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Electrochemical enzyme-based blood uric acid biosensor: new insight into the enzyme immobilization on the surface of electrode via poly-histidine tag

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Abstract

In a new approach, we considered the special affinity between Ni and poly-histidine tags of recombinant urate oxidase to utilize Ni-MOF for immobilizing the enzyme. In this study, a carbon paste electrode (CPE) was modified by histidine-tailed urate oxidase (H-UOX) and nickel-metal–organic framework (Ni-MOF) to construct H-UOX/Ni-MOF/CPE, which is a rapid, sensitive, and simple electrochemical biosensor for UA detection. The use of carboxy-terminal histidine-tailed urate oxidase in the construction of the electrode allows the urate oxidase enzyme to be positioned correctly in the electrode. This, in turn, enhances the efficiency of the biosensor. Characterization was carried out by X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), Brunauer–Emmett–Teller (BET), and field emission scanning electron microscopy (FE-SEM). At optimum conditions, the biosensor provided a short response time, linear response within 0.3–10 µM and 10–140 µM for UA with a detection limit of 0.084 µM, repeatability of 3.06%, and reproducibility of 4.9%. Furthermore, the biosensor revealed acceptable stability and selectivity of UA detection in the presence of the commonly coexisted ascorbic acid, dopamine, L-cysteine, urea, and glucose. The detection potential was at 0.4 V vs. Ag/AgCl.

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Acknowledgements

The authors would like to dedicate the present dissertation to the late Mr. Alireza Afzalipour and Mrs. Fakhereh Saba, the founders of the University of Kerman, because of their foresight and generosity in establishing this institution to train future generations of scientists. In addition, the authors acknowledge their appreciation to the late Dr. Parviz Dabiri for his generous support in establishing our research laboratory.

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Authors and Affiliations

  1. Department of Chemistry, Shahid Bahonar University of Kerman, Kerman, Iran

    Amineh Mashkoori, Ali Mostafavi & Tayebeh Shamspur

  2. Young Researchers Society, Shahid Bahonar University of Kerman, Kerman, Iran

    Amineh Mashkoori

  3. Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran

    Masoud Torkzadeh-Mahani

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  1. Amineh Mashkoori
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  2. Ali Mostafavi
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Correspondence to Masoud Torkzadeh-Mahani.

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Mashkoori, A., Mostafavi, A., Shamspur, T. et al. Electrochemical enzyme-based blood uric acid biosensor: new insight into the enzyme immobilization on the surface of electrode via poly-histidine tag. Microchim Acta 189, 326 (2022). https://doi.org/10.1007/s00604-022-05408-0

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