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RESEARCH PRODUCT
Expression and characterization of the recombinant juvenile hormone epoxide hydrolase (JHEH) from Manduca sexta.
Christophe MorisseauBruce D. HammockGlenn D. PrestwichTonya SeversonLi FengHubert WojtasekStéphane Debernardsubject
mechanismGene ExpressionBiochemistryPolymerase Chain ReactionSubstrate SpecificityManduca sextaManducaHydrolaseAnimalsEpoxide hydrolaserecombinant enzymeMolecular BiologyDNA Primerschemistry.chemical_classificationEpoxide HydrolasesbiologyMolecular massBase Sequencejuvenile hormoneInsect cell cultureHydrogen-Ion Concentrationbiology.organism_classificationMolecular biologyRecombinant Proteinsepoxide hydrolaseJuvenile HormonesEnzymechemistryBiochemistryManduca sextaInsect ScienceJuvenile hormoneManducaBaculoviridaedescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1998-05-01 | Insect biochemistry and molecular biology |