Samenvatting
The conversion of a series of pharmaceutical compounds was examined with three variants of cytochrome P450BM3 fused to phosphite dehydrogenase to enable cofactor recycling. Conditions for enzyme production were optimized and the purified PTDH-P450BM3 variants were tested against 32 commercial drugs using rapid UPLC-MS analysis. The sets of mutations (R47L/F87V/L188Q and R47L/F87V/L188Q/E267V/G415S) improved conversion for all compounds and a variety of products were detected. Product analysis showed that reaction types included C-hydroxylation, N-oxidation, demethylation, and aromatization. Interestingly, enzymatic aromatization could occur independent of the addition of reducing coenzyme. These results identified new conversions catalyzed by P450BM3 variants and show that a small set of mutations in the oxygenase domain can broaden both substrate range and reaction type.
Originele taal-2 | English |
---|---|
Artikelnummer | cbic.201700470 |
Pagina's (van-tot) | 326-337 |
Aantal pagina's | 12 |
Tijdschrift | ChemBioChem |
Volume | 19 |
Nummer van het tijdschrift | 4 |
Vroegere onlinedatum | 27-nov.-2017 |
DOI's | |
Status | Published - 16-feb.-2018 |