BIONATURE

The study explores the kinetic behavior of urease immobilized on alumina membranes with three different pore diameters (20, 100, and 200 nm). The goal is to elucidate how pore size affects enzyme activity, substrate accessibility, and overall urea conversion efficiency. The study aims to influence of Membrane Pore Size on Immobilized Urease Biocatalytic Activity. Membranes with varying pore sizes were used as urease immobilization substrates to examine the effect of structural confinement on enzyme kinetics. The reduction in pore diameter led to uneven substrate distribution, which affected the local urea conversion rate and limited overall catalytic performance. A 50 U/mL urease solution prepared in 20 mM MES buffer (pH 6.0) was applied to each activated membrane and incubated at 25°C for 3 hours to allow covalent attachment via amide bond formation. The findings of this study provide insights into the optimal design of biocatalytic membranes for efficient urea removal in water purification systems and enzymatic reactors. Understanding the pore size effect aids in designing high-sensitivity, stable biocatalytic interfaces for such sensors. In bioprocesses involving urea as a reactant or byproduct, immobilized urease systems could be integrated into closed-loop reactors for continuous detoxification, minimizing waste accumulation and improving sustainability. These results provide valuable design principles for biocatalytic membranes in water purification and biosensor applications. Future work should focus on integrating multi-scale porous structures and exploring enzyme stabilization stabilization through nanostructured coatings.