Immobilization of Aspergillus oryzae β galactosidase on concanavalin A-layered calcium alginate-cellulose beads and its application in lactose hydrolysis in continuous spiral bed reactors

Abstract

In this study, Aspergillus oryzae β galactosidase was immobilized on concanavalin A layered calcium alginate-cellulose beads as a bioaffinity support. Immobilized enzyme showed a remarkable broadening in temperature-activity profiles as compared to the native enzyme and exhibited 65% activity in the presence of 5% galactose. Michaelis constant (K m) was 2.57 mM and 5.38 mM for the free and the immobilized β galactosidase, respectively. Crosslinked β galactosidase showed greater catalytic activity in the presence of Mg 2+ and was more stable during storage at 4°C for 6 weeks. Immobilized enzyme hydrolyzed 67% lactose in milk in 8 h and 85% lactose in whey in 9 h in the stirred batch process at 50°C. The continuous hydrolysis of lactose by crosslinked β galactosidase in spiral bed reactor exhibited 93% and 88% hydrolysis of lactose at flow rate of 20 ml/h and 30 ml/h, after 1 month operation, respectively.