Site-specific mutagenesis high temperature resistant phytase gene TP and expression vector and application thereof

A high-temperature-resistant phytase and site-directed mutation technology, which can be used in plant gene improvement, application, genetic engineering, etc., can solve the problem of phytase enzyme activity damage, phytase heat-resistant effect is not ideal, and thermal stability is not very good Solve problems such as well to achieve the effect of improving thermal stability, considerable economic and social benefits, and good heat resistance
CN102943083AActive Publication Date: 2013-02-27TIANJIN CHIATAI FEED TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
TIANJIN CHIATAI FEED TECH
Publication Date
2013-02-27

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Abstract

The invention provides site-specific mutagenesis high temperature resistant phytase gene TP and an expression vector and application thereof. Amino acid in a specific area in a phytase amino acid sequence is mutated into praline and arginine so as to enable the phytase to have a special stable protein structure. Simultaneously, a gene sequence is optimized to synthesize a phytase gene according to pichia pastoris codon preference and GC content, recombinant plasmids are built and transferred to pichia pastoris, and positive converters are obtained through screening to conduct induction expression to obtain the high temperature resistant phytase. Experiments prove that the phytase obtained through expression of the pichia pastoris can resist high temperature plasmids with the temperature higher than 85 DEG C, and survival rate reaches 85%. Popularization and application of the high temperature resistant phytase have substantial economical benefit and social benefit on development of feed animal husbandry of our country, and simultaneously great ecological benefit can be obtained.
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Description

technical field

[0001] The invention belongs to the field of molecular biology feed additives, and in particular relates to a site-directed mutation high-temperature-resistant phytase gene TP, an expression carrier and application thereof. Background technique

[0002] Phytic acid (phytic acid) is the main storage form of phosphorus in plants, and phytic acid and its salts are the main storage form of phosphorus in crops such as grains, beans, and oilseeds. Due to the lack of corresponding enzymes in the stomach of monogastric animals, phytic acid cannot be hydrolyzed and utilized, which causes a waste of phosphorus sources, and the phosphorus in the unusable phytic acid is directly excreted, resulting in phosphorus pollution of soil and water. In addition, phytic acid can also chelate with mineral elements and protein, so that these nutrients cannot be effectively used, resulting in a reduction in the nutritional value of plant feed. Therefore, phytic acid is also generally...

Claims

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