A fluorometric fiber-optic biosensor for dual analysis of glucose and fructose using glucose-fructose-oxidoreductase isolated from Zymomonas mobilis

authored by
Sei Jin Lee, Mohammed Saleemuddin, Thomas Scheper, Heidi Loos, Hermann Sahm
Abstract

A fluorometric fiber-optic biosensor was used to determine the content of glucose and fructose according to the measurement principle of flow injection analysis (FIA). The enzyme glucose-fructose-oxidoreductase (GFOR) isolated from Zymomonas mobilis was confined in a measurement cell behind an ultrafiltration membrane. The GFOR catalyzes the oxidation of glucose to gluconolactone and the reduction of fructose to sorbitol according to the ping-pong mechanism. This unique enzyme contains NADPH tightly confined in the enzyme complex. The change of NADPH fluorescence was detected by a fluorosensor. The stability could be increased remarkably by crosslinking with glutaraldehyde. The detection range for glucose was within a concentration range of 0.055-55.5 mM and that for fructose within 0.278-331 mM. The response time for glucose and fructose was in the range of 40 s and 1 min, respectively. Steady-state was achieved for glucose injection in 2 min and fructose injection in 6.5 min. Sampling time for glucose could be reduced to 37% by including fructose in excess to the sample. This also makes the regenerating step unnecessary. The specificity of this biosensor was tested for different sugars. The results showed that this biosensor system is highly specific, sensitive and suitable for dual analysis of glucose and fructose.

External Organisation(s)
University of Münster
Forschungszentrum Jülich
Type
Article
Journal
Journal of biotechnology
Volume
36
Pages
39-44
No. of pages
6
ISSN
0168-1656
Publication date
29.07.1994
Publication status
Published
Peer reviewed
Yes
ASJC Scopus subject areas
Biotechnology, Bioengineering, Applied Microbiology and Biotechnology
Electronic version(s)
https://doi.org/10.1016/0168-1656(94)90021-3 (Access: Unknown)