ABSTRACT Small enzymes (10–50 kDa) encounter challenges of unstable enzyme‐carrier interactions and low activity retention during immobilization, with existing strategies lacking specificity. Unlike previous industrial enzyme‐focused studies, a rigid‐flexible layered immobilization strategy integrating networked metal–organic frameworks (MOFs) into flexible hydrogels is proposed. This innovative strategy stabilizes enzyme conformation, strengthens enzyme‐matrix interactions, and prevents enzyme leakage via a dual‐pore system. This synergistic system, composed of MOF micropores (50–150 nm, confinement) and hydrogel macropores (800–900 nm, mass transfer), resolves the stability‐accessibility trade‐off. Molecular docking shows MOFs reduce substrate‐enzyme distance by 34.2% and enhance rigidity. In industrial biocatalysis, immobilized horseradish peroxidase maintains robust conversion efficiency for the reaction of 1 M o ‐phenylenediamine over 40 cycles, effectively promoting the production efficiency of related chemical products. In the biomedical field, immobilizing thrombin reduces murine wound bleeding by 55%, providing a new solution to wound hemostasis. This scalable platform integrates nanoscale precision with macroscale responsiveness, holding great promise for advancing sustainable biocatalysis and biomedicine.
Zhu et al. (Tue,) studied this question.