HUBÁLEK, Jaromír, Jan HRADECKÝ, Vojtěch ADAM, Olga KRYŠTOFOVÁ, Dalibor HÚSKA, Michal MASAŘÍK, Libuše TRNKOVÁ, Aleš HORNA, Kateřina KLOSOVÁ, Martin ADÁMEK, Josef ZEHNÁLEK and René KIZEK. Spectrometric and voltammetric analysis of urease. Nickel nanoelectrode as an electrochemical sensor. Sensors. Basel, Switzerland: MDPI, 2007, vol. 7, No 7, p. 1238-1255, 17 pp. ISSN 1424-8220.
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Basic information
Original name Spectrometric and voltammetric analysis of urease. Nickel nanoelectrode as an electrochemical sensor
Name in Czech Spektrometrická a voltametrická analýza ureázy. Niklová nanoelektroda jako elektrochemický sensor
Authors HUBÁLEK, Jaromír (203 Czech Republic), Jan HRADECKÝ (203 Czech Republic), Vojtěch ADAM (203 Czech Republic), Olga KRYŠTOFOVÁ (203 Czech Republic), Dalibor HÚSKA (203 Czech Republic), Michal MASAŘÍK (203 Czech Republic), Libuše TRNKOVÁ (203 Czech Republic), Aleš HORNA (203 Czech Republic), Kateřina KLOSOVÁ (203 Czech Republic), Martin ADÁMEK (203 Czech Republic), Josef ZEHNÁLEK (203 Czech Republic) and René KIZEK (203 Czech Republic, guarantor).
Edition Sensors, Basel, Switzerland, MDPI, 2007, 1424-8220.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10405 Electrochemistry
Country of publisher Switzerland
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 1.573
RIV identification code RIV/00216224:14110/07:00019452
Organization unit Faculty of Medicine
UT WoS 000248319500013
Keywords in English urease; electrochemical methods; nanotechnology; nanotube; nickel electrode; hanging mercury drop electrode; spectrometry
Tags electrochemical methods, hanging mercury drop electrode, Nanotechnology, nanotube, nickel electrode, Spectrometry, urease
Tags Reviewed
Changed by Changed by: prof. RNDr. Michal Masařík, Ph.D., učo 21142. Changed: 25/6/2009 10:45.
Abstract
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.
Abstract (in Czech)
Ureáza je enzym katalyzující přeměnu močoviny na oxid uhličitý a amoniak. Ureáza je substrátově specifická, což znamená, že katalyzuje přeměnu pouze ureázy. Tato schopnost je základním diagnostickým kritériem používaným pro stanovení mnoha bakteriálních druhů. Metoda, která se využívá pro stanovení ureázy je založena na detekci její aktivity, intenzity hydrolýzy močoviny. Cílem této práce je nepřímé stanovení ureázy pomocí spektrometrických a elektrochemických metod.
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IAA401990701, research and development projectName: Studium vazby platinových cytostatik do struktury DNA; Vliv metalothioneinu
MSM0021630503, plan (intention)Name: Nové trendy v mikroelektronických systémech a nanotechnologiích (MIKROSYN)
1QS201710508, research and development projectName: Impedimetrické chemické mikrosenzory s nanomechanizovaným povrchem elektrod
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