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Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase

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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 is 684 ± 32 A·M−1·m−2, and the apparent Michaelis-Menten constant (KM app) is 0.31 ± 0.05 mM. The calibration plot is linear over the range 3–200 μM L-Arg, the limit of detection is 1 μM, 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 (> 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.

Schematic of an amperometric biosensor for L-arginine based on a Nafion/polyaniline (PANi) composite at a platinum screen-printed electrode (Pt-SPE) using a novel recombinant arginine deiminase (ADI) from Mycoplasma hominis.

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Acknowledgements

This work was partially funded by the “SMARTCANCERSENS” project from the European Community Seventh Framework Program under the Grant Agreement PIRSES-GA-2012-318053 and by the NATO Science for Peace (SFP) Project CBP.NUKR.SFPP 984173.

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Authors and Affiliations

  1. Laboratory of Biomolecular Electronics, Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Zabolotnogo Str. 150, Kyiv, 03680, Ukraine

    Mykhailo T. Zhybak & Yaroslav I. Korpan

  2. Institute of Cell Biology, NAS of Ukraine, Drahomanov Str. 14/16, Lviv, 79005, Ukraine

    Lyubov Y. Fayura, Mykhailo V. Gonchar & Andriy A. Sibirny

  3. Lviv State University of Physical Culture, DBH and RI, 11, Kostushko str, Lviv, 79007, Ukraine

    Yuriy R. Boretsky

  4. Department of Biotechnology and Microbiology, Rzeszow University, Zelwerowicza 4, 35-601, Rzeszow, Poland

    Andriy A. Sibirny

  5. Centre for Research in Electroanalytical Technologies, Department of Science, ITT Dublin, Tallaght, Dublin, 24, Ireland

    Eithne Dempsey

  6. Biosensors & Bioelectronics Centre, Linköping University, 58183, Linköping, Sweden

    Anthony P. F. Turner

Authors
  1. Mykhailo T. Zhybak
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  2. Lyubov Y. Fayura
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  3. Yuriy R. Boretsky
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  4. Mykhailo V. Gonchar
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  5. Andriy A. Sibirny
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  6. Eithne Dempsey
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  7. Anthony P. F. Turner
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  8. Yaroslav I. Korpan
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Correspondence to Mykhailo T. Zhybak or Yaroslav I. Korpan.

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Zhybak, M.T., Fayura, L.Y., Boretsky, Y.R. et al. Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase. Microchim Acta 184, 2679–2686 (2017). https://doi.org/10.1007/s00604-017-2290-4

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  • DOI: https://doi.org/10.1007/s00604-017-2290-4

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