His6-OPH enzyme-based bio-hybrid material for organophosphate detection
- 191 Downloads
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.
KeywordsEnzyme 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.
- 1.Chambers JE, Levi PE (1992) Organophosphates: chemistry, fate, and effects. Academic, New York, pp 3–8Google Scholar
- 2.Trojanowicz M, Hitchman ML (1996) Trends Anal Chem 15:38–45Google Scholar
- 10.Efremenko EN, Sergeeva VS (2001) Russian Chem Bull (Int Ed) 50:1825–1832Google Scholar
- 18.Lev O, Tsionsky M, Rabinovich L, Glezer V, Sampath S, Pankratov I, Gun J (1995) Anal Chem 67:22A–30AGoogle Scholar
- 20.Efremenko EN, Votchitseva YV, Aliev TK, Varfolomeyev SD (2005) Patent RU no. 2255975Google Scholar
- 21.Price NC (1999) Fundamentals of enzymology: an introduction to enzyme kinetics, 3rd edn. Oxford University Press, London, pp 118–153Google Scholar
- 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
- 28.EU Water Framework Directive. Available at: http://europa.eu/environment/water/water-framework/index_en.html