Studies on the preparation of a sufficient carrier from egg protein and carrageenan for cellulase with optimization and application

Abstract

Abstract Egg protein (EP) concentration, pH, and glutaraldehyde (GA) concentration were optimized using Box Behnken design (BBD) to prepare GA-EP-Carr (Carrageenan) beads as a carrier for Aspergillus niger MK981235 cellulase. It was recommended that the concentrations of GA, and EP be set at 11.21% (w/v), 8% (w/w), and pH 3, respectively. It was determined that 60 °C and 2% for free form and 60 °C and 3% for im-cellulase were the optimum temperature and CMC concentration parameters for maximum enzyme activity. Free and im-cellulase were determined to have Km and Vmax of 2.22 mg.ml-1 and 1.76 µmol.ml-1.min-1, and 4.55 mg.ml-1 and 3.33 µmol.ml-1.min-1, respectively. Covalent coupling of A. niger cellulase to GA- EP- Carr beads improved its thermodynamic parameters T1/2 and D-values by 2.48, 2.01, and 2.36 times at 40, 50, and 60 °C, respectively. GA- EP- Carr im-cellulase was 100% active for 60 days at 4 °C and can be used for CMC hydrolysis for 20 successive cycles. GA- EP- Carr im-cellulase showed remarkable efficiency in the clarification of mango, peach, grape, and orange juices emphasized by TSS (total soluble solids), turbidity, and reducing sugar measurements for 3 successive cycles. GA- EP- Carr im-cellulase can be applied with high efficiency in juice industry.