The enhancement of enzyme cascading via tetrahedral DNA framework modification

Enzyme clustering is widely used in many organisms to increase the catalytic efficiency of cascade reactions. Inspired by nature, organizing enzymes within a cascade reaction also draws much attention in both basic research and industrial processes. An important step for organizing enzymes precisely in vitro is enzyme modification. However, modifying enzymes without sacrificing their activity remains challenging until now. For example, labeling enzymes with DNA, one of the well-established enzyme modification methods, has been shown to significantly reduce the enzymatic activity. Herein we report an enzyme conjugation method that can rescue the reduction of enzymatic activity caused by DNA labeling. We demonstrate that immobilizing DNA-modified enzymes on the vertex of TDNs (tetrahedral DNA nanostructures) enhances the enzymatic activity compared with their unmodified counterparts. Using this strategy, we have further developed an ultra-sensitive and high-throughput electrochemical biosensor for sarcosine detection, which holds great promise for prostate cancer screening.

Automated summary

Enzyme clustering is widely used to improve catalytic efficiency in cascade reactions. Organizing enzymes within a cascade reaction is of great interest in basic research and industrial processes. Enzyme modification is a crucial step for precise organization of enzymes in vitro, but preserving enzyme activity during modification is challenging. Here, we present an enzyme conjugation method that rescues the reduction of enzymatic activity caused by DNA labeling. This method involves immobilizing DNA-modified enzymes on tetrahedral DNA nanostructures, resulting in enhanced enzymatic activity. We demonstrate the utility of this method by developing an ultra-sensitive and high-throughput electrochemical biosensor for sarcosine detection, which could be valuable for prostate cancer screening.