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. 2015 May 28;15(6):12668-81.
doi: 10.3390/s150612668.

An optical biosensor from green fluorescent Escherichia coli for the evaluation of single and combined heavy metal toxicities

Dedi Futra  1 , Lee Yook Heng  2   3 , Asmat Ahmad  4 , Salmijah Surif  5 , Tan Ling Ling  6
Affiliations

An optical biosensor from green fluorescent Escherichia coli for the evaluation of single and combined heavy metal toxicities

Dedi Futra et al. Sensors (Basel). .

Abstract

A fluorescence-based fiber optic toxicity biosensor based on genetically modified Escherichia coli (E. coli) with green fluorescent protein (GFP) was developed for the evaluation of the toxicity of several hazardous heavy metal ions. The toxic metals include Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(III). The optimum fluorescence excitation and emission wavelengths of the optical biosensor were 400 ± 2 nm and 485 ± 2 nm, respectively. Based on the toxicity observed under optimal conditions, the detection limits of Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(III) that can be detected using the toxicity biosensor were at 0.04, 0.32, 0.46, 2.80, 100, 250, 400, 720 and 2600 μg/L, respectively. The repeatability and reproducibility of the proposed biosensor were 3.5%-4.8% RSD (relative standard deviation) and 3.6%-5.1% RSD (n = 8), respectively. The biosensor response was stable for at least five weeks, and demonstrated higher sensitivity towards metal toxicity evaluation when compared to a conventional Microtox assay.

Keywords: GFP E. coli; hazardous heavy metal; optical fiber biosensor; whole cell biosensor.

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Figures

Figure 1
Figure 1
The emission spectra of the immobilized autoclaved green fluorescent protein (GFP) E. coli and the whole cell biosensor before and after exposure to the Cu(II) (1.0 µg/L) metal ion.
Figure 2
Figure 2
Selectivity performance of the immobilized GFP E. coli by Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(III).
Figure 3
Figure 3
Effect of GFP E. coli cell loading (A), alginate concentration (B), and toxicants solution pH (C), on the microbial toxicity biosensor response.
Figure 4
Figure 4
The stability profile of GFP E. coli biosensor at 0.8 Abs OD (600 nm) cell concentration, 4% alginate and 0.15 M CaCl2.
See this image and copyright information in PMC

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