Due to their high chemo-, stereo-, and regio-selectivity, enzymes have become an important tool for selective C–H bond functionalization and offer orthogonal reactivity compared to small molecule catalysts. Engineering of enzymes and substrates provides an excellent platform to broaden substrate scope and improve activity and selectivity. Cytochrome P450 biocatalysts have proven to be effective for selective oxidation of unactivated C–H bonds. The Sherman group reported characterization of TamI, a versatile and iterative cytochrome P450 monooxygenase, which acts co-dependently with flavin adenine dinucleotide-dependent oxidase, TamL, to catalyze regio- and stereo-selective hydroxylations and epoxidation to yield potent bioactive molecules tirandamycin A (TirA) and tirandamycin B (TirB). Based on the co-crystal structure (2.7Å resolution) of TamI with TirC, several important interactions were discovered that potentially mediate substrate binding. We aim to computationally study the various transformations performed by TamI utilizing quantum mechanical (QM) calculations in parallel with molecular dynamics (MD) simulations to develop mechanistic insights about the stereo- and regio-selectivity of C10 hydroxylation, C11/C12 epoxidation, and C18 hydroxylation. These computational results will provide insights for the Sherman and Montgomery group to engineer TamI to alter the regio- and stereo-selectivity and increase substrate scope. In addition, they aim to engineer the substrate by attaching readily removing anchoring groups to a panel of unnatural substrates to undergo selective C–H oxidations to assess the degree of promiscuity of TamI. The goal is to development a general biocatalytic tool for its use in selective C–H oxidations on a variety of complex substrates.

Investigation of TamI, an iterative P450 monoxygenase, for engineering as a biocatalyst for selective C–H oxidation

256th National Meeting (2018)

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