Quinoprotein glucose dehydrogenase and its application in an amperometric glucose sensor.
Glucose dehydrogenase (GDH), one of the recently discovered NAD(P)+-independent 'quinoprotein' class of oxidoreductase enzymes, was purified from Acinetobacter calcoaceticus LMD 79.41 and immobilised on a 1,1'-dimethylferrocene-modified graphite foil electrode. The second-order rate constant (ks) for the transfer of electrons between GDH and ferrocenemonocarboxylic acid (FMCA) in a homogeneous system, determined using direct current (DC) cyclic voltammetry, was found to be 9.4 x 10(6) litres mol-1 s-1. This value of ks for GDH was more than 40 times greater than that for the flavoprotein glucose oxidase (GOD) under identical conditions. Such high catalytic activities were also observed when GDH was immobilised in the presence of an insoluble ferrocene derivative; a biosensor based on GDH was found to produce more than twice the current density of similar GOD-based electrodes. The steady-state current produced by the GDH-based electrode was limited by the enzymic reaction since methods which increased the enzyme loadings elevated the upper limit of glucose detection from 5 mM to 15 mM. The temperature, pH, stability and response characteristics of the GDH-based glucose sensor illustrate its potential usefulness for a variety of practical applications. In particular, the high catalytic activity and oxygen insensitivity of this biosensor make it suitable for in vivo blood glucose monitoring in the management of diabetes.
D'Costa
E J
EJ
Biotechnology Centre, Cranfield Institute of Technology, Bedford, Great Britain.
Higgins
I J
IJ
Turner
A P
AP
D000150:Acinetobacter / Q000201:enzymology*; D001786:Blood Glucose / Q000032:analysis*; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005947:Glucose / Q000032:analysis*; D005948:Glucose Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008991:Monitoring, Physiologic / Q000379:methods
D001786:Blood Glucose; D004800:Enzymes, Immobilized; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone); D005947:Glucose