High-temperature acid-resistant alpha-amylase mutant strain and construction method thereof

A construction method and amylase technology, applied in the field of bioengineering, can solve problems such as incompatibility and loss of enzyme activity, and achieve the effects of stable enzyme activity, saving industrial grain, good thermal stability and acid stability

Active Publication Date: 2008-11-05
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0007] It can be seen that the optimum temperature of acid α-amylase in China is lower than 70°C at present. Because the enzyme activity will be greatly lost when

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  • High-temperature acid-resistant alpha-amylase mutant strain and construction method thereof
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  • High-temperature acid-resistant alpha-amylase mutant strain and construction method thereof

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[0030]Below in conjunction with the examples, the present invention is further described, the following examples are illustrative, not limiting, and the protection scope of the present invention cannot be limited by the following examples.

[0031] In the present invention, the obtained high-temperature acid-resistant α-amylase (acid-resistant transformation of α-amylase and its secretory expression in Bacillus subtilis. Food and Fermentation Industry, 2005, 31 (10): 33-36. Acid-resistant and resistant High-temperature α-amylase and its preparation method, patent application number: 200510013865.2), in order to improve the high-temperature-resistant α-amylase isolated from Bacillus licheniformis (China Industrial Microorganism Culture Collection Management Center, CICC, preservation number, 10181) The acid-resistant stability of the enzyme is a mutant gene obtained by changing the L-leucine at the 134th position of the mature peptide of the gene to L-arginine, and the L-serine ...

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Abstract

The invention relates to a mutant strain of a high temperature acid-proof Alpha-amylase and a method for producing the mutant strain. The high temperature acid-proof Alpha-amylase gene is obtained by mutating the Bacillus licheniformis high-temperature resistant Alpha-amylase gene (Leu134 Arg and Ser320 Ala); the high temperature acid-proof Alpha-amylase gene is integrated into the genom DNA of the parent strain Bacillus licheniformis by establishing a target carrier and by adopting the method of homologous recombination so that the Bacillus licheniformis mutate strain is obtained to realize the homology effective expression of the high temperature acid-proof Alpha-amylase gene. The Alpha-amylase produced by the mutate strain is resistant to high temperature and has the acid stability improved evidently, thereby providing good technique condition for the starch deep processing industry such as alcohol and mashing in China, lowering the cost, saving the energy and the industrial grain, and satisfying the requirements of the starch raw material liquefying technique on acid conditions.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a mutant strain of high-temperature acid-resistant alpha-amylase and a construction method thereof. Background technique [0002] α-amylase (α-1,4-glucan-glucanhydrolase EC 3.2.1.1), also known as liquefied amylase, can cut the α-1,4 bond arbitrarily from the inside of the starch molecule to produce maltose with a relatively small molecular weight dextrin. Under the action of α-amylase, the starch molecules degrade rapidly, the viscosity decreases, and the liquefaction is completed. α-amylase has considerable commercial value and is widely used in starch deep processing industry, alcohol industry, beer industry, citric acid industry, monosodium glutamate and starch saccharification industry, pharmaceutical industry and textile industry. α-amylase can be produced by microbial fermentation, and can also be extracted from plants and animals. At present, most of the industrial produc...

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Application Information

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IPC IPC(8): C12N1/21C12N15/56C12N15/75C12N15/90C12R1/10
Inventor 路福平杜连祥刘逸寒李玉王建玲王春霞
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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