Biocatalysis in flow systems has been shown to provide greater control over reagent addition, in-line product removal, and purification, and increased enzyme stability/lifespan. Continuous flow biocatalysis is projected to rapidly expand as additional enzymes become commercially accessible and the need for greener and more affordable biotransformation rises. While continuous flow biocatalysis has been extensively investigated by the pharmaceutical and food industries over the last decade, much of its potential remains unexplored. Immobilization as an integral step in biocatalysis traditionally employs physical adsorption, ionic interactions, or covalent bonding with amino groups present in the enzyme. In this study, we explore the possibility of carrying out biocatalysis in an automated flow system for the synthesis of gluconic acid using glucose oxidase. Glucose oxidase was immobilized using different techniques. Silica support using sodium polystyrene sulfonate (PSS) to bind to the support, chitosan to coordinate with the PSS, and finally glutardialdehyde to form azomethine bonds with the amines on the enzymes. Also used was epoxy-resin support that can form azomethine bonds with the enzymes. The reactions were monitored using an in-line Raman flow cell offering a non-invasive characterization of product formation in real-time. The characterization of reaction products using Raman helps in optimizing enzyme kinetics towards maximum efficiency.

Flow biocatalysis using glucose oxidase with in-line monitoring through Raman spectroscopy

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