Expression and Characterization of Mammalian Carotenoid Cleavage Dioxygenases.

Carotenoid cleavage dioxygenases (CCDs) are nonheme iron enzymes that catalyze double bond processing of carotenoids and their apocarotenoid metabolites. Mammalian genomes encode three members of this protein family, namely BCO1, BCO2, and RPE65. Mutations and genetic polymorphism in the corresponding genes are associated with inherited blinding diseases, vitamin A deficiency, and high carotenoid plasma levels. Here we describe a method for the heterologous expression of mammalian BCO1 and BCO2 in E. coli and the biochemical characterization of these recombinant enzymes. Dissecting the enzymatic properties of CCDs will advance our knowledge of the biochemical processes that are govern by these disease-associated enzymes and may assist the design of interventions directed against these disease states.