Substrate promiscuity of RdCCD1, a carotenoid cleavage oxygenase from Rosa damascena.
Several of the key flavor compounds in rose essential oil are C(13)-norisoprenoids, such as beta-damascenone, beta-damascone, and beta-ionone which are derived from carotenoid degradation. To search for genes putatively responsible for the cleavage of carotenoids, cloning of carotenoid cleavage (di-)oxygenase (CCD) genes from Rosa damascena was carried out by a degenerate primer approach and yielded a full-length cDNA (RdCCD1). The RdCCD1 gene was expressed in Escherichia coli and recombinant protein was assayed for its cleavage activity with a multitude of carotenoid substrates. The RdCCD1 protein was able to cleave a variety of carotenoids at the 9-10 and 9'-10' positions to produce a C(14) dialdehyde and two C(13) products, which vary depending on the carotenoid substrates. RdCCD1 could also cleave lycopene at the 5-6 and 5'-6' positions to produce 6-methyl-5-hepten-2-one. Expression of RdCCD1 was studied by real-time PCR in different tissues of rose. The RdCCD1 transcript was present predominantly in rose flower, where high levels of volatile C(13)-norisoprenoids are produced. Thus, the accumulation of C(13)-norisoprenoids in rose flower is correlated to the expression of RdCCD1.
Huang
Fong-Chin
FC
Technische Universität München, Hochfeldweg 1, D-85354 Freising, Germany.
Horváth
Györgyi
G
Molnár
Péter
P
Turcsi
Erika
E
Deli
József
J
Schrader
Jens
J
Sandmann
Gerhard
G
Schmidt
Holger
H
Schwab
Wilfried
W
D002338:Carotenoids / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D035264:Flowers / Q000201:enzymology; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D027826:Rosa / Q000201:enzymology* / Q000235:genetics; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity
D010940:Plant Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D002338:Carotenoids; D010105:Oxygenases; C500451:carotenoid oxygenase