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Electroanalysis for Quantitative Assessment of Bacterial Activity

石木 健吾 大阪府立大学 DOI:info:doi/10.24729/00016954

2020.07.07

概要

As mentioned above, electrochemical techniques are suitable methods for quantification of cell activity with high sensitivity and in real-time. Therefore, electrochemical analysis was performed for quantitative assessment of bacterial activity, focusing on the redox species, protein, and electron transfer in intracellular/extracellular environment.

Chapter 1 is introduction, which described about bacteria, color-based analysis of metabolic activity, and electrochemical techniques in bioanalysis.

Chapter 2 shows a usefulness of microbial platform constructed by conductive polymers for observation of bacterial activity. Conducting polymers works as a biocompatible matrix for entrapping bacterial cells on an indium-tin-oxide (ITO)-coated electrode. The cell density and viability were optically evaluated by microscopy. The conducting polymers also facilitated electrochemical evaluation of the respiratory activity of bacterial cells.

Chapter 3 demonstrates a viable bacterial detection focused on the electrochemical property of tetrazolium salts, which was converted to an insoluble and redox active formazan compound in viable microbial cells. I focused on not colorimetric but electrochemical property of MTT. The insolubility of this formazan was effectively exploited as a surface-confined redox event. Bacterial suspensions that incubated with a tetrazolium salt was applied to ITO electrode and heat to dry for the adsorption. A distinctive voltammetric oxidation peak appeared, and the magnitude was correlated to the number of viable microbes in the suspension.

Chapter 4 describes the precious metal-ion reduction by S. oneidensis. I tracked the formation of gold nanoparticles (Au NPs) on the S. oneidensis cell surfaces, and investigated the roles of membrane proteins and extracellular polysaccharides in this process. In addition, I propose a simple method for the detection of metal ions in solution, focusing on the light-scattering characteristics of the metal nanoparticles formed on the cells.

Chapter 5 denotes a quantitative evaluation of electron generation based on individual enzyme reactions in S. oneidensis. By using potentiometric measurements, I have examined intracellular electron generation in bacterial suspensions of S. oneidensis supplemented with different carbon sources or ferricyanide. Real-time measurements by potentiometry in bacterial suspensions enabled precise quantification of the number of electrons generated by S. oneidensis based on the Nernst equation, because the [ferricyanide]/[ferrocyanide] ratio immediately changed during the incubation.

Chapter 6 summarized the whole results and conclusions of the thesis.

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