以芳香胺玻璃为载体的固定化的胆固醇脂酶和胆固醇氧化酶

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Cholesterol Esterase and Cholesterol Oxidase Immobilized onto Arylamine Glass Beads

Cholesterol esterase (CEase) from bovine pancreas and cholesterol oxidase (COD) from Bravibacterium recombinant type have been immobilized individually and co immobilized onto arylamine glass (pore diameter 55nm) through the process of diazotization. CEase and COD retained 92.65% and 85.54% of the initial activity with conjugation yields of 7.2 mg/g and 8 3 mg/g support respectively when immobilized individually on arylamine glass beads, but retained 89.58% of the initial activity with a conjugation yield of 2.9 mg/g support when co immobilized on the same support. The effects of pH, temperature, time of incubation, substrate concentration, serum inorganic salts & metabolites, thermal stability, storage stability in cold and reu sability on the immobilized enzymes were studied and compared with those of free enzymes. The analytic use of both individually immobilized and co immobilized enzymes in discrete analysis of total and free cholesterol in serum is demonstrated.