Immobilization of glucose oxidase on sepharose by UV-initiated graft copolymerization

Journal Article · Fri Jan 01 00:00:00 EST 1982 · Biotechnol. Bioeng. Symp.; (United States)

DOI:https://doi.org/10.1002/bit.260240117· OSTI ID:5240149

D'Angiuro, L ^[1]; Cremonesi, P

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1. Stazione Sperimentale per la Cellulosa, Milan, Italy

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/sub 3/) fixed on Sepharose used as polysaccharide support. Small quantities of glycidylmethacrylate (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/sub m/' = 2.0 x 10/sup -2/M) are quite similar to those of free enzyme (K/sub m/ = 1.93 x 10/sup -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.

• OSTI ID: 5240149
• Journal Information: Biotechnol. Bioeng. Symp.; (United States), Vol. 24:1
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
59 BASIC BIOLOGICAL SCIENCES
COPOLYMERIZATION
RADIOINDUCTION
IMMOBILIZED ENZYMES
OXIDOREDUCTASES
REACTION KINETICS
GLUCOSE
ULTRAVIOLET RADIATION
ALDEHYDES
CARBOHYDRATES
CHEMICAL REACTIONS
ELECTROMAGNETIC RADIATION
ENZYMES
HEXOSES
KINETICS
MONOSACCHARIDES
ORGANIC COMPOUNDS
POLYMERIZATION
RADIATIONS
SACCHARIDES
560111* - Radiation Effects on Biochemicals- In Vitro- (-1987)
550200 - Biochemistry