The mechanism(s) by which electricity-producing microorganisms interact with an electrode is poorly understood. Outer membrane cytochromes and conductive pili are being considered as possible players, but the available information does not concur to a consensus mechanism yet. In this work we demonstrate that Geobacter sulfurreducens cells are able to change the way in which they exchange electrons with an electrode as a response to changes in the applied electrode potential. After several hours of polarization at 0.1 V Ag/AgCl-KCl (saturated), the voltammetric signature of the attached cells showed a single redox pair with a formal redox potential of about -0.08 V as calculated from chronopotentiometric analysis. A similar signal was obtained from cells adapted to 0.4 V. However, new redox couples were detected after conditioning at 0.6 V. A large oxidation process beyond 0.5 V transferring a higher current than that obtained at 0.1 V was found to be associated with two reduction waves at 0.23 and 0.50 V. The apparent equilibrium potential of these new processes was estimated to be at about 0.48 V from programmed current potentiometric results. Importantly, when polarization was lowered again to 0.1 V for 18 additional hours, the signals obtained at 0.6 V were found to greatly diminish in amplitude, whereas those previously found at the lower conditioning potential were recovered. Results clearly show the reversibility of cell adaptation to the electrode potential and pointto the polarization potential as a key variable to optimize energy production from an electricity producing population.
Down to the wire: electro‐active bacteria that exchange electrons with solid electrodes are studied by electrochemical and infrared techniques. The approach allows the identification of cell‐surface molecules involved in the direct electron transfer to the electrode, a development that is crucial for future utilization of these electricity‐producing microorganisms.
Electroactive bacteria can use a polarized electrode as final electron acceptor, allowing the use of electrochemical techniques for a very accurate quantification of its respiration rate. Biofilm cell respiration has been recently demonstrated to continue after the interruption of electrode polarization since these bacteria can store electrons in the haem groups of exocytoplasmic cytochromes. Interestingly, it has been shown that when the electrode is connected again, stored electrons can be recovered as a current superimposed to the basal steady state current produced by biofilm respiration. This work presents a model for the biofilm-catalysed electron transfer mechanism that reproduces the current profile obtained upon electrode reconnection. The model allows the estimation of kinetic parameters for internalization of the reduced substrate by the cells and the subsequent reduction of cell internal cytochromes, the electron transfer to mediators in the exterior of the cell, charge transport across the biofilm matrix to the electrode through fixed mediators and, finally, the oxidation of cytochromes at the biofilm/electrode interface. Based on these estimates, the distribution of stored charge within the biofilm can also be calculated. The results indicate that the processes involved in electron transfer from acetate to internal cytochromes represent the main limitation to current production, showing that both electron transport through the matrix of cytochromes and interfacial electron transfer are orders of magnitude faster than this process. Stored charge, on the other hand, is an order of magnitude higher inside the cells compared with that in the conductive matrix, suggesting that internal cytochromes are approximately ten times more abundant inside the cells than in the conductive matrix.
Devices that exploit electricity produced by electroactive bacteria such as Geobacter sulfurreducens have not yet been demonstrated beyond the laboratory scale. The current densities are far from the maximum that the bacteria can produce because fundamental properties such as the mechanism of extracellular electron transport and factors limiting cell respiration remain unclear. In this work, a strategy for the investigation of electroactive biofilms is presented. Numerical modeling of the response of G. sulfurreducens biofilms cultured on a rotating disk electrode has allowed for the discrimination of different limiting steps in the process of current production within a biofilm. The model outputs reveal that extracellular electron transport limits the respiration rate of the cells furthest from the electrode to the extent that cell division is not possible. The mathematical model also demonstrates that recent findings such as the existence of a redox gradient in actively respiring biofilms can be explained by an electron hopping mechanism but not when considering metallic-like conductivities.
Wired‐up bacteria: Confocal Raman microscopy in combination with 3D structural analysis is used in the measurement of redox gradients in electricity‐producing biofilms in vivo. The approach provides new relevant information for the understanding of electron conduction mechanisms in these systems.
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