Abstract: With the objective of achieving practicable β-glucosidase immobilization, porous ceramic balls were taken as the carriers, and the conditions on β-glucosidase immobilization were studied. The optimum operation of β-glucosidase immobilization was as follows:5 g porous ceramic balls were put into 0.3 mL β-glucosidase solution(pH 4.8), in which the concentrations of β-glucosidase and glutaraldehyde were 36.5 μmol/(mL·min)and 0.5%, respectively. The mixture was shaken at 4 ℃ for 2 h and then dried under low temperature. The immobilized β-glucosidase was obtained after enveloped by a solution which contained 0.3 g oil-based adhesive. The optimum reaction pH of immobilized β-glucosidase was 4.8, whereas the reaction pH of free β-glucosidase was 4.2. The optimum reaction temperatures of immobilized and free β-glucosidase were 60 ℃. Immobilized β-glucosidase offered the better properties than free β-glucosidase in pH, thermal and storage tests. The K_m of immobilized and free β-glucosidase were 7.9 mmol/L and 0.73 mmol/L, respectively. A sugar mixture containing cellobiose and glucose was enzymatically hydrolyzed by the immobilized β-glucosidase. The yields of cellobiose conversion could be higher than 80% in 5 repeated batch operations.