Multienzymatic Cascade in a 2-Phase System

Biocatalysis is of increasing importance as an alternative production process in chemical industry. In this context multienzymatic cascades have the potential to meet the demand for complex synthesis routes with high selectivity and purity of the products. Especially in-vitro systems have the potential for high space-time yields. They are less complex, comparatively simple to describe mathematically and therefor good to optimize. This project is about the biotechnological synthesis of cinnamyl cinnamate starting from cinnamaldehyde. Using a biomimetic Claisen-Tischtschenko reaction a sustainable production route of the ester is guaranteed. Isolated enzymes are used in a multiphasic reaction system. The aldehyde will be converted in the corresponding alcohol and acid by NAD(H) coupled sytnhesis. To ensure a high yield, these intermediates will be extracted to an organic phase. Within this phase the alcohol and acid are converted into the ester by a lipase. Removal of the ester will be carried out reaction integrated to ensure a continuous reaction system. The process will be realized in a recycle reactor with a stirred tank reactor for water phase reaction, a fixed bed reactor for organic phase reaction and a continuous operating centrifuge in-between. The present reaction system provides the innovative and novel use of two different in-situ product removal steps within one process. This provides a high potential for a continuous process with high space-time yield.This project aims to contribute to the development of a platform technology to combine process steps, which are usually individually performed. This research is about the continuous production of an in-vitro enzyme cascade with reaction integrated intermediate and reaction integrated product removal in a two phase system. The system will be described mathematical and modelled. The three enzymatic reactions and the mass transfer for the in-situ removal steps are described completely. In the end a whole process design will be given, based on the individual process steps, to give a valuable assistance for comparable processes.

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