Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta

Insect Biochem Mol Biol. 1998 May-Jun;28(5-6):409-19. doi: 10.1016/s0965-1748(98)00014-9.

Abstract

The cDNA of the microsomal Juvenile Hormone Epoxide Hydrolase (JHEH) from Manduca sexta was expressed in vitro in the baculovirus system. In insect cell culture, the recombinant enzyme (Ms-JHEH) was produced at a high level (100 fold over background EH catalytic activity). As expected, Ms-JHEH was localized in the microsomal fraction with a molecular mass of approximately 50 kDa. Ms-JHEH showed a substrate and inhibitor spectrum similar to the wild type JHEH isolated from eggs of M. sexta. Its enzymatic activity was the highest for Juvenile Hormone III. Ms-JHEH hydrolyzed several trans-epoxides faster than cis-epoxides. A putative hydroxyl-acyl enzyme intermediate was isolated suggesting a catalytic mechanism of Ms-JHEH similar to the mammalian EHs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Base Sequence
  • DNA Primers / genetics
  • Epoxide Hydrolases / genetics*
  • Epoxide Hydrolases / isolation & purification
  • Epoxide Hydrolases / metabolism
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Juvenile Hormones / metabolism
  • Manduca / enzymology*
  • Manduca / genetics*
  • Polymerase Chain Reaction
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Substrate Specificity

Substances

  • DNA Primers
  • Juvenile Hormones
  • Recombinant Proteins
  • Epoxide Hydrolases
  • juvenile hormone epoxide hydrolase