Improving the Bioactivity and Stability of Embedded Enzymes by Covalent Organic Frameworks

Abstract

De novo embedding enzymes within reticular chemistry materials have shown the enhancement of physical and chemical stability for versatile catalytic reactions. Compared to metal–organic frameworks (MOFs), covalent organic frameworks (COFs) are usually considered to be the more superior host of enzymes because of their large channels with low diffusion barriers, outstanding chemical/thermal stability, and metal-free nature. However, detailed investigations on the comparison of COFs and MOFs in enhancing biocatalytic performance have not been explored. Here, we de novo encapsulated enzymes within two COFs via a mechanochemical strategy, which avoided the extreme synthetic conditions of COFs and highly maintained the biological activities of the embedded enzymes. The enzymes@COFs biocomposites exhibited a much higher activity (3.4–14.7 times higher) and enhanced stability than those in MOFs (ZIF-8, ZIF-67, HKUST-1, MIL-53, and CaBDC), and the rate parameter (k_cat/K_m) of enzyme@COFs was 41.3 times higher than that of enzyme@ZIF-8. Further explorations showed that the conformation of enzymes inside MOFs was disrupted, owing to the harmful interfacial interactions between enzymes and metal ions as confirmed by ATR-FTIR, fluorescence spectroscopy, and XPS data. In contrast, enzymes that were embedded in metal-free COFs highly preserved the natural conformation of free enzymes. This study provides a better understanding of the interfacial interactions between reticular supports and enzymes, which paves a new road for optimizing the bioactivities of immobilized enzymes.