Abstract
The present study demonstrates the immobilization of Aspergillus oryzae beta galactosidase on native zinc oxide (ZnO) and zinc oxide nanoparticles (ZnO-NP) by simple adsorption mechanism. The binding of enzyme on ZnO-NP was confirmed by Fourier transform-infrared spectroscopy and atomic force microscopy. Native ZnO and ZnO-NP showed 60% and 85% immobilization yield, respectively. Soluble and immobilized enzyme preparations exhibited similar pH-optima at pH 4.5. ZnO-NP bound beta galactosidase retained 73% activity at pH 7.0 while soluble and ZnO adsorbed enzyme lost 68% and 53% activity under similar experimental conditions, respectively. There was a marked broadening in temperature-activity profile for ZnO-NP adsorbed beta galactosidase; it showed no difference in temperature-optima between 50 degrees C and 60 degrees C. Moreover, ZnO-NP adsorbed beta galactosidase retained 53% activity after 1 h incubation with 5% galactose while the native ZnO- and solubler beta galactosidase exhibited 35% and 28% activity under similar exposure, respectively. Native ZnO and ZnO-NP adsorbed beta galactosidase retained 61% and 75% of the initial activity after seventh repeated use, respectively. It was noticed that 54%, 63% and 71% milk lactose was hydrolyzed by soluble, ZnO adsorbed and ZnO-NP adsorbed beta galactosidase in batch process after 9 h while whey lactose was hydrolyzed to 61%, 68% and 81% under similar experimental conditions, respectively. In view of its easy production, improved stability against various denaturants and excellent reusability, ZnO-NP bound beta galactosidase may find its applications in constructing enzyme-based analytical devices for clinical, environmental and food technology. (C) 2011 Elsevier B.V. All rights reserved.