Abstract
A flow injection method for the determination of fructose has been developed using immobilized hexokinase, glucose phosphate isomerase and glucose-6-phosphate dehydrogenase. The produced NADH was monitored amperometrically with a graphite electrode modified by an adsorbed Nile-blue derivative. The response was linear from the detection limit, 1μM to 3 mMD-fructose for a sample volume of 25μl. Glucose, the only interfering substrate, was eliminated with a reactor containing immobilized glucose oxidase, mutarotase and catalase. The experiments indicate that hexokinase is selective for theβ-fructo-furanose isomer. The other isomers will spontaneously isomerize toβ-fructo-furanose with time, if the latter is consumed. The fructose response factor of an enzyme reactor or biosensor will therefore depend on the isomer distribution in the original sample solution. The effect of pH and temperature can be controlled, but complexing ligands like borate interfere in a complex way and should be absent.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Methods of Enzymatic Analysis (H. U. Bergmeyer, ed.), Verlag Chemie, Weinheirn, 1984, p. 321.
G. G. Guilbault,Analytical Uses of Immobilized Enzymes, Marcel Dekker, New York, 1984.
F. Schubert, D. Kirstein, F. Scheller, M. Abraham, L. Boross,Anal. Lett. 1986,19, 2155.
P. Linares, M. D. L. de Castro, M. Valcarcel,Anal Chim. Acta 1987,202, 199.
M. Hikuma, H. Obana, T. Yasuda, I. Karube, S. Suzuki,Enzyme Microb. Technol. 1980,2, 234.
K. Matsumoto, O. Hamada, H. Usheda, Y. Osajima,Anal. Chem. 1986,58, 2732.
K. Matsumoto, H. Kamikado, H. Matsubara, Y. Osajima,Anal. Chem. 1988,60, 147.
C. A. Swindlehurst, T. A. Nieman,Anal. Chim. Acta 1988,205, 195.
C. Olive, H. R. Levy,Biochem. 1967,6, 730.
L. Gorton, A. Torstensson, H. Jaegfeldt, G. Johansson,J. Electroanal. Chem. 1984,161, 103.
M. Polasek, L. Gorton, R. Appelqvist, G. Marko-Varga, G. Johansson, in: R. Appelqvist,Thesis, Lund, 1987.
L. Gorton, G. Johansson, A. Torstensson,J. Electroanal. Chem. 1985,196, 81.
R. Appelqvist, G. Marko-Varga, L. Gorton, A. Torstensson, G. Johansson,Anal. Chim. Acta 1985,169, 237.
H. H. Weetall, in:Methods in Enzymology, Vol. XLIV (K. Mosbach, ed.), Academic Press, New York, 1976, p. 134.
B. Olsson, B. Stålbom, G. Johansson,Anal Chim. Acta 1986,179, 203.
W. Pigman, H. S. Isbell,Advan. Carbohyd. Chem. 1968,23, 11;1969,24, 13.
R. S. Shallenberger,Pure and Appl. Chem. 1978,50, 1409.
W. Goux,J. Am. Chem. Soc. 1985,107, 4320.
M. Makkee, A. P. G. Kieboom, H. van Bekkum,Recl. Trav. Chim. Pays-Bas 1985,104, 230.
M. Cockman, D. G. Kubler, A. S. Oswald, L. Wilson,J. Carbohydr. Chem. 1987,6, 181.
R. K. Crane,The Enzymes, Vol. 6 (2nd Ed.), (P. D. Boyer, H. Lardy, K. Myrbäck, eds.), Academic Press, New York, 1962, pp. 60–66.
M. Skoog, G. Johansson (manuscript).
S. Takagi, G. A. Jeffrey,Acta Crystallogr. 1977,B33, 3510.
K. Bock, M. Meldal, B. Meyer, L. Wiebe,Acta Chem. Scand. 1983,B37, 101.
M. Makkee, A. P. G. Kieboom, H. van Bekkum,Recl. Trav. Chim. Pays-Bas 1984,103, 361.
M. D. Lilly, W. E. Hornby, E. M. Crook,Biochem. J. 1966,100, 718.
W. E. Morf,Mikrochim. Acta [Wien] 1980,II, 317.
G. Johansson, L. Ögren, B. Olsson,Anal. Chim. Acta 1983,145, 71.
H. Huck, A. Schelter-Graf, H.-L. Schmidt,Bioelectrochem. Bioenerg. 1984,13, 199.
A. Sols, G. De la Fuente, C. Villar-Palasi, C. Asensio,Biochim. Biophys. Acta 1958,30, 92.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Skoog, M., Johansson, G., Olsson, B. et al. Fructose determination using immobilized enzymes in a flow system with special emphasis on the effect of isomerism. Mikrochim Acta 96, 131–142 (1988). https://doi.org/10.1007/BF01236098
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01236098