On-pot and cell-free biocatalysis using coimmobilized enzymes on advanced materials

Complex synthetic schemes catalyzed by multienzyme systems immobilized on solid materials are gaining momentum in chemical biomanufacturing. These systems harness the high chemo-, regio-, and stereoselectivity of the enzymes and the recyclability of the heterogeneous catalysts. Moreover, when the enzymes become part of a solid material, they can be easily integrated into packed-bed reactor for continuous biotransformations. Advances in nanotechnology have enabled to precisely control the spatial distribution of multienzyme systems on the surfaces of advanced materials. This chapter summarizes the challenges and opportunities of using immobilized multienzyme systems in step-wise chemical transformations. A plethora of examples given herein illustrates how the nature and the physicochemical properties of the materials as well as the engineering of the protein surfaces impact on the overall functionality of immobilized multienzyme systems to perform cascade reactions. Furthermore, some insights are provided to orchestrate both stabilities and activities of the enzymes in these systems. Immobilized multienzyme systems are an excellent opportunity to increase the robustness and expand the application scope of cell-free biocatalytic systems for synthetic, analytical, medical, and environmental chemistry.