An Optical Biosensor from Green Fluorescent Escherichia coli for the Evaluation of Single and Combined Heavy Metal Toxicities
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. GFP-Modified E. coli Bacteria
2.3. GFP E. coli Bacterial Cell Culture
2.4. Development of Microbial Biosensor
2.5. Selectivity Study
2.6. Optimization of Whole Cell Biosensor Response
2.7. Reproducibility and Long Term Stability of the Biosensor
2.8. Evaluation of Single and Combined Metal Toxicities via Biosensor and Microtox Assay
3. Results and Discussion
3.1. Characteritics of the Whole Cell Biosensor Response
3.2. Selectivity Study
3.3. Optimization of Experimental Conditions
3.4. Reproducibility and Long Term Stability Studies
3.5. Biosensor Response towards Various Concentrations of Single Toxicants
Toxicants | Dynamic Range (µg/L) | LOD (µg/L) | Slopes (%RFU/µg·L−1) | EC50 (µg/L) | R2 |
---|---|---|---|---|---|
GFP cell biosensor, 2 min (n = 3) | |||||
Cu(II) | (0.05–1) | 0.04 | −45.081 | 0.9 | 0.98 |
Cd(II) | (0.50–10) | 0.32 | −5.015 | 8.9 | 0.99 |
Pb(II) | (0.70–20) | 0.46 | −2.564 | 17.4 | 0.99 |
Zn(II) | (5–100) | 2.80 | −0.506 | 84.4 | 0.99 |
Cr(VI) | (0.10–5) × 103 | 1.00 × 102 | −0.009 | 4.5 × 103 | 0.98 |
Co(II) | (0.50–7) × 103 | 2.50 × 102 | −0.006 | 6.8 × 103 | 0.99 |
Ni(II) | (0.70–10) × 103 | 4.00 × 102 | −0.005 | 9.0 × 103 | 0.98 |
Ag(II) | (1.00–20) × 103 | 7.20 × 102 | −0.002 | 2.0 × 104 | 0.99 |
Fe(III) | (5.00–70) × 103 | 2.60 × 103 | 0.001 | 6.4 × 104 | 0.99 |
Microtox assay, 15 min (n = 3) | |||||
Cu(II) | (0.03–2) × 103 | 10.12 | 38.836× 10−3 * | 1.1 × 103 | 0.98 |
Cd(II) | (0.5–80) × 103 | 0.42 × 103 | 0.953 × 10−3 * | 5.6 × 104 | 0.99 |
Pb(II) | (0.5–80) × 103 | 0.45 × 103 | 0.810 × 10−3 * | 6.1 × 103 | 0.99 |
Zn(II) | (0.5–100) × 103 | 0.46 × 103 | 0.703 × 10−3 * | 6.2 × 104 | 0.99 |
Cr(VI) | (1–150) × 103 | 0.50 × 103 | 0.446 × 10−3 * | 9.0 × 104 | 0.98 |
Co(II) | (10–150) × 103 | 5.60 × 103 | 0.500 × 10−3 * | 9.8 × 104 | 0.99 |
Ni(II) | (1–120) × 103 | 0.65 × 103 | 0.549 × 10−3 * | 6.5 × 104 | 0.98 |
Ag(II) | (1–120) × 103 | 0.52 × 103 | 0.601 × 10−3 * | 6.3 × 104 | 0.96 |
Fe(III) | (15–150) × 103 | 10.25 × 103 | 0.504 × 10−3 * | 9.5 × 104 | 0.97 |
Analyte | Biological Component | Immobilization Matrix | Dynamic Range (μg/L) | LOD (μg/L) | Time (min) | References |
---|---|---|---|---|---|---|
Cu(II) | GFP E. coli | Alginate film | (0.05–1) | 4.0 × 10−2 | 2 | This work |
H. crispa | Agar gel | (0.01–672) × 103 | 6.0 | 20 | [37] | |
A. fischeri | Alginate microspheres | (0.1–2.0) × 102 | 6.4 | 6 | [19] | |
Cd(II) | GFP E. coli | Alginate film | (0.5–10) | 0.32 | 2 | This work |
S. cereviceae | Agar gel | (4.6–45.8) × 103 | 1.83 × 103 | 8 | [37] | |
A. fischeri | Alginate microspheres | (0.2–5.0) × 103 | 1.6 × 102 | 6 | [19] | |
Pb(II) | GFP E. coli | Alginate film | (0.7–20) | 0.36 | 2 | This work |
A. fischeri | Alginate microspheres | (0.5–7) × 102 | 0.5 × 102 | 6 | [19] | |
Zn(II) | GFP E. coli | Alginate film | (5–100) | 2.80 | 2 | This work |
A. fischeri | Alginate microspheres | (0.5–7) × 102 | 0.32 × 103 | 6 | [19] | |
Cr(VI) | GFP E. coli | Alginate film | (0.1–5) × 103 | 1.0 × 102 | 2 | This work |
A.ferrooxidans | Cellulose membrane | (0.02–118) × 103 | 18 | 1 | [36] | |
A. fischeri | Alginate microspheres | (0.1–2 ) × 104 | 1.0 × 103 | 6 | [19] | |
Co(II) | GFP E. coli | Alginate film | (0.5–7) × 103 | 2.5 × 102 | 2 | This work |
A. fischeri | Alginate microspheres | (0.5–5.0) × 104 | 1.7 × 103 | 6 | [19] | |
Ni(II) | GFP E. coli | Alginate film | (0.7–10) × 103 | 4.2 × 102 | 2 | This work |
B. sphaericus | Whatman membrane | (2–40) | 0.02 | 2 | [38] | |
A. fischeri | Alginate microspheres | (0.5–7) × 104 | 2.8 × 103 | 6 | [19] | |
Ag(I) | GFP E. coli | Alginate film | (0.1–2) ×104 | 7.2 × 102 | 2 | This work |
A. fischeri | Alginate microspheres | (0.2–7) × 104 | 1.8 × 103 | 6 | [19] | |
Fe(III) | GFP E. coli | Alginate film | (5.0–70) × 103 | 2.60 × 103 | 2 | This work |
A. fischeri | Alginate microspheres | (0.5–7) × 104 | 0.31 × 104 | 6 | [19] |
Heavy Metals and Incubation Times | This Work | Futra et al. [19] |
---|---|---|
Times (min) | 2 | 6 |
Cu(II) (μg/L) | 0.9 | 1.7 × 102 |
Cd(II) (μg/L) | 8.9 | 6.3 × 103 |
Pb(II) (μg/L) | 17.4 | 0.7 × 103 |
Zn(II) (µg/L | 84.4 | 6.0 × 102 |
Cr(VI) (μg/L) | 4.5 × 103 | 1.8 × 104 |
Co(II) (μg/L) | 6.8 × 103 | 6.6 × 104 |
Ni(II) (μg/L) | 9.0 × 103 | 6.6 × 104 |
Ag(I) (μg/L) | 2.0 × 104 | 6.0 × 104 |
Fe(III) (µg/L) | 6.4 × 104 | 7.0 × 104 |
3.6. Biosensor Response towards Combined Metals
Toxicant Mixture | AI | Toxicity Rate |
---|---|---|
(1:1 w/w) | ||
Pb(II) + Zn(II) | −0.41 | Antagonistic |
Cu(II) + Zn(II) | −3.73 | Antagonistic |
Cu(II) + Pb(II) | −1.72 | Antagonistic |
Cd(II) +Zn(II) | −1.62 | Antagonistic |
Cd(II) + Pb(II) | −2.94 | Antagonistic |
Cd(II) + Cu(II) | −1.97 | Antagonistic |
(2:1 w/w) | ||
Pb(II) + Zn(II) | −0.27 | Antagonistic |
Cu(II) + Zn(II) | −1.47 | Antagonistic |
Cu(II) + Pb(II) | −1.79 | Antagonistic |
Cd(II) +Zn(II) | −0.72 | Antagonistic |
Cd(II) + Pb(II) | −2.07 | Antagonistic |
Cd(II) + Cu(II) | −1.87 | Antagonistic |
(1:2 w/w) | ||
Pb(II) + Zn(II) | −0.878 | Antagonistic |
Cu(II) + Zn(II) | −1.64 | Antagonistic |
Cu(II) + Pb(II) | −2.00 | Antagonistic |
Cd(II) +Zn(II) | −0.75 | Antagonistic |
Cd(II) + Pb(II) | −1.58 | Antagonistic |
Cd(II) + Cu(II) | −2.88 | Antagonistic |
(1:1:1 to 1:1:1:1 w/w) | ||
Cu(II) + Cd(II) + Pb(II) | −1.984 | Antagonistic |
Cu(II) + Cd(II) + Zn(II) | −1.990 | Antagonistic |
Cd(II) + Pb(II) + Zn(II) | −3.186 | Antagonistic |
Cu(II) + Cd(II) + Pb(II) + Zn(II) | −6.033 | Antagonistic |
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Futra, D.; Heng, L.Y.; Ahmad, A.; Surif, S.; Ling, T.L. An Optical Biosensor from Green Fluorescent Escherichia coli for the Evaluation of Single and Combined Heavy Metal Toxicities. Sensors 2015, 15, 12668-12681. https://doi.org/10.3390/s150612668
Futra D, Heng LY, Ahmad A, Surif S, Ling TL. An Optical Biosensor from Green Fluorescent Escherichia coli for the Evaluation of Single and Combined Heavy Metal Toxicities. Sensors. 2015; 15(6):12668-12681. https://doi.org/10.3390/s150612668
Chicago/Turabian StyleFutra, Dedi, Lee Yook Heng, Asmat Ahmad, Salmijah Surif, and Tan Ling Ling. 2015. "An Optical Biosensor from Green Fluorescent Escherichia coli for the Evaluation of Single and Combined Heavy Metal Toxicities" Sensors 15, no. 6: 12668-12681. https://doi.org/10.3390/s150612668