A New Farnesyl Diphosphate Synthase Gene fromTaxus mediaRehder: Cloning, Characterization and Functional Complementation

Abstract

Farnesyl diphosphate synthase (FPS; EC 2.5.1.10) catalyzes the production of 15-carbon farnesyl diphosphate which is a branch-point intermediate for many terpenoids. This reaction is considered to be a ratelimiting step in terpenoid biosynthesis. Here we report for the first time the cloning of a new full-length cDNA encoding farnesyl diphosphate synthase from a gymnosperm plant species, Taxus media Rehder, designated as TmFPS1. The full-length cDNA of TmFPS1 (GenBank accession number: AY461811) was 1 464 bp with a 1 056-bp open reading frame encoding a 351-amino acid polypeptide with a calculated molecular weight of 40.3 kDa and a theoretical pI of 5.07. Bioinformatic analysis revealed that TmFPS1 contained all five conserved domains of prenyltransferases, and showed homology to other FPSs of plant origin. Phylogenetic analysis showed that farnesyl diphosphate synthases can be divided into two groups: one of prokaryotic origin and the other of eukaryotic origin. TmFPS1 was grouped with FPSs of plant origin. Homologybased structural modeling showed that TmFPS1 had the typical spatial structure of FPS, whose most prominent structural feature is the arrangement of 13 core helices around a large central cavity in which the catalytic reaction takes place. Our bioinformatic analysis strongly suggests that TmFPS1 is a functional gene. Southern blot analysis revealed that TmFPS1 belongs to a small FPS gene family in T. media. Northern blot analysis indicated that TmFPS1 is expressed in all tested tissues, including the needles, stems and roots of T. media. Subsequently, functional complementation with TmFPS1 in a FPS-deficient mutant yeast demonstrated that TmFPS1 did encode farnesyl diphosphate synthase, which rescued the yeast mutant. This study will be helpful in future investigations aiming at understanding the detailed role of FPS in terpenoid biosynthesis flux control at the molecular genetic level.

Differential expression of farnesyl diphosphate synthase gene from Withania somnifera in different chemotypes and in response to elicitors

“…FPPS catalyzed reaction occurs in two consecutive steps; condensation of isopentanyl diphosphate (IPP) with dimethylallyl diphosphate (DMAPP) to form 10-C intermediate geranyl diphosphate (GPP) and condensation of GPP with another molecule of IPP which results into 15-C FPP. This gene has been characterized from variety of plant species including Arabidopsis (Closa et al 2010), Artemisia (Matsushita et al 1996), Hevea (Takaya et al 2003), maize (Cervantes et al 2006), rice (Sanmiya et al 1997) and Taxus media (Liao et al 2006). It has been shown that overexpression of ginseng farnesyl diphosphate synthase in Centella asiatica hairy roots enhanced phytosterol and triterpene biosynthesis .…”

Engineering the MEP pathway enhanced ajmalicine biosynthesis

“…roseus improved catharanthine production . The plastidial MEP pathway provides the general acyclic 5‐carbon isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) for biosynthesis of terpenoids, especially monoterpenoids and diterpenoids . Ajmalicine belongs to the family of monoterpene indole alkaloids, so the MEP pathway also provides its precursors theoretically.…”

Molecular Cloning and Characterisation of Farnesyl Pyrophosphate Synthase from Tripterygium wilfordii

“…FPS plays a key role [ 30 – 32 ] in the catalytic reaction in isoprenoid biosynthesis; this step is considered rate-limiting. In the present study, we reported the molecular characterisation of two FPS genes from T .…”

“…wilfordii . The analysis of FPS gene expression patterns in various plants demonstrated not only that the gene expression is tissue-specific [ 33 – 34 ] but also that the isoprenoid derivative contents increase [ 20 , 30 , 35 – 38 ]. The gene expression analysis showed that FPS genes could increase the isoprenoid substance contents in the plants [ 39 – 42 ].…”