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Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor

Urease is the enzyme catalyzing the hydrolysis of urea into carbon dioxide and ammonia. This enzyme is substrate-specific, which means that the enzyme catalyzes the hydrolysis of urea only. This feature is a basic diagnostic criterion used in the determination of many bacteria species. Most of the m...

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Autores principales: Hubalek, Jaromir, Hradecky, Jan, Adam, Vojtech, Krystofova, Olga, Huska, Dalibor, Masarik, Michal, Trnkova, Libuse, Horna, Ales, Klosova, Katerina, Adamek, Martin, Zehnalek, Josef, Kizek, Rene
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923183/
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author Hubalek, Jaromir
Hradecky, Jan
Adam, Vojtech
Krystofova, Olga
Huska, Dalibor
Masarik, Michal
Trnkova, Libuse
Horna, Ales
Klosova, Katerina
Adamek, Martin
Zehnalek, Josef
Kizek, Rene
author_facet Hubalek, Jaromir
Hradecky, Jan
Adam, Vojtech
Krystofova, Olga
Huska, Dalibor
Masarik, Michal
Trnkova, Libuse
Horna, Ales
Klosova, Katerina
Adamek, Martin
Zehnalek, Josef
Kizek, Rene
author_sort Hubalek, Jaromir
collection PubMed
description Urease is the enzyme catalyzing the hydrolysis of urea into carbon dioxide and ammonia. This enzyme is substrate-specific, which means that the enzyme catalyzes the hydrolysis of urea only. This feature is a basic diagnostic criterion used in the determination of many bacteria species. Most of the methods utilized for detection of urease are based on analysis of its enzyme activity – the hydrolysis of urea. The aim of this work was to detect urease indirectly by spectrometric method and directly by voltammetric methods. As spectrometric method we used is called indophenol assay. The sensitivity of detection itself is not sufficient to analyse the samples without pre-concentration steps. Therefore we utilized adsorptive transfer stripping technique coupled with differential pulse voltammetry to detect urease. The influence of accumulation time, pH of supporting electrolyte and concentration of urease on the enzyme peak height was investigated. Under the optimized experimental conditions (0.2 M acetate buffer pH 4.6 and accumulation time of 120 s) the detection limit of urease evaluated as 3 S/N was 200 ng/ml. The activity of urease enzyme depends on the presence of nickel. Thus the influence of nickel(II) ions on electrochemical response of the enzyme was studied. Based on the results obtained the interaction of nickel(II) ions and urease can be determined using electrochemical methods. Therefore we prepared Ni nanoelectrodes to measure urease. The Ni nanoelectrodes was analysed after the template dissolution by scanning electron microscopy. The results shown vertically aligned Ni nanopillars almost covered the electrode surface, whereas the defect places are minor and insignificant in comparison with total electrode surface. We were able to not only detect urease itself but also to distinguish its native and denatured form.
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spelling pubmed-39231832014-02-13 Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor Hubalek, Jaromir Hradecky, Jan Adam, Vojtech Krystofova, Olga Huska, Dalibor Masarik, Michal Trnkova, Libuse Horna, Ales Klosova, Katerina Adamek, Martin Zehnalek, Josef Kizek, Rene Sensors (Basel) Full Paper Urease is the enzyme catalyzing the hydrolysis of urea into carbon dioxide and ammonia. This enzyme is substrate-specific, which means that the enzyme catalyzes the hydrolysis of urea only. This feature is a basic diagnostic criterion used in the determination of many bacteria species. Most of the methods utilized for detection of urease are based on analysis of its enzyme activity – the hydrolysis of urea. The aim of this work was to detect urease indirectly by spectrometric method and directly by voltammetric methods. As spectrometric method we used is called indophenol assay. The sensitivity of detection itself is not sufficient to analyse the samples without pre-concentration steps. Therefore we utilized adsorptive transfer stripping technique coupled with differential pulse voltammetry to detect urease. The influence of accumulation time, pH of supporting electrolyte and concentration of urease on the enzyme peak height was investigated. Under the optimized experimental conditions (0.2 M acetate buffer pH 4.6 and accumulation time of 120 s) the detection limit of urease evaluated as 3 S/N was 200 ng/ml. The activity of urease enzyme depends on the presence of nickel. Thus the influence of nickel(II) ions on electrochemical response of the enzyme was studied. Based on the results obtained the interaction of nickel(II) ions and urease can be determined using electrochemical methods. Therefore we prepared Ni nanoelectrodes to measure urease. The Ni nanoelectrodes was analysed after the template dissolution by scanning electron microscopy. The results shown vertically aligned Ni nanopillars almost covered the electrode surface, whereas the defect places are minor and insignificant in comparison with total electrode surface. We were able to not only detect urease itself but also to distinguish its native and denatured form. Molecular Diversity Preservation International (MDPI) 2007-07-16 /pmc/articles/PMC3923183/ Text en © 2007 by MDPI (http://www.mdpi.org). Reproduction is permitted for noncommercial purposes.
spellingShingle Full Paper
Hubalek, Jaromir
Hradecky, Jan
Adam, Vojtech
Krystofova, Olga
Huska, Dalibor
Masarik, Michal
Trnkova, Libuse
Horna, Ales
Klosova, Katerina
Adamek, Martin
Zehnalek, Josef
Kizek, Rene
Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title_full Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title_fullStr Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title_full_unstemmed Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title_short Spectrometric and Voltammetric Analysis of Urease – Nickel Nanoelectrode as an Electrochemical Sensor
title_sort spectrometric and voltammetric analysis of urease – nickel nanoelectrode as an electrochemical sensor
topic Full Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923183/
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