Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Construction of two lux-tagged Hg2+-specific biosensors and their luminescence performance

Abstract

Two Hg2+-specific biosensors were constructed using bacterial luciferase as reporter gene and plasmid-free Pseudomonas putida X4 and Enterobacter aerogenes NTG-01 as host strains. The performance of X4 biosensor was compared with that of NTG-01 biosensor in the same assay conditions. The maximum bioluminescence for X4 (pmerRluxCDABE-Kan) biosensor was found during the midexponential phase and that for NTG-01 (pmerRluxCDABE-Kan) was at the late exponential phase. The shortest induction time of two biosensors was 30 min. The maximum light signal output for NTG-01 and X4 sensors was observed at the incubation time of 5 and 4 h, respectively. The lowest detectable concentration of mercury by the two biosensors were both of 100 pM at 28°C, pH 7 and an initial cell number of 106 CFU ml−1. Cd2+, Zn2+, Co2+, Cu2+, and Pb2 + ions at nanomolar level did not interfere with the measurement by the biosensors. These results show that the sensitivity of the two biosensors is sufficient for the detection of Hg2+ under most contaminated environments.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Bontidean I, Mortari A, Suzanne S, Brown NL, Karlson U, Larsen MM, Vangronsveld J, Corbisier P, Csoregi E (2004) Biosensors for detection of mercury in contaminated soils. Environ Pollut 131:255–262

  2. Christopher R, Raina MM (2003) Issues underlying use of biosensor to measure metal bioavailability. Ecotox Environ Safe 56:140–147

  3. Farrell RE, Germida JJ, Huang PM (1993) Effects of chemical speciation in growth media on the toxicity of mercury (II). Appl Environ Microbiol 59:1507–1514

  4. Gibson TJ (1984) Studies on the epstein-barr virus genome. Cambridge University PhD Thesis

  5. Hakkila K, Maksimow M, Karp M, Virta M (2002) Reporter genes lucFF, luxCDABE, gfp, and dsred have different characteristics in whole-cell bacterial sensors. Anal Biochem 301:235–242

  6. Hansen LH, Sørensen SJ (2000) Versatile biosensor vectors for detection and quantification of mercury. FEMS Microbiol Lett 193:123–127

  7. Harkins M, Porter AR, Paton GI (2004) The role of host organism, transcriptional switches and reporter mechanisms in the performance of Hg-induced biosensors. J Appl Microbiol 97:1192–1200

  8. Ivask A, Virta M, Kahru 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 34:1439–1447

  9. Karp MT, Korpela MT, Kurittu JS, Karvinen JT (1998) A recombinant Escherichia coli sensor strain for the detection of tetracyclines. Anal Chem 70:4457–4462

  10. Kohler S, Belkon S, Schmid RD (2000) Reporter gene bioassays in environmental analysis. Fresenius J Anal Chem 366:769–779

  11. Pan YY, Chen WL, Huang QY (2005) Isolation, identification and 16S rDNA sequence analysis of a bacterial resistant to copper and cadmium. Microbiology (In Chinese with English abstract) 32:68–72

  12. Pan-Hou M, Kiyono M, Omurn T (2004) Development of a specific and sensitive bacteria sensor for detection of mercury at picomolar lever in environment. J Health Sci 50:379–384

  13. Petänen T, Romantschuk M, Virta M, Karp M (2001) Construction and use of broad host range mercury and arsenite sensor plasmid in the soil bacterium Pseudomonas fluorescens OS8. Microbial Ecol 41:360–368

  14. Rasmussen LD, Turner RR, Barkay T (1997) Cell-density-dependent sensitivity of a mer-lux bioassay. Appl Environ Microbiol 63:3291–3293

  15. Sambrook J, Russell DW (2001) A laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Woodbury

  16. Selifonova O, Burlage R, Barkay T (1993) Bioluminescent sensors for detection of bioavailable Hg (II) in the environment. Appl Environ Microbiol 59:3083–3090

  17. Vandermeer J, Malik KA, Daunert S, Feliciano J, Harms H, Gsell M, Balluch D, Stocker J (2003) Development of a set of simple bacterial biosensors for quantitative and rapid measurements of arsenite and arsenate in potable water. Env Sci Tec 37:4747–4750

Download references

Acknowledgements

We thank Kaisa Hakkila for providing plasmid pmerRluxCDABE.

Author information

Correspondence to Qiao-Yun Huang.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Fu, Y., Chen, W. & Huang, Q. Construction of two lux-tagged Hg2+-specific biosensors and their luminescence performance. Appl Microbiol Biotechnol 79, 363–370 (2008). https://doi.org/10.1007/s00253-008-1442-1

Download citation

Keywords

  • LuxCDABE
  • Biosensor
  • Mercury
  • Luminescence