IMMOBILIZATION OF BOVINE CATALASE ONTO MAGNETIC NANOPARTICLES

Abstract

The scope of this study is to achieve carrier-bound immobilization of catalase onto magnetic particles (Fe3O4 and Fe2O3NiO2 center dot H2O) to specify the optimum conditions of immobilization. Removal of H2O2 and the properties of immobilized sets were also investigated. To that end, adsorption and then cross-linking methods onto magnetic particles were performed. The optimum immobilization conditions were found for catalase: immobilization time (15 min for Fe3O4; 10 min for Fe2O3NiO2 center dot H2O), the initial enzyme concentration (1 mg/mL), amount of magnetic particles (25 mg), and glutaraldehyde concentration (3%). The activity reaction conditions (optimum temperature, optimum pH, pH stability, thermal stability, operational stability, and reusability) were characterized. Also kinetic parameters were calculated by Lineweaver-Burk plots. The optimum pH values were found to be 7.0, 7.0, and 8.0 for free enzyme, Fe3O4-immobilized catalases, and Fe2O3NiO2 center dot H2O-immobilized catalases, respectively. All immobilized catalase systems displayed the optimum temperature between 25 and 35 degrees C. Reusability studies showed that Fe3O4-immobilized catalase can be used 11 times with 50% loss in original activity, while Fe2O3NiO2 center dot H2O-immobilized catalase lost 67% of activity after the same number of uses. Furthermore, immobilized catalase systems exhibited improved thermal and pH stability. The results transparently indicate that it is possible to have binding between enzyme and magnetic nanoparticles.