Immobilization of Cellulase on Zinc Oxide Deposited on Zeolite Pellets for Enzymatic Saccharification of Cellulose

The consumption of fossil fuels to fulfill the global energy demand can cause global warming issues. Renewable energy, i.e., bioethanol, from lignocellulosic biomass, is a promising source of alternative energy to fossil fuels. The conversion of lignocellulosic biomass into bioethanol requires the release of fermentable sugars during the saccharification process using cellulase. However, the utilization of this enzyme on an industrial scale is not feasible due to its difficult separation, instability, and high cost. Here, we present a method for cellulase immobilization on functionalized zinc oxide prepared from either zinc nitrate hexahydrate (ZnO(I)) or zinc acetate dihydrate (ZnO(II)) solutions on zeolite (ZEO) pellets. The immobilized cellulase on ZnO-ZEO structures was characterized by scanning electron microscopy, Xray diffraction spectroscopy, and Fourier transform infrared spectroscopy. The immobilization efficiencies of immobilized cellulase either on ZnO(I)-ZEO or ZnO(II)-ZEO were determined as 58.17 ± 0.75% and 55.51 ± 0.81%, respectively. The immobilized cellulase on ZnO-ZEO was capable of catalyzing microcrystalline cellulose breakdown, releasing reducing sugars. The immobilized cellulase on these structures could be recycled up to four repetitive runs. Based on kinetic data, both the Michaelis constants (Km) and maximum reaction velocity (Vmax) of the immobilized cellulase on the ZnO-ZEO structures were lower than those of free cellulase. This suggests that immobilized cellulase has a higher affinity toward the substrate, but a lower reaction rate than the free enzyme