Cayron Julien, Prudent Elsa, Escoffier Camille, Gueguen Erwan, Mandrand-Berthelot Marie-Andrée, Pignol David, Garcia Daniel, Rodrigue Agnès
Université de Lyon, Lyon, 69003, France.
INSA de Lyon, Villeurbanne, 69621, France.
Environ Sci Pollut Res Int. 2017 Jan;24(1):4-14. doi: 10.1007/s11356-015-5580-6. Epub 2015 Oct 24.
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), which makes its use compatible with the norms governing the drinking water quality.
由于镍对生物体有害,因此检测水中的镍至关重要。检测镍的一种方法是使用全细胞生物传感器。本研究的目的是构建一种用于检测镍的发光细菌生物传感器,该传感器具有高特异性和低检测限的特性。为此,所实施的调控电路依赖于RcnR镍/钴金属调节因子及其与lux报告基因融合的rcnA天然靶启动子。为了使RcnR能够特异性检测镍,测试了几种突变并保留了C35A。删除镍外排泵rcnA并引入编码几种镍摄取系统的基因降低了检测阈值。当在几种大肠杆菌菌株中检测这些构建体时,发现检测阈值变化很大。TD2158野生型大肠杆菌产生的生物传感器比其W3110大肠杆菌K12等效物的活性和灵敏度高十倍。这种生物传感器能够可靠地检测低至80 nM(4.7 μg l)的镍浓度,这使得其使用符合饮用水质量标准。