Analytical and Bioanalytical Chemistry

, Volume 400, Issue 4, pp 1083–1092 | Cite as

A new bacterial biosensor for trichloroethylene detection based on a three-dimensional carbon nanotubes bioarchitecture

  • Mouna Hnaien
  • Florence LagardeEmail author
  • Joan Bausells
  • Abdelhamid Errachid
  • Nicole Jaffrezic-Renault
Original Paper


Trichloroethylene (TCE), a suspected human carcinogen, is one of the most common volatile groundwater contaminants. Many different methodologies have already been developed for the determination of TCE and its degradation products in water, but most of them are costly, time-consuming and require well-trained operators. In this work, a fast, sensitive and miniaturised whole cell conductometric biosensor was developed for the determination of trichloroethylene. The biosensor assembly was prepared by immobilising Pseudomonas putida F1 bacteria (PpF1) at the surface of gold interdigitated microelectrodes through a three-dimensional alkanethiol self-assembly monolayer/carbon nanotube architecture functionalised with Pseudomonas antibodies. The biosensor response was linear from 0.07 to 100 μM of TCE (9–13,100 μg L−1). No significant loss of the enzymatic activity was observed after 5 weeks of storage at 4 °C in the M457 pH 7 defined medium (two or three measurements per week). Ninety-two per cent of the initial signal still remained after 7 weeks. The biosensor response to TCE was not significantly affected by cis-1,2-dichloroethylene and vinyl chloride and, in a limited way, by phenol. Toluene was the major interference found. The bacterial biosensor was successfully applied to the determination of TCE in spiked groundwater samples and in six water samples collected in an urban industrial site contaminated with TCE. Gas chromatography–mass spectrometric analysis of these samples confirmed the biosensor measurements.


General view of the interdigitated microelectrodes


Trichloroethylene Whole cell biosensor Pseudomonas putida F1 Interdigitated microelectrodes Single-wall carbon nanotubes Self-assembly monolayer 



This work was supported by the French National Agency for Research (EVASOL project ANR-07-ECOT 005-03). Joan Bausells also acknowledges the Spanish Ministry of Science and Innovation for its support through the Nanoselect project (CSD2007-00041) of the Consolider-Ingenio 2010 Programme.

Many thanks to Dr. Rebecca Parales from the University of California (Davis) for providing P. putida F1 mutant defective in todC1 gene, Eric Dugat-Bony from the Laboratoire Microorganismes: Génome et Environnement of Clermont-Ferrand for his precious advices, and Jean-Yves Richard and C. Curver from Sita Remediation (Suez group) for providing field water samples and HS-GC/MS results.

Supplementary material

216_2010_4336_MOESM1_ESM.pdf (281 kb)
ESM 1 (PDF 280 kb)


  1. 1.
    Moran MJ, Zogorski JS, Squillage PJ (2007) Chlorinated solvents in groundwater of the United States. Environ Sci Technol 41:74–81CrossRefGoogle Scholar
  2. 2.
    Morioka Y (1996) Control of pollution of groundwater contaminated by toxic chemicals. J Jpn Soc Water Environ 19:529–533Google Scholar
  3. 3.
    Pooley K, Blessing M, Schmidt TC, Haderlein SB, Mac Quarrie KTB, Prommer H (2009) Aerobic biodegradation of chlorinated ethenes in a fractured bedrock aquifer: quantitative assessment by compound-specific isotope analysis (CSIA) and reactive transport modeling. Environ Sci Technol 43:7458–7464CrossRefGoogle Scholar
  4. 4.
    Takeuchia M, Nanbab K, Iwamotoc H, Nireid H, Kusudae T, Kazaokae O, Owakid M, Furuyaa K (2005) In situ bioremediation of a cis-dichloroethylene-contaminated aquifer utilizing methane-rich groundwater from an uncontaminated aquifer. Water Res 39:2438–2444CrossRefGoogle Scholar
  5. 5.
    Kim S, Kim D, Pollack GM, Collins LB, Rusyn I (2009) Pharmacokinetic analysis of trichloroethylene metabolism in male B6C3F1 mice: formation and disposition of trichloroacetic acid, dichloroacetic acid, S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-l-cysteine. Toxicol Appl Pharmacol 238:90–99CrossRefGoogle Scholar
  6. 6.
    Magnuson JK, Romine MF, Burris DR, Kingsley MT (2000) Trichloroethene reductive dehalogenase from Dehalococcoides ethenogenes: sequence of tceA and substrate range characterization. Appl Environ Microbiol 66:5141–5147CrossRefGoogle Scholar
  7. 7.
    Delinsky AD, Bruckner JV, Bartlett MG (2005) A review of analytical methods for the determination of trichloroethylene and its major metabolites chloral hydrate, trichloroacetic acid and dichloroacetic acid. Biomed Chromatogr 19:617–639CrossRefGoogle Scholar
  8. 8.
    Niri VH, Bragg L, Pawliszyn J (2008) Fast analysis of volatile organic compounds and disinfection by-products in drinking water using solid-phase microextraction–gas chromatography/time-of-flight mass spectrometry. J Chromatogr A 1201:222–227CrossRefGoogle Scholar
  9. 9.
    Han TS, Kim YC, Sasaki S, Yano K, Ikebukuro K, Kitayama A, Nagamune T, Karube I (2001) Microbial sensor for trichloroethylene determination. Anal Chim Acta 431:225–230CrossRefGoogle Scholar
  10. 10.
    Han TS, Sasaki S, Yano K, Ikebukuro K, Atsushi K, Nagamune T, Karube I (2002) Flow injection microbial trichloroethylene sensor. Talanta 57:271–276CrossRefGoogle Scholar
  11. 11.
    Han TS, Sasaki S, Yano K, Ikebukuro K, Atsushi K, Nagamune T, Karube I (2003) Development of a reactor type bio-sensor for trichloroethylene. Anal Lett 36:539–547CrossRefGoogle Scholar
  12. 12.
    Gibson DT, Koch JR, Kallio RE (1968) Oxidative degradation of aromatic hydrocarbons by microorganisms. I. enzymatic formation of catechol from benzene. Biochemistry 7:2653–2662CrossRefGoogle Scholar
  13. 13.
    Spain JC, Gibson DT (1988) Oxidation of substituted phenols by Pseudomonas putida Fl and Pseudomonas sp. strain JS6. Appl Environ Microbiol 54:1399–1404Google Scholar
  14. 14.
    Wackett LP, Gibson DT (1988) Degradation of trichloroethylene by toluene dioxygenase in whole-cell studies with Pseudomonas putida Fl. Appl Environ Microbiol 54:1703–1708Google Scholar
  15. 15.
    Li S, Wackett LP (1992) Trichloroethylene oxidation by toluene dioxygenase. Biochem Biophys Res Commun 185:443–451CrossRefGoogle Scholar
  16. 16.
    Hori K, Mii J, Morono Y, Tanji Y, Unno H (2005) Kinetic analyses of trichloroethylene cometabolism by toluene-degrading bacteria harboring a tod homologous gene. Biochem Eng J 26:59–64CrossRefGoogle Scholar
  17. 17.
    Finette BA, Subramanian V, Gibson DT (1984) Isolation and characterization of Pseudomonas putida PpF1 mutants defective in the toluene dioxygenase enzyme system. J Bacteriol 160:1003–1009Google Scholar
  18. 18.
    Guedri H, Durrieu C (2008) A self-assembled monolayers based conductometric algal whole cell biosensor for water monitoring. Microchim Acta 163:179–184CrossRefGoogle Scholar
  19. 19.
    Chouteau C, Dzyadevych S, Durrieu C, Chovelon JM (2005) A bi-enzymatic whole cell conductometric biosensor for heavy metal ions and pesticides detection in water samples. Biosens Bioelectron 21:273–281CrossRefGoogle Scholar
  20. 20.
    Kumar S, Kundu S, Pakshirajan K, Dasu VV (2008) Cephalosporins determination with a novel microbial biosensor based on permeabilized Pseudomonas aeruginosa whole cells. Appl Biochem Biotechnol 151:653–664CrossRefGoogle Scholar
  21. 21.
    Chu YF, Hsu CH, Soma PK, Lo YM (2009) Immobilization of bioluminescent Escherichia coli cells using natural and artificial fibers treated with polyethyleneimine. Bioresour Technol 100:3167–3174CrossRefGoogle Scholar
  22. 22.
    Jha SK, Kanungo M, Nath A, D’Souza SF (2009) Entrapment of live microbial cells in electropolymerized polyaniline and their use as urea biosensor. Biosens Bioelectron 24:2637–2642CrossRefGoogle Scholar
  23. 23.
    Yoo SK, Lee JH, Yun SS, Gu MB, Lee JH (2007) Fabrication of a bio-MEMS based cell-chip for toxicity monitoring. Biosens Bioelectron 22:1586–1592CrossRefGoogle Scholar
  24. 24.
    D’Souza SF (2001) Microbial biosensors. Biosens Bioelectron 16:337–353CrossRefGoogle Scholar
  25. 25.
    Lei Y, Chen W, Mulchandani A (2006) Microbial biosensors. Anal Chim Acta 568:200–210CrossRefGoogle Scholar
  26. 26.
    Teles FRR, Fonseca LP (2008) Applications of polymers for biomolecule immobilization in electrochemical biosensors. Mat Sci Eng C 28:1530–1543CrossRefGoogle Scholar
  27. 27.
    Chaki NK, Vijayamohanan K (2002) Self-assembled monolayers as a tunable platform for biosensor applications. Biosens Bioelectron 17:1–12CrossRefGoogle Scholar
  28. 28.
    Mendes RK, Carvalhal RF, Kubota LT (2008) Effects of different self-assembled monolayers on enzyme immobilization procedures in peroxidase-based biosensor development. J Electroanal Chem 612:164–172CrossRefGoogle Scholar
  29. 29.
    Jung SK, Namgung MO, Oh SY, Oh BK (2009) Fabrication of self-assembled oligophenylethynylenethiol monolayer for electrochemical glucose biosensor. Ultramicroscopy 109:911–915CrossRefGoogle Scholar
  30. 30.
    Baldrich E, Laczka O, Del Campo FJ, Muñoz FX (2008) Gold immuno-functionalisation via self-assembled monolayers: study of critical parameters and comparative performance for protein and bacteria detection. J Immunol Methods 336:203–212CrossRefGoogle Scholar
  31. 31.
    Chen H, Heng CK, Puiu PD, Zhou XD, Lee AC, Lim TM, Tan SN (2005) Detection of Saccharomyces cerevisiae immobilized on self-assembled monolayer (SAM) of alkanethiolate using electrochemical impedance spectroscopy. Anal Chim Acta 554:52–59CrossRefGoogle Scholar
  32. 32.
    Hong SR, Choi SJ, Do Jeong H, Hong S (2009) Development of QCM biosensor to detect a marine derived pathogenic bacteria Edwardsiella tarda using a novel immobilisation method. Biosens Bioelectron 24:1635–1640CrossRefGoogle Scholar
  33. 33.
    Laczka O, Baldrich E, del Campo FJ, Muñoz FX (2008) Immunofunctionalisation of gold transducers for bacterial detection by physisorption. Anal Bioanal Chem 391:2825–2835CrossRefGoogle Scholar
  34. 34.
    Pyun JC, Kim SD, Chung JW (2005) New immobilization method for immunoaffinity biosensors by using thiolated proteins. Anal Biochem 347:227–233CrossRefGoogle Scholar
  35. 35.
    Gooding JJ (2005) Nanostructuring electrodes with carbon nanotubes: a review on electrochemistry and applications for sensing. Electrochim Acta 50:3049–3060CrossRefGoogle Scholar
  36. 36.
    Kim SN, Rusling JF, Papadimitrakopoulos F (2007) Carbon nanotubes for electronic and electrochemical detection of biomolecules. Adv Mater 19:3214–3228CrossRefGoogle Scholar
  37. 37.
    Zylstra GJ, Mc Combie WR, Gibson DT, Finette BA (1988) Toluene degradation by Pseudomonas putida F1: genetic organization of the tod operon. Appl Environ Microbiol 54:1498–1503Google Scholar
  38. 38.
    Jaffrezic-Renault N, Dzyadevych SV (2008) Conductometric microbiosensors for environmental monitoring. Sensors 8:2569–2588CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Mouna Hnaien
    • 1
  • Florence Lagarde
    • 1
    Email author
  • Joan Bausells
    • 2
  • Abdelhamid Errachid
    • 1
  • Nicole Jaffrezic-Renault
    • 1
  1. 1.Laboratoire des Sciences AnalytiquesUniversité de LyonVilleurbanne CédexFrance
  2. 2.Centre Nacional de Microelectrònica (IMB-CSIC), Campus UABBarcelonaSpain

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