Whole-cell biosensors based on reporter genes allow detection of toxic metals in water with high selectivity and sensitivity under laboratory conditions; nevertheless, their transfer to a commercial inline water analyzer requires specific adaptation and optimization to field conditions as well as economical considerations. We focused here on both the influence of the bacterial host and the choice of the reporter gene by following the responses of global toxicity biosensors based on constitutive bacterial promoters as well as arsenite biosensors based on the arsenite-inducible Pars promoter. We observed important variations of the bioluminescence emission levels in five different Escherichia coli strains harboring two different lux-based biosensors, suggesting that the best host strain has to be empirically selected for each new biosensor under construction. We also investigated the bioluminescence reporter gene system transferred into Deinococcus deserti, an environmental, desiccation- and radiation-tolerant bacterium that would reduce the manufacturing costs of bacterial biosensors for commercial water analyzers and open the field of biodetection in radioactive environments. We thus successfully obtained a cell survival biosensor and a metal biosensor able to detect a concentration as low as 100 nM of arsenite in D. deserti. We demonstrated that the arsenite biosensor resisted desiccation and remained functional after 7 days stored in air-dried D. deserti cells. We also report here the use of a new near-infrared (NIR) fluorescent reporter candidate, a bacteriophytochrome from the magnetotactic bacterium Magnetospirillum magneticum AMB-1, which showed a NIR fluorescent signal that remained optimal despite increasing sample turbidity, while in similar conditions, a drastic loss of the lux-based biosensors signal was observed.
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This work was funded by the French national research agency ANR on the call-for-project ECOTECH (project COMBITOX, ANR-11-ECOT-0009) and also supported by the Centre National de la Recherche Scientifique and the Commissariat à l’Énergie Atomique et aux Énergies Alternatives (program NRBC). We thank all the members of the COMBITOX project for fruitful discussions and interactions. We also thank Dr. Eric Giraud for the kind gift of the PPΦ3295 plasmid.
Ansaldi M, Bazin I, Cholat P et al (2015) Toward inline multiplex biodetection of metals, bacteria, and toxins in water networks: the COMBITOX project. Environ Sci Pollut Res Int. doi:10.1007/s11356-015-5582-4Google Scholar
Cayron J, Prudent E, Escoffier C et al (2015) Pushing the limits of nickel detection to nanomolar range using a set of engineered bioluminescent Escherichia coli. Environ Sci Pollut Res Int. doi:10.1007/s11356-015-5580-6Google Scholar
de Groot A, Chapon V, Servant P et al (2005) Deinococcus deserti sp. nov., a gamma-radiation-tolerant bacterium isolated from the Sahara Desert. Int J Syst Evol Microbiol 55:2441–2446. doi:10.1099/ijs.0.63717-0CrossRefGoogle Scholar
de Groot A, Roche D, Fernandez B et al (2014) RNA sequencing and proteogenomics reveal the importance of leaderless mRNAs in the radiation-tolerant bacterium Deinococcus deserti. Genome Biol Evol 6:932–948. doi:10.1093/gbe/evu069CrossRefGoogle Scholar
Guzman LM, Belin D, Carson MJ, Beckwith J (1995) Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 177:4121–4130CrossRefGoogle Scholar
Hayashi K, Morooka N, Yamamoto Y et al (2006) Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110. Mol Syst Biol 2:2006.0007. doi:10.1038/msb4100049CrossRefGoogle Scholar
Hynninen A, Virta M (2010) Whole-cell bioreporters for the detection of bioavailable metals. Adv Biochem Eng Biotechnol 118:31–63. doi:10.1007/10_2009_9Google Scholar
Matsunaga T, Okamura Y, Fukuda Y et al (2005) Complete genome sequence of the facultative anaerobic magnetotactic bacterium Magnetospirillum sp. strain AMB-1. DNA Res Int J Rapid Publ Rep Genes Genomes 12:157–166. doi:10.1093/dnares/dsi002Google Scholar
Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26:101–106CrossRefGoogle Scholar
Peng Z, Yan Y, Xu Y et al (2010) Improvement of an E. coli bioreporter for monitoring trace amounts of phenol by deletion of the inducible σ54-dependent promoter. Biotechnol Lett 32:1265–1270. doi:10.1007/s10529-010-0317-6CrossRefGoogle Scholar
Sun J-Z, Peter Kingori G, Si R-W et al (2015) Microbial fuel cell-based biosensors for environmental monitoring: a review. Water Sci Technol J Int Assoc Water Pollut Res 71:801–809. doi:10.2166/wst.2015.035CrossRefGoogle Scholar
Vinay M, Franche N, Grégori G et al (2015) Phage-based fluorescent biosensor prototypes to specifically detect enteric bacteria such as E. coli and Salmonella enterica Typhimurium. PLoS One 10:e0131466. doi:10.1371/journal.pone.0131466CrossRefGoogle Scholar
Winson MK, Swift S, Hill PJ et al (1998) Engineering the luxCDABE genes from Photorhabdus luminescens to provide a bioluminescent reporter for constitutive and promoter probe plasmids and mini-Tn5 constructs. FEMS Microbiol Lett 163:193–202CrossRefGoogle Scholar
Yagur-Kroll S, Belkin S (2014) Molecular manipulations for enhancing luminescent bioreporters performance in the detection of toxic chemicals. Adv Biochem Eng Biotechnol 145:137–149. doi:10.1007/978-3-662-43619-6_4Google Scholar