A Sensitive Voltammetric Biosensor for Escherichia Coli Detection Using an Electroactive Substrate for [Formula Omitted]-D-Glucuronidase

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 [Formula Omitted]-D-glucuronidase. This enzyme was induced by adding methyl-[Formula Omitted]-D-glucuronide s...

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Bibliographic Details
Published in:IEEE sensors journal 2019-01, Vol.19 (18), p.7789
Main Authors: Zuser, Karen, Ettenauer, Jorg, Kellner, Karlheinz, Posnicek, Thomas, Mazza, Giulia, Brandl, Martin
Format: Article
Language:English
Subjects:
Bacteria
Biosensors
Cleavage
E coli
Enzyme activity
Enzymes
Hydroxyquinoline
Sodium salts
Substrates
Voltammetry
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Summary: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 [Formula Omitted]-D-glucuronidase. This enzyme was induced by adding methyl-[Formula Omitted]-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 [Formula Omitted]-D-glucuronidase 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.
ISSN:1530-437X
1558-1748
DOI:10.1109/JSEN.2019.2917883