Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution

Org Lett. 2004 Jan 22;6(2):177-80. doi: 10.1021/ol035898m.

Abstract

[reaction: see text] The epoxide hydrolase (EH) from Aspergillus niger, which shows a selectivity factor of only E = 4.6 in the hydrolytic kinetic resolution of glycidyl phenyl ether, has been subjected to directed evolution for the purpose of enhancing enantioselectivity. After only one round of error-prone polymerase chain reaction (epPCR), enantioselectivity was more than doubled (E = 10.8). The improved mutant enzyme contains three amino acid exchanges, two of which are spatially far from the catalytically active center.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus niger / enzymology
  • Catalysis
  • Crystallography, X-Ray
  • Directed Molecular Evolution*
  • Epoxide Hydrolases / chemistry*
  • Kinetics
  • Molecular Structure
  • Phenyl Ethers / chemistry
  • Protein Conformation
  • Stereoisomerism

Substances

  • Phenyl Ethers
  • phenylglycidyl ether
  • Epoxide Hydrolases