title: Cloning and characterization of farnesyl pyrophosphate synthase from the highly branched isoprenoid producing diatom Rhizosolenia setigera.
abstract: The diatom Rhizosolenia setigera Brightwell produces highly branched isoprenoid (HBI) hydrocarbons that are ubiquitously present in marine environments. The hydrocarbon composition of R. setigera varies between C25 and C30 HBIs depending on the life cycle stage with regard to auxosporulation. To better understand how these hydrocarbons are biosynthesized, we characterized the farnesyl pyrophosphate (FPP) synthase (FPPS) enzyme of R. setigera. An isolated 1465-bp cDNA clone contained an open reading frame spanning 1299-bp encoding a protein with 432 amino acid residues. Expression of the RsFPPS cDNA coding region in Escherichia coli produced a protein that exhibited FPPS activity in vitro. A reduction in HBI content from diatoms treated with an FPPS inhibitor, risedronate, suggested that RsFPPS supplies precursors for HBI biosynthesis. Product analysis by gas chromatography-mass spectrometry also revealed that RsFPPS produced small amounts of the cis-isomers of geranyl pyrophosphate and FPP, candidate precursors for the cis-isomers of HBIs previously characterized. Furthermore, RsFPPS gene expression at various life stages of R. setigera in relation to auxosporulation were also analyzed. Herein, we present data on the possible role of RsFPPS in HBI biosynthesis, and it is to our knowledge the first instance that an FPPS was cloned and characterized from a diatom.
authors.affiliation: 1] Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan [2] Institute of Aquaculture, University of the Philippines Visayas, Philippines.
authors.forename: Victor Marco Emmanuel N
authors.initials: VM
authors.lastname: Ferriols
authors.affiliation: Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Yaginuma
authors.affiliation: Faculty of Agriculture, Kochi University, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Adachi
authors.affiliation: Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Takada
authors.affiliation: Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Matsunaga
authors.affiliation: Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Okada
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D017377:Diatoms / Q000145:classification / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D051228:Geranyltranstransferase / Q000235:genetics / Q000378:metabolism*; D007536:Isomerism; D008018:Life Cycle Stages; D008969:Molecular Sequence Data; D010802:Phylogeny; D011106:Polyisoprenyl Phosphates / Q000032:analysis / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D016415:Sequence Alignment; D012717:Sesquiterpenes / Q000032:analysis / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D011106:Polyisoprenyl Phosphates; D011994:Recombinant Proteins; D012717:Sesquiterpenes; D013729:Terpenes; C015234:geranyl pyrophosphate; C004808:farnesyl pyrophosphate; D051228:Geranyltranstransferase