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Spatial lactobacillus plantarum SS18-5 for reducing alpha-glucosidase activity and application thereof

A technology of Lactobacillus plantarum and glucosidase, applied in the field of Lactobacillus plantarum SS18-5 in space

Active Publication Date: 2018-05-08
FULLARTON BIOENG TECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no domestic and foreign invention patents and literature reports on the determination method of α-glucosidase inhibitor derived from Lactobacillus plantarum, especially the optimization of the reaction system for α-glucosidase activity inhibition test.

Method used

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  • Spatial lactobacillus plantarum SS18-5 for reducing alpha-glucosidase activity and application thereof
  • Spatial lactobacillus plantarum SS18-5 for reducing alpha-glucosidase activity and application thereof
  • Spatial lactobacillus plantarum SS18-5 for reducing alpha-glucosidase activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, the optimization of α-glucosidase inhibitor screening method

[0067] In this embodiment, a single factor multilevel test is adopted to determine a better α-glucosidase activity inhibition test reaction system, and all test reactions are carried out in a CS016-0096 type 96-well cell culture microplate (350 μL per well). Determination of OD by BioTek microplate reader 405nm value.

[0068] In following steps 1 to 5, substrate solution, 0.1mol / L phosphate buffer saline, α-glucosidase solution and 0.1mol / L Na 2 CO 3 The total volume of the aqueous solution was 210 μL.

[0069] 1. Optimization of substrate concentration

[0070] Carry out the α-glucosidase activity inhibition experiment according to the following steps successively:

[0071] (1) Mix 50 μL of substrate solution and 10 μL of 0.1mol / L phosphate buffer (pH 6.8) evenly, then let it stand at 37°C for 10 minutes;

[0072] (2) Add 50 μL of α-glucosidase solution (enzyme activity 20 U / mL), mix we...

Embodiment 2

[0107] Embodiment 2, the establishment of α-glucosidase inhibitor screening method

[0108] Based on the optimization results of Example 1, the screening method for alpha-glucosidase inhibitors is established, and the specific methods are as follows:

[0109] 1. Carry out the α-glucosidase activity inhibition experiment according to the following steps in turn:

[0110] (1) Mix 50 μL substrate solution (PNPG concentration is 0.2mmol / L), 40 μL 0.1mol / L phosphate buffer (pH6.8) and 20 μL fermentation supernatant of the strain to be tested, and then stand at 37°C Preheat for 10 minutes;

[0111] (2) Add 40 μL α-glucosidase solution (enzyme activity 20U / mL), mix well, and then stand at 37°C for 40 minutes;

[0112] (3) Add 60μL 0.1mol / L Na 2 CO 3 The aqueous solution was used to terminate the reaction, and the absorbance value (OD value) was measured at a wavelength of 405 nm using a BioTek microplate reader (model ELx808, American Gene Co., Ltd.).

[0113] At the same time, ...

Embodiment 3

[0121] Example 3, Screening of Lactobacillus plantarum strains with higher inhibitory rate to α-glucosidase activity

[0122] 1. Isolation and purification of bacterial strains

[0123] Lactobacillus plantarum GS18 on the ground was carried back by Tiangong-2 and Shenzhou-11 spacecraft to obtain space-mutated strains, from which 120 strains were isolated and purified, and the strain codes were respectively marked as SS18-1 to SS18-120.

[0124] 2. Determination of the inhibitory rate of the strain on α-glucosidase activity

[0125] The 120 strains obtained in Step 1 were tested for the inhibition rate of α-glucosidase activity according to the method in Example 2, and Lactobacillus plantarum (Lactobacillus plantarum) GS18 was used as a control, and the results are shown in Table 2.

[0126] Table 2 Space Lactobacillus plantarum to α-glucosidase activity inhibitory rate result

[0127]

[0128]

[0129] The results in Table 2 show that compared with the original ground ...

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Abstract

The invention discloses spatial lactobacillus plantarum SS18-5 for reducing alpha-glucosidase activity and application thereof, and particularly provides lactobacillus plantarum Fullarton-SS18-5 the preservation number of which is CGMCC NO.14917. With original lactobacillus plantarum GS18 on the ground to serve as a contrast, the spatial lactobacillus plantarum Fullarton-SS18-5 which has a prettyhigh resistance rate on the alpha-glucosidase activity, namely high yield of alpha-glucosidase inhibitors, is screened out; the spatial lactobacillus plantarum SS18-5 is excellent in capability of tolerating inverse environment of a gastrointestinal tract and capable of successfully exerting a healthcare efficacy through the gastrointestinal tract, and can become the most potential bacterial strain for preventing and treating diabetes.

Description

technical field [0001] The invention relates to a space lactobacillus plantarum SS18-5 which reduces the activity of α-glucosidase and application thereof. Background technique [0002] Diabetes Mellitus (DM) is a metabolic disorder disease caused by insufficient insulin secretion or impaired insulin secretion function, resulting in persistent high blood sugar. It has become a recognized disease that endangers human health. At present, chemical drugs for the prevention and treatment of diabetes, because of their large side effects, make the research on safe and natural diabetes drugs become a hot spot. Screening research on 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 as the first-line drugs for lowering postprandial blood sugar by the third Asia-Pacific diabetes treatment drug guideline. The screening method for α-glucosidase inhi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04C12Q1/34A61K35/747A61P3/10C12R1/25
CPCA61K2035/115C12N1/20C12P1/04C12Q1/34C12N1/205C12R2001/25
Inventor 郝红炜张红星刘慧谢远红
Owner FULLARTON BIOENG TECH BEIJING CO LTD
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