Stoichiometric Regeneration of ATP by A NAD(P)/CoA-free and Phosphate-balanced In Vitro Synthetic Enzymatic Biosystem

摘要

The regeneration of adenosine triphosphate is crucial for establishing a cost-effective biosystem in which energy-dependent processes are involved. Here, an invitro synthetic enzymatic biosystem was designed to utilize low-cost starch-based material as energy source for the stoichiometric regeneration of ATP. This system required neither NAD(P) nor coenzyme A, and theoretically produced 3 ATPs per glucose equivalent of starch. When integrated with the L-theanine-producing enzymatic reaction which consumes 1 ATP to generate 1 L-theanine molecule, our system resulted in a near-theoretical yield of 2.94 L-theanine per glucose equivalent. Furthermore, inorganic phosphate concentration of the system was nearly constant throughout the reaction period, suggesting the homeostatic reaction environment for enzymatic processes. This work provides a useful strategy for cost-effective ATP regeneration which facilitates the production of ATP-dependent value-added chemicals.