Analytical and Bioanalytical Chemistry

, Volume 398, Issue 5, pp 1891–1898

Development of an enzymatic fiber-optic biosensor for detection of halogenated hydrocarbons

  • Sarka Bidmanova
  • Radka Chaloupkova
  • Jiri Damborsky
  • Zbynek Prokop
Original Paper

DOI: 10.1007/s00216-010-4083-z

Cite this article as:
Bidmanova, S., Chaloupkova, R., Damborsky, J. et al. Anal Bioanal Chem (2010) 398: 1891. doi:10.1007/s00216-010-4083-z

Abstract

An enzyme-based biosensor was developed by co-immobilization of purified enzyme haloalkane dehalogenase (EC 3.8.1.5) and a fluorescence pH indicator on the tip of an optical fiber. Haloalkane dehalogenase catalyzes hydrolytic dehalogenation of halogenated aliphatic hydrocarbons, which is accompanied by a pH change influencing the fluorescence of the indicator. The pH sensitivity of several fluorescent dyes was evaluated. The selected indicator 5(6)-carboxyfluorescein was conjugated with bovine serum albumin and its reaction was tested under different immobilization conditions. The biosensor was prepared by cross-linking of the conjugate in tandem with haloalkane dehalogenase using glutaraldehyde vapor. The biosensor, stored for 24 h in 50 mM phosphate buffer (pH 7.5) prior to measurement, was used after 15 min of equilibration, the halogenated compound was added, and the response was monitored for 30 min. Calibration of the biosensor with 1,2-dibromoethane and 3-chloro-2-(chloromethyl)-1-propene showed an excellent linear dependence, with detection limits of 0.133 and 0.014 mM, respectively. This biosensor provides a new tool for continuous in situ monitoring of halogenated environmental pollutants.

https://static-content.springer.com/image/art%3A10.1007%2Fs00216-010-4083-z/MediaObjects/216_2010_4083_Figa_HTML.gif
Figure

Fiber-optic biosensor composed of purified enzyme haloalkane dehalogenase (HLD) and fluorescence pH indicator 5(6)-caboxyfluorescein conjugated with bovine serum albumin (BSA)

Keywords

Enzyme-based biosensorHaloalkane dehalogenaseHalogenated hydrocarbonsOptical biosensorpH indicator

Abbreviations

BSA

bovine serum albumin

CF

5(6)-carboxyfluorescein

CF-BSA

conjugates of 5(6)-carboxyfluorescein and albumin

CLEA

cross-linked enzyme aggregate

EDC

1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

HEPES

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

ISE

ion-selective electrode

NHS

N-hydroxysuccinimide

PVA

poly(vinyl alcohol)

Supplementary material

216_2010_4083_MOESM1_ESM.pdf (58 kb)
ESM 1(PDF 57.9 kb)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Sarka Bidmanova
    • 1
  • Radka Chaloupkova
    • 1
  • Jiri Damborsky
    • 1
  • Zbynek Prokop
    • 1
  1. 1.Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment, Faculty of ScienceMasaryk UniversityBrnoCzech Republic