Development of reaction systems for ATP regeneration from the inexpensive bulk chemical ethylene glycol
In this project, enzyme systems for the cost-effective regeneration of ATP in technical reaction systems will be developed. The project is planned as a joint project of the working groups of Prof. Thomas Walther (Professorship of Bioprocess Engineering, TU Dresden) and of Prof. Andreas Liese (Chair of Technical Biocatalysis, TU Hamburg), where AG Walther is responsible for the design and construction of the ATP regenerating enzyme cascades as well as for the optimization of individual enzyme activities, and the AG Liese will work on the reaction engineering and optimization of the reaction systems as well as transfer to larger scales.ATP regenerating reaction systems have a variety of industrial applications. Examples include the synthesis of phosphorylated fine chemicals, syntheses with phosphorylation as an intermediate step and cell-free protein expression. A major obstacle to the commercialization of ATP-dependent, cell-free, product syntheses is the sometimes very high price of the substrates and cofactors used for ATP regeneration.To solve this technical problem, the development of reaction systems is proposed, which allow an ATP regeneration of the low-cost and potentially "green" bulk chemical ethylene glycol. The implementation of the designed enzyme cascades requires the enhancement of enzyme activity on a non-natural substrate. This increase in activity is to be achieved through a combination of rational and evolutionary protein design. In addition, a reaction-technical analysis and optimization of the transfer of the various systems on a technical scale is examined.The enzyme cascades for the regeneration of ATP will be constructed from isolated enzymes as well as be implemented in whole-cell biocatalysts. In each case, the ATP-regenerating systems are coupled to the synthesis of an ATP-consuming reaction for the synthesis of a phosphorylated or non-phosphorylated end product in order to determine advantages and limits of both design variants.The reaction cascades described have the potential to be developed into a new universal ATP regeneration system, which can also be used on an industrial scale.