Cholesterol Esterase and Cholesterol Oxidase Immobilized onto Arylamine Glass Beads
10.3321/j.issn:1000-3061.2002.02.006
- VernacularTitle: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
- Author:
V.Malik
1
;
S.Singh
;
C.S.Pundir
Author Information
1. M.D.University
- Keywords:
cholesterol;
cholesterol esterase;
cholesterol oxidase;
immobilization;
arylamine glass;
serum;
cholesterol determination
- From:
Chinese Journal of Biotechnology
2002;18(2):155-161
- CountryChina
- Language:Chinese
-
Abstract:
Cholesterol esterase (Cease) from bovine pancreas and cholesterol oxidasc (COD) from Bravibacterium recombinant type have been innobilized 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 8.3mg/g support respectively when imnobilized 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 reusability 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.
