Evaluation of different enzyme immobilisation methods for multi-step synthesis in microreactors

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The aim of this project is to establish a framework for systematic comparison of different immobilisation methods (entrapment, affinity, and covalent binding) for three different enzyme-catalysed processes (inulinase, transketolase and transaminase) in batch and continuous flow systems under defined reaction conditions. To achieve this, the performance of the different immobilisation methods will be compared in terms of enzyme activity, productivity, and stability. Initially, reactions will be performed in batch mode with inulinase. Then the most promising immobilisation methods based on enzyme stability and productivity will be applied to the second enzyme, transketolase, in order to determine if the methods can be transferred directly. Based on data from the batch reactions, two methods will be chosen to immobilise first transketolase and then transaminase and carry out reactions under continuous flow in microreactors. The aim is to determine if the immobilised enzymes perform similar in batch and continuous flow systems. The findings of this project will provide a framework for the selection of suitable immobilisation methods for rapid testing of enzymes at the microscale thereby reducing time and cost of biocatalyst development.