Entrapment of Glucose Oxidase by 2-Hydroxyethyl Methacrylate

Abstract

Glucose oxidase (β -D-Glucose: oxygen oxidoreductase E. C.1.1.3.4) was entrapped in polymer matrix, cross-linked poly-2-hydroxyethyl methacrylate, to obtain insoluble enzyme. The optimum time for polymerization in entrapping the enzyme was found to be 4 hr at 20° C. The enzymic activity of the insoluble one was independent of polymerization initiator, riboflavin, ammonium peroxodisulfate, potassium peroxodisulfate and of carrier, 2-hydroxyethyl methacrylate. However, the particle size of the insoluble one and concentration of cross-linking agent had an influence on enzymic activity. The concentration of cross-linking agent that gave the highest activity was 15 mol percent (N, N'-methylene bisacrylamide in a 0.1 mole of the monomer). The behaviors of the enzyme entrapped in various polymers were investigated by measuring the values derived from elution of enzyme from polymers, from dynamics and from kinetics. Both the enzyme and co-enzyme (FAD) were not eluted from the particles with low concentrations of the cross-linking agent (0-10 mol%). However, Michaelis constants depended on the particle size. On the other hand, it was observed that the enzyme and coenzyme (FAD) were eluted from the particles which were polymerized with high cross-linking agent (10-20 mol %). The Michaelis constants remained constant in these particles. Interactions between synthetic polymers and enzyme were observed to be strong in the particles with low concentration of crosslinking agent.