Cloning and characterization of farnesyl pyrophosphate synthase from the highly branched isoprenoid producing diatom Rhizosolenia setigera

• Published in: Scientific reports 2015-05, Vol.5 (1), p.10246-10246, Article 10246
• Main Authors: Ferriols, Victor Marco Emmanuel N., Yaginuma, Ryoko, Adachi, Masao, Takada, Kentaro, Matsunaga, Shigeki, Okada, Shigeru
• Format: Article
• Language: English
• Subjects: 101/58; 38/22; 38/39; 38/77; 631/45/147; 631/45/607/1172; 82/80; 82/83; Amino Acid Sequence; Amino acids; Biosynthesis; Bisphosphonates; Cloning; Cloning, Molecular; Diatoms - classification; Diatoms - metabolism; DNA, Complementary - metabolism; E coli; Escherichia coli - metabolism; Farnesyl pyrophosphate synthase; Gas chromatography; Gas Chromatography-Mass Spectrometry; Gene expression; Geranyltranstransferase - genetics; Geranyltranstransferase - metabolism; Humanities and Social Sciences; Hydrocarbons; Isomerism; Isomers; Life Cycle Stages; Life cycles; Marine environment; Mass spectrometry; Mass spectroscopy; Molecular Sequence Data; multidisciplinary; Phylogeny; Plankton; Polyisoprenyl Phosphates - analysis; Polyisoprenyl Phosphates - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Risedronic acid; Science; Sequence Alignment; Sesquiterpenes - analysis; Sesquiterpenes - metabolism; Terpenes - chemistry; Terpenes - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep10246

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 C 25 and C 30 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 Rs FPPS 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.