Recombinant pichia pastoris strain for regulating and controlling transglutaminase expression of zea mays through TEF1 promoter and construction method

Abstract

The invention relates to a recombinant pichia pastoris strain for regulating and controlling transglutaminase expression of zea mays through a TEF1 promoter. The recombinant pichia pastoris strain isa genetically engineered strain obtained in the mode that transglutaminase from the zea mays is transferred to pichia pastoris. According to the recombinant pichia pastoris strain, an alcohol oxidasepromoter PAOX1 in a pPIC9K plasmid is replaced by a promoter PTEF1 of a translation extension factor 1-alpha, the promoter PTEF1 is connected with a transglutaminase gene to form a loop, and a recombinant expression vector is constructed. The recombinant strain is easy to culture, high in secretion capacity and beneficial for mass transglutaminase expression, the enzyme activity can reach 479 mU / mL, and the recombinant pichia pastoris strain has the advantages of being safe and reliable and has important application prospects.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a recombinant Pichia pastoris strain and a construction method using a TEF1 promoter to regulate the expression of glutamine transaminase in glutinous corn, more specifically the construction of the recombinant plasmid pTEF9k-tgz and the use of the promoter P TEF1 Regulating the expression of sticky maize transglutaminase in Pichia pastoris. Background technique [0002] Transglutaminase (TGase, EC 2.3.2.13) is an enzyme that catalyzes the acyl transfer reaction, which can catalyze the γ-carboxamide group (amino acceptor) of glutamine residues and the conversion of lysine residues Acyl transfer reactions between ε-amino groups or other primary amino groups (amino donors). The catalysis of TGase changes the conformation of the protein and realizes the intramolecular and intermolecular cross-linking of the protein, thereby improving the functional properties of the prot...

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Problems solved by technology

In the 1950s, guinea pig liver TGase (Guinea pig transglutaminase, GTG) was the main source of commercial enzymes, but its source was rare, and the separation and purification process was complicated, resulting in high enzyme prices, making its application in the food industry face certain challenges. difficulty

[0005] Through the search, no patent publications related to the patent application of the present invention have been found

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