The asymmetric Michael addition reaction is an important reaction for synthesizing chiral compounds.Traditionally,chiral metal complexes are used as catalysts for chiral induction,but they are complex in structure and difficult to prepare.Artificial metalloenzymes,which utilize biomacromolecules to replace transition metal chiral catalysts,have become a research hotspot.In this study,two histidines and a carboxylate facial triple motif were rationally introduced on the original metal binding motif of TM1459 protein scaffold to coordinate Cu2+,and artificial Cu-TM1459 metalloenzyme was prepared.It was applied to catalyze the asymmetric Michael addition reaction,and the Cu-H52A/H58E variant metalloenzyme exhibited moderate reaction activity and high enantioselectivity(e.e.value up to 58％).Further studies on molecular docking and catalytic mechanism led to site-directed mutations of key residues near the metal binding site.The I108A/C106V/K24E mutant catalyzed the reaction with a yield of 99％and an e.e.value of 93％.
关键词
人工金属酶/不对称迈克尔加成反应/理性设计/蛋白质/生物催化/催化剂
Key words
artificial metalloenzymes/asymmetric Michael addition reaction/rational design/protein/biocatalysis/catalyst
维普
万方数据