Ye_2016_Catal.Commun_87_32

To improve the enantioconvergence of an epoxide hydrolase from Phaseolus vulgaris (PvEH1) towards styrene epoxides, its directed modification was performed based on the rational design by using the molecule docking simulation and the multiple alignment. The single- and multi-site mutation genes of pveh1 were constructed as designed theoretically by PCR, and expressed in E. coli BL21(DE3), respectively. Among all PvEH1 mutants tested, a three-site mutant, PvEH1 L105I/M160A/M175I, was selected having the highest activity of 10.66 U/g wet cell and the best regioselectivity (alphaS > 99%, betaR = 86.4%), by which rac-1a was transformed into (R)-1b with 87.8% enantiomeric excess (eep), much higher than that (33.6% eep) by PvEH1. Furthermore, it completely hydrolyzed rac-2a5a into (R)-2b5b with 52.370.9% eep.