An in vivo cytochrome P450(cin) (CYP176A1) catalytic system for metabolite production

Slessor, Kate E., Hawkes, David B., Farlow, Anthony, Pearson, Andrew G., Stok, Jeanette E. and De Voss, James J. (2012) An in vivo cytochrome P450(cin) (CYP176A1) catalytic system for metabolite production. Journal of Molecular Catalysis B: Enzymatic, 79 15-20. doi:10.1016/j.molcatb.2012.03.007

Author Slessor, Kate E.
Hawkes, David B.
Farlow, Anthony
Pearson, Andrew G.
Stok, Jeanette E.
De Voss, James J.
Title An in vivo cytochrome P450(cin) (CYP176A1) catalytic system for metabolite production
Formatted title
An in vivo cytochrome P450 cin (CYP176A1) catalytic system for metabolite production
Journal name Journal of Molecular Catalysis B: Enzymatic   Check publisher's open access policy
ISSN 1381-1177
Publication date 2012-07
Sub-type Article (original research)
DOI 10.1016/j.molcatb.2012.03.007
Volume 79
Start page 15
End page 20
Total pages 6
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2013
Language eng
Formatted abstract
Cytochrome P450cin (CYP176A1) is a bacterial P450 isolated from Citrobacter braakii that catalyses the hydroxylation of 1,8-cineole to (1R)-6β-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)-6β-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.
Keyword Camphor
Cytochrome P 450
Escherichia Coli
In vivo
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Available online 27 March 2012.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Chemistry and Molecular Biosciences
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 5 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 6 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 13 Jun 2012, 09:51:39 EST by Lucy O'Brien on behalf of School of Chemistry & Molecular Biosciences