Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce.
Homology based PCRs with degenerate primers designed from the conserved sequences among the known oxidosqualene cylases (OSCs) have resulted in cloning of a triterpene synthase (KcMS) from the young roots of Kandelia candel (L.) Druce (Rhizophoraceae). KcMS consists of a 2286 bp open reading frame, which codes for 761 amino acids. The deduced amino acid sequence showed 79% homology to a lupeol synthase from Ricinus communis suggesting it to be a lupeol synthase of K. candel. KcMS was expressed in a lanosterol synthase deficient yeast with the expression vector pYES2 under the control of GAL1 promoter. GC-MS analysis showed that the transformant accumulated a mixture of lupeol, beta-amyrin and alpha-amyrin in a 2:1:1 ratio, indicating that KcMS encodes a multifunctional triterpene synthase, although it showed high sequence homology to a R. communis lupeol synthase. This is the first OSC cloning from mangrove tree species.
Basyuni
Mohammad
M
United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
Oku
Hirosuke
H
Inafuku
Masashi
M
Baba
Shigeyuki
S
Iwasaki
Hironori
H
Oshiro
Keichiro
K
Okabe
Takafumi
T
Shibuya
Masaaki
M
Ebizuka
Yutaka
Y
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D043982:Rhizophoraceae / Q000201:enzymology* / Q000235:genetics*; D014315:Triterpenes / Q000737:chemistry / Q000378:metabolism*
D018076:DNA, Complementary; D010940:Plant Proteins; D014315:Triterpenes