Creation of Bioorthogonal Redox Systems Depending on Nicotinamide Flucytosine Dinucleotide

摘要

Many enzymes catalyzing biological redox chemistry depend on the omnipresent cofactor, nicotinamide adenine dinucleotide (NAD). NAD is also involved in various nonredox processes. It remains challenging to disconnect one particular NAD-dependent reaction from all others. Here we present a bioorthogonal system that catalyzes the os abiotic cofactor, nicotinamide flucytosine dinucleotide (NFCD). By screen NAD-dependent malic enzyme (ME), we identified the mutant ME-L; NFCD, yet marginal activity with NAD. We found that another synthetic also displayed similar activity with the ME mutants. Inspired by these (DLDH) and malate dehydrogenase (MDH) to DLDH-V152R and MD active with NFCD. When coupled with DLDH-V152R, ME-L310R/Q_4 convert L-malate. Our results opened the window to engineer bioorthogoi systems biology and synthetic biology.