Microorganism for treatment or prevention of corpulence and diabetes mellitus, and pharmaceutical composition containing the same
a microorganism and diabetes mellitus technology, applied in the direction of biocide, plant growth regulator, biochemistry apparatus and processes, etc., can solve the problems of imposing a substantial cost on the public as well as on patients, losing control of blood glucose levels, late complications, etc., and achieve the effect of reducing the amount of oligosaccharide absorbed
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example 1
Selecting of Microorganism which Produces Extracellular Polysaccharide from Samples
[0251] In order to isolate microorganisms which produce dietary fibers, samples were collected from glucose factory sewage and other locations. 10 g of the mixture thus collected were disrupted and suspended in 90 ml of physiological saline solution(0.85% NaCl). The said suspended samples were diluted to 10−2, 10−4, and 10−6 in physiological saline solution. These diluted samples then smeared on MRS agar medium containing 1 mg of cycloheximide per 100 ml medium(1% Peptone, 1% beef extract, 0.5% yeast extract, 2% glucose, 0.1% Tween-80, 0.2% Citric Acid Ammonium, 0.5% Sodium Acetate, 0.01% MgSO4, 0.005% MnSO4, 0.2% Sodium Phosphate pH6.5) and on BSH agar medium(2% glucose, 0.5% Peptone, 0.5% yeast extract, 0.27% Na2HPO4, 0.115% Citric Acid pH 5.0)(Hestirin and Schramm, J. Gen. Microbiol., 11:123,1954) and cultured in 30° C. for 72 hours. Approximately 2,000 colonies were selected and were initially i...
example 2
The Morphological Determination and Characteristics of the Selected BC-Y009 and BC-Y058
[0253] Microorganisms which show high polysaccharide productivity selected from the Example 1 were BC-YO09, BC-Y002, BC-Y015, BC-Y026, BC-Y058, BC-Y112, BC-Y130, and BC-Y201. Upon observing partial DNA sequences, BC-YO09, BC-Y002, BC-Y015 and BC-Y026 were microorganisms of Lactobacillus genus, and BC-Y058, BC-Y112, BC-Y130 and BC-Y201 were microorganisms of Acetobacter genus.
[0254] Among these bacteria, BC-Y009 and BC-Y058 which show high polysaccharide productivity were inoculated in MRS and BSH liquid mediums at 30° C. for 72 hours and cultured in suspension. Cultured mediums were centrifuged at 6,000 rpm in 4° C. to obtain microorganisms and the nucleic acids thereof were isolated by means of using the CTAB / NaCl method. By using 16 s rRNA consensus primer, 16 s rRNA was amplified by means of PCR method, and the sequence thus obtained, was determined. BLAST analysis (NCBI, USA) on the sequenc...
example 3
The Degree of Decomposition of Extracellular Polysacchride (Dietary Fiber) bV Intestinal Digestive Enzymes
[0260] In order to determine whether or not dietary fiber produced by said microorganisms is decomposed by intestinal digestive enzyme, 1 g of porcine pancreatin that shows the activity of 3×U.S.Pharmacopia (manufactured by Sigma) and comprises amylase, lipase, protease and nuclease, was suspended in buffer solution (pH7.5) of 1 g of dried dietary fiber. This suspension was incubated for 7 days at 40° C. and the suspension was collected once a day and the glucose therein was analyzed quantitatively by using DNS(3,5-dinitrosalicylic acid). The result thereof showed that dietary fibers has never been decomposed at all.
[0261] Therefore, it was confirmed that the dietary fibers produced by the microorganisms of the present invention do not decompose within the intestine.
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