Selective functionalization of unactivated C-H bonds using molecular dioxygen as the sole oxidant still remains synthetically challenging. The Cytochrome P450 superfamily of heme-thiolate enzymes catalyze a myriad of oxidation reactions often with high regio and stereo selectivity using molecular dioxygen as the source of oxygen atom and two reducing equivalents. Our laboratory has developed light-driven P450 biocatalysts by taking advantage of the photochemical properties of Ru(II)-diimine complexes covalently attached to non-native single cysteine residue of P450 heme domain mutants. We recently demonstrated using a model substrate that this light-activated approach is becoming valuable by achieving high total turnover numbers and reaction rates for such hydroxylation. We are currently immobilizing the hybrid enzymes on various solid support beads using the photosensitizer as an anchor. This approach presents the advantages to control the immobilization of the hybrid enzymes and prevent undesired misfolding and inactivation. Our current efforts have focused on the assembly and characterization of the hybrid enzyme functionalized beads using microplate readers. We will then assess their ability to hydroxylate various substrates using our previously developed photocatalytic procedures.

Towards the immobilization of light-driven P450 BM3 hybrid enzymes on solid support beads

248th National Meeting (2014)

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