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

该项目的目的是建立一个框架，系统比较三种不同酶催化过程（菊粉酶、转酮醇酶和转氨酶）在批处理和连续流动系统中的不同固定方法（包埋、亲和和共价结合）。为了实现这一点，不同的固定化方法的性能将在酶活性，生产力和稳定性方面进行比较。最初，反应将与菊粉酶分批进行。然后，基于酶稳定性和生产力的最有前途的固定化方法将应用于第二种酶转酮醇酶，以确定这些方法是否可以直接转移。根据批反应的数据，选择两种方法，先固定化转酮醇酶，再固定化转氨酶，在微反应器中连续流动进行反应。目的是确定固定化酶在批处理和连续流系统中的表现是否相似。该项目的研究结果将为选择合适的固定化方法提供一个框架，用于在微尺度上快速测试酶，从而减少生物催化剂开发的时间和成本。

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.