Diverging Mechanisms: Cytochrome-P450-Catalyzed Demethylation and gamma-Lactone Formation in Bacterial Gibberellin Biosynthesis

摘要

Biosynthesis of the gibberellin (GA) plant hormones evolved independently in plants and microbes, but the pathways proceed by similar transformations. The combined demethylation and gamma-lactone ring forming transformation is of significant mechanistic interest, yet remains unclear. The relevant CYP112 from bacteria was probed by activity assays and O-18(2)-labeling experiments. Notably, the ability of tert-butyl hydroperoxide to drive this transformation indicates use of the ferryl-oxo (Compound I) from the CYP catalytic cycle for this reaction. Together with the confirmed loss of C20 as CO2, this necessitates two catalytic cycles for carbon-carbon bond scission and gamma-lactone formation. The ability of CYP112 to hydroxylate the delta-lactone form of GA(15), shown by the labeling studies, is consistent with the implied use of a further oxygenated heterocycle in the final conversion of GA(24) into GA(9), with the partial labeling of GA(9), thus demonstrating that CYP112 partitions its reactants between two diverging mechanisms.