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Selective Oxidation of Vitamin D3 Enhanced by Long-Range Effects of a Substrate Channel Mutation in Cytochrome
Wenyu Chen1,2, Jamie N C Lynch1, Claudia Bustamante1
1Department of Chemistry, University of Oxford Inorganic Chemistry Laboratory, South Parks Road, Oxford, OX1 3QR, UK.
Abstract:
Vitamin D deficiency affects nearly half the population, with many requiring or opting for supplements with vitamin D3 (VD3), the precursor of vitamin D (1α,25-dihydroxyVD3). 25-HydroxyVD3, the circulating form of vitamin D, is a more effective supplement than VD3 but its synthesis is complex. We report here the engineering of cytochrome P450BM3 (CYP102A1) for the selective oxidation of VD3 to 25-hydroxyVD3. Long-range effects of the substrate-channel mutation Glu435Ile promoted binding of the VD3 side chain close to the heme, enhancing VD3 oxidation activity that reached 6.62 g of 25-hydroxyVD3 isolated from a 1-litre scale reaction (69.1 % yield; space-time-yield 331 mg/L/h).
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