Cloning and functional characterization of the germacradienol synthase (spterp13) from Streptomyces peucetius ATCC 27952.
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.
Ghimire
Gopal Prasad
GP
Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, SunMoon University, Asan, Korea.
Oh
Tae-Jin
TJ
Lee
Hei Chan
HC
Kim
Byung-Gee
BG
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009284:Naphthols / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013302:Streptomyces / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D009284:Naphthols; D011994:Recombinant Proteins; C515287:germacradienol