Amperometric Glucose Sensor Using Thermostable Co-Factor Binding Glucose Dehydrogenase

Abstract

A thermostable mediator-type enzyme glucose sensor was constructed. The electrode was fabricated using chemically cross-linked thermostable co-factor binding glucose dehydrogenase (GDH) from thermophilic bacteria in carbon paste matrix. The electrode responded directly proportional to D-glucose concentration from 0.01 mM to 3 mM in stirred buffer containing 1 mM 1-methoxyphenazinemethosulfate as a mediator with the steady-state mode. The storage stability was examined by incubating the enzyme electrode at 50°C during the measurement. The cross-linked GDH immobilized electrode showed good storage stability. Ninety percent of its initial response was retained after incubation in buffer solution for 9 days at 50°C. The flow injection analysis (FIA) glucose sensing system was also constructed by immobilizing the cross-linked GDH and ferrocene as a mediator in the carbon paste matrix. The FIA system was able to measure 600 samples for 100 h. Keywords : glucose dehydrogenase, thermostability, cross-link, co-factor binding, glucose sensor, FIA system. © 2003, The Institute of Electrical Engineers of Japan. All rights reserved.