Reaction Engineering and Comparison of Electroenzymatic and Enzymatic ATP Regeneration Systems

Abstract

Adenosine‐5’‐triphosphate (ATP) plays a crucial role in many biocatalytic reactions and its regeneration can influence the performance of a related enzymatic reaction significantly. Here, we established electrochemically coupled ATP regeneration by pyruvate oxidase and acetate kinase (ACK) for the phosphorylation of mevalonate catalyzed by mevalonate kinase. A yield of 84 % for the product mevalonate phosphate was reached and a total turnover number for ADP of 68. These metrics are promising for the development of an economic feasible bioprocess and surpass many other enzymatic ATP regeneration systems. A comparison was made to polyphosphate kinases (PPKs), ACK, pyruvate kinase, and creatine kinase in terms of the phosphate donor properties and biocatalytic metrics of exemplary reactions. Furthermore, our system was expanded by a PPK that enables the phosphorylation of AMP, which can broaden the spectrum of applications even further.

Development of a multi-enzyme cascade for 2′3′-cGAMP synthesis from nucleosides

“…11,12 Electrochemically coupled ATP regeneration by a pyruvate oxidase and an AcK has in addition already been demonstrated. 13 In recent years the use of polyphosphate kinases 2 (PPK2) has gained interest due to the use of low-cost polyphosphate (polyP). 12 They have been successfully used for ATP regeneration in enzyme cascades for methyltransferases, 14 multigram-scale aldehyde synthesis, 15 formation of nucleotide sugars, 16 and in the monoacylation of symmetrical diamines.…”