8-hydroxyquinoline-glucuronide Sodium Salt Used as Electroactive Substrate for a Sensitive Voltammetric Detection of Escherichia Coli in Water Samples

Ettenauer, Jörg; Zuser, Karen; Kellner, Karlheinz; Posnicek, Thomas

doi:10.1016/j.proeng.2016.11.179

Cited by 3 publications

(12 citation statements)

References 4 publications

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“…As a preliminary test prior to application of the assay to living E. coli cells, the ability of the β-D-glucuronidase enzyme to cleave the specific substrate 8-hydroxyquinoline glucuronide was studied [30]. Therefore, different concentrations of enzyme ranging from 2.5, 25 and 125 units were tested.…”

Section: F Electrochemical Verification Of Enzymatic 8-hqg Cleavagementioning

confidence: 99%

“…After verification of the general principle of the assay by testing a simple enzymatic solution, the developed method was tested using live E. coli cells [30]. Therefore, aliquots (450 μL) of overnight cultures of E. coli and C. freundii, as negative control, were supplemented with 7.9 mM methylβ-D-glucuronide sodium salt (MetGlu) to induce production of the enzyme GUS.…”

Section: G Voltammetric Detection Of E Coli and Temperature Evaluationmentioning

confidence: 99%

“…To detect very low concentrations of E. coli, a dilution series of an overnight culture (OD 0.5) was prepared [30]. Dilutions 10 −7 -10 −9 were used for the experiment.…”

Section: H Detection Of Low E Coli Concentrationsmentioning

confidence: 99%

“…In order to validate the designed assay for cross reactivity and false-positive results, various bacterial strains (Bacillus atrophaeus, Brevundimonas diminuta, Citrobacter freundii, Pseudomonas putida, and Pseudomonas stutzeri) were tested [30]. For this purpose, fresh cultures of each strain were incubated overnight and then diluted to an optical density of 0.5.…”

Section: Cross Reactivity With Other Bacterial Strainsmentioning

confidence: 99%

“…In this study, we report a novel protocol incorporated into a biosensor prototype for the voltammetric detection of E. coli. Certain parts of the presented data are pre-published in [30]- [33]. Here, we show in detail a) the design, construction and evaluation of a new, sensitive potentiostat, b) the proof of principle of the proposed methodology with GUS enzyme solutions in different concentrations, c) followed by its verification and optimization with living E. coli cells (e.g.…”

Section: Introductionmentioning

confidence: 99%

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A Sensitive Voltammetric Biosensor forEscherichia ColiDetection Using an Electroactive Substrate for $\beta$ -D-Glucuronidase

et al. 2019

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In this paper, we developed a sensitive and simple electrochemical method for the rapid detection of Escherichia coli in water samples. The general principle of the assay utilizes the enzyme β-D-glucuronidase. This enzyme was induced by adding methyl-β-D-glucuronide sodium salt and its activity promoted the cleavage of 8-hydroxyquinoline glucuronide to the electroactive compound 8-hydroxyquinoline. This cleavage product was further oxidized on the working electrode of a potentiostat using cyclic voltammetry. The obtained current output signal in a specific voltage range (400 to 600 mV) indicated enzyme activity and subsequently was an evidence for E. coli cells in the sample. For our experiment, we designed a low-cost potentiostat and show an evaluation of this instrument. First, the β-Dglucuronidase assay was tested with various concentrations of enzyme solutions before living E. coli cells were investigated. Our presented method allowed a clear and a sensitive identification of 1 colony-forming unit of E. coli without any interference from other investigated bacterial strains. Comprising only few working steps (filtration, incubation, and voltammetric analysis), the method allowed incorporation into an automated prototype that delivered results similar to those obtained from samples treated in laboratory.

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Section: F Electrochemical Verification Of Enzymatic 8-hqg Cleavagementioning

confidence: 99%

Section: G Voltammetric Detection Of E Coli and Temperature Evaluationmentioning

confidence: 99%

“…To detect very low concentrations of E. coli, a dilution series of an overnight culture (OD 0.5) was prepared [30]. Dilutions 10 −7 -10 −9 were used for the experiment.…”

Section: H Detection Of Low E Coli Concentrationsmentioning

confidence: 99%

Section: Cross Reactivity With Other Bacterial Strainsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Sensitive Voltammetric Biosensor forEscherichia ColiDetection Using an Electroactive Substrate for $\beta$ -D-Glucuronidase

et al. 2019

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Electroanalytical properties of chlorophenol red at disposable carbon electrodes: Implications for Escherichia coli detection

Casimero

Bigham

McGlynn

et al. 2019

Bioelectrochemistry

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The use of coliforms and Escherichia coli as indicator species for assessing the quality of water is well established and a large variety of methods based on -galactosidase (B-GAL) activity, inherent to the microbes within this classification, have arisen to enable their detection and enumeration.Chlorophenol red (CPR) is widely used as a chromogenic label, but its capacity for translation to electroanalytical devices has yet to be fully explored. The CPR moiety is capable of undergoing oxidation at carbon substrates (+0.7 V) giving rise to a variety of phenolic intermediates.Electrochemical, XPS and enzymatic techniques were employed to characterise the underpinning chemistry and the intermediate identified as a 1,2-quinone derivative in which the chlorine substituent is retained. The latter was found to accumulate at the electrode and, in contrast to the parent CPR, was found to be detected at a significantly less positive potential (+0.3V). Bacterial hydrolysis of a CPR labelled substrate was demonstrated with the 1,2-quinone oxidation product found to accumulate at the electrode and detected using square wave voltammetry. Proof of concept for the efficacy of the alternative electrode pathway was established through the detection of E.coli after an incubation time of 2.5 h with no interference from the labelled substrates.

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Novel sensor platform for rapid detection and quantification of coliforms on food contact surfaces

Badalyan

Díaz

Bücking

et al. 2018

Journal of Microbiological Methods

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8-hydroxyquinoline-glucuronide Sodium Salt Used as Electroactive Substrate for a Sensitive Voltammetric Detection of Escherichia Coli in Water Samples

Cited by 3 publications

References 4 publications

A Sensitive Voltammetric Biosensor forEscherichia ColiDetection Using an Electroactive Substrate for $\beta$ -D-Glucuronidase

A Sensitive Voltammetric Biosensor forEscherichia ColiDetection Using an Electroactive Substrate for $\beta$ -D-Glucuronidase

Electroanalytical properties of chlorophenol red at disposable carbon electrodes: Implications for Escherichia coli detection

Novel sensor platform for rapid detection and quantification of coliforms on food contact surfaces

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