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His6-OPH enzyme-based bio-hybrid material for organophosphate detection

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Abstract

In this work, we report on the development of a bio-sensing film for the detection of organophosphorous compounds using sol–gel technology. A novel sol–gel immobilization method employing tetraethoxysilane/3-glycidoxypropyltrimethoxysilane/water hybrid material was developed and used to immobilize the hexahistidine-tagged organophosphorous hydrolase enzyme (His6-OPH). Bio-sensing layers with encapsulated His6-OPH of various structures (water/silane, precursor ratios) have been prepared. The optimal (P = 5:1, R = 188) bio-sensing layers retained 90% of the initial enzyme activity. Furthermore, the bio-sensing layer prepared by this method was able to maintain its activity at or above 80% of its initial activity for 2 weeks. The bio-hybrid film also showed excellent reusability and improved activity at neutral pH in comparison to the same enzyme in solution.

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Acknowledgments

This work was supported by grants from Slovenian Research Agency (ARRS) for Young researchers (1000-08-310045) and international Slovenia-Russia Cooperation in Science, 2010–2011, the project no. BI-RU/10-11-017.

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Authors and Affiliations

  1. Faculty of Mechanical Engineering, Centre of Sensor Technology, University of Maribor, Smetanova 17, 2000, Maribor, Slovenia

    Nina Frančič, Aljoša Košak & Aleksandra Lobnik

  2. Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119992, Russia

    Ilya Lyagin & Elena N. Efremenko

Authors
  1. Nina Frančič
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  2. Aljoša Košak
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  3. Ilya Lyagin
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  4. Elena N. Efremenko
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  5. Aleksandra Lobnik
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Correspondence to Aleksandra Lobnik.

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Frančič, N., Košak, A., Lyagin, I. et al. His6-OPH enzyme-based bio-hybrid material for organophosphate detection. Anal Bioanal Chem 401, 2631–2638 (2011). https://doi.org/10.1007/s00216-011-5336-1

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  • DOI: https://doi.org/10.1007/s00216-011-5336-1

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