25-Hydroxyvitamin D3 synthesis by enzymatic steroid side chain hydroxylation with water

MARKUS WARNKE; TOBIAS JUNG; JURI DERMER; KARIN HIPP; NICO JEHMLICH; MARTIN VON BERGEN; SASCHA FERLAINO; ALEXANDER FRIES; MICHAEL MÜLLER; MATTHIAS BOLL

Angewandte Chemie

WILEY-V C H VERLAG GMBH

Lugar: Weinheim; Año: 2015

1433-7851

The hydroxylation of vitamin D3 (VD3, cholecalciferol) side chains into 25-hydroxyvitamin D3 (25OHVD3) is a crucial reaction in the formation of the circulating and biologically active forms of VD3. It is usually catalyzed by cytochrome P450 monooxygenases that depend on complex electron donor systems. Here, we employ cell- free extracts and a purified Mo-enzyme from a bacterium anaerobically grown with cholesterol for the regioselective, ferricyanide-dependent hydroxylation of VD3 and proVD3 (7- dehydrocholesterol) into the corresponding tertiary alcohols with >99% yield. Hydroxylation of VD3 strictly depended on a cyclodextrin-assisted isomerization of VD3 into preVD3, the actual enzymatic substrate. The facile and robust method developed for 25OHVD3 synthesis is a novel example for the concept of substrate- engineered catalysis and offers an attractive alternative to chemical or O2/electron donor-dependent enzymatic procedures.