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Amperometric L-argi...
Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase
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- Zhybak, Mykhailo T. (author)
- National Academic Science Ukraine, Ukraine
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- Fayura, Lyubov Y. (author)
- NAS Ukraine, Ukraine
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- Boretsky, Yuriy R. (author)
- Lviv State University of Phys Culture, Ukraine
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- Gonchar, Mykhailo V. (author)
- NAS Ukraine, Ukraine
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- Sibirny, Andriy A. (author)
- NAS Ukraine, Ukraine; Rzeszow University, Poland
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- Dempsey, Eithne (author)
- ITT Dublin, Ireland
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- Turner, Anthony (author)
- Linköpings universitet,Biosensorer och bioelektronik,Tekniska fakulteten
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- Korpan, Yaroslav I. (author)
- National Academic Science Ukraine, Ukraine
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(creator_code:org_t)
- 2017-05-02
- 2017
- English.
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In: Microchimica Acta. - : SPRINGER WIEN. - 0026-3672 .- 1436-5073. ; 184:8, s. 2679-2686
- Related links:
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https://liu.diva-por... (primary) (Raw object)
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http://mural.maynoot...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- The authors describe an amperometric biosensor for the amino acid L-arginine (L-Arg). It is based on the use of a Nafion/Polyaniline (PANi) composite on a platinum screen-printed electrode (Pt-SPE) using a novel recombinant arginine deiminase isolated from Mycoplasma hominis. The protein was over-expressed, purified and employed as a biorecognition element of the sensor. Enzymatic hydrolysis of L-Arg leads to the formation of ammonium ions which diffuse into the Nafion/PANi layer and induce the electroreduction of PANi at a potential of -0.35 V (vs Ag/AgCl). L-Arg sensitivity is 684 +/- 32 A.M-1.m(-2), and the apparent Michaelis-Menten constant K-M(app)) is 0.31 +/- 0.05 mM. The calibration plot is linear over the range 3-200 mu M L-Arg, the limit of detection is 1 mu M, and the response time (for 90% of the total signal change to occur) is 15 s. The sensor is selective and exhibits good storage stability (amp;gt; 1 month without loss in signal). The biosensor was applied to the analysis of L-Arg in pharmaceutical samples and of ammonium and L-Arg in spiked human plasma obtained from blood of healthy volunteers and those with a hepatic disorder. Data generated were found to be in good agreement with a reference fluorometric enzymatic assay.
Subject headings
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Keyword
- L-arginine; Ammonium ions; Amperometry; Nafion; Polyaniline; Recombinant enzyme; Plasma samples; Pharmaceuticals
Publication and Content Type
- ref (subject category)
- art (subject category)
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