The development of immobilized enzymes is a growing field of interest within the industrial biotechnology community. Glucose oxidase (GOx) is an important commercial enzyme used in biofuel cells, food and fermentation processes, medical applications, and glucose monitoring. We have explored the efficacy of Amberzyme oxirane resin as a support for GOx immobilization. The resin beads are rigid acrylic porous polymeric spheres that have been functionalized with epoxide groups. These epoxide groups on the resin surface provide for a one-step process for reaction with available nucleophiles, such as the epsilon amino groups of lysines found on the enzyme surface. This creates a covalent linkage between an enzyme and the functionalized resin, allowing immobilization reactions to proceed under milder conditions, with fewer processing steps and with reduced hazardous chemical waste, than achievable with typical multistep immobilization protocols. We have used the native and deglycosylated GOx enzyme to determine the impact of glycosylation on attachment efficiency. Enzyme loading and enzyme activity measurements for each were compared as a function of increasing enzyme-immobilization solution concentration. It was determined that the Amberzyme oxirane resin beads were an effective substrate for GOx enzyme immobilization and that, although higher loading levels can be achieved with the deglycosylated GOx, there is no corresponding increase in specific enzyme activity. © Copyright 2008, Mary Ann Liebert, Inc.
