Use of lux-based biosensors for rapid diagnosis of pollutants in arable soils

Palmer, Gabrielle; McFadzean, Ross; Killham, Ken; Sinclair, A. H.; Paton, Graeme I.

doi:10.1016/s0045-6535(97)10225-9

Cited by 42 publications

(25 citation statements)

References 25 publications

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“…In this report we have demonstrated, using hybrid A-domains of DmpR and XylR, that the amino acid residues 110 to 186 are intimately involved in mediating the distinct effector profiles of these two regulators. The aromatic-sensing properties of natural regulators of catabolic pathways have been used in biosensor applications (2,9,11,13,19,33). The identification of a subregion of the A-domain that is primarily responsible for determining effector specificity provides a target for directed mutagenesis to modify sensitivity and expand the effector specificity of this class of regulators for greater utility in such biosensor systems.…”

Section: ])mentioning

confidence: 99%

Identification of an Effector Specificity Subregion within the Aromatic-Responsive Regulators DmpR and XylR by DNA Shuffling

et al. 2000

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The Pseudomonas derived 54 -dependent regulators DmpR and XylR control the expression of genes involved in catabolism of aromatic compounds. Binding to distinct, nonoverlapping groups of aromatic effectors controls the activities of these transcriptional activators. Previous work has derived a common mechanistic model for these two regulators in which effector binding by the N-terminal 210 residues (the A-domain) of the protein relieves repression of an intrinsic ATPase activity essential for its transcription-promoting property and allows productive interaction with the transcriptional apparatus. Here we dissect the A-domains of DmpR and XylR by DNA shuffling to identify the region(s) that mediates the differences in the effector specificity profiles. Analysis of in vivo transcription in response to multiple aromatic effectors and the in vitro phenolbinding abilities of regulator derivatives with hybrid DmpR/XylR A-domains reveals that residues 110 to 186 are key determinants that distinguish the effector profiles of DmpR and XylR. Moreover, the properties of some mosaic DmpR/XylR derivatives reveal that high-affinity aromatic effector binding can be completely uncoupled from the ability to promote transcription. Hence, novel aromatic binding properties will only be translated into functional transcriptional activation if effector binding also triggers release of interdomain repression.

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Section: ])mentioning

confidence: 99%

Identification of an Effector Specificity Subregion within the Aromatic-Responsive Regulators DmpR and XylR by DNA Shuffling

et al. 2000

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“…Expression of the reporter gene provides a measurable response when the appropriate transcription activator protein interacts with an effector molecule to signal a particular environmental condition. Bacterial biosensors have been created to detect naphthalene (10), benzene derivatives, including toluene and xylene (1,11,20,41); and certain toxic metals (20,25).…”

mentioning

confidence: 99%

“…The Pseudomonas putida XylR protein has served as the detection component for a number of biosensors (1,11,20,41) based on the ability of this protein to activate transcription in response to xylene and toluene. DmpR, the product of the Pseudomonas sp.…”

mentioning

confidence: 99%

Generation of Novel Bacterial Regulatory Proteins That Detect Priority Pollutant Phenols

Wise

Kuske

2000

Appl Environ Microbiol

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The genetic systems of bacteria that have the ability to use organic pollutants as carbon and energy sources can be adapted to create bacterial biosensors for the detection of industrial pollution. The creation of bacterial biosensors is hampered by a lack of information about the genetic systems that control production of bacterial enzymes that metabolize pollutants. We have attempted to overcome this problem through modification of DmpR, a regulatory protein for the phenol degradation pathway of Pseudomonas sp. strain CF600. The phenol detection capacity of DmpR was altered by using mutagenic PCR targeted to the DmpR sensor domain. DmpR mutants were identified that both increased sensitivity to the phenolic effectors of wild-type DmpR and increased the range of molecules detected. The phenol detection characteristics of seven DmpR mutants were demonstrated through their ability to activate transcription of a lacZ reporter gene. Effectors of the DmpR derivatives included phenol, 2-chlorophenol, 2,4-dichlorophenol, 4-chloro-3-methylphenol, 2,4-dimethylphenol, 2-nitrophenol, and 4-nitrophenol.

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“…The choice of the biosensor was appropriate due to its robust, wide pH ranges (3.0-10.00) and sensitivity in measuring the toxicity of the final leachate collected (Palmer et al, 1998). To ascertain that the application of the ameliorants to the lead spiked soils had a toxicity reduction effect on the soil sample leachate, a toxicity test based on a lux-biosensor was used (Fig.…”

Section: Biosensor Based Toxicity Test Of the Leachatementioning

confidence: 99%

Determination of adsorption efficiency based on cation exchange capacity related to red earth, bone meal and pulverised fly ash as ameliorants to lead contaminated soils

Gatima

Mwinyihija²,

Killham

2006

Int. J. Environ. Sci. Technol.

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Efficient treatment strategies to reduce the toxicity of metal contaminated soil using cost effective techniques such as naturally available ameliorants and industrial waste have emerged. Three easily available amendments were determined: Bone meal, red earth/mud and pulverised fly ash (PFA). The application of ameliorants offered a possible alternative in situ remediation of contaminated sites without disruption to the ecosystem profile. In comparison to other ameliorants Red earth/mud was found to be efficient in intercepting lead leaching from soil amended with different lead compounds based on CEC (Cmol/g). This was associated with the heterogeneous adsorbency principle in red mud which is associated with its ability to bind metal ions (M 2+ ) onto one or two types of surface sites at pH< 6.0. However areas that need to be studied and assessed (for public health concerns) critically for wide spread application of all the ameliorants include off-site effects of the ameliorants.

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Use of lux-based biosensors for rapid diagnosis of pollutants in arable soils

Cited by 42 publications

References 25 publications

Identification of an Effector Specificity Subregion within the Aromatic-Responsive Regulators DmpR and XylR by DNA Shuffling

Identification of an Effector Specificity Subregion within the Aromatic-Responsive Regulators DmpR and XylR by DNA Shuffling

Generation of Novel Bacterial Regulatory Proteins That Detect Priority Pollutant Phenols

Determination of adsorption efficiency based on cation exchange capacity related to red earth, bone meal and pulverised fly ash as ameliorants to lead contaminated soils

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