Journal
ANALYTICA CHIMICA ACTA
Volume 693, Issue 1-2, Pages 41-46Publisher
ELSEVIER
DOI: 10.1016/j.aca.2011.03.029
Keywords
Nitrite biosensor; Cytochrome cd(1); Cytochrome c(552); Photopolymerizable PVA; Screen printed electrodes
Categories
Funding
- Fundacao para a Ciencia e Tecnologia [SFRH/BD/22800/2005, SFRH/BD/28921/2006, POCI/QUI/58026/2004]
- Basque Government [MV-2010-1-12]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/28921/2006, SFRH/BD/22800/2005, POCI/QUI/58026/2004] Funding Source: FCT
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In this work, a novel enzymatic biosensor for determination of nitrites constructed on an electrochemical transducing platform is proposed. The sensor is based on cytochrome-cd(1) (cyt-cd(1)) nitrite reductase from Marinobacter hydrocarbonoclasticus strain 617 as biological recognition element, and its putative physiological redox partner cytochrome-c(552) (cyt-c(552)), as electron mediator. The proteins were co-immobilized using a photopolymerizable polyvinyl alcohol (PVA) derivative, onto carbon paste screen printed electrodes (CPSPEs); the optimal modification conditions were 100 mu M cyt-cd(1)/100 mu M cyt-c(552) and 50% PVA, after a 48 h polymerization time. Electrochemical characterization of the mediator was carried out by cyclic voltammetry. The one-electron exchange between cyt-c(552) and the working electrode is a quasi-reversible process, without mass transport limitations. The formal potential of the mediator is 254 +/- 2 mV vs NHE and the intermolecular electron transfer rate constant between cytochromes c(552) and cd(1) is 9.9 x 10(3) M-1 s(-1). The analytical parameters of the biosensor response to nitrite as assessed by amperometric measurements were: linear range from 10 to 200 mu M; detection and quantification limits of 7 and 24 mu M, respectively; sensitivity of 2.49 +/- 0.08 A mol(-1) cm(2) mu M-1. Catalytic profiles in the presence of possible interfering species were also investigated. The interference from competitive enzymatic reduction of dissolved oxygen could be overcome by tuning the cyclic voltammograms for faster sweep rates. (C) 2011 Elsevier B.V. All rights reserved.
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