title: A bioluminescent arsenite biosensor designed for inline water analyzer.
abstract: Whole-cell biosensors based on the reporter gene system can offer rapid detection of trace levels of organic or metallic compounds in water. They are well characterized in laboratory conditions, but their transfer into technological devices for the surveillance of water networks remains at a conceptual level. The development of a semi-autonomous inline water analyzer stumbles across the conservation of the bacterial biosensors over a period of time compatible with the autonomy requested by the end-user while maintaining a satisfactory sensitivity, specificity, and time response. We focused here on assessing the effect of lyophilization on two biosensors based on the reporter gene system and hosted in Escherichia coli. The reporter gene used here is the entire bacterial luciferase lux operon (luxCDABE) for an autonomous bioluminescence emission without the need to add any substrate. In the cell-survival biosensor that is used to determine the overall fitness of the bacteria when mixed with the water sample, lux expression is driven by a constitutive E. coli promoter PrpoD. In the arsenite biosensor, the arsenite-inducible promoter Parsinvolved in arsenite resistance in E. coli controls lux expression. Evaluation of the shelf life of these lyophilized biosensors kept at 4 °C over a year evidenced that about 40 % of the lyophilized cells can be revived in such storage conditions. The performances of the lyophilized biosensor after 7 months in storage are maintained, with a detection limit of 0.2 μM arsenite for a response in about an hour with good reproducibility. These results pave the way to the use in tandem of both biosensors (one for general toxicity and one for arsenite contamination) as consumables of an autonomous analyzer in the field.
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Prévéral
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Brutesco
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: Elodie C T
authors.initials: ECT
authors.lastname: Descamps
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Escoffier
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: David
authors.initials: D
authors.lastname: Pignol
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France. nicolas.ginet@cea.fr.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France. nicolas.ginet@cea.fr.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France. nicolas.ginet@cea.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ginet
authors.affiliation: CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France.|CNRS, UMR 7265 Biol Veget and Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France.|Aix-Marseille Université, BVME UMR7265, Marseille, F-13284, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Garcia
mesh_terms: D018053:Arsenites / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D049453:Luciferases, Bacterial / Q000235:genetics / Q000378:metabolism*; D009876:Operon; D011401:Promoter Regions, Genetic; D015203:Reproducibility of Results
chemical_list: D018053:Arsenites; D049453:Luciferases, Bacterial; C015001:arsenite
keywords: Bioluminescence; Biosensors; Lyophilization