Host: The Japanese Society of Plant Physiologists
Pages 0040
Latex of a succulent shrub Euphorbia tirucalli includes triterpens, sterol, or other secondary metabolites, and can be processed into biofuel, or medicinal and insecticidal compounds. To isolate a homologue gene for a mevalonate pathway key enzyme, squalene synthase, we performed RT- and genomic PCRs to reveal putative ORF encodes 411 residues of amino-acid sequence, in which 380-residues of N-terminal region exhibits high homology with those from other organisms, while 31-residues of C-terminal hydrophobic region was less homologous. The cDNA fragment which encodes the N-terminal 380-residue sequence was subcloned into a bacterial expression vector, pET32b, and the resultant vector was overexpressed in E. coli BL21(DE3). Using soluble fraction of the bacterial proteins, FPP and NADPH, in vitro squalene synthase activity was being examined by GC-MS. This work was partly performed as one of the technology development projects of the "Green Biotechnology Program" in NEDO.
