Cloning and functional characterization of the germacradienol synthase (spterp13) from Streptomyces peucetius ATCC 27952

J Microbiol Biotechnol. 2008 Jul;18(7):1216-20.

Abstract

Sequence analysis of the metabolically rich genome of Streptomyces peucetius ATCC 27952 revealed a 2,199 bp sesquiterpene alcohol (germacradienol) synthase-encoding gene from the germacradienol synthase/terpene cyclase gene cluster. The gene was named spterp13, and its putative function is as a germacradienol synthase/terpene cyclase. The amino acid sequence of Spterp13 shows 66% identity with SAV2163 (GeoA) from S. avermitilis MA- 4680 and 65% identity with SCO6073 from S. coelicolor A3(2), which produces germacradienol/geosmin. The fulllength recombinant protein was heterologously expressed as a his-tagged fusion protein in Escherichia coli, purified, and shown to catalyze the Mg2+-dependent conversion of farnesyl diphosphate to the germacradienol, which was verified by gas chromatography/mass spectrometry.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism
  • Cloning, Molecular*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Molecular Sequence Data
  • Molecular Weight
  • Naphthols / metabolism*
  • Phylogeny
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Streptomyces / chemistry
  • Streptomyces / classification
  • Streptomyces / enzymology*
  • Streptomyces / genetics

Substances

  • Bacterial Proteins
  • Naphthols
  • Recombinant Proteins
  • germacradienol

Associated data

  • GENBANK/ABY50951