Whole-cell fluorescent bacterial bioreporter for arsenic detection in water

  • E. Elcin
  • H. A. ÖktemEmail author
Original Paper


Microbial whole-cell bioreporters have been developed for environmental monitoring of arsenic contamination. Despite the great interest in bacterial bioreporters for arsenite detection, relatively few studies reported their response to arsenate levels. In this study, green fluorescent protein-based whole-cell Escherichia coli bioreporter was constructed for the measurement of both bioavailable arsenite and arsenate in water. The developed bacterial bioreporter has much higher sensitivity toward arsenate in inorganic phosphate-limiting medium compared to minimal medium. Both arsenite and arsenate were detected at 10 µg/l within 2-h induction time period. Furthermore, the bioreporter was able to detect and estimate arsenic levels in groundwater sample. The results demonstrate that the constructed bacterial arsenic bioreporter is applicable in determining the concentrations of the most abundant two ionic forms of arsenic as opposed to analytical methods which gives total content of arsenic. Thus, a comparison of bioavailability of arsenic and its form with its total content would give an insight about the risk of toxicity and also planning of removal processes.


Arsenic Bacterial bioreporter Green fluorescent protein Contaminated groundwater 



We would like to thank METU Central Laboratory Molecular Biology and Biotechnology Section for technical support. This work was supported by ÖYP-YÖK Research Capacity Development Funds to EE (Budget No. and by Nanobiz Technology Inc. EE was funded by TUBITAK BIDEB 2211 fellowship. Authors declare no conflict of interest.


  1. Agency for Toxic Substances and Disease Registry (ATSDR) U.S. Department of Health and Human Services, PHS (2017) Priority list of hazardous substances. Accessed 10 Aug 2018
  2. Aide M, Beighley D, Dunn D (2016) Arsenic in the soil environment: a soil chemistry review. Int J Appl Agric Res 11:1–28CrossRefGoogle Scholar
  3. Akter A, Ali MH (2011) Arsenic contamination in groundwater and its proposed remedial measures. Int J Environ Sci Technol 8(2):433–443. CrossRefGoogle Scholar
  4. Cai J, DuBow MS (1997) Use of a luminescent bacterial biosensor for biomonitoring and characterization of arsenic toxicity of chromated copper arsenate (CCA). Biodegradation 8:105–111. CrossRefGoogle Scholar
  5. Carlin A, Shi W, Dey S, Rosen BP (1995) The ars operon of Escherichia coli confers arsenical and antimonial resistance. J Bacteriol 177(4):981–986. CrossRefGoogle Scholar
  6. Chen CM, Mobley HLT, Rosen BP (1985) Separate resistances to arsenate and arsenite (antimonate) encoded by the arsenical resistance operon of R factor R773. J Bacteriol 161(2):758–763Google Scholar
  7. Crameri A, Whitehorn EA, Tate E, Stemmer WPC (1996) Improved green fluorescent protein by molecular evolution using DNA shuffling. Nat Biotechnol 14:315–319. CrossRefGoogle Scholar
  8. Del Razo LM, Styblo M, Cullen WR, Thomas DJ (2001) Determination of trivalent methylated arsenicals in biological matrices. Toxicol Appl Pharmacol 174:282–293CrossRefGoogle Scholar
  9. Diesel E, Schreiber M, van Der Meer JR (2009) Development of bacteria-based bioassays for arsenic detection in natural waters. Anal Bioanal Chem 394:687–693. CrossRefGoogle Scholar
  10. Fendorf S, Michael HA, van Geen A (2010) Spatial and temporal variations of groundwater arsenic in South and Southeast Asia. Science 328:1123–1127CrossRefGoogle Scholar
  11. Fisher AT, López-Carrillo L, Gamboa-Loira B, Cebrián ME (2017) Standards for arsenic in drinking water: implications for policy in Mexico. J Public Health Policy 38(4):395–406. CrossRefGoogle Scholar
  12. Hynninen A, Virta M (2010) Whole-cell bioreporters for the detection of bioavailable metals. In: Gu MB, Belkin S (eds) Whole cell sensing systems II: applications, vol 118. Springer, Heidelberg, pp 31–63Google Scholar
  13. Kaur H, Kumar R, Babu JN, Mittal S (2015) Advances in arsenic biosensor development—a comprehensive review. Biosens Bioelectron 63:533–545. CrossRefGoogle Scholar
  14. Li L, Liang J, Hong W, Zhao Y, Sun S, Yang X, Xu A, Hang H, Wu L, Chen S (2015) Evolved bacterial biosensor for arsenite detection in environmental water. Environ Sci Technol 49:6149–6155. CrossRefGoogle Scholar
  15. 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(7):1624–1631. CrossRefGoogle Scholar
  16. Majumdar KK, Ghose A, Ghose N, Biswas A, Mazumder DNG (2014) Effect of safe water on arsenicosis: a follow-up study. J Fam Med Primary Care 3:124–128. CrossRefGoogle Scholar
  17. Merulla D, Buffi N, Beggah S, Truffer F, Geiser M, Renaud P, van der Meer JR (2013) Bioreporters and biosensors for arsenic detection. Biotechnological solutions for a world-wide pollution problem. Curr Opin Biotechnol 24(3):534–541. CrossRefGoogle Scholar
  18. Olaniran AO, Balgobind A, Pillay B (2013) Bioavailability of heavy metals in soil: impact on microbial biodegradation of organic compounds and possible improvement strategies. Int J Mol Sci 14:10197–10228. CrossRefGoogle Scholar
  19. T.C. Orman ve Su İşleri Bakanlığı (2015) Yerüstü su kalitesi yönetmeliğinde değişiklik yapilmasina dair yönetmelik. RG No. 29327Google Scholar
  20. Panagiotaras D, Nikolopoulos D (2015) Arsenic occurrence and fate in the environment; a geochemical perspective. J Earth Sci Clim Change 6:4. Google Scholar
  21. Prévéral S, Brutesco C, Descamps ECT, Escoffier C, Pignol D, Ginet N, Garcia D (2017) A bioluminescent arsenite biosensor designed for inline water analyzer. Environ Sci Pollut Res 24:25–32. CrossRefGoogle Scholar
  22. Rahman MM, Dong Z, Naidu R (2015) Concentrations of arsenic and other elements in groundwater of Bangladesh and West Bengal, India: potential cancer risk. Chemosphere 139:54–64. CrossRefGoogle Scholar
  23. Roberto FF, Barnes JM, Bruhn DF (2002) Evaluation of a GFP reporter gene construct for environmental arsenic detection. Talanta 58:181–188. CrossRefGoogle Scholar
  24. Rothert A, Deo SK, Millner L, Puckett LG, Madou MJ, Daunert S (2005) Whole-cell-reporter-gene-based biosensing systems on a compact disk microfluidics platform. Anal Biochem 342:11–19. CrossRefGoogle Scholar
  25. Shankar S, Shanker U, Shikha (2014) Arsenic contamination of groundwater: a review of sources, prevalence, health risks, and strategies for mitigation. Sci World J. Google Scholar
  26. Singh R, Singh S, Parihar P, Singh VP, Prasad SM (2015) Arsenic contamination, consequences and remediation techniques: a review. Ecotoxicol Environ Saf 112:247–270. CrossRefGoogle Scholar
  27. Suzuki K, Wakao N, Kimura T, Sakka K, Ohmiya K (1998) Expression and regulation of the arsenic resistance operon of Acidiphilium multivorum AIU 301 plasmid pKW301 in Escherichia coli. Appl Environ Microbiol 64(2):411–418Google Scholar
  28. Tani C, Inoue K, Tani Y, Harun-ur-Rashid M, Azuma N, Ueda S, Yoshida K, Maeda I (2009) Sensitive fluorescent microplate bioassay using recombinant Escherichia coli with multiple promoter-reporter units in tandem for detection of arsenic. J Biosci Bioeng 108(5):414–420. CrossRefGoogle Scholar
  29. T.C. Sağlık Bakanlığı. Türkiye Halk Sağlığı Kurumu (2013) İnsani tüketim amaçli sular hakkinda yönetmelikte değişiklik yapilmasina dair yönetmelik. RG No:28580Google Scholar
  30. U.S. EPA (2002) Arsenic treatment technologies for soil, waste, and water. EPA/National Service Center for Environmental Publications, Cincinnati, U.SGoogle Scholar
  31. Van Der Meer JR, Belkin S (2010) Where microbiology meets microengineering: design and applications of reporter bacteria. Nat Rev Microbiol 8:511–522. CrossRefGoogle Scholar
  32. WHO (2011) Arsenic in drinking-water. Background document for preparation of WHO Guidelines for drinking-water quality. Geneva, World Health Organization. https://www.WHO/SDE/WSH/03.04/75/Rev/1. Accessed 10 Aug 2018
  33. Willsky GR, Malamy MH (1980) Effect of arsenate on inorganic phosphate transport in Escherichia coli. J Bacteriol 144:366–374Google Scholar
  34. Wu J, Rosen BP (1993) Metalloregulated expression of the ars operon. J Biol Chem 268(1):52–58Google Scholar
  35. Yang HC, Fu HL, Lin YF, Rosen BP (2012) Pathways of arsenic uptake and efflux. Curr Top Membr 69:325–358. CrossRefGoogle Scholar

Copyright information

© Islamic Azad University (IAU) 2018

Authors and Affiliations

  1. 1.Department of BiotechnologyMiddle East Technical UniversityAnkaraTurkey
  2. 2.Department of Biological SciencesMiddle East Technical UniversityAnkaraTurkey
  3. 3.Nanobiz Technology Inc.AnkaraTurkey

Personalised recommendations