Collect. Czech. Chem. Commun. 1990, 55, 2568-2574

Preparation of enzyme electrode for D-glucose determination by immobilization of glucose oxidase on collagen membrane

Eva Vrbová and Miroslav Marek

Biotechnology Center, department of Biochemistry and Microbiology, Prague Institute of Chemical Technology, 166 28 Prague 6


An enzyme electrode for the determination of D-glucose was prepared by immobilization of glucose oxidase (EC on an activated collagen membrane using glutaraldehyde and the Ugi reaction resp. and by subsequent fixation of the membrane to an oxygen sensor of the Clark type. Two different procedures for the modification of the support, the composition of the reaction mixture and the immobilization time were examined. The electrode prepared was tested as regards the effect of pH and temperature on the magnitude of the response. The range of the linear dependence of the sensor response on substrate concentration (1.7 . 10-5 - 2.0 . 10-3), the apparent Michaelis constant of the immobilized enzyme (KM(app.) = 4.16 . 10-3 mol dm-3) and the stability of the biosensor as function of storage mode and number of assays performed with one electrode were determined. In view of the high stability and linear range of the concentration dependence the enzyme electrode is suitable for the determination of D-glucose in samples analyzed in agricultural and food laboratories.