High efficiency preparation method of high temperature alpha-amylase and mutant thereof

Abstract

A high temperature alpha-amylase and a high efficiency preparation method of the mutant thereof belongs to the fermentation engineering field. The invention provides a method for high efficiently secreting expressing a high temperature alpha-amylase and mutant thereof with alpha-amylase activity through a bacillus, especially a bacillus licheniformis mutant strain as a host cell. The synthesis and secretion level of the high temperature alpha-amylase or mutant thereof of the recombinant strain of the invention reaches 18-25mg / mL during fed-batch fermentation and is 3-4.5 times of the originalstrain high temperature alpha-amylase synthesis level. The high temperature alpha-amylase of the invention is useful for reducing the fermentation manufacture cost of the industrial enzyme, simplifying the fermentation manufacture process and reducing fermentation industrial environment pressure.

Description

A high-efficiency preparation method of high-temperature α-amylase and its mutants technical field The invention is a method for efficiently preparing high-temperature alpha-amylase with starch hydrolysis activity and mutants thereof by using bacillus host cells. The high-temperature α-amylase with starch hydrolysis activity in the present invention is derived from the bacterial strain of Bacillus licheniformis, and the mutant of the high-temperature α-amylase is derived from the artificial mutant of α-amylase of the bacterial strain of Bacillus licheniformis. Further, the present invention uses a specific expression vector to efficiently secrete, express and prepare high-temperature α-amylase and its mutants in Bacillus, especially Bacillus licheniformis host cells. The invention belongs to the field of fermentation engineering. Background technique Starch is a class of polymer substances formed by the polymerization of glucose. Starch consists of two parts, amylose and...

AI Technical Summary

Problems solved by technology

At present, the enzyme production level of BLA industrial production strains is maintained at about 6-10 mg / mL under industrial production conditions, and there is still a large space for the protein secretion potential of more than 20 mg / mL (Veithetal.JMolMicrobiolBiotechnol, 2004)

Images