LACTOBACILLUS BREVIS JB PML-131 STRAIN HAVING ANTI-DIABETIC AND ANTI-HYPERLIPIDEMIA ACTIVITY, AND USE OF SAME

The present purpose of the invention to achieve, the present invention refers to hypoglycemic and antihypertensive dyslipidemia activity (Lactobacillus brevis) JB PML-131 Lactobacillus tide by spraying provides strains.

In the present invention the n bit parallel data inputted separation lactic acid bacterium to from fermented soybeans, dyslipidemia antihypertensive and hypoglycemic at excellent active corresponding advertisement based on the shown list identifying strains, JB PML-131 same strain Lactobacillus tide by spraying (Lactobacillus brevis) is provided to have.

(Lactobacillus brevis) Lactobacillus tide by spraying said JB PML-131 dielectric agricultural Institute with agriculture National strains user 22 May 2014 filed under access the resource center (strain: KACC91950P).

Furthermore, culture of the present invention refers to said strain or a unit having a hypoglycemic or antihypertensive stanching bleeding symptoms compositions for the.

In the composition according to one embodiment of the present invention, said composition is preferably food, a food additive or pharmaceuticals and may be in the form, said a dairy food (milk, soybean milk, milk), fermented milk (liquid yogurt, yogurt arcuate), abundant, meat, sausage, bread, chocolate, candy current, [...] , confectionery, pizza, ramyon, gum type , ice cream current, sprocket, beverage, alcoholic drinks and vitamin complex the group consisting of but may be selected from, is at.

The food of the present invention may contain functional foods containing these, said active ingredient of the present invention the functional food in addition and an auxiliary component is changed to an enlarging mode can further. When functional foods have functional components of the present invention, vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid (folic acid), vitamin C, vitamin D3, and vitamin such as vitamin E, copper, calcium, iron, magnesium, potassium, zinc, and various minerals including such as or Lactobacillus lactic may include a.

Functional in addition of the present invention in food, health drink, as well as the typical drinks and such as various flavor furthermore, the number of carbohydrate or natural number can be comprising. Martin TAU zero flavor, Stevia extract and such as natural sweetness system or , saccharin, aspartame sweetener synthesis such as as to the aromatic hydrocarbon. Natural carbohydrate include glucose, oligosaccharides such as monosaccharide, maltose, disaccharide such as sucrose, dextrin, cyclodextrin polysaccharide such as, xylitol, sorbitol, erythro such as sugar alcohol which as to the aromatic hydrocarbon [...].

Culturing strain of the present invention obtained in the culture of said strain or additive the food of a signal transmitted from the video signal input, said culture of strain or other it has been deposited by either adding a speed of n times, and for preparing foods or food component and may be used in conjunction, conventional method suitably according to can be used. A use purpose thereof tilted mixing of active ingredient (prevention, health or therapeutic treatment of) according to can be determined for.

The present invention refers to in addition, culture of or strain of the present invention provides including pharmaceuticals as an active ingredient. Dyslipidemia antihypertensive and which has a hypoglycemic said pharmaceuticals active the second scattered reflection, for preventing or treating diabetes and hyperlipidemia can be. Preferably said strain is Lactobacillus tide by spraying, more preferably Lactobacillus is JB PML-131 tide by spraying.

Furthermore, the present invention refers to said dispersed by a blender before being inoculated strain including hypoglycemic and antihypertensive stanching bleeding symptoms microorganism preparation provides method for producing. Culturing strain the method of the present invention, which are used conventional art of the incubation the method, preferably L-cis reel person culture media added a character generator, is at.

stanching bleeding symptoms antihypertensive and hypoglycemic of the present invention as the active ingredient a microorganism preparation made by using JB PML-131 Lactobacillus tide by spraying can be. Microorganism preparation stanching bleeding symptoms antihypertensive and hypoglycemic the present invention according to a solution, powder, suspension, dispersion, emulsion, planetary dispersion, paste, dust, [...] can be degraded and compositions or granular material, are not limited to.

Hereinafter, . as further described and/or at least two different embodiment of the present invention. Stage, embodiment examples of the present invention relate to a is, aspect of the present invention embodiment a content is not limited.

In the embodiment 1: search lactic acid derived fermented soybeans

1) strain separation

Present fermented soybeans to separate for lactic acid bacterium which allows both commercially and feeds them to a fermented soybeans, so that a large amount uniform fermented soy beans each 0.85% sodium chloride added sterile physiological saline by mixing the meter after dilution by dilution the smears the culture medium to which agar BCP. BCP agar manner in a medium free of lactic acid generated a single colony are separated off and the songs changing L (Sigma) is added 0.05% lactic acid-Γ-butyrolactone (Difco Laboratories, Detroit, mi, USA) MRS agar medium (1.5% agar, w/v) the pure water into a separated the lactic acid bacterium to by culturing.

Collar of Korean jacket using telephone -2) 16S rRNA bio-sequence listing

Originated from foods lactic acid bacterium to procedures for molecular biological method for separating a the 16S rRNA bio-sequence listing of identifying is performed for all the using. DNA separation Wizard Genomic DNA Prep in cultured cells. Kit (GeneAll Biotechnology, Korea) separating chromosome DNA using corresponding advertisement based on the shown list, 16S rDNA to amplify the forward primer (27F) : (5 '-AGA GTT TGA TCC TGG CTC AG-3' : seq ID no 2) and another for the reverse primer (1492R) : (5 '-TAC GGT TAC CTT GTT ACG ACT T-3' : seq ID no:3) PCR (polymerase chain reaction) is performed for all the using. 2% agarose gel electrophoresis PCR reaction product of a combination of a winding vibration corresponding advertisement based on the shown list the laser diode according to the embodiment, gene arranged to improve the crystal quality (quality) GeneAllR Gel SV kit using (GeneAll Biotechnology, Korea), the third to eo [...] preparation method. Automatic sequencing (ABI 3130) analyzer bio-sequence listing DNA using the amplified PCR product of bacteria bio-sequence listing of 16S rRNA 27F, the an analyte using a primer 1492R. Lactic acid bacteria which are isolated by a vacuum line obtained analysis result showing a homology of DNA sequence of seq ID no 1 NCBI BLAST N analysis program (http://www.ncbi.nlm. nihgov/blast/)performed using corresponding advertisement based on the shown list, Lactobacillus tide by spraying (Lactobacillus brevis) and a 100% been exhibiting homologous, JB PML-131 tide by spraying was referred to as Lactobacillus same.

In the embodiment 2: safety of evaluation

Cytotoxic the a human kidney epithelial cell lines (Human kidney epithelial cell line) in 293T cells in an when the transformed cell it is found out that an optical output peak of the acid in higher cell survival. Lactobacillus tide by spraying derived in plants (Lactobacillus brevis) to make sure that safety of JB PML-131 normal cell line (Human kidney epithelial cell line) in 293T human elongated epithelial cell lines in 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS, promega, USA) prolonged periods of ischemia through assay method it was determined that rate. Culture of supernatant, and JB PML-131 0.1%, 0.3%, 0.5% concentration 293T cells useful in inhibiting and treating the high concentration strain result the toxicity until concentration 0.5% cytotoxic effect the identified not high safety when the transformed cell been judged (also 1).

In the embodiment 3: hypoglycemic functional estimation

1) α-glucosidase (AGI) inhibitory activity

Lactobacillus lactic acid originated from foods in the experiments present JB PML-131 tide by spraying to make sure that active AGI of strain culture supernatant by using a liquid-based assay for assessing activity set up. Strain culture flowed, the third to eo after filter m micro 0.22. 0.2 U/mL α-glucosidase enzyme solution and 100 mm potassium phosphate buffer (pH 7.0) and senses a rotation velocity of the disk buffer NPG -2.5 mm ρ (ρ-nitrophenyl a-glucopyranoside) by the application of 20 minutes in 37 °C in reacting a 405 nm it was determined that activity measuring absorbance. As a result, soldiers contrast benign the culture broth JB PML-131 tide by spraying Lactobacillus acarbose (acarbose) the much higher than AGI been found to have an activity (also 2).

2) absorbent 2-NBDG muscle to cells

Number 2 an insulin diabetes type absorbent and secretion by muscle, fat a cell-containing body sample, insulin resistance in peripheral tissues this is attained absorbent blood glucose levels in peripheral tissues raise of the level of blood glucose of the failure is especially. Insulin receptor is insulin cells is adapted to engage and a coplanar signal delivery cascade GLUT4 (Glucose transporter-4) through (signal transduction cascade) for guiding the constitution: an inlet into cells glucose. Muscles along cells in blood glucose in C2C12 for assessing the absorption 2-NBDG into cells to for processing amount of glucose is introduced into the muscle cells it was determined that fluorescence (fluorescence).

Lactobacillus tide by spraying of a metal oxide fiber JB PML-131 absorbing blood glucose in cells to make sure that culture of strain by using a liquid supernatant was assessed for absorbance blood glucose. Muscle cells in C2C12 cell lines cells well plate 96 and frequency divides the 37 °C, 5% CO₂ incubator in then grown on time 24, free glucose medium (starving) the probing star starting to write is delayed medium (no glucose). Herein and supernatant culture strain 2-N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino-2-deoxy-d-glucose (2-NBDG) good by adding. Culture a medium by etching are removed and then washed with PBS at 100 micro l PBS (fluorescence micro reader) for dividing a washing tub by isolating a micro reader EX/EM = 488/535 nm using factor have been measured by conditions. Control strains Lactobacillus experiments (Control) JB PML-131 tide by spraying amount glucose in cells than to a terminal is further increased (also 3).

In the embodiment 4: antihypertensive hemostatic functional estimation

1) BSH (Bile salt hydrolase) assay

Upon application of a voltage while cholesterol blood for storage of, the assay is attached under and into the BSH (Bile salt hydrolase) since in similar to an in, native source anticholesterol for storage of, separates an analytical measuring function decomposition bile composite method by cylinder to have assay.

The control group (PC) benign assay BSH standard strain antistatic (Lactobacillus acidophilus) KCTC 3164, voice the control group (NC) in gram-negative bacteria (Escherichia coli) e.coli ACTC 8739, and native source, separates lactic acid bacteria Lactobacillus tide by spraying using JB PML-131 BSH (Lactobacillus brevis) it was determined that activity. Disk diffusion by lactic acid bacteria (Lactobacilli) MRS (Control), lactic acid bacteria (Lactobacilli) MRS to MRSBA-T including TDCA (Taurodeoxycholic acid), lactic acid bacteria (Lactobacilli) including GDCA (Glycodeoxycholic acid) to MRS 37 °C the culture medium to which flat MRSBA-G in which culturing time 72, disk formed around the electric precipitation [...] granular white and opaque or ring for determining the diameter of a the of the result of comparison between the Figure 4 and a equal. As a result, voice the control group (NC) of a precipitation ring [...] granular white and opaque or any out-of-not observed implying, positive the control group (PC) in the case of two forms of composite bile GDCA TDCA and a photo-reactive components decomposing both where only a ring precipitation was observed to aspects. TDCA (Taurodeoxycholic acid) Lactobacillus tide by spraying with a view to presenting a clear the JB PML-131 GDCA (Glycodeoxycholic acid) implying the decomposition activity was shown activity decomposition only. In comparison with the control group positive in particular forming precipitation annularity 1.5 or a large ring precipitation a massage cream, an essence, capable of confirming the granular white and opaque around the formed for sustain [...] it has been confirmed.

2) measuring effectiveness of removing cholesterol

Preparing 1, 2, 3, last on the order of several tens of years included in the cholesterol in a human body and fermented milk found reduction effect (Lactobacilli) after lactic acid (Bifidobacteria) bifidobacteria resistant to oxygen and disclosed is a lipid metabolism to a beneficial effect and certification been progress studies to be, that for a conventional studies to partially turned through reducing cholesterol in host microorganism it has been reported that using the.

Fermented milk for reducing cholesterol direct effect but no known for substantially it is small , to restrict dynamic absorption of cholesterol by bacteria by the action in vitro (in vitro) cholesterol in he report decreases. These vivo as to be similar to an environmental a in the presence of bile when cultured antistatic (Lactobacillus acidophilus) cholesterol absorption by was achieved through strong and long-lasting is generated. For determining effects removing cholesterol cholesterol reduction effect with a standard strain antistatic (Lactobacillus acidophilus) KCTC 3164, separates source native and a lactic acid bacteria Lactobacillus tide by spraying by cylinder to have JB PML-131 (Lactobacillus brevis). Biliary cholesterol and soluble (oxgall) (Lactobacilli) lactic acid in the MRS liquid the culture medium to which 1% glucose 24 time culture nor post-incubation, culture supernatant in extracted cholesterol. Using spectrophotometer OD₅₅₀ standards in, water-soluble cholesterol and liquid sample to corresponding advertisement based on the shown list, Figure 5 and a equal result. If positive result of the control group (PC) using the reduced cholesterol 25%. Lactobacillus tide by spraying in comparison with the control group positive and the JB PML-131 cholesterol reduced using the 0.4 times.