Flavonoids are common colorants in plants and have long-established biomedicinal properties. Anthocyanidin synthase (ANS), a 2-oxoglutarate iron-dependent oxygenase, catalyzes the penultimate step in the biosynthesis of the anthocyanin class of flavonoids. The crystal structure of ANS reveals a multicomponent active site containing metal, cosubstrate, and two molecules of a substrate analog (dihydroquercetin). An additional structure obtained after 30 min exposure to dioxygen is consistent with the oxidation of the dihydroquercetin to quercetin and the concomitant decarboxylation of 2-oxoglutarate to succinate. Together with in vitro studies, the crystal structures suggest a mechanism for ANS-catalyzed anthocyanidin formation from the natural leucoanthocyanidin substrates involving stereoselective C-3 hydroxylation. The structure of ANS provides a template for the ubiquitous family of plant nonhaem oxygenases for future engineering and inhibition studies.

Dioxygenases are nonheme iron-containing enzymes important in the biosynthesis of plant signaling compounds such as abscisic acid, gibberellins, and ethylene and also of secondary metabolites, notably flavonoids and alkaloids. Plant dioxygenases fall into two classes: lipoxygenases and 2-oxoacid-dependent dioxygenases. The latter catalyze hydroxylation, epoxidation, and desaturation reactions; some enzymes catalyze more than one type of reaction in successive steps in a biosynthetic pathway. This review highlights recent discoveries on both enzyme groups, particularly in relation to gibberellin biosynthesis, in vivo activity of 1-aminocyclopropane-1-carboxylate oxidase, and molecular structure/function relationships. Similarities between the roles of monooxygenases and dioxygenases are also discussed.

SummaryThe intensity and pattern of anthocyanin biosynthesis in Antirrhinum flowers is controlled by several genes. We have isolated six cDNA clones encoding enzymes in the pathway committed to flavonoid biosynthesis and used these to assay how the regulatory genes that modify colour pattern affect the expression of biosynthetic genes. The biosynthetic genes of the later part of the pathway appear to be co-ordinately regulated by two genes, Delila (Del), and Eluta ( N ) , while the early steps (which also lead to flavone synthesis) are controlled differently. This division of control is not the same as control of anthocyanin biosynthesis by the regulatory genes R(S) and C1 in maize aleurone, and may result from the adaptive significance of different flavonoids in flowers and seeds, reflecting their attractiveness to insects and mammals respectively. €1 and de/ are probably involved in transcriptional control and both genes appear to be able to repress expression of some biosynthetic genes and activate expression of others.