Dynamic Exchange Assembly of Enzymes with Single-Crystal Covalent Organic Frameworks

Employing single-crystal porous materials to study their assembly with biomacromolecules is a powerful method for uncovering the fundamental principles behind biocomposite formation and advancing their practical applications, but it remains a formidable challenge. Herein, we develop a dynamic exchange strategy to investigate the assembly process of enzymes with covalent organic frameworks (COFs). To demonstrate the proof of concept, we design and synthesize a single-crystal three-dimensional (3D) microporous COF, whose structure is fully solved by single-crystal X-ray diffraction, enabling precise characterization of its interactions with enzymes. It is found that the assembly of COFs with enzymes is driven by the dynamic, reversible imine linkage in COFs, and this dynamic exchange strategy is versatile, allowing it to be applied to various enzymes and COFs. Additionally, the controlled distribution of enzymes within the crystals enables a highly efficient cascade of large enzymes to catalyze large substrates. This study offers an innovative platform for biocatalysis and establishes guiding principles for developing biohybrid materials, opening new avenues for the assembly of biomacromolecules and porous materials.