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Immobilization of glucose oxidase on sepharose by UV‐initiated graft copolymerization
Author(s) -
D'angiuro L.,
Cremonesi P.
Publication year - 1982
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.260240117
Subject(s) - glucose oxidase , thermostability , sepharose , immobilized enzyme , chemistry , copolymer , enzyme , covalent bond , oxidase test , catalysis , polymer , polymer chemistry , chromatography , combinatorial chemistry , biochemistry , organic chemistry
The performance of a new method of enzyme immobilization based on photochemically initiated direct graft copolymerization was recently investigated. The immobilization reaction can be carried out in a simple way and by carefully selecting the reaction conditions, the enzyme‐graft copolymer can be obtained as the main reaction product. Coupling efficiency of glucose oxidase has been found to depend only on the amount of photocatalyst (FeCl 3 ) fixed on Sepharose used as polysaccharide support. Small quantities of glycidymethacrylate (GMA) (0.25 g/g dry Sepharose) are sufficient but necessary to achieve the best enzyme coupling efficiency (20–40%). Enzyme immobilization occurs very rapidly and the entire reaction occurs within 60 min. Reaction patterns and physicochemical characteristics of the obtained enzyme‐graft copolymers exclude the glucose oxidase entrapment: therefore a covalent attachment mechanism may be proposed. The kinetic parameters of immobilized glucose oxidase ( K m ′ = 2.0 × 10 −2 M ) are quite similar to those of free enzyme ( K m = 1.93 × 10 −2 M ), and no diffusion limitation phenomena are evidenced in samples having different enzyme or polymer content. Lyophilization, thermostability, and long‐term continuous operation also have been investigated. The advantages of this method over that using vinylenzyme copolymerization are discussed.