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


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.


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



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 (4.9 mb)
Supplementary material, approximately 1.96 MB.


  1. 1.
    Chambers JE, Levi PE (1992) Organophosphates: chemistry, fate, and effects. Academic, New York, pp 3–8Google Scholar
  2. 2.
    Trojanowicz M, Hitchman ML (1996) Trends Anal Chem 15:38–45Google Scholar
  3. 3.
    Palchetti I, Cagnini A, Del Carlo M, Coppi C, Mascini M, Turner APF (1997) Anal Chim Acta 337:315–321CrossRefGoogle Scholar
  4. 4.
    Joshi KA, Tang J, Haddon R, Wang J, Chen W, Mulchandani A (2005) Electroanalysis 17(1):54–58CrossRefGoogle Scholar
  5. 5.
    Mulchandani A, Chen W, Mulchandani P, Wang J, Rogers KM (2001) Biosens Bioelectron 16:225–230CrossRefGoogle Scholar
  6. 6.
    Mulchandani A, Chen W, Mulchandani P, Wang J, Chen L (1999) Anal Chem 71:2246–2249CrossRefGoogle Scholar
  7. 7.
    Mulchandani A, Pan S, Chen W (1999) Biotechnol Progr 15:130–134CrossRefGoogle Scholar
  8. 8.
    Zourob M, Siomnian A, Wild J, Mohr S, Xuding Fan, Abdulhalim I, Goddard NJ (2007) Analyst 132:114–120CrossRefGoogle Scholar
  9. 9.
    Ramanathan M, Simonian AL (2007) Biosens Bioelectron 22:3001–3007CrossRefGoogle Scholar
  10. 10.
    Efremenko EN, Sergeeva VS (2001) Russian Chem Bull (Int Ed) 50:1825–1832Google Scholar
  11. 11.
    Votchitseva YA, Efremenko EN, Aliev TK, Volfomeyev SD (2006) Biocemistry (Moscow) 71:167–172CrossRefGoogle Scholar
  12. 12.
    Pierre AC (2004) Biocatal Biotransform 22:145–170CrossRefGoogle Scholar
  13. 13.
    Kandimalla VB, Tripathi VS, Ju H (2006) Crit Rev Anal Chem 36:73–106CrossRefGoogle Scholar
  14. 14.
    Avnir D, Coradin T, Lev O, Livage J (2005) J Mater Chem 16:1013–1030CrossRefGoogle Scholar
  15. 15.
    Jerónimo PCA, Araújo AN, Conceicao M, Montenegro BSM (2007) Talanta 72:13–27CrossRefGoogle Scholar
  16. 16.
    Gupta R, Chaudhury NK (2007) Biosens Bioelectron 22:2387–2399CrossRefGoogle Scholar
  17. 17.
    Dave BC, Dunn B, Valentine JS, Zink JI (1994) Anal Chem 66:1120A–1127ACrossRefGoogle Scholar
  18. 18.
    Lev O, Tsionsky M, Rabinovich L, Glezer V, Sampath S, Pankratov I, Gun J (1995) Anal Chem 67:22A–30AGoogle Scholar
  19. 19.
    Wang J (1999) Anal Chim Acta 399:21–27CrossRefGoogle Scholar
  20. 20.
    Efremenko EN, Votchitseva YV, Aliev TK, Varfolomeyev SD (2005) Patent RU no. 2255975Google Scholar
  21. 21.
    Price NC (1999) Fundamentals of enzymology: an introduction to enzyme kinetics, 3rd edn. Oxford University Press, London, pp 118–153Google Scholar
  22. 22.
    Brinker CJ, Sehgal R, Hietala SL, Deshpande R, Smith DM, Loy D, Ashley CS (1994) J Membrane Sci 94:85–102CrossRefGoogle Scholar
  23. 23.
    Chu L, Tejedor-Tejedor MI, Anderson MA (1997) Microporous Mater 8:207–213CrossRefGoogle Scholar
  24. 24.
    Sakka S (2005) Handbook of sol-gel science and technology, processing, characterization and applications; volume II: characterization of sol-gel materials and products. Kluwer, Norwell, pp 65–89Google Scholar
  25. 25.
    Chen JP, Lin WS (2003) Enzyme Microb Technol 32:801–811CrossRefGoogle Scholar
  26. 26.
    Reetz MT, Zonta A, Simpelkamp J (1996) Biotechnol Bioeng 49:527–534CrossRefGoogle Scholar
  27. 27.
    Scholze H (1985) J Non-Cryst Solids 73:669–680CrossRefGoogle Scholar
  28. 28.
    EU Water Framework Directive. Available at:

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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