Phytobacterium GBE48, application thereof and method for alpha-glucosidase activity inhibition quantitative detection

A technology of glucosidase and activity inhibition, applied in the field of microorganisms, can solve the problems of no research and reports, no research on screening methods for strains producing α-glucosidase inhibitors, etc., and achieve a strong ability to withstand adverse environmental conditions in the gastrointestinal tract Effect

Inactive Publication Date: 2018-07-17
BEIJING UNIV OF AGRI
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  • Abstract
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Problems solved by technology

[0006] However, in the existing patents, there is no detailed research on the screening method for the strains producing α-glucosidase inhibitors
Although there are two related literature reports in China, such as "Screening of probiotic lactic acid bacteria with α-glucosidase inhibition" published by Lu Jiali et al. and "Screening of a probiotic strain with potential hypoglycemic effect" published by Chen Pei et al., However, these two articles are all about the screening of α-glucosidase-inhibiting probiotic lactic acid bacteria, and the assay methods for α-glucosidase inhibitors derived from Lactobacillus plantarum, especially for the α-glucosidase activity inhibition test reaction system optimization, but there is no relevant research and report in the prior art

Method used

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  • Phytobacterium GBE48, application thereof and method for alpha-glucosidase activity inhibition quantitative detection
  • Phytobacterium GBE48, application thereof and method for alpha-glucosidase activity inhibition quantitative detection
  • Phytobacterium GBE48, application thereof and method for alpha-glucosidase activity inhibition quantitative detection

Examples

Experimental program
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Effect test

experiment example 2

[0089] Experimental Example 2 Determination of the inhibitory rate of α-glucosidase activity by Lactobacillus plantarum GBE48

[0090] The total reaction system in each well is 220 μL, in 50 μL, 1.5 mmol / L PNPG substrate solution and pH 6.8, 0.1mol / L 15 μL phosphate buffer, add 15 μL of the fermentation supernatant of the strain to be tested, and the mixture After reacting at 37°C for 10 minutes, add 70 μL of α-glucosidase solution with an enzyme activity of 20 U / mL, continue to react at 37°C for 50 minutes, and then add 0.1 mol / L Na 2 CO 3 70 μL of the solution was used to stop the reaction. Measure the absorbance of the reaction solution at 405 nm. The absorbance value is directly proportional to the free amount of p-nitrophenol and inversely proportional to the inhibitory rate of α-glucosidase activity.

[0091] In the reaction system, 15 μL of phosphate buffer solution with pH 6.8 and 0.1mol / L was used as the blank control of the α-glucosidase solution and the sample to...

experiment example 3

[0098] Experimental example 3 Lactobacillus plantarum GBE48 resistance to gastrointestinal adverse environment test

[0099] Preparation of artificial simulated gastric juice: NaCl 0.20%, pepsin (enzyme activity: 250 U / mg, purchased from Sigma, USA, product number: P7000-25G) 0.30%, HCl to adjust pH to 2.5, filter and sterilize for later use.

[0100] Artificially simulated intestinal juice: trypsin (enzyme activity: 1655U / mg, purchased from Sigma, USA, product number: 93615-5G) was dissolved in 0.1mol / L phosphate buffer solution with pH 8.0 to make the final concentration 1g / L, Add 0.3% ox bile salt and filter through a 0.22 μm filter membrane to prepare simulated intestinal fluid.

[0101] Cultivate 3 tubes of 10mL MRS culture solution of 2 strains (Lactobacillus plantarum subsp. plantarum and Lactobacillus rhamnosus) at 37℃, repeat 3 times for each group, centrifuge at 6000r / min for 10min, discard the supernatant, and wash with 0.85 After centrifugation and washing with 10...

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Abstract

The invention provides a phytobacterium GBE48, application thereof and a method for alpha-glucosidase activity inhibition quantitative detection. The phytobacterium GBE48 (CGMCCNo.15130) has the highgastrointestinal tract inverse environment resistance, has the high alpha-glucosidase activity inhibition rate and is the most potential bacterial strain for treating diabetes. Meanwhile, according tothe method, by building and optimizing an experiment reaction system, measurement of different bacterial strains for alpha-glucosidase activity inhibition can be achieved, screening of alpha-glucosidase inhibitors from lactobacillus plantarum can be achieved, and a gap of the related research field is filled.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a method for quantitative detection of lactobacillus plantarum GBE48 and its application and α-glucosidase activity inhibition. Background technique [0002] Diabetes Mellitus (DM) is a metabolic disorder with persistent high blood sugar caused by insufficient insulin secretion or impaired insulin secretion function. It has become a recognized disease that endangers human health. At present, the drugs for the prevention and treatment of diabetes are still mainly chemical agents. Due to the large side effects of chemical agents, the research on safe and natural drugs has become a hot spot. [0003] The screening of enzyme inhibitors derived from microorganisms has become a new research hotspot in the field of diabetes prevention and treatment. Among them, α-glucosidase inhibitors have been recommended by the third Asian-Pacific region diabetes treatment drug guideline as the first-li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/34A61K35/747A61P3/10C12R1/25
CPCA61K2035/115C12N1/20C12Q1/34C12N1/205C12R2001/25
Inventor 谢远红刘慧张红星高秀芝熊利霞金君华
Owner BEIJING UNIV OF AGRI
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