Advances in cofactor immobilization for enhanced continuous-flow biocatalysis

The merging of biocatalysis with continuous-flow chemistry opens up new opportunities for sustainable and efficient chemical synthesis. Cofactor-dependent enzymes are essential for various industrially attractive biocatalytic reactions. However, implementing these enzymes and biocatalytic reactions in industry remains challenging due to the inherent cost of cofactors and the requirement for their external supply in significant quantities. The development of efficient, low cost, simple and versatile methods for cofactor immobilization can address this important obstacle for biocatalysis in flow. This review explores recent progress in cofactor immobilization for biocatalysis by analyzing advantages and current limitations of the available methods that comprise covalent tethering, ionic adsorption, physical entrapment, and hybrid variations thereof. Moreover, this review analyzes all these immobilization techniques specifically for their utilization in continuous-flow chemistry and provides a perspective for future work in this area. This review will serve as a guide for steering the field towards more sustainable and economically viable continuous-flow biocatalysis.