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Alpha-amylase inhibitor producing strain and screening method thereof

An amylase inhibitor and screening method technology, which is applied in the field of alpha-amylase inhibitor-producing bacteria and its screening, and can solve the problems reported in the screening method of microbial-derived alpha-amylase inhibitor and difficult to measure inhibitor concentration and enzyme activity. relationship, the inability of high-throughput screening, etc., to achieve the effect of reducing sample processing, improving detection efficiency, and high-throughput screening

Active Publication Date: 2011-07-20
ZHEJIANG UNIV OF TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in this reaction, the influence of soluble starch and α-amylase on the background Abs546 value of the measurement system is close to 0.2, and the background has a greater influence on the measurement interval of 0.4-0.7, and it is difficult to determine the relationship between inhibitor concentration and enzyme activity
At the same time, the color development of the reaction requires a boiling water bath, which cannot be carried out on a 96-well plate, and cannot be used for high-throughput screening using a microplate reader.
At present, there is no efficient, sensitive, and rapid screening method for microbial-sourced α-amylase inhibitors reported.

Method used

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  • Alpha-amylase inhibitor producing strain and screening method thereof
  • Alpha-amylase inhibitor producing strain and screening method thereof

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Experimental program
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Effect test

Embodiment 1

[0034] (1) Preparation of medium:

[0035] Preparation of plate medium and slant medium: the medium components are as follows (g / L): sucrose 25g / l, peptone 2g / l, tyrosine 1g / l, K 2 HPO 4 ·3H 2 O 0.5g / l, KCl 0.5g / l, MgSO 4 ·7H 2 O 0.5g / l, FeSO 4 ·7H 2 O 0.1g / l, agar 20g / l, solvent is water, initial pH 7.0; when preparing, add 20g agar strips to 1000mL water, stir while heating until completely dissolved, then add 25g sucrose, 2g peptone, 1g tyrosine , K 2 HPO 4 ·3H 2 O 0.5g, KCl 0.5g, MgSO 4 ·7H 2 O 0.5g, FeSO 4 ·7H 2 O0.1g, and stir until completely dissolved, then replenish water to 1000ml.

[0036] Pour 5ml into a test tube with a volume of 20ml, sterilize at 121°C for 20min, place a slant, and cool to obtain a slant medium; at the same time, cool the sterilized medium to 60°C, Pour it into a sterilized plate and cool down to obtain a medium plate. Fermentation medium preparation: Add 80g of maltose, 20g of glucose, 10g of soybean cake powder, 5g of calcium ca...

Embodiment 2

[0045] (1) Preparation of medium:

[0046] Preparation of slant medium, plate medium, seed medium, and fermentation medium: slant medium and plate medium are mediums with the same composition, and are prepared as follows: add 20g agar strips to 1000mL water, and stir while heating until Completely dissolve, then add 30g of sucrose, 2g of peptone, 1g of tyrosine, K 2 HPO 4 ·3H 2 O 1g, KCl 0.5g, MgSO 4 ·7H 2 O 0.5g, FeSO 4 ·7H 2 O 0.1g, and stir until completely dissolved, then add water to 1000ml, adjust pH to 7.0. According to the amount of each 5ml, pour it into a test tube with a volume of 20ml, sterilize at 121°C for 20min, place a slope, and cool it to obtain a slope medium; at the same time, cool the sterilized medium to 60°C, under sterile conditions Pour into a sterilized petri dish and cool down to obtain a dish culture medium;

[0047] Fermentation medium: Add 40g of maltose, 40g of glucose, 15g of soybean cake powder, 5g of calcium carbonate, 5g of glycerin, ...

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Abstract

The invention discloses an alpha-amylase inhibitor producing strain, i.e. Actinoplanes sp.ZJB-08196 preserved in the China Center for Type Culture Collection located in Wuhan university, Wuhan Province, China on Feb. 16th, 2009 with the preservation number of CCTCC No. M209022. The alpha-amylase inhibitor is acarbose. The invention also discloses a screening method of the alpha-amylase inhibitor producing strain. The screening method provided by the invention is simple and effective, can screen a great amount of samples in a short time, has high sensitivity, reduces the long processes for processing and analyzing samples through HPLC (High-Performance Liquid Chromatography), and the like and improves detection efficiency.

Description

(1) Technical field [0001] The present invention particularly relates to α-amylase inhibitor producing bacteria and its screening method. (2) Background technology [0002] Diabetes is a major multiple disease that is prevalent worldwide. Due to the huge change in lifestyle and the lack of people's awareness of health care, there are currently 92.4 million diabetics in my country, including 50.2 million males and 42.2 million females, surpassing India and becoming the country with the largest number of diabetics in the world. The incidence rate (9.7%) ) is second only to the United States (11%), in which 93.7% of patients with type II diabetes. Currently commonly used drugs for the treatment of type II diabetes include: biguanides that increase insulin sensitivity—Metformin; sulfonylureas that promote insulin secretion—Gliclazide (trade name Tang Bingkang); α-glucosidase inhibitor Drugs—Acarbose, Miglitol (trade name Aotianping), Voglibose (trade name Beixin). Among them, ...

Claims

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Application Information

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IPC IPC(8): C12N1/00C12P19/00C12R1/045
Inventor 郑裕国王远山冯志华薛亚平王亚军沈寅初
Owner ZHEJIANG UNIV OF TECH
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