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dc.contributor.authorSlessor, Kate E
dc.contributor.authorHawkes, David B
dc.contributor.authorFarlow, Anthony
dc.contributor.authorPearson, Andrew G
dc.contributor.authorStok, Jeanette E
dc.contributor.authorDe Voss, James J
dc.contributor.editorY. Asano, J.D. Stewart, F. Hollmann
dc.date.accessioned2017-05-03T13:03:37Z
dc.date.available2017-05-03T13:03:37Z
dc.date.issued2012
dc.date.modified2013-06-11T04:32:55Z
dc.identifier.issn1381-1177
dc.identifier.doi10.1016/j.molcatb.2012.03.007
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.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.format.extent309340 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglish
dc.language.isoeng
dc.publisherElsevier
dc.publisher.placeNetherlands
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom15
dc.relation.ispartofpageto20
dc.relation.ispartofjournalJournal of Molecular Catalysis B: Enzymatic
dc.relation.ispartofvolume79
dc.rights.retentionY
dc.subject.fieldofresearchBiologically active molecules
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchcode340401
dc.subject.fieldofresearchcode3101
dc.titleAn in vivo cytochrome P450cin (CYP176A1) catalytic system for metabolite production
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.rights.copyright© 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.
gro.date.issued2012
gro.hasfulltextFull Text
gro.griffith.authorPearson, Andrew G.


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