Development of a Specific and Sensitive Bacteria Sensor for Detection of Mercury at Picomolar Levels in Environment

Omura, Tomoko; Kiyono, Masako; Pan-Hou, Hidemitsu

doi:10.1248/jhs.50.379

Cited by 17 publications

(9 citation statements)

References 17 publications

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“…Escherichia coli is an enteric Gram-negative bacterium with the sequenced genome [ 25 ], well developed methods for genetic modification [ 26 ] and various laboratory strains available. Although the relevance of this bacterium for environmental testing is questionable, it has been widely used in the construction of various bacterial sensors [ 9 , 10 , 12 , 14 , 17 ], mainly due to its compatibility with the chemical-response elements used.…”

Section: Resultsmentioning

confidence: 99%

“…Since then, a number of papers on bacterial strains "sensing" inorganic substances (mainly heavy metals or metalloids) but also organic compounds (e.g., benzene and its derivatives, naphthalene, polychlorinated biphenyls) have been published (reviewed in [ 4 , 7 ]). Currently, recombinant bacterial "lights-on" heavy metal sensors expressing bacterial bioluminescence system as a reporter include sensors for Cd, Pb [ 8 ], Hg [ 8 - 12 ], Cr [ 8 ], Ni, Co [ 13 ], Zn, Cu [ 8 , 14 , 15 ] and As [ 16 ]. In most of these sensors, the bioluminescence-encoding genes from Vibrio fischeri (that may become labile in temperatures above 30°C [ 4 ] and thus, restrict testing) has been used.…”

Section: Introductionmentioning

confidence: 99%

“…In most of these sensors, the bioluminescence-encoding genes from Vibrio fischeri (that may become labile in temperatures above 30°C [ 4 ] and thus, restrict testing) has been used. Most of these papers describe just a few sensors that are often based on Escherichia coli [ 9 , 10 , 12 , 14 ] orsome other Gram-negative bacterium from the genera Pseudomonas or Alcaligenes [ 8 , 11 , 13 , 15 ] as a host. Thus, Gram-positive bacteria have rarely been used as hosts and no Gram-positive metal sensors expressing luxCDABE as a reporter have been constructed so far.…”

Section: Introductionmentioning

confidence: 99%

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A suite of recombinant luminescent bacterial strains for the quantification of bioavailable heavy metals and toxicity testing

2009

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Background: Recombinant whole-cell sensors have already proven useful in the assessment of the bioavailability of environmental pollutants like heavy metals and organic compounds. In this work 19 recombinant bacterial strains representing various Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (Escherichia coli, Pseudomonas fluorescens) bacteria were constructed to express the luminescence encoding genes luxCDABE (from Photorhabdus luminescens) as a response to bioavailable heavy metals ("lights-on" metal sensors containing metalresponse elements, 13 strains) or in a constitutive manner ("lights-off" constructs, 6 strains).

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A suite of recombinant luminescent bacterial strains for the quantification of bioavailable heavy metals and toxicity testing

2009

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“…Furthermore, possible cross-reactions in water samples with multiple analytes have to be critically investigated in future experiments. However, mercury biosensors based on the same biological elements have been found to be highly specific in previous studies [56, 57]. Moreover, as biosensors with different specificities can easily be used in parallel in our proposed system, cross-reactivities of individual sensing elements could potentially be compensated for.…”

Section: Discussionmentioning

confidence: 99%

A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs

et al. 2019

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Biosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically modified organisms and are therefore limited in shelf life, usability and biosafety. We addressed these issues by the construction of an extensible, cell-free biosensor. Storage is possible through freeze drying on paper. Following the addition of an aqueous sample, a highly efficient cell-free protein synthesis (CFPS) reaction is initiated. Specific allosteric transcription factors modulate the expression of ‘superfolder’ green fluorescent protein (sfGFP) depending on the presence of the substance of interest. The resulting fluorescence intensities are analyzed with a conventional smartphone accompanied by simple and cheap light filters. An ordinary differential equitation (ODE) model of the biosensors was developed, which enabled prediction and optimization of performance. With an optimized cell-free biosensor based on the Shigella flexneri MerR transcriptional activator, detection of 6 μg/L Hg(II) ions in water was achieved. Furthermore, a completely new biosensor for the detection of gamma-hydroxybutyrate (GHB), a substance used as date-rape drug, was established by employing the naturally occurring transcriptional repressor BlcR from Agrobacterium tumefaciens .

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“…The structure of integral membrane protein merT is yet to be solved and is currently been attempted by X-ray crystallography (Stroud et al 2009) and ssNMR (Steele and Opella 1997). The structural elucidation of merT can help us to understand the mechanism of mercury and organomercurials transport into bacteria (Kiyono and Pan-Hou 2006;Rossy et al 2004) and further help in development of novel protein based mercury biosensors (Omura et al 2004). These techniques require high concentration of pure protein in soluble form, which has proven to be a major challenge.…”

Section: Introductionmentioning

confidence: 99%

Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli

Senthil

Pennathur

2010

Biotechnol Lett

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The mercury transporter, merT, from Cupriavidus metallidurans was cloned into pRSET-C and expressed in various E. coli hosts. Expression of merT gene failed in common expression hosts like E. coli BL21(DE3), E. coli BL21(DE3)pLysS and E. coli GJ1158 due to expression induced toxicity. The protein was successfully expressed in E. coli C43(DE3) as inclusion bodies. The inclusion bodies were solubilized with Triton X-100 detergent. The detergent solubilized protein with N-terminal His-tag was purified in a single-step by immobilized metal affinity chromatography with a yield of 8 mg l(-1).

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Development of a Specific and Sensitive Bacteria Sensor for Detection of Mercury at Picomolar Levels in Environment

Cited by 17 publications

References 17 publications

A suite of recombinant luminescent bacterial strains for the quantification of bioavailable heavy metals and toxicity testing

A suite of recombinant luminescent bacterial strains for the quantification of bioavailable heavy metals and toxicity testing

A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs

Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli

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