基于生物计算策略定点突变改造亮氨酸脱氢酶的底物专一性

he method for synthesizing chiral amines by amine dehydrogenase has the advantages of environmental friendliness, high stereoselectivity, and mild reaction conditions. However, natural amine dehydrogenase has low catalytic activity, which greatly limits its application in the synthesis of chiral amines. In this study, a novel amino acid dehydrogenase was used as a template. Based on a biocomputing strategy, conservative analysis, homology modeling, and molecular docking analysis were performed to select mutation sites, and mutants L52Ser and T143Cys were obtained. The enzyme activities of the two mutant strains to 2-pentanone were 1.55 U/mg and 2.06 U/mg, respectively. The enzyme activity of mutant 143Cys was 188% higher than that of the original strain, and the Tm value was increased by 2.55 C. It is used for efficient preparation for chiral amines laid the foundation.