Molecular cloning and functional analysis of the gene encoding geranylgeranyl diphosphate synthase from Jatropha curcas

Abstract

Geranylgeranyl diphosphate (GGPP) synthase (GGPPS, EC: 2.5.1.29) catalyzes the condensation of isopentenyl diphosphate (IPP) with allylic diphosphates to yield (all-E)-GGPP. GGPP is one of the key
precursors in the biosynthesis of biologically significant isoprenoid compounds. Here we report for the first time the cloning of a full-length cDNA encoding GGPPS (Jc-GGPPS) from Jatropha curcas L. The full-length cDNA was 1414 base pair (bp), with an 1110-bp open reading frame (ORF) encoding a 370- amino-acids polypeptide. Bioinformatic analysis revealed that Jc-GGPPS is a member of the polyprenyltransferases with two highly conserved aspartate-rich motifs, and showed high homology to other plant GGPPSs. Phylogenetic tree analysis indicated that plant GGPPSs could be classified into two groups, angiosperm and gymnosperm, while Jc-GGPPS had closer relationship with angiosperm plant GGPPSs. Functional analysis of Jc-GGPPS in GGPPS-deficient mutant plasmid pAC-BETA (³ crtE)
demonstrated that Jc-GGPPS mediated the biosynthesis of carotenoid and provided the general precursor for diterpenes biosynthesis.