Zhybak, Mykhailo T. and Fayura, Lyubov Y. and Boretsky, Yuriy R. and Gonchar, Mykhailo V. and Sibirny, Andriy A. and Dempsey, Eithne and Turner, Anthony P.F. and Korpan, Yaroslav I. (2017) Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase. Microchimica Acta, 184 (8). pp. 2679-2686. ISSN 1436-5073

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Abstract

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 was 684±32 A⋅M-1⋅m-2, and the apparent Michaelis-Menten constant (KMapp) 0.31±0.05 mM. The calibration plot was linear over the range 3 - 200 μM L-Arg, the limit of detection was 1 μM, and the response time (for 90% of the total signal change to occur) 15 s. The sensor was selective and exhibited good storage stability (> 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 was found to be in good agreement with a reference fluorometric enzymatic assay.

Item Type:	Article
Additional Information:	This is the preprint version of the published article, which is available at: Zhybak, M.T., Fayura, L.Y., Boretsky, Y.R. et al. Microchim Acta (2017) 184: 2679. https://doi.org/10.1007/s00604-017-2290-4
Keywords:	L-Arginine; ammonium ions; amperometry; nafion; polyaniline; recombinantenzyme; plazma samples; pharmaceuticals;
Academic Unit:	Faculty of Science and Engineering > Chemistry
Item ID:	11437
Identification Number:	https://doi.org/10.1007/s00604-017-2290-4
Depositing User:	Eithne Dempsey
Date Deposited:	21 Oct 2019 17:04
Journal or Publication Title:	Microchimica Acta
Publisher:	Springer
Refereed:	Yes
URI:	https://link.springer.com
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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