Hydrophobic Covalent Organic Frameworks as a Platform for Instantaneous and High‐Capacity Immobilization of Diutina ( Candida ) rugos

Covalent Organic Frameworks (COFs) are emerging as promising platforms for enzyme immobilization; however, the impact of their structural and chemical properties on adsorption and enzymatic activity remains underexplored. Here, Diutina rugosa lipase (DrL) was immobilized on four COFs (RIO‐12, RIO‐13, RIO‐54, and RIO‐55) synthesized from simple, commercially available precursors. The effects of COF chemical composition and textural properties on adsorption dynamics and catalytic performance were evaluated. Immobilization occurred almost instantaneously, reaching completion in less than 10 s regardless of the COF used, with loading efficiencies of 99.8%–99.9%, protein binding capacities of approximately 44 mg g ⁻¹ of support, and enzyme leaching below 0.3% per washing cycle for all composites. Our results show that fully organic COF surfaces, rich in aromatic and nitrogen‐containing functional groups, exhibit a strong affinity for lipases, enabling rapid immobilization primarily driven by hydrophobic interactions, whereas pore size and surface area play a secondary role. In contrast, enzymatic activity was strongly influenced by surface area, likely through a positive partition effect that increases the local substrate concentration near the enzyme. Notably, RIO‐12 exhibited a remarkable enhancement in activity, achieving a specific activity 14.8‐fold higher than that of the free enzyme (408.5 vs. 27.7 U mg ⁻¹ ).