School of Chemistry and Chemical Engineering, and Institute of Molecular Medicine, Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China.
Nano Lett. 2024 Apr 17;24(15):4682-4690. doi: 10.1021/acs.nanolett.4c01137. Epub 2024 Apr 2.
Multienzyme assemblies mediated by multivalent interaction play a crucial role in cellular processes. However, the three-dimensional (3D) programming of an enzyme complex with defined enzyme activity in vitro remains unexplored, primarily owing to limitations in precisely controlling the spatial topological configuration. Herein, we introduce a nanoscale 3D enzyme assembly using a tetrahedral DNA framework (TDF), enabling the replication of spatial topological configuration and maintenance of an identical edge-to-edge distance akin to natural enzymes. Our results demonstrate that 3D nanoscale enzyme assemblies in both two-enzyme systems (glucose oxidase (GOx)/horseradish peroxidase (HRP)) and three-enzyme systems (amylglucosidase (AGO)/GOx/HRP) lead to enhanced cascade catalytic activity compared to the low-dimensional structure, resulting in ∼5.9- and ∼7.7-fold enhancements over homogeneous diffusional mixtures of free enzymes, respectively. Furthermore, we demonstrate the enzyme assemblies for the detection of the metabolism biomarkers creatinine and creatine, achieving a low limit of detection, high sensitivity, and broad detection range.
多价相互作用介导的多酶组装在细胞过程中起着至关重要的作用。然而，在体外具有明确酶活性的酶复合物的三维（3D）编程仍然没有得到探索，主要是由于在精确控制空间拓扑结构方面存在局限性。在此，我们使用四面体 DNA 框架（TDF）引入了纳米级 3D 酶组装，从而能够复制空间拓扑结构并保持与天然酶相同的边缘到边缘的距离。我们的结果表明，与低维结构相比，两种酶系统（葡萄糖氧化酶（GOx）/辣根过氧化物酶（HRP））和三种酶系统（淀粉葡萄糖苷酶（AGO）/GOx/HRP）中的 3D 纳米级酶组装导致级联催化活性增强，分别约为游离酶均相扩散混合物的 5.9 倍和 7.7 倍。此外，我们还展示了用于检测代谢生物标志物肌酸酐和肌酸的酶组装，实现了低检测限、高灵敏度和宽检测范围。
