Immobilization of β-glucosidase by self-catalysis and compared to crosslinking with glutaraldehyde

Nanoparticles have been successfully used for immobilization of different enzymes. The enzyme immobilized by nanomaterials has promising application in the biotechnological industry. The β-glucosidase is very important in industrial field. The immobilization of β-glucosidase by nanomaterials increases its activity and reusability. In this work, the β-glucosidase was extracted from fruiting bodies of Agrocybe aegirit and was purified by anion exchange. The β-glucosidase was immobilized by SiO2 nanoparticles (nano-SiO2) crosslink with glutaraldehyde (GA). On the other hand, the β-glucosidase firstly immobilized by process adsorption and then crosslink with genipin (GP) which was produced by the hydrolysis of geniposide by β-glucosidase. Then we compared the properties of both immobilized GA-crosslinked β-glucosidase and GP-crosslinked β-glucosidase, their immobilization yields were 83.34% and 96.29% respectively. The GA-crosslinked β-glucosidase and free β-glucosidase revealed optimal pH at 6.0. The optimal pH of GP-crosslinked β-glucosidase was ranging between 4.5 and 7.5. The GA-crosslinked β-glucosidase and GP-crosslinked β-glucosidase revealed the optimal temperature at 50 °C, 70 °C respectively. For the free β-glucosidase, the optimal temperature was 55 °C. Furthermore, the GA-crosslinked β-glucosidase and GP-crosslinked β-glucosidase were characterized by scanning electron microscope (SEM). The geniposide consumption in the reaction system of GA-crosslinked β-glucosidase was analyzed by high performance liquid chromatography (HPLC).