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

Stéphane Debernard, Christophe Morisseau, Tonya F. Severson, Li Feng, Hubert Wojtasek, Glenn D. Prestwich, Bruce D. Hammock

Research output: Contribution to journalArticle

34 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)409-419
Number of pages11
JournalInsect Biochemistry and Molecular Biology
Volume28
Issue number5-6
DOIs
StatePublished - May 1998

Fingerprint

epoxide hydrolase
Manduca
Manduca sexta
juvenile hormones
Epoxy Compounds
epoxides
Baculoviridae
Molecular mass
Enzymes
juvenile hormone epoxide hydrolase
Cell culture
Hydroxyl Radical
Eggs
enzymes
Insects
catalytic activity
Catalyst activity
Complementary DNA
Cell Culture Techniques
cell culture

Keywords

  • Epoxide Hydrolase
  • Juvenile Hormone
  • Manduca sexta
  • Mechanism
  • Recombinant enzyme

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry

Cite this

Debernard, S., Morisseau, C., Severson, T. F., Feng, L., Wojtasek, H., Prestwich, G. D., & Hammock, B. D. (1998). Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta. Insect Biochemistry and Molecular Biology, 28(5-6), 409-419. https://doi.org/10.1016/S0965-1748(98)00014-9

Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta. / Debernard, Stéphane; Morisseau, Christophe; Severson, Tonya F.; Feng, Li; Wojtasek, Hubert; Prestwich, Glenn D.; Hammock, Bruce D.

In: Insect Biochemistry and Molecular Biology, Vol. 28, No. 5-6, 05.1998, p. 409-419.

Research output: Contribution to journalArticle

Debernard, S, Morisseau, C, Severson, TF, Feng, L, Wojtasek, H, Prestwich, GD & Hammock, BD 1998, 'Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta', Insect Biochemistry and Molecular Biology, vol. 28, no. 5-6, pp. 409-419. https://doi.org/10.1016/S0965-1748(98)00014-9
Debernard, Stéphane ; Morisseau, Christophe ; Severson, Tonya F. ; Feng, Li ; Wojtasek, Hubert ; Prestwich, Glenn D. ; Hammock, Bruce D. / Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta. In: Insect Biochemistry and Molecular Biology. 1998 ; Vol. 28, No. 5-6. pp. 409-419.
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