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KMID : 0545120080180071216
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 7 p.1216 ~ p.1220
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Cloning and Functional Characterization of the Germacradienol Synthase (spterp13) from Streptomyces peucetius ATCC 27952
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Ghimire Gopal Prasad
Oh Tae-Jin
Lee Hei-Chan
Kim Byung-Gee
Sohng Jae-Kyung
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Abstract
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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.
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KEYWORD
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Geosmin, germacradienol, heterologous expression, sesquiterpene cyclase, Streptomyces peucetius.
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