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. 2021 Sep 13;11(9):332.
doi: 10.3390/bios11090332.

Use of PEDOT:PSS/Graphene/Nafion Composite in Biosensors Based on Acetic Acid Bacteria

Yulia Plekhanova  1 , Sergei Tarasov  1 , Anatoly Reshetilov  1
Affiliations

Use of PEDOT:PSS/Graphene/Nafion Composite in Biosensors Based on Acetic Acid Bacteria

Yulia Plekhanova et al. Biosensors (Basel). .

Abstract

Immobilization of the biocomponent is one of the most important stages in the development of microbial biosensors. In this study, we examined the electrochemical properties of a novel PEDOT:PSS/graphene/Nafion composite used to immobilize Gluconobacter oxydans bacterial cells on the surface of a graphite screen-printed electrode. Bioelectrode responses to glucose in the presence of a redox mediator 2,6-dichlorophenolindophenol were studied. The presence of graphene in the composite reduced the negative effect of PEDOT:PSS on cells and improved its conductivity. The use of Nafion enabled maintaining the activity of acetic acid bacteria at the original level for 120 days. The sensitivity of the bioelectrode based on G. oxydans/PEDOT:PSS/graphene/Nafion composite was shown to be 22 μA × mM-1 × cm-2 within the linear range of glucose concentrations. The developed composite can be used both in designing bioelectrochemical microbial devices and in biotechnology productions for long-term immobilization of microorganisms.

Keywords: Gluconobacter oxydans; Nafion; PEDOT:PSS; glucose biosensors; graphene; microbial immobilization; screen-printed electrodes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Biosensor formation protocols.
Figure 2
Figure 2
Cyclic voltammograms of a bioelectrode depending on the order of applying the mixture components. 1, Layer-by-layer application of a PEDOT:PSS/graphene mixture, then of a mixture of cells with Nafion; 2, a mixture of PEDOT:PSS/graphene and Nafion, then cells; 3, a mixture of Nafion with cells, then the mixing with PEDOT:PSS/graphene.
Figure 3
Figure 3
Cyclic voltammograms of electrodes at various biosensor-formation stages: 1, SPE/PEDOT; 2, SPE/PEDOT/graphene; 3, SPE/PEDOT/graphene/Nafion; 4, SPE/PEDOT/graphene/Nafion/G. oxydans; 5, SPE/PEDOT/graphene/Nafion/G. oxydans + 1 mM glucose. Measurements were carried out in the presence of 42 µM DCPIP; scan rate, 3 mV/s.
Figure 4
Figure 4
Nyquist diagrams for various composites: 1, SPE/PEDOT:PSS; 2, SPE/PEDOT: PSS/graphene; 3, SPE/PEDOT:PSS/graphene/Nafion; 4, SPE/PEDOT:PSS/graphene/Nafion/G. oxydans; 5, SPE/PEDOT/graphene/Nafion/G. oxydans at an addition of 1 mM glucose. Measurements were carried out in the presence of 5 mM [Fe(CN)6]3-/4-.
Figure 5
Figure 5
Dependence of the biosensor signal on the introduction of 0.3 mM (1) and 1 mM (2) glucose on the concentration of bacterial cells on the surface of an electrode modified with PEDOT:PSS and graphene.
Figure 6
Figure 6
Calibration curves of glucose biosensors with various compositions of cells and Nafion on the surface of an electrode modified with PEDOT:PSS and graphene. Concentration of cells on the electrode surface, 0.3 mg/mm2: (a) day 1; (b) day 15.
Figure 7
Figure 7
Effect of pH (a) and NaCl concentration (b) on the response of microbial biosensor. Concentration of glucose in the measuring cell, 0.3 mM. Concentration of background buffer solution, 25 mM.
Figure 8
Figure 8
Calibration curves of glucose biosensors with various compositions of composite material on the electrode surface: 1, SPE/chitosan/G. oxydans; 2, SPE/Nafion/G. oxydans; 3, SPE/PEDOT:PSS/Nafion/G. oxydans; 4, SPE/PEDOT:PSS/graphene/Nafion/G. oxydans. Concentration of cells on the electrode surface, 0.3 mg/mm2.
Figure 9
Figure 9
Operational stability of SPE/PEDOT:PSS/graphene/Nafion/G. oxydans biosensor. Concentration of glucose in the measuring cell, 0.3 mM.
Figure 10
Figure 10
Long-term stability of SPE/PEDOT:PSS/graphene/Nafion/G. oxydans biosensor. Concentration of glucose in the measuring cell, 1 mM.
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