Inhibition of immobilized acetylcholinesterase by aflatoxin B1 in a potentiometric biosensor

Authors

  • K. V. Stepurska Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022; University Claude Bernard Lyon 1, Institute of Analytical Sciences, UMR5280 CNRS/UCBL/ENS, 5, rue de la Doua, Villeurbanne, France, 69100 Author
  • M. I. Korobko Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author
  • V. M. Arkhypova Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author
  • O. O. Soldatkin Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author
  • F. Lagarde University Claude Bernard Lyon 1, Institute of Analytical Sciences, UMR5280 CNRS/UCBL/ENS, 5, rue de la Doua, Villeurbanne, France, 69100 Author
  • A. P. Soldatkin Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author
  • S. V. Dzyadevych Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680; Institute of High Technologies, Taras Shevchenko National University of Kyiv 2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022 Author

DOI:

https://doi.org/10.7124/bc.000929

Keywords:

biosensor, immobilized acetylcholinesterase, type of inhibition, aflatoxin B1, potentiometric transducer

Abstract

Aim. To identify a type of inhibition of immobilized acetylcholinesterase by aflatoxin B1. Methods. A bioselective element of the potentiometric biosensor was created using acetylcholinesterase, which was covalently immobilized on the surface of the pH-FET sensor by glutaraldehyde crosslinking with bovine serum albumin. Results. Optimal conditions for the potentiometric biosensor operation such as pH-optimum of the enzyme action and its inhibition were defined. An apparent Michaelis constant, as well as a maximum initial reaction rate of immobilized acetylcholinesterase as a part of the biosensor were determined. The type of reversible inhibition of immobilized acetylcholinesterase by aflatoxin B1 in potentiometric biosensor was identified by using a new graphical “degree of inhibition” method and the obtained result was confirmed with one of the tradi-tional methods, such as the Lineweaver-Burk plot. Conclusions. This study helps to understand the mechanisms of enzyme inhibition in biosensors and brings the biosensor implementation closer.

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Published

2016-09-30

Issue

Vol. 32 No. 4 (2016)

Section

Molecular and Cell Biotechnologies