Kinetics of Intracellular Electron Generation in Shewanella oneidensis MR‑1

• Published in: Analytical chemistry (Washington) 2019-11, Vol.91 (22), p.14401-14406
• Main Authors: Ishiki, Kengo, Shiigi, Hiroshi
• Format: Article
• Language: English
• Subjects: Bacteria; Biochemical fuel cells; Carbon; Carbon sources; Cell viability; Chemistry; Coenzyme A; Cytochrome; Cytochromes; Dehydrogenase; Dehydrogenases; Electrons; Ferricyanide; Ferrocyanide; Fuel technology; Intracellular; Iron cyanides; L-Lactate dehydrogenase; Lactate dehydrogenase; Lactic acid; Materials recovery; Membranes; Metabolism; Metal industry wastewaters; Metals; Microorganisms; Outer membranes; Purification; Pyruvic acid; Quantitative analysis; Reaction kinetics; Shewanella oneidensis; Tricarboxylic acid cycle; Wastewater treatment
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.9b02900

Efficient utilization of bacterial bioresources requires quantitative evaluation of metabolic activity in living bacterial cells. Shewanella oneidensis MR-1 transfers electrons generated within the cell to the extracellular environment via the cytochrome complex in the inner/outer membranes and is one of the most useful bacteria for the recovery of metals, treatment of wastewater, and preparation of microbial fuel cells. Here, we performed a quantitative evaluation of electron generation based on individual enzyme reactions in S. oneidensis MR-1. By using potentiometric measurements, we have examined intracellular electron generation in bacterial suspensions of S. oneidensis supplemented with different carbon sources (formate, lactate, pyruvate, or acetyl coenzyme A) or ferricyanide, which was almost completely reduced to ferrocyanide during the incubation without affecting bacterial cell viability. The amount of electron generation strongly depended on the nature of the carbon source. Analysis of the obtained kinetic parameters of intracellular electron generation demonstrated that formate was the most effective carbon source, as it enabled 2.5-fold faster electron generation rate than other sources. We established that the respective contributions of lactate dehydrogenase, pyruvate dehydrogenase/pyruvate-formate-lyase, and tricarboxylic acid cycle to lactate metabolism were 62%, 31%, and 7.4%, correspondingly. Furthermore, we clarified that electrons may be generated at 1.6 × 10–12 A s–1 by ideal metabolism in a single living cell. These findings establish the basis for biological strategies of electron production and facilitate the utilization of S. oneidensis as a bioresource in practical applications, including energy production, environmental purification, and recovery of useful materials.