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dc.contributor.authorE. Slessor, Kateen_US
dc.contributor.authorB. Hawkes, Daviden_US
dc.contributor.authorFarlow, Anthonyen_US
dc.contributor.authorG. Pearson, Andrewen_US
dc.contributor.authorE. Stok, Jeanetteen_US
dc.contributor.authorJ. De Voss, Jamesen_US
dc.contributor.editorY. Asano, J.D. Stewart, F. Hollmannen_US
dc.date.accessioned2017-05-03T13:03:37Z
dc.date.available2017-05-03T13:03:37Z
dc.date.issued2012en_US
dc.date.modified2013-06-11T04:32:55Z
dc.identifier.issn13811177en_US
dc.identifier.doi10.1016/j.molcatb.2012.03.007en_US
dc.identifier.urihttp://hdl.handle.net/10072/47670
dc.description.abstractCytochrome P450cin (CYP176A1) is a bacterial P450 isolated from Citrobacter braakii that catalyses the hydroxylation of 1,8-cineole to (1R)-6 -hydroxycineole. P450cin uses two redox partners in vitro for catalysis: cindoxin, its physiological FMN-containing redox partner, and Escherichia coli flavodoxin reductase. Here we report the construction of a tricistronic plasmid that expresses P450cin, cindoxin and E. coli flavodoxin reductase and a bicistronic plasmid that encodes only P450cin and cindoxin. E. coli transformed with the bicistronic vector effectively catalysed the oxidation of 1,8-cineole, with the endogenous E. coli flavodoxin reductase presumably acting as the terminal electron transfer protein. This in vivo system was capable of producing enantiomerically pure (1R)-6 -hydroxycineole in yields of ~1 g/L culture, thus providing a simple, one-step synthesis of this compound. In addition, the metabolism of (1R)- and (1S)-camphor, structural homologues of 1,8-cineole was also evaluated in order to investigate the ability of this in vivo system to produce compounds for mechanistic studies. Significant quantities of five of the six possible secondary alcohols arising from methylene oxidation of both (1R)- and (1S)-camphor were isolated and structurally characterised. The similarity of the (1R)- and (1S)-camphor product profiles highlight the importance of the inherent reactivity of the substrate in determining the regiochemistry of oxidation in the absence of any specific enzyme-substrate binding interactions.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_US
dc.format.extent309340 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglishen_US
dc.language.isoen_US
dc.publisherElsevieren_US
dc.publisher.placeNetherlandsen_US
dc.relation.ispartofstudentpublicationNen_US
dc.relation.ispartofpagefrom15en_US
dc.relation.ispartofpageto20en_US
dc.relation.ispartofjournalJournal of Molecular Catalysis B: Enzymaticen_US
dc.relation.ispartofvolume79en_US
dc.rights.retentionYen_US
dc.subject.fieldofresearchBiologically Active Moleculesen_US
dc.subject.fieldofresearchcode030401en_US
dc.titleAn in vivo cytochrome P450cin (CYP176A1) catalytic system for metabolite productionen_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.rights.copyrightCopyright 2012 Elsevier. This is the author-manuscript version of this paper. Reproduced in accordance with the copyright policy of the publisher. Please refer to the journal's website for access to the definitive, published version.en_US
gro.date.issued2012
gro.hasfulltextFull Text


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