Synthesis of [4S‐2H]NADH, [4R‐2H]NADH, [4‐2H2]NADH and [4‐2H]NAD+ cofactors through heterogeneous biocatalysis in heavy water

This practitioner protocol describes the synthesis of a family of deuterated nicotinamide cofactors: [4S‐2H]NADH, [4R‐2H]NADH, [4‐2H2]NADH and [4‐2H]NAD+. The application of a recently developed H2‐driven heterogeneous biocatalyst enables the cofactors to be prepared with high (>90%) 2H‐incorporation with 2H2O as the only isotope source.