Multi‐Functional Oxidase Activity of CYP102A1 (P450BM3) in the Oxidation of Quinolines and Tetrahydroquinolines

Abstract

Tetrahydroquinoline,q uinoline,a nd dihydroquinolinone are common core motifs in drug molecules.Screening of a4 8-variant library of the cytochrome P450 enzyme CYP102A1 (P450BM3), followed by targeted mutagenesis based on mutation-selectivity correlations from initial hits,has enabled the hydroxylation of substituted tetrahydroquinolines, quinolines,a nd 3,4-dihydro-2-quinolinones at most positions around the two rings in good to high yields at synthetically relevant scales (1.5 gL À1 day À1 ). Other oxidase activities,s uch as CÀCb ond desaturation, aromatization, and CÀCb ond formation, were also observed. The enzyme variants,w ith mutations at the key active site residues S72, A82, F87, I263, E267, A328, and A330, provide direct and sustainable routes to oxy-functionalized derivatives of these building blockm olecules for synthesis and drug discovery.Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

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Oxidative Diversification of Steroids by Nature-Inspired Scanning Glycine Mutagenesis of P450BM3 (CYP102A1)

“…The relevant plasmid was then transformed into chemically competent E. coli BL21 (DE3) for enzyme variant production and subsequent purification as described previously. 29,40,41 Screening scale reactions. Steroidal substrates were dissolved in an organic solvent (DMSO, EtOH or MeOH) and added as a stock, typically at 100 mM concentration.…”

A Promiscuous Bacterial P450: The Unparalleled Diversity of BM3 in Pharmaceutical Metabolism

“…F87A/V, A328I and I263G favored C5 and C6 oxidations and also promoted metabolite dimer formation, with F87 variants displaying activity towards aromatization. The range of biotransformations performed by different variants in this study highlighted BM3’s role as a selective multi-functional oxidase [ 94 ].…”