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Monooxygenase, Peroxidase and Peroxygenase Properties and Mechanisms of Cytochrome P450 pp 319–339Cite as

Cytochrome P450cin (CYP176A1)

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 851))

Abstract

Cytochrome P450cin (P450cin) (CYP176A1) is a bacterial P450 enzyme that catalyses the enantiospecific hydroxylation of 1,8-cineole to (1R)-6β-hydroxycineole when reconstituted with its natural reduction-oxidation (redox) partner cindoxin, E. coli flavodoxin reductase, and NADPH as a source of electrons. This catalytic system has become a useful tool in the study of P450s as not only can large quantities of P450cin be prepared and rates of oxidation up to 1,500 min−1 achieved, but it also displays a number of unusual characteristics. These include an asparagine residue in P450cin that has been found in place of the usual conserved threonine residue observed in most P450s. In general, this conserved threonine controls oxygen activation to create the potent ferryl (Fe(IV=O) porphyrin cation radical required for substrate oxidation. Another atypical characteristic of P450cin is that it utilises an FMN-containing redoxin (cindoxin) rather than a ferridoxin as is usually observed with other bacterial P450s (e.g. P450cam). This chapter will review what is currently known about P450cin and how this enzyme has provided a greater understanding of P450s in general.

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Notes

  1. 1.

    Abbreviations: P450 or CYP cytochrome P450, Fld E. coli flavodoxin, FdR E. coli flavodoxin reductase, GC/MS gas chromatography/mass spectrometry, Pdx putidaredoxin, PdR putidaredoxin reductase, CPR NADPH-cytochrome P450 oxidoreductase, CdR cindoxin reductase, Cdx cindoxin, HPLC high-performance liquid chromatography, E 1 quinone/semiquinone, E 2 semiquinone/hydroquinone, EPR electron paramagnetic resonance, ENDOR electron nuclear double resonance.

  2. 2.

    There is no consistency for naming hydroxycineoles in the literature. In order to discuss stereochemistry we use descriptors α and β. Employing a plane that passes through C5, C6, C7 and C8 of the molecule (See Fig. 12.2), we term any substituents that lie below the plane α and any substituents that are above this plane β. Three new stereogenic centres at C1, C4 and C6 are created following the hydroxylation of the meso cineole at either one of the carbons that lie adjacent to the C1 bridgehead. To differentiate the entantiomers formed following oxidation we have described the pro-R carbon as the carbon atom that leads to the R-C1 isomer and the pro-S carbon as that leading to the S-C1 isomer.

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Acknowledgements

The authors would like to acknowledge that this work was supported in part by ARC Grants DP110104455 and DP140103229.

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Authors and Affiliations

  1. School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, Brisbane, 4072, Australia

    Jeanette E. Stok, Kate E. Slessor, Anthony J. Farlow, David B. Hawkes & James J. De Voss

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  1. Jeanette E. Stok
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Correspondence to James J. De Voss .

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Editors and Affiliations

  1. Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, British Columbia, Canada

    Eugene G. Hrycay

  2. Fac. of Pharmaceutical Sciences, The University of British Columbia , Vancouver, British Columbia, Canada

    Stelvio M. Bandiera

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Stok, J.E., Slessor, K.E., Farlow, A.J., Hawkes, D.B., De Voss, J.J. (2015). Cytochrome P450cin (CYP176A1). In: Hrycay, E., Bandiera, S. (eds) Monooxygenase, Peroxidase and Peroxygenase Properties and Mechanisms of Cytochrome P450. Advances in Experimental Medicine and Biology, vol 851. Springer, Cham. https://doi.org/10.1007/978-3-319-16009-2_12

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