Cloning and functional expression of cycloartenol synthases from mangrove species Rhizophora stylosa Griff. and Kandelia candel (L.) Druce.
To obtain cDNAs encoding oxidosqualene cyclase (OSC), we cloned two cDNAs, KcCAS and RsCAS, from roots of Kandelia candel (L.) Druce and leaves of Rhizophora stylosa Griff. by homology based PCR method respectively. The deduced amino acid sequences of both OSCs showed 82% homology to cycloartenol synthases from Lotus japonicus (OSC5) and Ricinus cummunis (RcCAS), suggesting that these are cycloartenol synthases of K. candel and R. stylosa. The genes obtained were expressed in a lanosterol synthase deficient Saccharomyces cerevisiae (ERG7) strain, GIL77. GC-MS analysis identified the accumulated reaction product in the yeast transformant to be cycloartenol, indicating that both KcCAS and RsCAS encode cycloartenol synthase.
Basyuni
Mohammad
M
United Graduate School of Agricultural Sciences, Kagoshima University, Korimoto, Kagoshima, Japan.
Oku
Hirosuke
H
Tsujimoto
Etsuko
E
Baba
Shigeyuki
S
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D019751:Intramolecular Transferases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D043982:Rhizophoraceae / Q000201:enzymology* / Q000235:genetics; D012441:Saccharomyces cerevisiae
D019751:Intramolecular Transferases; C021923:cycloartenol synthase