween-80 0.05%, magnesium sulfate 0.02%, manganese sulfate 0.005% 0.2% of salt, 0.001% of thiamine, 0.2% of K2PO4, 0.01% of calcium carbonate, 0.005% of pyridoxal-5-phosphate, 1% of amino acid, 0.006% of bromocresol purple, 2% in distilled water, pH adjusted to 5.3], and incubated at 37 ° C for 24 to 48 hours to determine the color change to purple.

The bromocresol purple in the decarbonated enzyme medium is yellow at pH 5.2 but turns purple as the pH rises to 6.8. Therefore, the production of bio-amines can be confirmed by using a substance that turns purple when the pH rises due to the formation of bio-amines.

As a result of analyzing biosurfactant activity of the strain Lactobacillus plantarum HAC07 of the present invention, tyrosine, histidine, ornithine and lysine were all found to be negative as shown in Table 2 below. From these results, it was confirmed that the strain of the present invention has no bio-amine generating ability capable of inducing an irritable immune response.

From the amino acid precursor, Amine  Whether to generate Strain name Tyrosine Histidine Ornithine Lysine L. plantarum HAC07 - - - -

Example  3. Lactobacillus Flora Room HAC07  Antibiotic resistance of the strain

The above isolates for erythromycin, gentamycin, ampicillin, tetracycline, chloramphenicol, streptomycin, ciprofloxacin, and penicillin G, The antibiotic resistance of the strain was confirmed.

Specifically, each antibiotic was stair-diluted (1: 2) and mixed in MRS solid medium at a concentration of 0.025 to 64 / / ml. The minimum inhibitory concentration (MIC) was measured by inoculating Lactobacillus plantarum strain cultured for 18 hours into a solid medium containing the above antibiotic at a concentration of 1 x 10 &lt; ⁵ &gt; CFU, and culturing at 37 DEG C for 24 hours.

As a result, as shown in Table 3, 8 antibiotics such as erythromycin, gentamycin, ampicillin, tetracycline, chloramphenicol, streptomycin, ciprofloxacin and penicillin were confirmed to be resistant against the minimum inhibitory concentration Respectively.

Strain Minimum inhibitory concentration (/ / mL) Em Gm Am Te Ch Sm Ci Pe L. plantarum HAC07 <0.25 32 <0.25 16 4 8 32 <0.25 Reference value 4 128 4 64 16 > 256 > 32 4 * Em: erythromycin; Gm: Gentamicin; Am: Ampicillin; Te: tetracycline; Ch: chloramphenicol; Sm: streptomycin; Ci: ciprofloxacin; Pe: Penicillin G
* Reference value: Lactobacillus flutarium antibiotic resistance determination standard value according to Danielsen and Wind (2003)

Example  4. Lactobacillus Flora Room HAC07  Strain Acid resistance  And My bile  analysis

When lactobacillus is ingested with food, it stays for about one hour on the secretion of strong gastric acid. After staying in high-bile bile of the duodenum for about 2 hours, it moves to the intestines. Lactobacillus can survive in this extreme environment and enter the intestines so that it can stick to the intestines and provide the host with good nutrition and serve as probiotics. Therefore, the SSDP (simulated stomach duodenum passage) method, which is similar to the human (gastroduodenal and duodenal) environment, was performed to confirm the acid resistance and biliary bite property of the lactic acid bacteria isolated in the present invention.

Specifically, the isolated Lactobacillus plantarum strain was cultured in MRS liquid medium for 16 hours at 37 ° C, and 1 ml of the strain was placed in a sterilized disposable tube and centrifuged at 12,000 xg for 5 minutes at 4 ° C Afterwards, the supernatant was discarded and the cell pellet was washed twice with physiological saline. Then, physiological saline was removed, and the precipitated bacteria were mixed with 10 ml of MRS liquid medium adjusted to pH 3, and then mixed well. 1 ml of the suspension was diluted stepwise to 10-fold, plated on MRS solid medium, and the number of initial cells (CFU / ml) was counted. The remaining 9 ml was incubated at 37 ° C for one hour, and then 4 ml of bile acid (10 g oxgall was mixed with 100 ml of distilled water and autoclaved) and 17 ml duodenum juice (6.4 g / L NaHCO ₃ , 0.239 g / L KCl and 1.28 g / L NaCl were mixed well in distilled water, pH adjusted to 7.4, and autoclaved). The cells were cultured in a 37 ° C incubator for 2 hours or more, diluted 10-fold in a stepwise manner, and plated on a MRS solid medium to count the number of surviving bacteria. (CFU / ml) and the number of viable cells remaining (CFU / ml) after 1 hour and 3 hours of culture were compared to the survival rates (Table 4).

The Lactobacillus plantarum HAC07 strain of the present invention has a survival rate of about 19%, which is superior to the comparative strain in acid resistance and biliary properties. Biliary properties.

Initial cell number
(log CFU / mL) After 1 hour at pH 3.0
(log CFU / mL) After more than 2 hours at 3.25% OXGAL
(log CFU / mL) Total survival (%) L. plantarum HAC03 9.11 + 0.047 8.81 + 0.061 7.55 + 0.089 2.76 + 0.135 L. rhamnosus GG 9.12 ± 0.311 8.54 0.281 6.82 + 0.064 0.54 + - 0.293

Example  5. Lactobacillus Flora Room  Effect of short-chain fatty acid production of strain

Intestinal lactic acid bacteria interact with the human body as a host by producing metabolites using intestinal materials. Intestinal bacteria take up and decompose carbohydrates to produce short chain fatty acids (SCFAs), and the produced short chain fatty acids have several beneficial functions in the intestines. Short chain fatty acids, including acetate, propionate and butyrate, increase the motility of the intestines and interfere with the establishment of pathogens. They act as an energy source for intestinal epithelial cells, And prevent fat accumulation, which can help prevent obesity. In addition, when short-chain fatty acids are absorbed in the intestine, they lower the cholesterol synthesis, lowering the blood cholesterol concentration and inhibiting glucose uptake, thus helping prevent and treat metabolic diseases such as hyperlipidemia and diabetes.

The intestinal productivity of strains can be investigated relatively simply by measuring the amount of short chain fatty acid changes by co-culturing mouse cecum sample with each lactic acid bacteria strain. In the present invention, an animal experiment was conducted to identify short-chain fatty acids produced in the intestines of the isolated Lactobacillus plantarum strain.

Specifically, 7-week-old male C57BL / 6J mice (specific pathogen-free) were purchased and grown in an environment of 23 ° C, 55 ± 10% humidity and 12 hours / cancer cycle. The mouse cecum was separated and inoculated with the Lactobacillus plantarum HAC07 strain or the comparative strain ( Lactobacillus plantraum 299v) of the present invention, and then cultured in an anaerobic chamber for 6 hours. After fermentation in an anaerobic environment, the amount of short chain fatty acids in each sample was analyzed by gas chromatography (Shimadzu GC2010). As a control, the same amount of sterilized PBS was inoculated instead of the strain, and the results were compared. By comparing the amount of short chain fatty acids in the culture broth, it was possible to confirm the short chain fatty acid production by the probiotics.

Gas chromatography was performed using a Shimadzu GC2010 instrument. The standard curve and peak retention time were analyzed using a volatile fatty acid mixture (ultrapure) from Sigma (Supelco). Short chain fatty acids were extracted from the feces and compared with the standard curve .

Short chain fatty acid extraction is described by Schwiertz et. Al., (2009) were used. Briefly, an extract containing oxalic acid (0.1 mole / L) and sodium azide (40 mmole / L) was added to a sample (up to 80 mg) and incubated for 1 hour at room temperature with shaking incubation ) And centrifuged at 16,000 xg for 5 minutes at 24 DEG C, and the resulting supernatant was analyzed using an FID instrument. The column was analyzed using a HP INNO-WAS 30 m x 32 mm with a splitter temperature of 260 ° C and an FID of 260 ° C, and a column of 27.1 psi at 100 ° C to 180 ° C at a rate of 25 ° C / m.

As a result, as shown in FIG. 1, the lactobacillus plantarum HAC07 of the present invention significantly increased the production of propionic acid (FIG. 1B) and the total production of short chain fatty acids (FIG. 1D) Was significantly higher than that of Lactobacillus plantarum 299v as well as the control group.

Claims (10)

Lactobacillus plantarum HAC07 strain (accession number: KCTC13243BP) which has an effect of preventing or improving anti-inflammatory and metabolic diseases.
The method according to claim 1,
The metabolic disease is at least one selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis, fatty liver and cardiovascular disease.
The method according to claim 1,
Wherein the strain is resistant to at least one antibiotic selected from the group consisting of erythromycin, gentamicin, ampicillin, tetracycline, chloramphenicol, streptomycin, ciprofloxacin and penicillin.
The method according to claim 1,
Wherein said strain does not produce a biological amine from one or more amino acid precursors selected from the group consisting of tyrosine, histidine, ornithine, and lysine.
The method according to claim 1,
Wherein said strain has the effect of preventing or improving anti-inflammatory and metabolic diseases by generating short chain fatty acids. &Lt; RTI ID = 0.0 &gt;&lt; / RTI &gt;
A probiotic formulation comprising at least one selected from the group consisting of a strain of any one of claims 1 to 5, a culture of the strain, a concentrate of the culture, and a dried product as an active ingredient.
A method for preventing or ameliorating an anti-inflammatory and metabolic disease comprising at least one selected from the group consisting of a strain of any one of claims 1 to 5, a culture of the strain, a concentrate of the culture, Functional food composition.
8. The method of claim 7,
Wherein the metabolic disease is at least one selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis, fatty liver and cardiovascular disease.
A pharmaceutical composition for the prevention or treatment of an anti-inflammatory and metabolic disease comprising as an active ingredient at least one selected from the group consisting of the strain of any one of claims 1 to 5, the culture of the strain, the concentrate of the culture and the dried product Gt;
10. The method of claim 9,
Wherein the metabolic disease is at least one selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, atherosclerosis, fatty liver and cardiovascular disease.

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