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A Novel Electrochemical Biosensor Based on Tetx2 Monooxygenase Immobilized on a Nano-Porous Glassy Carbon Electrode for Tetracycline Residue Detection Publisher Pubmed



Besharati M1, 2 ; Hamedi J1, 2 ; Hosseinkhani S3 ; Saber R4
Authors
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Authors Affiliations
  1. 1. Department of Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, 14155-6455, Iran
  2. 2. Microbial Technology and Products Research Center, University of Tehran, Tehran, Iran
  3. 3. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
  4. 4. Research Center of Medical Science, Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, Iran

Source: Bioelectrochemistry Published:2019


Abstract

Different carbon-based nanostructures were used to investigate direct electron transfer (DET) of TetX2 monooxygenase (TetX2), and an enzyme-based biosensor for sensitive determination of tetracycline (TC) also fabricated. A polyethyleneimine (PEI) with positive charge groups was used for immobilization of TetX2 on modified glassy carbon electrodes. Cyclic voltammetry (CV) was employed to study the electrochemical characteristics of the immobilized enzyme and the performance of the proposed biosensor. Amongst multiple carbon-modified electrodes, nano-porous glassy carbon electrode (NPGCE) was selected because of its amplified signal response for flavin adenine dinucleotide (FAD) and superior electrocatalytic behavior toward oxygen reduction. The cyclic voltammogram of PEI/TetX2/NPGCE showed two couple of well-defined and quasi-reversible redox peaks of FAD, consistent with the realization of DET. The prepared electrode was then successfully introduced as a biosensing interface based on the oxygen reduction peak current, resulting in a linear range response from 0.5 to 5 μM with a good detection limit of 18 nM. The as-fabricated electrode demonstrates a fast response and excellent stability for the detection of TC. The results indicate that this simple, rapid, eco-friendly and economic strategy of PEI/TetX2/NPGCE preparation has potential for the fabrication of an enzyme-based biosensor for the practical detection of TC in food products. © 2019 Elsevier B.V.