File Download
There are no files associated with this item.
Links for fulltext
(May Require Subscription)
- Publisher Website: 10.1073/pnas.0337625100
- Scopus: eid_2-s2.0-0037452696
- PMID: 12556563
- WOS: WOS:000181073000021
- Find via
Supplementary
- Citations:
- Appears in Collections:
Article: Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis
Title | Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis |
---|---|
Authors | |
Keywords | Secondary metabolism Sesquiterpene synthase Streptomyces gene |
Issue Date | 2003 |
Publisher | National 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? |
Abstract | The 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 Identifier | http://hdl.handle.net/10722/154230 |
ISSN | 2023 Impact Factor: 9.4 2023 SCImago Journal Rankings: 3.737 |
ISI Accession Number ID | |
References |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Cane, DE | en_US |
dc.contributor.author | Watt, RM | en_US |
dc.date.accessioned | 2012-08-08T08:24:05Z | - |
dc.date.available | 2012-08-08T08:24:05Z | - |
dc.date.issued | 2003 | en_US |
dc.identifier.citation | Proceedings Of The National Academy Of Sciences Of The United States Of America, 2003, v. 100 n. 4, p. 1547-1551 | en_US |
dc.identifier.issn | 0027-8424 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/154230 | - |
dc.description.abstract | The 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.language | eng | en_US |
dc.publisher | National Academy of Sciences. The Journal's web site is located at http://www.pnas.org | en_US |
dc.relation.ispartof | Proceedings of the National Academy of Sciences of the United States of America | en_US |
dc.subject | Secondary metabolism | - |
dc.subject | Sesquiterpene synthase | - |
dc.subject | Streptomyces gene | - |
dc.subject.mesh | Amino Acid Sequence | en_US |
dc.subject.mesh | Base Sequence | en_US |
dc.subject.mesh | Dna Primers | en_US |
dc.subject.mesh | Gas Chromatography-Mass Spectrometry | en_US |
dc.subject.mesh | Ligases - Chemistry - Genetics - Metabolism | en_US |
dc.subject.mesh | Molecular Sequence Data | en_US |
dc.subject.mesh | Naphthols - Metabolism | en_US |
dc.subject.mesh | Polymerase Chain Reaction | en_US |
dc.subject.mesh | Streptomyces - Enzymology - Metabolism | en_US |
dc.title | Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis | en_US |
dc.type | Article | en_US |
dc.identifier.email | Watt, RM:rmwatt@hku.hk | en_US |
dc.identifier.authority | Watt, RM=rp00043 | en_US |
dc.description.nature | link_to_subscribed_fulltext | en_US |
dc.identifier.doi | 10.1073/pnas.0337625100 | en_US |
dc.identifier.pmid | 12556563 | - |
dc.identifier.scopus | eid_2-s2.0-0037452696 | en_US |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-0037452696&selection=ref&src=s&origin=recordpage | en_US |
dc.identifier.volume | 100 | en_US |
dc.identifier.issue | 4 | en_US |
dc.identifier.spage | 1547 | en_US |
dc.identifier.epage | 1551 | en_US |
dc.identifier.isi | WOS:000181073000021 | - |
dc.publisher.place | United States | en_US |
dc.identifier.scopusauthorid | Cane, DE=7103058020 | en_US |
dc.identifier.scopusauthorid | Watt, RM=7102907536 | en_US |
dc.identifier.issnl | 0027-8424 | - |