Outer Membrane C-type Cytochromes Research Articles

The catalytic effect of TiO2 nanosheets on extracellular electron transfer of Shewanella loihica PV-4.

Electron transfer kinetics of Shewanella loihica PV-4 at the up-growing TiO2 nanosheet (TiO2-NS) modified carbon paper (CP) electrode was investigated. The effect of TiO2-NSs, which speeds up the interfacial electron transfer of outer membrane c-type cytochromes (OMCs), was revealed for the first time. TiO2-NSs with a polar surface modified hydrophobic CP into super-hydrophilic TiO2-NS/CP. The favorable interaction between PV-4 and TiO2-NSs not only enhanced microbial adhesion, but also altered the redox nature of OMCs. The mid-point potential of OMCs at TiO2-NS/CP was shifted to a more negative potential, indicating a higher thermodynamic driving force for the protein to release electrons. Moreover, electron transfer from OMCs to TiO2-NSs was also benefited from the positive shift of flat-band potential Vfb owing to reduced pH at the electrode/microorganism interface, as well as good electrical conductivity of TiO2-NSs. As a result, the electron transfer rate constant ket of OMCs at the TiO2-NS/CP anode was about three times faster than that at the CP anode. The accelerated electron transfer kinetics as well as 15% increase of biomass together accounted for a 97% increase of the maximum output power density in the MFC. The result expanded our knowledge about the role of a designed TiO2 nanostructure in microbial electron transfer that can be applied in other bio-electrochemical systems.

Ferrous ion regulated extracellular electron transfer: towards self-suppressed microbial iron(III) oxide reduction.

By using an electrochemical strategy, we demonstrated that ferrous ions are capable of decreasing bacterial EET activity in a certain potential range where the conduction-band edge of natural abundant iron(III) oxides is located. It is proposed that ferrous ions enable alteration of the formal potential of outer membrane c-type cytochromes, a crucial protein involved in the EET process.

Conduction-band edge dependence of carbon-coated hematite stimulated extracellular electron transfer of Shewanella oneidensis in bioelectrochemical systems

Bacteria-based bioelectrochemical systems (BESs) are promising technologies used for alternative energy generation, wastewater treatment, and environmental monitoring. However, their practical application is limited by the bioelectrode performance, mainly resulting from low extracellular electron transfer (EET) efficiency. In this study, a carbon-coated hematite (C/Hematite) electrode was successfully obtained by a green and solvent-free route, that is, heat treatment in an oxygen-rich environment using solid ferrocene as the precursor. The as-prepared C/Hematite electrode was evaluated as a high-performance electrode material in a Shewanella oneidensis-inoculated BES. The maximum biocurrent density of the Shewanella-attached C/Hematite electrode reached 0.22±0.01mAcm−2, which is nearly 6-times higher than that of a bare carbon cloth (CC) electrode (0.036±0.005mAcm−2). Electrochemical measurements revealed that the enhanced conductivity and better energy matching between the outer membrane c-type cytochromes of S. oneidensis and the electrode contributed to the improved EET efficiency. The results of this study demonstrated that the semiconductive properties of iron oxides play important roles for the involved bacterial extracellular respiration activities.

Nitrogen doped carbon nanoparticles enhanced extracellular electron transfer for high-performance microbial fuel cells anode

Nitrogen doped carbon nanoparticles (NDCN) were applied to modify the carbon cloth anodes of microbial fuel cells (MFCs) inoculated with Shewanella oneidensis MR-1, one of the most well-studied exoelectrogens. Experimental results demonstrated that the use of NDCN increased anodic absorption of flavins (i.e., the soluble electron mediator secreted by S. oneidensis MR-1), facilitating shuttle-mediated extracellular electron transfer. In addition, we also found that NDCN enabled enhanced contact-based direct electron transfer via outer-membrane c-type cytochromes. Taken together, the performance of MFCs with the NDCN-modified anode was enormously enhanced, delivering a maximum power density 3.5 times’ higher than that of the MFCs without the modification of carbon cloth anodes.

Coupled mercury-cell sorption, reduction, and oxidation on methylmercury production by Geobacter sulfurreducens PCA.

G. sulfurreducens PCA cells have been shown to reduce, sorb, and methylate Hg(II) species, but it is unclear whether this organism can oxidize and methylate dissolved elemental Hg(0) as shown for Desulfovibrio desulfuricans ND132. Using Hg(II) and Hg(0) separately as Hg sources in washed cell assays in phosphate buffered saline (pH 7.4), we report how cell-mediated Hg reduction and oxidation compete or synergize with sorption, thus affecting the production of toxic methylmercury by PCA cells. Methylation is found to be positively correlated to Hg sorption (r = 0.73) but negatively correlated to Hg reduction (r = -0.62). These reactions depend on the Hg and cell concentrations or the ratio of Hg to cellular thiols (-SH). Oxidation and methylation of Hg(0) are favored at relatively low Hg to cell-SH molar ratios (e.g., <1). Increasing Hg to cell ratios from 0.25 × 10(-19) to 25 × 10(-19) moles-Hg/cell (equivalent to Hg/cell-SH of 0.71 to 71) shifts the major reaction from oxidation to reduction. In the absence of five outer membrane c-type cytochromes, mutant ΔomcBESTZ also shows decreases in Hg reduction and increases in methylation. However, the presence of competing thiol-binding ions such as Zn(2+) leads to increased Hg reduction and decreased methylation. These results suggest that the coupled cell-Hg sorption and redox transformations are important in controlling the rates of Hg uptake and methylation by G. sulfurreducens PCA in anoxic environments.

A trans-outer membrane porin-cytochrome protein complex for extracellular electron transfer by Geobacter sulfurreducens PCA.

The multi-heme, outer membrane c-type cytochrome (c-Cyt) OmcB of Geobacter sulfurreducens was previously proposed to mediate electron transfer across the outer membrane. However, the underlying mechanism has remained uncharacterized. In G. sulfurreducens, the omcB gene is part of two tandem four-gene clusters, each is predicted to encode a transcriptional factor (OrfR/OrfS), a porin-like outer membrane protein (OmbB/OmbC), a periplasmic c-type cytochrome (OmaB/OmaC) and an outer membrane c-Cyt (OmcB/OmcC) respectively. Here, we showed that OmbB/OmbC, OmaB/OmaC and OmcB/OmcC of G. sulfurreducens PCA formed the porin-cytochrome (Pcc) protein complexes, which were involved in transferring electrons across the outer membrane. The isolated Pcc protein complexes reconstituted in proteoliposomes transferred electrons from reduced methyl viologen across the lipid bilayer of liposomes to Fe(III)-citrate and ferrihydrite. The pcc clusters were found in all eight sequenced Geobacter and 11 other bacterial genomes from six different phyla, demonstrating a widespread distribution of Pcc protein complexes in phylogenetically diverse bacteria. Deletion of ombB-omaB-omcB-orfS-ombC-omaC-omcC gene clusters had no impact on the growth of G. sulfurreducens PCA with fumarate but diminished the ability of G. sulfurreducens PCA to reduce Fe(III)-citrate and ferrihydrite. Complementation with the ombB-omaB-omcB gene cluster restored the ability of G. sulfurreducens PCA to reduce Fe(III)-citrate and ferrihydrite.

Flavin redox bifurcation as a mechanism for controlling the direction of electron flow during extracellular electron transfer.

The iron-reducing bacterium Shewanella oneidensis MR-1 has a dual directional electronic conduit involving 40 heme redox centers in flavin-binding outer-membrane c-type cytochromes (OM c-Cyts). While the mechanism for electron export from the OM c-Cyts to an anode is well understood, how the redox centers in OM c-Cyts take electrons from a cathode has not been elucidated at the molecular level. Electrochemical analysis of live cells during switching from anodic to cathodic conditions showed that altering the direction of electron flow does not require gene expression or protein synthesis, but simply redox potential shift about 300 mV for a flavin cofactor interacting with the OM c-Cyts. That is, the redox bifurcation of the riboflavin cofactor in OM c-Cyts switches the direction of electron conduction in the biological conduit at the cell-electrode interface to drive bacterial metabolism as either anode or cathode catalysts.

Cell-secreted flavins bound to membrane cytochromes dictate electron transfer reactions to surfaces with diverse charge and pH.

The variety of solid surfaces to and from which microbes can deliver electrons by extracellular electron transport (EET) processes via outer-membrane c-type cytochromes (OM c-Cyts) expands the importance of microbial respiration in natural environments and industrial applications. Here, we demonstrate that the bifurcated EET pathway of OM c-Cyts sustains the diversity of the EET surface in Shewanella oneidensis MR-1 via specific binding with cell-secreted flavin mononucleotide (FMN) and riboflavin (RF). Microbial current production and whole-cell differential pulse voltammetry revealed that RF and FMN enhance EET as bound cofactors in a similar manner. Conversely, FMN and RF were clearly differentiated in the EET enhancement by gene-deletion of OM c-Cyts and the dependency of the electrode potential and pH. These results indicate that RF and FMN have specific binding sites in OM c-Cyts and highlight the potential roles of these flavin-cytochrome complexes in controlling the rate of electron transfer to surfaces with diverse potential and pH.

Uptake of self-secreted flavins as bound cofactors for extracellular electron transfer in Geobacter species

Geobacter cells utilize self-secreted riboflavin as a bound-cofactor in outer-membrane c-type cytochromes to enhance the rate of bacterial electron transport.

Influence of outer membrane c‐type cytochromes on particle size and activity of extracellular nanoparticles produced by Shewanella oneidensis

The metal-reducing bacterium Shewanella oneidensis is capable of reducing various metal(loid)s and produces nanoparticles (NPs) extracellularly, in which outer membrane c-type cytochromes (OMCs) have been suggested to play important roles. The objective of this study was to investigate the influence of the OMCs, that is, MtrC and OmcA, on the size and activity of the extracellular silver NPs (AgNPs) and silver sulfide NPs (Ag(2)S NPs) produced by S. oneidensis MR-1. We found that (i) the lack of OMCs on S. oneidensis cell surface decreased the particle size of the extracellular biogenic AgNPs and Ag(2)S NPs; (ii) the biogenic AgNPs from the mutant lacking OMCs showed higher antibacterial activity; and (iii) the biogenic Ag(2)S NPs from the mutant lacking OMCs exhibited higher catalytic activity in methylviologen reduction. The results suggest that it may be possible to control particle size and activity of the extracellular biogenic NPs via controlled expression of the genes encoding surface proteins. In addition, we also reveal that in extracellular biosynthesis of NPs the usually neglected non-cell-associated NPs could have high catalytic activity, highlighting the need of novel methods that can efficiently retain extracellular NPs in the biosynthesis processes.

Proteins involved in electron transfer to Fe(III) and Mn(IV) oxides by Geobacter sulfurreducens and Geobacter uraniireducens

Whole-genome microarray analysis of Geobacter sulfurreducens grown on insoluble Fe(III) oxide or Mn(IV) oxide versus soluble Fe(III) citrate revealed significantly different expression patterns. The most upregulated genes, omcS and omcT, encode cell-surface c-type cytochromes, OmcS being required for Fe(III) and Mn(IV) oxide reduction. Other electron transport genes upregulated on both metal oxides included genes encoding putative menaquinol : ferricytochrome c oxidoreductase complexes Cbc4 and Cbc5, periplasmic c-type cytochromes Dhc2 and PccF, outer membrane c-type cytochromes OmcC, OmcG and OmcV, multicopper oxidase OmpB, the structural components of electrically conductive pili, PilA-N and PilA-C, and enzymes that detoxify reactive oxygen/nitrogen species. Genes upregulated on Fe(III) oxide encode putative menaquinol : ferricytochrome c oxidoreductase complexes Cbc3 and Cbc6, periplasmic c-type cytochromes, including PccG and PccJ, and outer membrane c-type cytochromes, including OmcA, OmcE, OmcH, OmcL, OmcN, OmcO and OmcP. Electron transport genes upregulated on Mn(IV) oxide encode periplasmic c-type cytochromes PccR, PgcA, PpcA and PpcD, outer membrane c-type cytochromes OmaB/OmaC, OmcB and OmcZ, multicopper oxidase OmpC and menaquinone-reducing enzymes. Genetic studies indicated that MacA, OmcB, OmcF, OmcG, OmcH, OmcI, OmcJ, OmcM, OmcV and PccH, the putative Cbc5 complex subunit CbcC and the putative Cbc3 complex subunit CbcV are important for reduction of Fe(III) oxide but not essential for Mn(IV) oxide reduction. Gene expression patterns for Geobacter uraniireducens were similar. These results demonstrate that the physiology of Fe(III)-reducing bacteria differs significantly during growth on different insoluble and soluble electron acceptors and emphasize the importance of c-type cytochromes for extracellular electron transfer in G. sulfurreducens.

Mechanisms of long-distance extracellular electron transfer of metal-reducing bacteria mediated by nanocolloidal semiconductive iron oxides

Although nanocrystalline Fe(III) oxides are abundant in natural minerals, few studies have noticed their biological significance from the view point of semiconductive properties. To examine the roles of semiconductive Fe(III) oxides in the respiratory activity of microorganisms, we cultured the Fe(III)-reducing bacterium Shewanella loihica PV-4 in an electrochemical cell and examined the influence of cell surface-associated Fe-oxide nanocolloids on the extracellular electron transfer (EET) efficiency. It was found that the respiratory current was greatly improved (over 40 fold) when α-Fe2O3 or α-FeOOH was supplemented into the cell cultures. In contrast, only a small improvement (less than 4 fold) was observed after the addition of Fe3O4 and γ-Fe2O3. It was also found that the outer-membrane c-type cytochromes (c-Cyts) of Shewanella cells had a midpoint potential close to the conduction-band edge of α-Fe2O3 and α-FeOOH, whereas a large energy separation existed for γ-Fe2O3 and Fe3O4. These observations support the recently proposed model for bacterial long-distance EET (R. Nakamura, et al., Angew. Chem., Int. Ed., 2009, 48, 508–511), in which c-Cyts serve as electrical linkages between nanocolloids, allowing the cell population to exploit semiconductor-mediated electron-hopping. Support for this model was also provided by the mutational experiments of outer-membrane c-Cyts and the clear percolation behavior on the formation of conductive networks with respect to the amount of loaded colloids. Thus the EET mediated by semiconductive nanocolloids highlighted the microbial ability to take advantage of semiconductor-based electrochemistry for respiratory activities.

Electron transfer mechanisms between microorganisms and electrodes in bioelectrochemical systems

Microbes have been shown to naturally form veritable electric grids in which different species acting as electron donors and others acting as electron acceptors cooperate. The uptake of electrons from cells adjacent to them is a mechanism used by microorganisms to gain energy for cell growth and maintenance. The external discharge of electrons in lieu of a terminal electron acceptor, and the reduction of external substrates to uphold certain metabolic processes, also plays a significant role in a variety of microbial environments. These vital microbial respiration events, viz. extracellular electron transfer to and from microorganisms, have attracted widespread attention in recent decades and have led to the development of fascinating research concerning microbial electrochemical sensors and bioelectrochemical systems for environmental and bioproduction applications involving different fuels and chemicals. In such systems, microorganisms use mainly either (1) indirect routes involving use of small redox-active organic molecules referred to as redox mediators, secreted by cells or added exogenously, (2) primary metabolites or other intermediates, or (3) direct modes involving physical contact in which naturally occurring outer-membrane c-type cytochromes shuttle electrons for the reduction or oxidation of electrodes. Electron transfer mechanisms play a role in maximizing the performance of microbe–electrode interaction-based systems and help very much in providing an understanding of how such systems operate. This review summarizes the mechanisms of electron transfer between bacteria and electrodes, at both the anode and the cathode, in bioelectrochemical systems. The use over the years of various electrochemical approaches and techniques, cyclic voltammetry in particular, for obtaining a better understanding of the microbial electrocatalysis and the electron transfer mechanisms involved is also described and exemplified.

Biotic and Abiotic Reduction and Solubilization of Pu(IV)O2•xH2O(am) as Affected by Anthraquinone-2,6-disulfonate (AQDS) and Ethylenediaminetetraacetate (EDTA)

This study measured reductive solubilization of plutonium(IV) hydrous oxide (Pu(IV)O(2)·xH(2)O((am))) with hydrogen (H(2)) as electron donor, in the presence or absence of dissimilatory metal-reducing bacteria (DMRB), anthraquinone-2,6-disulfonate (AQDS), and ethylenediaminetetraacetate (EDTA). In PIPES buffer at pH 7 with excess H(2), Shewanella oneidensis and Geobacter sulfurreducens both solubilized <0.001% of 0.5 mM Pu(IV)O(2)·xH(2)O((am)) over 8 days, with or without AQDS. However, Pu((aq)) increased by an order of magnitude in some treatments, and increases in solubility were associated with production of Pu(III)((aq)). The solid phase of these treatments contained Pu(III)(OH)(3(am)), with more in the DMRB treatments compared with abiotic controls. In the presence of EDTA and AQDS, PuO(2)·xH(2)O((am)) was completely solubilized by S. oneidensis and G. sulfurreducens in ∼24 h. Without AQDS, bioreductive solubilization was slower (∼22 days) and less extensive (∼83-94%). In the absence of DMRB, EDTA facilitated reductive solubilization of 89% (without AQDS) to 98% (with AQDS) of the added PuO(2)·xH(2)O((am)) over 418 days. An in vitro assay demonstrated electron transfer to PuO(2)·xH(2)O((am)) from the S. oneidensis outer-membrane c-type cytochrome MtrC. Our results (1) suggest that PuO(2)·xH(2)O((am)) reductive solubilization may be important in reducing environments, especially in the presence of complexing ligands and electron shuttles, (2) highlight the environmental importance of polynuclear, colloidal Pu, (3) provide additional evidence that Pu(III)-EDTA is a more likely mobile form of Pu than Pu(IV)-EDTA, and (4) provide another example of outer-membrane cytochromes and electron-shuttling compounds facilitating bioreduction of insoluble electron acceptors in geologic environments.

Long-range electron conduction of Shewanella biofilms mediated by outer membrane C-type cytochromes

We investigated the role of c-type cytochromes (c-Cyts) in electron conduction across biofilms of Shewanella oneidensis MR-1 and the relevance of the electron conductivity for biological current generation. Following the formation of monolayer and multilayer biofilms on indium-tin oxide electrodes, we quantified the c-Cyts that were electrically wired with the electrode surface using whole-cell voltammetry. A multilayer biofilm with a thickness of 16μm exhibited a redox peak with an 8-fold larger coulombic area than that of a monolayer biofilm (about 0.5-μm thickness), indicating an abundance of c-Cyts that are able to perform redox-cycling reactions with the distant electrode surface. To determine if this electron conduit of c-Cyts participated in biological current generation, we conducted slow-scan voltammetry for multilayer biofilms. A large anodic current of c-Cyts caused by microbial lactate oxidization was observed during the slow-potential scans, demonstrating the transport of respiratory electrons via the sequential redox cycling of c-Cyts. Experiments with deletion mutants deficient in outer-membrane (OM) c-Cyts (ΔmtrC/ΔomcA, ΔpilD), and the biosynthetic protein of capsular polysaccharide (ΔSO3177) suggested that cell-surface-bound c-Cyts, but those located on pili or extracellular polymeric substrates, play a predominant role in the long-range electron conduction in the biofilm of S. oneidensis MR-1.

Manganese sulfide formation via concomitant microbial manganese oxide and thiosulfate reduction

The dissimilatory metal-reducing bacterium, Shewanella oneidensis MR-1 produced γ-MnS (rambergite) nanoparticles during the concurrent reduction of MnO₂ and thiosulfate coupled to H₂ oxidation. To investigate effect of direct microbial reduction of MnO₂ on MnS formation, two MR-1 mutants defective in outer membrane c-type cytochromes (ΔmtrC/ΔomcA and ΔmtrC/ΔomcA/ΔmtrF) were also used and it was determined that direct reduction of MnO₂ was dominant relative to chemical reduction by biogenic sulfide generated from thiosulfate reduction. Although bicarbonate was excluded from the medium, incubations of strain MR-1 with lactate as the electron donor produced MnCO₃ (rhodochrosite) as well as MnS in nearly equivalent amounts as estimated by micro X-ray diffraction (micro-XRD) analysis. It was concluded that carbonate released from lactate metabolism promoted MnCO₃ formation and that Mn(II) mineralogy was strongly affected by carbonate ions even in the presence of abundant sulfide and weakly alkaline conditions expected to favour the precipitation of MnS. Formation of MnS, as determined by a combination of micro-XRD, transmission electron microscopy, energy dispersive X-ray spectroscopy, and selected area electron diffraction analyses was consistent with equilibrium speciation modelling predictions. Biogenic manganese sulfide may be a manganese sink in the Mn biogeochemical cycle in select environments such as deep anoxic marine basins within the Baltic Sea.

Contribution of Extracellular Polymeric Substances from Shewanella sp. HRCR-1 Biofilms to U(VI) Immobilization

The goal of this study was to quantify the contribution of extracellular polymeric substances (EPS) to U(VI) immobilization by Shewanella sp. HRCR-1. Through comparison of U(VI) immobilization using cells with bound EPS (bEPS) and cells with minimal EPS, we show that (i) bEPS from Shewanella sp. HRCR-1 biofilms contribute significantly to U(VI) immobilization, especially at low initial U(VI) concentrations, through both sorption and reduction; (ii) bEPS can be considered a functional extension of the cells for U(VI) immobilization and they likely play more important roles at lower initial U(VI) concentrations; and (iii) the U(VI) reduction efficiency is dependent upon the initial U(VI) concentration and decreases at lower concentrations. To quantify the relative contributions of sorption and reduction to U(VI) immobilization by EPS fractions, we isolated loosely associated EPS (laEPS) and bEPS from Shewanella sp. HRCR-1 biofilms grown in a hollow fiber membrane biofilm reactor and tested their reactivity with U(VI). We found that, when reduced, the isolated cell-free EPS fractions could reduce U(VI). Polysaccharides in the EPS likely contributed to U(VI) sorption and dominated the reactivity of laEPS, while redox active components (e.g., outer membrane c-type cytochromes), especially in bEPS, possibly facilitated U(VI) reduction.

Quantitative Analysis of Cell Surface Membrane Proteins Using Membrane-Impermeable Chemical Probe Coupled with18O Labeling

We report a mass spectrometry-based strategy for quantitative analysis of cell surface membrane proteome changes. The strategy includes enrichment of surface membrane proteins using a membrane-impermeable chemical probe followed by stable isotope (18)O labeling and LC-MS analysis. We applied this strategy for enriching membrane proteins expressed by Shewanella oneidensis MR-1, a Gram-negative bacterium with known metal-reduction capability via extracellular electron transfer between outer membrane proteins and extracellular electron receptors. LC/MS/MS analysis resulted in the identification of about 400 proteins with 79% of them being predicted to be membrane localized. Quantitative aspects of the membrane enrichment were shown by peptide level (16)O and (18)O labeling of proteins from wild-type and mutant cells (generated from deletion of a type II secretion protein, GspD) prior to LC-MS analysis. Using a chemical probe labeled pure protein as an internal standard for normalization, the quantitative data revealed reduced abundances in Delta gspD mutant cells of many outer membrane proteins including the outer membrane c-type cytochromes OmcA and MtrC, in agreement with a previous report that these proteins are substrates of the type II secretion system.

Anode Biofilm Transcriptomics Reveals Outer Surface Components Essential for High Density Current Production in Geobacter sulfurreducens Fuel Cells

The mechanisms by which Geobacter sulfurreducens transfers electrons through relatively thick (>50 µm) biofilms to electrodes acting as a sole electron acceptor were investigated. Biofilms of Geobacter sulfurreducens were grown either in flow-through systems with graphite anodes as the electron acceptor or on the same graphite surface, but with fumarate as the sole electron acceptor. Fumarate-grown biofilms were not immediately capable of significant current production, suggesting substantial physiological differences from current-producing biofilms. Microarray analysis revealed 13 genes in current-harvesting biofilms that had significantly higher transcript levels. The greatest increases were for pilA, the gene immediately downstream of pilA, and the genes for two outer c-type membrane cytochromes, OmcB and OmcZ. Down-regulated genes included the genes for the outer-membrane c-type cytochromes, OmcS and OmcT. Results of quantitative RT-PCR of gene transcript levels during biofilm growth were consistent with microarray results. OmcZ and the outer-surface c-type cytochrome, OmcE, were more abundant and OmcS was less abundant in current-harvesting cells. Strains in which pilA, the gene immediately downstream from pilA, omcB, omcS, omcE, or omcZ was deleted demonstrated that only deletion of pilA or omcZ severely inhibited current production and biofilm formation in current-harvesting mode. In contrast, these gene deletions had no impact on biofilm formation on graphite surfaces when fumarate served as the electron acceptor. These results suggest that biofilms grown harvesting current are specifically poised for electron transfer to electrodes and that, in addition to pili, OmcZ is a key component in electron transfer through differentiated G. sulfurreducens biofilms to electrodes.

Harnessing self-organizing bacterial networks for microbial fuel cells

Present concerns about the depletion of natural resources have encouraged studies to exploit microbial ingenuity to harness renewable sources of energy.1 Microbes have evolved diverse metabolic processes to inhabit a broad range of natural environments and can performmost chemical reactions that are thermodynamically possible to generate energy. In anaerobic environments, such as soils, sediments, and the deep sea, some microbes employ a unique metabolic strategy to respire on the surface of iron-based minerals.2 Metal-reducing bacteria possess a significant quantity of c-type cytochromes (c-Cyts) in their outer membrane (OM) and transfer electrons to the attached Fe(III) oxides during metabolism: see Figure 1(a). These bacteria produce pilus-like appendages—called bacterial nanowires, several tens of microns long—that are thought to transport electrons to Fe(III) oxides located at relatively long distances from the cell surface. Using extracellular metal oxides for respiration is essential to eliminate excess electrons generated by the oxidation of organic matter, which would otherwise inhibit energy production. The significance of this unique microbial metabolic strategy in anaerobic ecosystems has been argued extensively.2 Recent reports demonstrated that metal-reducing bacteria can also use various artificial solid materials such as graphite, gold, and indium-tin-oxide (ITO) electrodes as a terminal-electron acceptor.3 This important result highlights the potential to harvest electricity by collecting waste electrons from bacteria through the electrode in microbial fuel cells (MFCs), a type of electrochemical fuel cell. A great deal of research has focused on identifying the OM proteins responsible for extracellular electron transfer and improving the MFC power density. However, the specific mechaFigure 1. (a) Bacterial extracellular electron transfer mediated by outer membrane c-type cytochromes (OM c-Cyts). (b) Cyclic voltammetry of Shewanella loihica in the absence (trace 1) and presence (trace 2) of α-Fe2O3 colloids, the α-Fe2O3 colloidal solution lacking cells (trace 3), and the cell-free supernatant (trace 4). SHE: Standard hydrogen electrode. (c) Proposed model for the bacterial electron-transfer process mediated byα-Fe2O3. V.B, C.B: Valence, conduction bands.