Selective oxidation of vitamin D3 enhanced by long-range effects of a substrate channel mutation in cytochrome P450BM3 (CYP102A1)

Journal article

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 P450_BM3 (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).

Authors: Chen, W; Lynch, JNC; Bustamante, C; Zhang, Y; Wong, LL

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Wiley
• Journal: Chemistry - A European Journal
• Volume: 30
• Issue: 51
• Article number: e202401487
• Place of publication: Germany
• Publication date: 2024-08-22
• Acceptance date: 2024-07-04
• DOI: 10.1002/chem.202401487
• EISSN: 1521-3765
• ISSN: 0947-6539
• Pmid: 38963680
• Language: English
• Keywords: protein engineering; vitamin D3 oxidation; P450 enzymes; monooxygenases cC-H activation