Pushing the limits of nickel detection to nanomolar range using a set of engineered bioluminescent Escherichia coli.
The detection of nickel in water is of great importance due to its harmfulness for living organism. A way to detect Ni is the use of whole-cell biosensors. The aim of the present work was to build a light-emitting bacterial biosensor for the detection of Ni with high specificity and low detection limit properties. For that purpose, the regulatory circuit implemented relied on the RcnR Ni/Co metallo-regulator and its rcnA natural target promoter fused to the lux reporter genes. To convert RcnR to specifically detect Ni, several mutations were tested and the C35A retained. Deleting the Ni efflux pump rcnA and introducing genes encoding several Ni-uptake systems lowered the detection thresholds. When these constructs were assayed in several Escherichia coli strains, it appeared that the detection thresholds were highly variable. The TD2158 wild-type E. coli gave rise to a biosensor ten times more active and sensitive than its W3110 E. coli K12 equivalent. This biosensor was able to confidently detect Ni concentrations as little as 80 nM (4.7 μg l-1), which makes its use compatible with the norms governing the drinking water quality.
Université de Lyon, Lyon, 69003, France.|INSA de Lyon, Villeurbanne, 69621, France.|CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, Université Lyon 1, 10 rue Dubois, 69622, Villeurbanne Cedex, France.
Julien
J
Cayron
Université de Lyon, Lyon, 69003, France.|INSA de Lyon, Villeurbanne, 69621, France.|CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, Université Lyon 1, 10 rue Dubois, 69622, Villeurbanne Cedex, France.
Elsa
E
Prudent
CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, France.|CNRS, UMR Biol Veget & Microbiol Environ, Aix-Marseille Université, Saint-Paul-lez-Durance, 13108, France.
Camille
C
Escoffier
Université de Lyon, Lyon, 69003, France.|INSA de Lyon, Villeurbanne, 69621, France.|CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, Université Lyon 1, 10 rue Dubois, 69622, Villeurbanne Cedex, France.
Erwan
E
Gueguen
Université de Lyon, Lyon, 69003, France.|INSA de Lyon, Villeurbanne, 69621, France.|CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, Université Lyon 1, 10 rue Dubois, 69622, Villeurbanne Cedex, France.
Marie-Andrée
MA
Mandrand-Berthelot
CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, France.|CNRS, UMR Biol Veget & Microbiol Environ, Aix-Marseille Université, Saint-Paul-lez-Durance, 13108, France.
David
D
Pignol
CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, France.|CNRS, UMR Biol Veget & Microbiol Environ, Aix-Marseille Université, Saint-Paul-lez-Durance, 13108, France.
Daniel
D
Garcia
Université de Lyon, Lyon, 69003, France. agnes.rodrigue@insa-lyon.fr.|INSA de Lyon, Villeurbanne, 69621, France. agnes.rodrigue@insa-lyon.fr.|CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, Université Lyon 1, 10 rue Dubois, 69622, Villeurbanne Cedex, France. agnes.rodrigue@insa-lyon.fr.
Agnès
A
Rodrigue
D015374:Biosensing Techniques; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000502:physiology*; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D009532:Nickel / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; C558390:RcnR protein, E coli; D012097:Repressor Proteins; D009532:Nickel
Bio-luminescence; Biosensor; Metal; Water control; Whole bacteria