Biopolym. Cell. 2018; 34(1):14-23.
http://dx.doi.org/10.7124/bc.00096C
Molecular and Cell Biotechnologies
Development of pyruvate oxidase-based amperometric biosensor for pyruvate determination
1, 2Knyzhnykova D. V., 1Topolnikova Ya. V., 1Kucherenko I. S., 1, 2Soldatkin O. O.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Institute of High Technologies,
    Taras Shevchenko National University of Kyiv
    2, korp.5, Pr. Akademika Hlushkova, Kyiv, Ukraine, 03022

Abstract

Aim. The development and optimization of the amperometric biosensor for pyruvate determina-tion. Methods. Immobilized pyruvate oxidase was used as a biorecognition element of the biosen-sor, a platinum disc electrode- as an electrochemical transducer. Results. Different variants of immobilization of pyruvate oxidase were tested and the optimal one was chosen for the creation of a biorecognition element of the biosensor. Optimal concentrations of cofactors for the best per-formance of the pyruvate oxidase-based biosensor were selected. The developed biosensor dem-onstrated a high sensitivity to pyruvate and wide linear range of work. High selectivity of the pro-posed biosensor towards electrically active substances and other substrates present in real samples was shown. The biosensor is characterized by high signal reproducibility and operational stability over two weeks. Conclusions. The highly selective amperometric biosensor for determination of pyruvate in biological samples has been developed. Its analytical characteristics are studied. The biosensor can be further used for the pyruvate analysis in blood serum.
Keywords: pyruvate, pyruvate oxidase, amperometric biosensor
Full text: (PDF, in English)

References

[1] Pundir CS, Narwal V, Batra B. Determination of lactic acid with special emphasis on biosensing methods: A review. Biosens Bioelectron. 2016;86:777-790. PubMed
[2] Bhat MA, Prasad K, Trivedi D, Rajeev BR, Battur H. Pyruvic acid levels in serum and saliva: A new course for oral cancer screening? J Oral Maxillofac Pathol. 2016;20(1):102-5.
[3] Gajovic N, Beinyamin G, Warsinke A, Scheller FW, Heller A. Operation of a miniature redox hydrogel-based pyruvate sensor in undiluted deoxygenated calf serum. Anal Chem. 2000;72(13):2963-8.
[4] Bhat A, Bhat M, Prasad K, Trivedi D, Acharya S. Estimation of Pyruvic acid in serum and saliva among healthy and potentially malignant disorder subjects - a stepping stone for cancer screening? J Clin Exp Dent. 2015;7(4):e462-5.
[5] Arai G, Noma T, Habu H, Yasumori I. Pyruvate sensor based on pyruvate oxidase immobilized in a poly(mercapto-p-benzoquinone) film. J Electroanal Chem. 1999; 464(2): 143–8.
[6] Akyilmaz E, Yorganci E. Construction of an amperometric pyruvate oxidase enzyme electrode for determination of pyruvate and phosphate. Electrochim Acta. 2007; 52(28): 7972–7.
[7] Topolnikova YV, Knyzhnykova DV, Kucherenko IS, Dzyadevych SV, Soldatkin OO. Development of am-perometric biosensor system for simultaneous determination of pyruvate and lactate. Sens Electron Microsyst Technol 2017; 14(4): 13–26.
[8] Kucherenko IS, Soldatkin OO, Kasap BO, Öztürk S, Akata B, Soldatkin AP, Dzyadevych SV. Elaboration of Urease Adsorption on Silicalite for Biosensor Creation. Electroanalysis. 2012; 24(6): 1380–5.
[9] Soldatkina OV, Kucherenko IS, Pyeshkova VM, Alekseev SA, Soldatkin OO, Dzyadevych SV. Improvement of amperometric transducer selectivity using nanosized phenylenediamine films. Nanoscale Res Lett. 2017;12(1):594.