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Analytical and Bioanalytical Chemistry

, Volume 401, Issue 8, pp 2631–2638 | Cite as

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

  • Nina Frančič
  • Aljoša Košak
  • Ilya Lyagin
  • Elena N. Efremenko
  • Aleksandra LobnikEmail author
Original Paper

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.

Keywords

Enzyme immobilization His6-OPH Sol–gel Organophosphate detection Bio-sensing layer 

Notes

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.

Supplementary material

216_2011_5336_MOESM1_ESM.zip (4.9 mb)
Supplementary material, approximately 1.96 MB.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Nina Frančič
    • 1
  • Aljoša Košak
    • 1
  • Ilya Lyagin
    • 2
  • Elena N. Efremenko
    • 2
  • Aleksandra Lobnik
    • 1
  1. 1.Faculty of Mechanical Engineering, Centre of Sensor TechnologyUniversity of MariborMariborSlovenia
  2. 2.Faculty of ChemistryLomonosov Moscow State UniversityMoscowRussia

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