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Article: Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis

TitleExpression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis
Authors
Issue Date2003
PublisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.org
Citation
Proceedings Of The National Academy Of Sciences Of The United States Of America, 2003, v. 100 n. 4, p. 1547-1551 How to Cite?
AbstractThe PCR has been used to amplify a 2,181-bp ORF from Streptomyces coelicolor A3(2), designated SC9B1.20 (= SCO6073), encoding a protein of 726 amino acids and showing significant sequence similarity at the deduced amino acid level in both the N-terminal and C-terminal halves to the known sesquiterpene synthase pentalenene synthase. The full-length recombinant protein was expressed at high levels in Escherichia coli and shown to catalyze the Mg2+-dependent conversion of farnesyl diphosphate to the sesquiterpene alcohol (4S, 7R)-germacra-1 (10)E, 5E-diene-11-ol. The enzymatic cyclization had a kcat of 6.2 ± 0.5 × 10-3 s-1 and a Km for farnesyl diphosphate of 62 ± 8 nM. Expression of the N-terminal (366 amino acids) domain of the SC9B1.20 protein also gave a fully functional cyclase which converted farnesyl diphosphate to the identical sesquiterpene alcohol with a slightly lower kcat of 3.2 ± 0.4 × 10-3 s-1 and a twofold greater km of 115 ± 14 nM. By contrast, the expressed C-terminal domain of SC9B1.20 had no farnesyl diphosphate cyclase activity. The formation of the germacradienol seems to be the committed step in the formation of geosmin, the characteristic odoriferous constituent of Streptomyces species.
Persistent Identifierhttp://hdl.handle.net/10722/154230
ISSN
2015 Impact Factor: 9.423
2015 SCImago Journal Rankings: 6.883
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorCane, DEen_US
dc.contributor.authorWatt, RMen_US
dc.date.accessioned2012-08-08T08:24:05Z-
dc.date.available2012-08-08T08:24:05Z-
dc.date.issued2003en_US
dc.identifier.citationProceedings Of The National Academy Of Sciences Of The United States Of America, 2003, v. 100 n. 4, p. 1547-1551en_US
dc.identifier.issn0027-8424en_US
dc.identifier.urihttp://hdl.handle.net/10722/154230-
dc.description.abstractThe PCR has been used to amplify a 2,181-bp ORF from Streptomyces coelicolor A3(2), designated SC9B1.20 (= SCO6073), encoding a protein of 726 amino acids and showing significant sequence similarity at the deduced amino acid level in both the N-terminal and C-terminal halves to the known sesquiterpene synthase pentalenene synthase. The full-length recombinant protein was expressed at high levels in Escherichia coli and shown to catalyze the Mg2+-dependent conversion of farnesyl diphosphate to the sesquiterpene alcohol (4S, 7R)-germacra-1 (10)E, 5E-diene-11-ol. The enzymatic cyclization had a kcat of 6.2 ± 0.5 × 10-3 s-1 and a Km for farnesyl diphosphate of 62 ± 8 nM. Expression of the N-terminal (366 amino acids) domain of the SC9B1.20 protein also gave a fully functional cyclase which converted farnesyl diphosphate to the identical sesquiterpene alcohol with a slightly lower kcat of 3.2 ± 0.4 × 10-3 s-1 and a twofold greater km of 115 ± 14 nM. By contrast, the expressed C-terminal domain of SC9B1.20 had no farnesyl diphosphate cyclase activity. The formation of the germacradienol seems to be the committed step in the formation of geosmin, the characteristic odoriferous constituent of Streptomyces species.en_US
dc.languageengen_US
dc.publisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.orgen_US
dc.relation.ispartofProceedings of the National Academy of Sciences of the United States of Americaen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshDna Primersen_US
dc.subject.meshGas Chromatography-Mass Spectrometryen_US
dc.subject.meshLigases - Chemistry - Genetics - Metabolismen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshNaphthols - Metabolismen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshStreptomyces - Enzymology - Metabolismen_US
dc.titleExpression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesisen_US
dc.typeArticleen_US
dc.identifier.emailWatt, RM:rmwatt@hku.hken_US
dc.identifier.authorityWatt, RM=rp00043en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1073/pnas.0337625100en_US
dc.identifier.pmid12556563-
dc.identifier.scopuseid_2-s2.0-0037452696en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037452696&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume100en_US
dc.identifier.issue4en_US
dc.identifier.spage1547en_US
dc.identifier.epage1551en_US
dc.identifier.isiWOS:000181073000021-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridCane, DE=7103058020en_US
dc.identifier.scopusauthoridWatt, RM=7102907536en_US

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