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Molecular Biology Reports

, Volume 39, Issue 12, pp 11225–11229 | Cite as

Development of a broad-spectrum fluorescent heavy metal bacterial biosensor

  • P. Gireesh-Babu
  • Aparna ChaudhariEmail author
Article

Abstract

Bacterial biosensors can measure pollution in terms of their actual toxicity to living organisms. A recombinant bacterial biosensor has been constructed that is known to respond to toxic levels of Zn2+, Cd2+ and Hg2+. The zinc regulatory gene zntR and zntA promoter from znt operon of E. coli have been used to trigger the expression of GFP reporter protein at toxic levels of these ions. The sensor was induced with 3–800 ppm of Zn2+, 0.005–4 ppm of Cd2+ and 0.001–0.12 ppm of Hg2+ ions. Induction studies were also performed in liquid media to quantify GFP fluorescence using fluorimeter. To determine the optimum culture conditions three different incubation periods (16, 20 and 24 h) were followed. Results showed an increased and consistent fluorescence in cells incubated for 16 h. Maximum induction for Zn2+, Cd2+ and Hg2+ was observed at 20, 0.005 and 0.002 ppm, respectively. The pPROBE-zntR-zntA biosensor reported here can be employed as a primary screening technique for aquatic heavy metal pollution.

Keywords

Bacterial biosensor Heavy metals Zinc Cadmium Mercury GFP 

Notes

Acknowledgments

The authors would like to acknowledge Dr. W. S. Lakra, Director, CIFE and Dr. Dilip Kumar, Former Director, CIFE for providing necessary facilities, ICAR for JRF fellowship to the first author, Dr. Kshitish Majumdar and Dr. S. Ayyappan for their valuable help, advice and support.

References

  1. 1.
    Mukhopadhyay MK, Konar SK (1985) Effects of copper, zinc and iron mixture on fish and aquatic ecosystem. Environ Ecol 3:58–64Google Scholar
  2. 2.
    Kumar A, Mathur RP (1991) Bioaccumulation kinetics and organ distribution of lead in a fresh water teleost, Colisa fasciatus. Environ Technol 12:731–735CrossRefGoogle Scholar
  3. 3.
    Mohan D, Choudhary A (1991) Zinc accumulation in a few tissues of fish, Puntius sophore (Ham.) after sublethal exposure. J Nat Conserv 3:205–208Google Scholar
  4. 4.
    Wepener V, van Vuren JHJ, du Preez HH (2001) Uptake and distribution of copper, iron and zinc mixture in gill, liver and plasma of a freshwater teleost, Tilapia sparrmanii. Water SA 27:99–108Google Scholar
  5. 5.
    Tecon R, Meer JR (2008) Bacterial biosensors for measuring availability of environmental pollutants. Sensors 8:4062–4080CrossRefGoogle Scholar
  6. 6.
    Rensing C, Mitra B, Rosen BP (1997) The zntA gene of Escherichia coli encodes a Zn(II)-translocating P-type ATPase. Proc Natl Acad Sci USA 94:14326–14331PubMedCrossRefGoogle Scholar
  7. 7.
    Beard SJ, Hashim R, Membrillo-Hernandez J, Hughes MN, Poole RK (1997) Zinc(II) tolerance in Escherichia coli K-12: evidence that the zntA gene (o732) encodes a cation transport ATPase. Mol Microbiol 25:883–891PubMedCrossRefGoogle Scholar
  8. 8.
    Blencowe DK, Samantha JM, Morby AP (1997) Preliminary characterization of zntA, a gene which encodes a Zn (II)/Cd (II)-export protein in Escherichia coli. Biotechnol et alia 2:1–6Google Scholar
  9. 9.
    Outten CE, Outten FW, O’Halloran TV (1999) DNA distortion mechanism for transcriptional activation by ZntR, a Zn (II)-responsive MerR homologue in Escherichia coli. J Biol Chem 274:37517–37524PubMedCrossRefGoogle Scholar
  10. 10.
    Ivask A, Virta M, Karhu A (2002) Construction and use of specific luminescent recombinant bacterial sensors for the assessment of bioavailable fraction of cadmium, zinc, mercury and chromium in the soil. Soil Biol Biochem 34:1439–1447CrossRefGoogle Scholar
  11. 11.
    Liao VHC, Ou KL (2005) Development and testing of a green fluorescent protein–based bacterial biosensor for measuring bioavailable arsenic in contaminated groundwater samples. Environ Toxicol Chem 24:1624–1631PubMedCrossRefGoogle Scholar
  12. 12.
    Siddiki MSR, Kawakami Y, Ueda S, Maeda I (2011) Solid phase biosensors for arsenic or cadmium composed of a trans factor and cis element complex. Sensors 11:10063–10073PubMedCrossRefGoogle Scholar
  13. 13.
    Wu CH (2009) Optimization of a whole-cell cadmium sensor with a toggle gene circuit. Biotechnol Prog 25:898–903PubMedCrossRefGoogle Scholar
  14. 14.
    Raja CE, Selvam GS (2011) Construction of green fluorescent protein based bacterial biosensor for heavy metal remediation. Int J Environ Sci Technol 8:793–798Google Scholar
  15. 15.
    Priyadarshi H, Alam A, Gireesh-Babu P, Das R, Kishore P, Kumar S, Chaudhari A (2012) A GFP-based bacterial biosensor with chromosomally integrated sensing cassette for quantitative detection of Hg(II) in environment. J Environ Sci 24:963–968CrossRefGoogle Scholar
  16. 16.
    Chakraborty T, Gireesh-Babu P, Alam A, Chaudhari A (2008) GFP expressing bacterial biosensor to measure lead contamination in aquatic environment. Curr Sci 94:800–805Google Scholar
  17. 17.
    Hillson NJ, Hu P, Andersen GL, Shapiro L (2007) Caulobacter crescentus as a whole-cell uranium biosensor. Appl Environ Microbiol 73:7615–7621PubMedCrossRefGoogle Scholar
  18. 18.
    Sambrook J, Russell DW, Janssen KA, Irwin N (2001) Molecular cloning: a laboratory manual. Cold Spring harbor Laboratory, Cold Spring HarborGoogle Scholar
  19. 19.
    Li YF, Li FY, Ho CL, Liao VHC (2008) Construction and comparison of fluorescence and bioluminescence bacterial biosensors for the detection of bioavailable toluene and related compounds. Environ Pollut 152:123–129PubMedCrossRefGoogle Scholar
  20. 20.
    Shaner NC, Steinbach PA, Tsien RY (2005) A guide to choosing fluorescent proteins. Nat Methods 2:905–909PubMedCrossRefGoogle Scholar
  21. 21.
    Binet MR, Poole RK (2000) Cd(II), Pb(II) and Zn(II) ions regulate expression of the metal-transporting P-type ATPase ZntA in Escherichia coli. FEBS Lett 473:67–70PubMedCrossRefGoogle Scholar
  22. 22.
    Gatti D, Mitra B, Rosen BP (2000) Escherichia coli soft metal ion translocating ATPases. J Biol Chem 275:34009–34012PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.Division of Fish Genetics and BiotechnologyCentral Institute of Fisheries EducationMumbaiIndia

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