Emergence of ADM-mediated bioconjugate to enhance longevity and catalytic efficiency of urease.

The versatile nature of the urease enzyme makes it a valuable asset in biological and industrial contexts. The creation of bioconjugates using enzyme-polymer combinations has extended the shelf life and stability of urease. A triblock copolymer, PAM-co-PDPA-co-PMAA@urease (ADM@urease), was synthesized using acrylamide (AM), 2,5-dioxopyrrolidin-1-ylacrylate (DPA), methacrylic acid (MAA), and urease via the RAFT-Grafting-To polymerization method. This polymeric interface stabilizes the enzyme and enhances substrate binding and product release, significantly boosting enzymatic efficiency. To enhance pH's influence on urease activity, three ADM grades were developed by adjusting pH-responsive MAA levels, confirmed by GPC analysis. ADM micellized at acidic pH values of 6.47 or lower, with a critical micelle concentration (CMC) of at least 0.125 mg/mL. Kinetic evaluations using Berthelot reagents at various pH levels and temperatures compared free enzyme and urease encapsulated in ADM@urease. The Michaelis-Menten constant (Km) values, derived from the Lineweaver-Burk plot, were similar for both forms. The ADM@urease demonstrated optimal stability and catalytic efficacy with a Km value of 1.18 and Vmax of 1.92 at pH 4. By improving the stability, efficiency, and performance of urease, this encapsulation technology offers potential for sustainable, eco-friendly industrial applications and advancements in biotechnology.