A Redox-Neutral, Two-Enzyme Cascade for the Production of Malate and Gluconate from Pyruvate and Glucose

A triple mutant of NADP(H)-dependent malate dehydrogenase from thermotolerant Thermococcus kodakarensis has an altered cofactor preference for NAD+, as well improved production compared to wildtype dehydrogenase. By combining with glucose Sulfolobus solfataricus and NAD+/NADH in a closed reaction environment, gluconate could be produced pyruvate glucose. After 3 h, the yield was 15.96 mM. These data demonstrate feasibility system capable regeneration platform chemicals.