title: Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce.
abstract: 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.
authors.affiliation: United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Basyuni
authors.forename: Hirosuke
authors.initials: H
authors.lastname: Oku
authors.forename: Masashi
authors.initials: M
authors.lastname: Inafuku
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Baba
authors.forename: Hironori
authors.initials: H
authors.lastname: Iwasaki
authors.forename: Keichiro
authors.initials: K
authors.lastname: Oshiro
authors.forename: Takafumi
authors.initials: T
authors.lastname: Okabe
authors.forename: Masaaki
authors.initials: M
authors.lastname: Shibuya
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Ebizuka
mesh_terms: 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*
chemical_list: D018076:DNA, Complementary; D010940:Plant Proteins; D014315:Triterpenes