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Exploring PTDH-P450BM3 variants for the synthesis of drug metabolites
- Source :
- ChemBioChem, 19(4):cbic.201700470, 326-337. WILEY-V C H VERLAG GMBH
- Publication Year :
- 2018
- Publisher :
- WILEY-V C H VERLAG GMBH, 2018.
-
Abstract
- The conversion of a series of pharmaceutical compounds was examined with three variants of cytochrome P450BM3 fused to phosphite dehydrogenase (PTDH) to enable cofactor recycling. Conditions for enzyme production were optimized, and the purified PTDH-P450BM3 variants were tested against 32 commercial drugs by 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 showed that a small set of mutations in the oxygenase domain could broaden both substrate range and reaction type.
- Subjects :
- 0301 basic medicine
Oxygenase
Cytochrome
030226 pharmacology & pharmacy
Biochemistry
Cofactor
Mass Spectrometry
Enzyme catalysis
03 medical and health sciences
0302 clinical medicine
Bacterial Proteins
Cytochrome P-450 Enzyme System
enzyme fusion
NADH, NADPH Oxidoreductases
Molecular Biology
Chromatography, High Pressure Liquid
Demethylation
NADPH-Ferrihemoprotein Reductase
chemistry.chemical_classification
biology
Molecular Structure
Chemistry
enzymatic conversion
Organic Chemistry
Aromatization
Genetic Variation
PTDH-P450 variants
drug metabolite synthesis
030104 developmental biology
Enzyme
Pharmaceutical Preparations
Mutation
biology.protein
Biocatalysis
Molecular Medicine
Drug metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 14397633 and 14394227
- Volume :
- 19
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- ChemBioChem
- Accession number :
- edsair.doi.dedup.....71cbe78bfe5fa8d61d30f5037bb65f4f