Cloning and Functional Characterization of the Germacradienol Synthase (spterp13) from Streptomyces peucetius ATCC 27952

  • Ghimire, Gopal Prasad (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, SunMoon University) ;
  • Oh, Tae-Jin (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, SunMoon University) ;
  • Lee, Hei-Chan (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, SunMoon University) ;
  • Kim, Byung-Gee (School of Chemical and Biological Engineering, Seoul National University) ;
  • Sohng, Jae-Kyung (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, SunMoon University)
  • Published : 2008.07.31

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 MA4680 and 65% identity with SCO6073 from S. coelicolor A3(2), which produces germacradienol/geosmin. The full-length recombinant protein was heterologously expressed as a his-tagged fusion protein in Escherichia coli, purified, and shown to catalyze the $Mg^{2+}$-dependent conversion of farnesyl diphosphate to the germacradienol, which was verified by gas chromatography/mass spectrometry.

Keywords

References

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