Abstract
This chapter focuses on the four main immobilization techniques used in the development of enzyme electrodes: adsorption, entrapment, covalent coupling, and cross-linking. The performance of the immobilization method depends on the enzyme and each method has both advantages and drawbacks. Loss of the activity results from the change in enzyme conformation depending on the method of immobilization as well as modification of the microenvironment (i.e., local pH of the enzyme). The activity of the immobilized enzyme decreases as a result of barrier of diffusion, which slows down or prevents the substrate from reaching the active site of the enzyme. The development and the characteristics of an acetylcholinesterase biosensor are described
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References
Chibata I. (1995) Immobilized Enzyme. Kodansha Ltd., Halsted Press, Tokyo, Japan, 1978.
Mesing R. Immobilized Enzymes for Industrial Reactors. Academic Press, New York, NY.
Taylor F. R. (1991) Protein Immobilization: Fundamentals and Applications (Taylor F. R., ed.) Marcel Dekker, New York, NY.
Wang B. and Dong S. (2000) Organic phase enzyme electrode for phenolic determination based on a functionalised sol-gel composite. J. Electroanal. Chem. 478, 45–50.
Wallace G. Smyth M. and Zhao H. (1999) Conducting electroactive polymerbased biosensors. Trends Anal. Chem. 18, 245–251.
Wang J., Nascimento V. B., Kane S. L., Rogers K., Smyth M. R. and Agnes L. (1996) Screen-printed tyrosinase-containing electrodes for biosensing of enzyme inhibitors. Talanta 43, 1903–1907.
Mello L. D. and Kubota L.T. (2002) Analytical, nutritional and clinical methods. Review of the use of biosensors as analytical tools in the food and drink industries. Food Chem. 77(2), 237–256.
Ferretti S., Paynter S., Russel D. A., Sapsford K. E., and Richardson D. J. (2000) Self-assembled monolayers: a versatile tool for the formulation of the biosurfaces. Trends Anal. Chem. 19, 530–540.
Ellman G. L., Courtney K. P., Andres V., and Fearstherstone R. M. (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7, 88–90.
Nunes G. S., Montesinos T., Marques P.B. O., Fournier D., and Marty J.-L. (2001). Acetylcholine enzyme sensor for determining methamidophos insecticide. Evaluation of some genetically modified acetylcholinesterases from Drosophila melanogaster. Anal. Chim. Acta 434, 1–8.
Andreescu S., Barthelmebs L., and Marty J.-L. (2002) Immobilization of acetylcholinesterase on screen-printed electrodes: comparative study between three immobilization methods and applications to the detection of organophosphorus insecticides. Anal. Chim. Acta 464, 171–180.
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© 2006 Humana Press Inc.
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Silva Nunes, G., Marty, JL. (2006). Immobilization of Enzymes on Electrodes. In: Guisan, J.M. (eds) Immobilization of Enzymes and Cells. Methods in Biotechnology™, vol 22. Humana Press. https://doi.org/10.1007/978-1-59745-053-9_21
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DOI: https://doi.org/10.1007/978-1-59745-053-9_21
Publisher Name: Humana Press
Print ISBN: 978-1-58829-290-2
Online ISBN: 978-1-59745-053-9
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