Enzymic synthesis of juvenile hormone in locust corpora allata: evidence for a microsomal cytochrome P-450 linked methyl farnesoate epoxidase.

Homogenates of corpora allata from adult Locusta migratoria in phosphate-buffered EDTA have been analysed by sucrose-density-gradient centrifugation. Succinate-cytochrome c reductase activity (mitochondrial) bands between d20/4 1.13-1.15, whereas NADPH-cytochrome c reductase and NADPH-dependent methyl farnesoate 10.11-epoxidase activities band identically between d20/4 1.06-1.12. We conclude that the methyl farnesoate epoxidase is exclusively microsomal. Farnesoic acid O-methyltransferase is an exclusively soluble enzyme which stoichiometrically transfers the S-methyl group from S-adenosylmethionine to farnesoic acid. No carboxyl esterase activity was found. Isolated microsomes were used to obtain an apparent Km = 7.7 X 10-6 M for the epoxidase, although substrate solubility limits the rate to 0.5 V. As expected, the product (juvenile hormone III) is chiral (10 R). The epoxidase is inhibited by excess NADP+ and oxidised cytochrome c, but neither inhibited nor synergised by NADH. NADH supports less than 10% of the NADPH rate of epoxidation. The epoxidase is inhibited by a carbon monoxide/oxygen atmosphere, half-maximal inhibition occurring at a CO/O2 ratio of 4.0. This inhibition is reversed by white-light irradiation.