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Galactose Oxidase: Applications of the covalently immobilized enzyme in a packed Bed configuration
Author(s) -
Dahodwala S. K.,
Weibel M. K.,
Humphrey A. E.
Publication year - 1976
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260181204
Subject(s) - immobilized enzyme , chemistry , covalent bond , glucose oxidase , lactose , reagent , chromatography , galactose oxidase , packed bed , enzyme , enzyme kinetics , kinetics , azide , combinatorial chemistry , organic chemistry , active site , physics , quantum mechanics
Galactose oxidase (E.G. 1.1.3.9) was covalently immobilized to chemically modified porous silica particles by reaction of the native enzyme with pendant benzoyl azide groups on the carrier. The enzyme loading on the carrier was 100–150 units per milliliter. The immobilized enzyme was incorporated into a hardware assembly suitable for the determination of galactose or lactose concentrations in complex biological fluids. The prototype instrument as described is suitable for continuous, on‐line monitoring or discrete sample analysis. Reaction conditions can be readily provided which maintain global first order kinetics within the reactor and strict linearity of the procedure over a wide range of sample concentrations. Auto‐inactivation of the immobilized enzyme can be prevented by K 3 Fe(CN) 6 and long‐term reactor stability can be achieved by the periodic application of the reagent to the enzyme reactor in situ .