Efficient-expression recombinant TEV enzyme with high activity and stability as well as preparation method, determination method and application of recombinant TEV enzyme

Abstract

The invention discloses an efficient-expression recombinant TEV enzyme with high activity and stability as well as a preparation method, a determination method and application of the recombinant TEV enzyme. The recombinant TEV enzyme comprises an amino acid sequence for coding TEV protease or a mutation sequence of the amino acid sequence, a CL7 tag is fused at the N end of the amino acid sequence for coding the TEV protease or the mutation sequence of the amino acid sequence, a polyarginine tag is fused at the C end of the amino acid sequence for coding the TEV protease or the mutant sequence of the amino acid sequence. Compared with existing recombinant TEV protease, the recombinant TEV protease not only can overcome the defect that the protein stability and activity are reduced due to the fact that the wild type TEV protease shears own specific sites in the expression process, but also has higher protein yield, better protein stability and higher activity, the recombinant TEV protease is more suitable for large-scale production and industrial use.

Description

technical field [0001] The invention relates to the technical field of recombinant proteins, in particular to a high-efficiency expression recombinant TEV enzyme with high activity and stability and its preparation, determination method and application. Background technique [0002] The expression of recombinant proteins is widely used in bioengineering, including protein purification, localization and functional analysis. According to different expression systems, it is mainly divided into E. coli expression system, yeast expression system, insect cell expression system and mammalian cell expression system. Among them, yeast expression system, insect expression system and mammalian expression system belong to eukaryotic expression system, and Escherichia coli expression system belongs to prokaryotic expression system. Compared with the eukaryotic expression system, the prokaryotic expression system has many advantages, including clear genetic background, fast reproduction,...

AI Technical Summary

Problems solved by technology

However, there are three problems in the expression and purification of wild-type TEV protease in E. coli: self-cleavage, rare codons and poor solubility

In general, although many studies have improved the expression of TEV protease by adding a fusion

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