Expression and purification method of high-stability double-base enzyme Kex2 mutant

The invention discloses an expression and purification method of a high-stability double-base enzyme Kex2 mutant, which comprises the following steps of: (1) providing a variant of a double-base enzyme Kex2, adding a His tag at the C end of the variant, and adding a signal peptide 1 or a signal peptide 2 or an alpha-factor guide peptide or an acid phosphatase PHO5 guide peptide or an Aga guide peptide at the N end of the variant; (2) recombinant expression plasmid; (3) construction of an expression system and expression of kex2 protease; and (4) purifying the Kex2 protease. According to the invention, efficient and high-stability secretory expression of Kex2 is realized, the protein has good biological activity, and an effective scheme is provided for solving the technical problem that Kex2 is difficult to express efficiently in the prior art. 本发明公开一种高稳定性双碱基酶Kex2突变体的表达纯化方法，包括步骤：(1)提供双碱基酶Kex2的变体，在变体C端加His tag标签，变体N端加信号肽1或信号肽2或α-factor引导肽或酸性磷酸酶PHO5引导肽或Aga引导肽；(2)重组表达质粒；(3)表达系统构建与kex2蛋白酶的表达；(4)Kex2蛋白酶纯化。本发明实现了Kex