Juan Pablo Busalmen

C-Type Cytochromes Wire Electricity-Producing Bacteria to Electrodes†

Fil: Busalmen, Juan Pablo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata. Instituto de Investigaciones en Ciencia y Tecnologia de Materiales. Universidad Nacional de Mar del Plata. Facultad de Ingenieria. Instituto de Investigaciones en Ciencia y Tecnologia de Materiales; Argentina

Spectroscopic Slicing to Reveal Internal Redox Gradients in Electricity-Producing Biofilms†

Fil: Robuschi, Luciana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Mar del Plata. Instituto de Investigacion en Ciencia y Tecnologia de Materiales (i); Argentina. Universidad Nacional de Mar del Plata. Facultad de Ingenieria; Argentina

Charge accumulation and electron transfer kinetics in Geobacter sulfurreducens biofilms

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.

ATR-SEIRAs characterization of surface redox processes in G. sulfurreducens.

In this work we report on the occurrence of at least two different redox pairs on the cell surface of the electrogenic bacteria Geobacter sulfurreducens adsorbed on gold that are expressed in response to the polarization potential. As previously reported on graphite (Environ. Sci. Technol. 42 (2008) 2445) a typical low potential redox pair is found centered at around -0.06 V when cells are polarized for a few hours at 0.2 V, while a new pair centered at around 0.38 V is expressed upon polarization at 0.6 V. Reversible changes in the IR band pattern of whole cells where obtained by Attenuated Total Reflection-Surface Enhanced Infrared Absorption Spectroscopy (ATR-SEIRAS) upon potential cycling around both redox pairs. Changes clearly resemble the electrochemical turnover of oxidized/reduced states in c-type cytochromes, thus evidencing the nature of the involved molecules. The expression of external cytochromes in response to the potential of the electron acceptor suggests the existence of alternative pathways of electron transport with different energy yield, though it remains to be demonstrated.

Limitations for current production in Geobacter sulfurreducens biofilms.

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.

Evaluation of potato-processing wastewater treatment in a microbial fuel cell.

Wastewaters from potato-processing industries have been traditionally treated by a sequence of steps that include the production of methane as the anaerobic one. This work explores the feasibility of replacing or supplementing methanogenesis with the emerging technology of microbial fuel cells (MFCs). Electricity producing biofilms have been enriched from a real anaerobic sludge, and the conversion of potato-processing wastewater into electricity has been studied. When tested as a single treatment step, MFCs were able to process the wastewater with high COD removal but with low energetic conversion efficiency. On the other hand, as a complimentary step for methanogenesis, they improved conversion efficiency and significantly reduced the organic matter load of the final effluent. These results point at the combination of the energetic yield of methanogenesis and the improved COD removal of the electricity producing treatment as the implementation choice.

New ceramic electrodes allow reaching the target current density in bioelectrochemical systems

Whereas most of the studies conducted nowadays to boost electrode performance in bioelectrochemical systems (BES) are focused on carbonaceous scaffolds, in this study we demonstrate that ice-templated titanium-based ceramics (ITTC) can provide a new alternative for this purpose. We combined the chemistry of titanium suboxides (Ti4O7) with an ice-templating technique (ISISA) to produce electrically conducting and highly porous (88% porosity) 3D architectures. The ITTC platforms were characterized by strongly aligned macrochannels that provided a direct path for substrate supply under a flow-through configuration, while supporting the growth of electroactive Geobacter sulfurreducens biofilms. This new electrode material is demonstrated to outperform graphite when used as an anode in bioelectrochemical reactors, providing volumetric current densities of 9500 A m−3, equating to projected current densities of 128.7 A m−2 and maximum power densities of 1.9 kW m−3. The performance of the ITTC scaffolds levels that of any of the available materials on the current state of research. The presented alternative may lead to the start of a branch into the exploration of conducting ITTC materials in the growing area of bioelectrochemical technologies.

A long way to the electrode: how do Geobacter cells transport their electrons?

The mechanism of electron transport in Geobacter sulfurreducens biofilms is a topic under intense study and debate. Although some proteins were found to be essential for current production, the specific role that each one plays in electron transport to the electrode remains to be elucidated and a consensus on the mechanism of electron transport has not been reached. In the present paper, to understand the state of the art in the topic, electron transport from inside of the cell to the electrode in Geobacter sulfurreducens biofilms is analysed, reviewing genetic studies, biofilm conductivity assays and electrochemical and spectro-electrochemical experiments. Furthermore, crucial data still required to achieve a deeper understanding are highlighted.

Opportunities behind the unusual ability of geobacter sulfurreducens for exocellular respiration and electricity production

The possibility to improve the connection of cells to the electrode is significant for microbial fuel cell technology. In this communication we demonstrate that an improved connection can be made by controlling the physiological state of electricity-harvesting bacteria as Geobacter sulfurreducens.

Physiological Stratification in Electricity-Producing Biofilms of Geobacter sulfurreducens

The elucidation of mechanisms and limitations in electrode respiration by electroactive biofilms is significant for the development of rapidly emerging clean energy production and wastewater treatment technologies. In Geobacter sulfurreducens biofilms, the controlling steps in current production are thought to be the metabolic activity of cells, but still remain to be determined. By quantifying the DNA, RNA, and protein content during the long-term growth of biofilms on polarized graphite electrodes, we show in this work that current production becomes independent of DNA accumulation immediately after a maximal current is achieved. Indeed, the mean respiratory rate of biofilms rapidly decreases after this point, which indicates the progressive accumulation of cells that do not contribute to current production or contribute to a negligible extent. These results support the occurrence of physiological stratification within biofilms as a consequence of respiratory limitations imposed by limited biofilm conductivity.