Abstract
Using SiO 2 nanoparticles as a carrier, a novel immobilized glucose oxidase (GOD) (EC1.1.3.4) was prepared via crosslinking with glutaraldehyde (GA). The optimal immobilization condition was achieved with 1% (v/v) GA, 2% (v/v) 3-aminopropiltrietoxysilane (APTS), 2.5 mg GOD (in 34 mg carrier) and solution pH of 6.5. The immobilized GOD showed maximal catalytic activity at pH 7.0 and 60 °C, and more than 85% of initial activity at the temperature from 20 °C to 80 °C. After immobilization, the enzyme exhibited improved thermal, storage and operation stability. The immobilized GOD still maintained 85% of its initial activity after the incubation at 45 °C for 360 min, whereas free enzyme had only 23% of initial activity after the same incubation. After kept at 4 °C for 30 days, the immobilized and free enzyme retained 84% and 60% of initial activity, respectively. The immobilized GOD also preserved 87% of its initial activity after six consecutive operations.
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