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Biotransformation, Using Recombinant CYP450-Expressing Baker's Yeast Cells, Identifies a Novel CYP2D6.10A122V Variant Which Is a Superior Metabolizer of Codeine to Morphine Than the Wild-Type Enzyme

Williams, Ibidapo S.; Gatchie, Linda; Bharate, Sandip B.; Chaudhuri, Bhabatosh

Authors

Ibidapo Williams Ibidapo.Williams@uwe.ac.uk
Research Fellow- Cell Cultural Monitoring Technology

Linda Gatchie

Sandip B. Bharate

Bhabatosh Chaudhuri



Abstract

© Copyright 2018 American Chemical Society. CYP2D6, a cytochrome P450 (CYP) enzyme, metabolizes codeine to morphine. Within the human body, 0-15% of codeine undergoes O-demethylation by CYP2D6 to form morphine, a far stronger analgesic than codeine. Genetic polymorphisms in wild-type CYP2D6 (CYP2D6-wt) are known to cause poor-to-extensive metabolism of codeine and other CYP2D6 substrates. We have established a platform technology that allows stable expression of human CYP genes from chromosomal loci of baker's yeast cells. Four CYP2D6 alleles, (i) chemically synthesized CYP2D6.1, (ii) chemically synthesized CYP2D6-wt, (iii) chemically synthesized CYP2D6.10, and (iv) a novel CYP2D6.10 variant CYP2D6-C (i.e., CYP2D6.10A122V) isolated from a liver cDNA library, were cloned for chromosomal integration in yeast cells. When expressed in yeast, CYP2D6.10 enzyme shows weak activity compared with CYP2D6-wt and CYP2D6.1 which have moderate activity, as reported earlier. Surprisingly, however, the CYP2D6-C enzyme is far more active than CYP2D6.10. More surprisingly, although CYP2D6.10 is a known low metabolizer of codeine, yeast cells expressing CYP2D6-C transform >70% of codeine to morphine, which is more than twice that of cells expressing the extensive metabolizers, CYP2D6.1, and CYP2D6-wt. The latter two enzymes predominantly catalyze formation of codeine's N-demethylation product, norcodeine, with >55% yield. Molecular modeling studies explain the specificity of CYP2D6-C for O-demethylation, validating observed experimental results. The yeast-based CYP2D6 expression systems, described here, could find generic use in CYP2D6-mediated drug metabolism and also in high-yield chemical reactions that allow the formation of regio-specific dealkylation products.

Citation

Williams, I. S., Gatchie, L., Bharate, S. B., & Chaudhuri, B. (2018). Biotransformation, Using Recombinant CYP450-Expressing Baker's Yeast Cells, Identifies a Novel CYP2D6.10A122V Variant Which Is a Superior Metabolizer of Codeine to Morphine Than the Wild-Type Enzyme. ACS Omega, 3(8), 8903-8912. https://doi.org/10.1021/acsomega.8b00809

Journal Article Type Article
Acceptance Date Aug 1, 2018
Online Publication Date Aug 9, 2018
Publication Date Aug 31, 2018
Journal ACS Omega
Electronic ISSN 2470-1343
Publisher American Chemical Society
Peer Reviewed Peer Reviewed
Volume 3
Issue 8
Pages 8903-8912
DOI https://doi.org/10.1021/acsomega.8b00809
Keywords Baker’s yeast, biotransformation, codeine, CYP2D6 variants, morphine, SNPs.
Public URL https://uwe-repository.worktribe.com/output/862217
Publisher URL http://dx.doi.org/10.1021/acsomega.8b00809
Related Public URLs https://cdn-pubs.acs.org
Additional Information Additional Information : This is the author's accepted manuscript. The final published version is available here: http://dx.doi.org/10.1021/acsomega.8b00809.

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