Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/120062
Citations
Scopus Web of Science® Altmetric
?
?
Type: Journal article
Title: Selective hydroxylation of 1,8- and 1,4-cineole using bacterial P450 variants
Author: Lee, J.
Wong, S.
Stok, J.
Bagster, S.A.
Beckett, J.
Clegg, J.
Brock, A.
De Voss, J.
Bell, S.
Citation: Archives of Biochemistry and Biophysics, 2019; 663:54-63
Publisher: Elsevier
Issue Date: 2019
ISSN: 0003-9861
1096-0384
Statement of
Responsibility: 
Joel H.Z. Lee, Siew Hoon Wong, Jeanette E. Stok, Sarah A. Bagster, James Beckett, Jack K. Clegg, Aidan J. Brock, James J. De Voss, Stephen G. Bell
Abstract: This study has evaluated the use of the P450 metalloenzymes CYP176A1, CYP101A1 and CYP102A1, together with engineered protein variants of CYP101A1 and CYP102A1, to alter the regioselectivity of 1,8- and 1,4-cineole hydroxylation. CYP176A1 was less selective for 1,4-cineole oxidation when compared to its preferred substrate, 1,8-cineole. The CYP102A1 variants significantly improved the activity over the WT enzyme for oxidation of 1,4- and 1,8-cineole. The CYP102A1 R47L/Y51F/A74G/F87V/L188Q mutant generated predominantly (1S)-6α-hydroxy-1,8-cineole (78% e.e.) from 1,8-cineole. Oxidation of 1,4-cineole by the CYP102A1 R47L/Y51F/F87A/I401P variant generated the 3α product in >90% yield. WT CYP101A1 formed a mixture metabolites with 1,8-cineole and very little product was generated with 1,4-cineole. In contrast the F87W/Y96F/L244A/V247L and F87W/Y96F/L244A variants of CYP101A1 favoured formation of 5α-hydroxy-1,8-cineole (>88%, 1S 86% e.e.) while the F87V/Y96F/L244A variant generated (1S)-6α-hydroxy-1,8-cineole in excess (90% regioselective, >99% e.e.). The CYP101A1 F87W/Y96F/L244A/V247L and F87W/Y96F/L244A mutants improved the oxidation of 1,4-cineole generating an excess of the 3α metabolite (1S > 99% e.e. with the latter). The CYP101A1 F87L/Y96F variant also improved the oxidation of this substrate but shifted the site of oxidation to the isopropyl group, (8-hydroxy-1,4-cineole). When this 8-hydroxy metabolite was generated in significant quantities desaturation of C8C9 to the corresponding alkene was also detected.
Keywords: Enzyme catalysis; Cytochrome P450; Cineole; Hydroxylation; C-H bond oxidation
Rights: © 2018 Elsevier Inc. All rights reserved.
RMID: 0030106390
DOI: 10.1016/j.abb.2018.12.025
Grant ID: http://purl.org/au-research/grants/arc/DP140103229
http://purl.org/au-research/grants/arc/FT140100355
Appears in Collections:Chemistry publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.