Preparation method and application of amylase mutant with high specific activity and strong ability to degrade raw starch

Abstract

The invention provides a high-specific-activity amylase mutant with good ability to degrade raw starch and a preparation method and application thereof, and relates to the field of gene engineering. Acid amylase derived from Talaromyces leycettanus JCM12802 is used as a female parent, and the wild type is subjected to site-directed mutagenesis by means of molecular biological techniques. Under the modification condition, the amylase mutant has significantly higher specific activity than the wild (before mutation) female parent. By using the scheme, it is possible to greatly increase the specific activity and acting conditions of amylase, and basis is provided for the amylase in industrial production field. The scheme is of important guidance for increasing the catalytic efficiency and specific activity of amylase and other enzymes.

Description

technical field

[0001] The invention relates to the field of genetic engineering and genetic engineering, in particular to a method for preparing a high specific activity amylase mutant and its application. Background technique

[0002] Amylase is a biocatalyst with a wide range of uses, which can be used in bread making industry, starch saccharification and liquefaction, textile desizing, paper making, cleaning agent industry, chemistry, clinical medicine analysis and pharmaceutical industry, etc. The amylase family includes alpha-amylases, beta-amylases and glucoamylases. α-amylase is an endonuclease, which cuts the α-1,4 glycosidic bonds inside the starch molecule in an irregular manner, and makes the starch generate dextrin and oligosaccharides. It is a calcium ion-dependent enzyme. β-amylase sequentially cuts maltose from the non-reducing end, which is an exonuclease. Glucoamylase is an exonuclease acting on α-1,4 glycosidic bonds, cleaving glucose molecules from non-...

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