Modification of the substrate specificity of leucine dehydrogenase by site-directed mutagenesis based on biocomputing strategies

Synthesis of chiral amines by amine dehydrogenase has the advantages environmental friendliness, high stereoselectivity, and mild reaction conditions. However, low catalytic activity, which greatly limits its application in large-scale synthesis amines. In this study, a novel was obtained modifying substrate specificity leucine via computer-aided protein engineering strategy. Furthermore, conservation analysis, homology modeling molecular docking analysis were carried out biocomputing strategy to select mutation sites, mutants L52S T143C obtained. The enzyme activities two 2-pentanone 1.55 U/mg 2.06 U/mg, respectively. activity latter 188% Tm value 2.55 °Chigher than those original mutant, laid foundation for efficient preparation using enzyme.