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 55 nm) 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 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.
Malik
V
V
Biochemistry Research Laboratory, Department of Bio-Sciences, M. D. University, Rohtak-124 001, Haryana, India.
Singh
S
S
Pundir
C S
CS
D000588:Amines; D000818:Animals; D002417:Cattle; D002784:Cholesterol / Q000097:blood; D002789:Cholesterol Oxidase / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D005898:Glass; D006361:Heating; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals; D010179:Pancreas / Q000201:enzymology; D012492:Salts; D002787:Sterol Esterase / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D000588:Amines; D004800:Enzymes, Immobilized; D008670:Metals; D012492:Salts; D002784:Cholesterol; D002789:Cholesterol Oxidase; D002787:Sterol Esterase