The carotenoid cleavage dioxygenase 1 enzyme has broad substrate specificity, cleaving multiple carotenoids at two different bond positions.
In many organisms, various enzymes mediate site-specific carotenoid cleavage to generate biologically active apocarotenoids. These carotenoid-derived products include provitamin A, hormones, and flavor and fragrance molecules. In plants, the CCD1 enzyme cleaves carotenoids at 9,10 (9',10') bonds to generate multiple apocarotenoid products. Here we systematically analyzed volatile apocarotenoids generated by maize CCD1 (ZmCCD1) from multiple carotenoid substrates. ZmCCD1 did not cleave geranylgeranyl diphosphate or phytoene but did cleave other linear and cyclic carotenoids, producing volatiles derived from 9,10 (9',10') bond cleavage. Additionally the Arabidopsis, maize, and tomato CCD1 enzymes all cleaved lycopene to generate 6-methyl-5-hepten-2-one. 6-Methyl-5-hepten-2-one, an important flavor volatile in tomato, was produced by cleavage of the 5,6 or 5',6' bond positions of lycopene but not geranylgeranyl diphosphate, zeta-carotene, or phytoene. In vitro, ZmCCD1 cleaved linear and cyclic carotenoids with equal efficiency. Based on the pattern of apocarotenoid volatiles produced, we propose that CCD1 recognizes its cleavage site based on the saturation status between carbons 7 and 8 (7' and 8') and carbons 11 and 12 (11' and 12') as well as the methyl groups on carbons 5, 9, and 13 (5', 9', and 13').
Vogel
Jonathan T
JT
Horticultural Sciences Department and the Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32611, USA.
Tan
Bao-Cai
BC
McCarty
Donald R
DR
Klee
Harry J
HJ
D001665:Binding Sites; D002338:Carotenoids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000737:chemistry / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D000077276:Lycopene; D008954:Models, Biological; D008956:Models, Chemical; D010940:Plant Proteins / Q000737:chemistry; D011487:Protein Conformation; D013379:Substrate Specificity; D003313:Zea mays / Q000201:enzymology
D010940:Plant Proteins; D002338:Carotenoids; C100185:(all-E) phytoene; D049308:Dioxygenases; C431939:carotenoid cleavage dioxygenase 1; D000077276:Lycopene