Urease immobilization on arylamine glass beads and its characterization.

Abstract

Jack bean urease has been immobilized on arylamine glass beads (200-400 mesh size, 75-100 Å pore size) and its properties compared with soluble enzyme. The binding of urease was 13.71 mg per gram beads. The Km for soluble and immobilized urease for urea was 4.20 mM and 8.81 mM, respectively. Vmax values of urease decreased from 200 to 43.48 μmol of ammonia formed per min per mg protein at 37°C on immobilization. Both pH and buffer ions influenced the activities of soluble as well as immobilized urease. Soluble urease exhibited pH optima at 5.5 and 8.0. However, immobilized urease showed one additional pH optimum at 6.5. In comparison to phosphate buffer, citrate buffer was inhibitory to urease activity. Immobilization of urease on arylamine glass beads resulted in improved thermal, storage and operational stability. Because of inertness of support and stability of immobilized urease, the preparation can find applications in ‘artificial kidney’ and urea estimation in biological fluids viz., blood, milk etc.