Ethylene biosynthesis: processing of a substrate analog supports a radical mechanism for the ethylene-forming enzyme

Background: The chemical mechanism of the final step of ethylene biosynthesis (the conversion of 1-aminocyclopropanecarboxylic acid, ACC, to ethylene by ACC oxidase, the ethylene-forming enzyme, EFE) is poorly understood. Two possibilities have been suggested: a radical mechanism and an N-hydroxylation mechanism. We investigated reaction pathways available to radical intermediates in this reaction using an ACC analog, 1-aminocyclobutanecarboxylic acid (ACBC) as a substrate. Results: ACBC was converted to dehydroproline (Delta(1)-pyrroline-2-carboxylic acid) by the EFE via a ring expansion process. The possibility that an N-hydroxyaminoacid (produced during two-electron oxidation) acts as an intermediate in this process was eliminated by control experiments. Chemical model reactions involving two-electron oxidants, such as a positive halogen (X+), which presumably generate N-halo derivatives, produce only decarboxylation products. Radical-based oxidants, in contrast, generate dehydroproline. Model reactions involving sequential single-electron transfer mechanisms also produce dehydroproline; thus our results support the proposal that the EFE-catalyzed step of ethylene biosynthesis proceeds using a radical-based mechanism. Conclusions: Our results provide support for a radical mechanism in the final step of ethylene biosynthesis and refute an alternative N-hydroxylation mechanism. This work extends the idea that the intrinsic chemical reactivity of a high energy iron-ore intermediate can account for the observed products in ethylene biosynthesis.