Nucleic Acid‐based Enzyme Cascades—Current Trends and Future Perspectives

The natural micro‐ and nanoscale organization of biomacromolecules is a remarkable principle within living cells, allowing for the control of cellular functions by compartmentalization, dimensional diffusion and substrate channeling. In order to explore these biological mechanisms and harness their potential for applications such as sensing and catalysis, molecular scaffolding has emerged as a promising approach. In the case of synthetic enzyme cascades, developments in DNA nanotechnology have produced particularly powerful scaffolds whose addressability can be programmed with nanometer precision. In this minireview, we summarize recent developments in the field of biomimetic multicatalytic cascade reactions organized on DNA nanostructures. We emphasize the impact of the underlying design principles like DNA origami, efficient strategies for enzyme immobilization, as well as the importance of experimental design parameters and theoretical modeling. We show how DNA nanostructures have enabled a better understanding of diffusion and compartmentalization effects at the nanometer length scale, and discuss the challenges and future potential for commercial applications. Nucleic acid nanostructures are capable of precisely arranging proteins at nanoscale distances and have thus proven to be powerful scaffolds for studying mechanistic effects in enzyme cascades. This minireview summarizes the current status of proposed underlying mechanistic effects, derives aspects for further systematic analysis, and discusses future perspectives for biocatalytic applications.