Biocatalysis and continuous process implementation are becoming key technologies for the sustainable production of substances. Immobilization of biocatalysts, which enables their stabilization and long-term use, and the application of miniaturized flow systems, which lead to process intensification, play a crucial role in this. This work focuses on immobilizing biocatalysts by trapping them in hydrogels further integrated into microfluidic reactors between two plates. Flow biotransformations with permeabilized yeast and isolated amine transaminase using a copolymer hydrogel in the microreactor were performed. To describe the biocatalytic process of converting fumaric acid to L-malic acid, a mathematical model was developed, which successfully predicted the performance of the continuous process in the microreactor with hydrogel and enabled its optimization. Besides, an innovative method for preparing homogeneous nanoscale enzyme aggregates of amine transaminase was developed, where the latter were immobilized in a flow microreactor to perform continuous transamination. In the microreactor between two plates, an all-enzyme hydrogel of phenolic acid decarboxylase was also generated. Additionally, bioreduction with yeast cells in a deep eutectic solvent was conducted, yielding an improved cofactor regeneration and the stability of the system. The results represent an important step in the development of biocatalytic processes and their implementation in industry.
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