Artificial Cu-TM1459 metalloenzyme catalyzes asymmetric Michael addition reaction
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％.
artificial metalloenzymesasymmetric Michael addition reactionrational designproteinbiocatalysiscatalyst
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