Cloning, expression and enantioselective hydrolytic catalysis of a microsomal epoxide hydrolase from a marine fish, Mugil cephalus

Biotechnol Lett. 2007 Feb;29(2):237-46. doi: 10.1007/s10529-006-9222-4. Epub 2006 Dec 7.

Abstract

The cDNA of a marine fish microsomal epoxide hydrolase (mEH) gene from Mugil cephalus was cloned by rapid amplification of cDNA ends (RACE) techniques. The homology model for the mEH of M. cephalus showed a characteristic structure of alpha/beta-hydrolase-fold main domain with a lid domain over the active site. The characteristic catalytic triad, consisting of Asp(238), His(444), and Glu(417), was highly conserved. The cloned mEH gene was expressed in Escherichia coli and the recombinant mEH exhibited (R)-preferred hydrolysis activity toward racemic styrene oxide. We obtained enantiopure (S)-styrene oxide with a high enantiopurity of more than 99% enantiomeric excess and yield of 15.4% by batch kinetic resolution of 20 mM racemic styrene oxide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Blotting, Western
  • Catalysis
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • Epoxide Hydrolases / chemistry
  • Epoxide Hydrolases / genetics*
  • Epoxide Hydrolases / metabolism
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Fishes
  • Gene Expression Regulation, Enzymologic
  • Humans
  • Hydrolysis
  • Molecular Sequence Data
  • Molecular Structure
  • Protein Structure, Secondary
  • Seawater
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Stereoisomerism
  • Substrate Specificity

Substances

  • DNA, Complementary
  • Fish Proteins
  • Epoxide Hydrolases