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PMID:25996801
| Citation |
Ferriols, VM, Yaginuma, R, Adachi, M, Takada, K, Matsunaga, S and Okada, S (2015) Cloning and characterization of farnesyl pyrophosphate synthase from the highly branched isoprenoid producing diatom Rhizosolenia setigera. Sci Rep 5:10246 |
|---|---|
| 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. |
| Links |
PubMed PMC4440519 Online version:10.1038/srep10246 |
| Keywords |
Amino Acid Sequence; Cloning, Molecular; DNA, Complementary/metabolism; Diatoms/classification; Diatoms/metabolism; Escherichia coli/metabolism; Gas Chromatography-Mass Spectrometry; Geranyltranstransferase/genetics; Geranyltranstransferase/metabolism; Isomerism; Life Cycle Stages; Molecular Sequence Data; Phylogeny; Polyisoprenyl Phosphates/analysis; Polyisoprenyl Phosphates/metabolism; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; Sequence Alignment; Sesquiterpenes/analysis; Sesquiterpenes/metabolism; Terpenes/chemistry; Terpenes/metabolism |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
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Contributes to |
GO:0102059: 2-cis,6-cis-farnesyl pyrophosphate synthase activity |
ECO:0000325: chromatography evidence |
F |
Figure 6. GC/MS total ion chromatogram of enzyme reaction products using IPP and DMAPP as substrates. Upper panels show prominent peaks corresponding to A) geraniol (GOH) and B) E,E-farnesol (E,E-FOH) with minor peaks for nerol (NOH) and Z,E-farnesol (Z,E-FOH) marked with asterisks (*). Lower panels are magnified images of the boxed areas in the upper panels. Other minor peaks in the upper panels are either contaminants or artefacts of treatment with alkaline phosphatase as determined in control alkaline phosphatase reactions using GPP or E,E-FPP standards. |
complete | |||
Notes
See also
References
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