Enhancement of extracellular electron transfer and bioelectricity output by synthetic porin

The microbial fuel cell (MFC), is a promising environmental biotechnology for harvesting electricity energy from organic wastes. However, low bacterial membrane permeability of electron shuttles is a limiting factor that restricts the electron shuttle‐mediated extracellular electron transfer (EET) from bacteria to electrodes, thus the electricity power output of MFCs. To this end, we heterologously expressed a porin protein OprF from Pseudomonas aeruginosa PAO1 into Escherichia coli, which dramatically increased its membrane permeability, delivering a much higher current output in MFCs than its parental strain (BL21). We found that the oprF‐expression strain showed more efficient EET than its parental strain. More strikingly, the enhanced membrane permeability also rendered the oprF‐expression strain an efficient usage of riboflavin as the electron shuttle, whereas its parental strain was incapable of. Our results substantiated that membrane permeability is crucial for the efficient EET, and indicated that the expression of synthetic porins could be an efficient strategy to enhance bioelectricity generation by microorganisms (including electrogenic bacteria) in MFCs. Biotechnol. Bioeng. 2013; 110: 408–416. © 2012 Wiley Periodicals, Inc.     

Metabolites produced by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. enable a Gram-positive bacterium to achieve extracellular electron transfer

Previous studies revealed the abundance of Pseudomonas sp. in the microbial community of a microbial fuel cell (MFC). These bacteria can transfer electrons to the electrode via self-produced phenazine-based mediators. A MFC fed with acetate where several Pseudomonas sp. were present was found to be rich in a Gram-positive bacterium, identified as Brevibacillus sp. PTH1. Remarkably, MFCs operated with only the Brevibacillus strain in their anodes had poor electricity generation. Upon replacement of the anodic aqueous part of Brevibacillus containing MFCs with the cell-free anodic supernatants of MFCs operated with Pseudomonas sp. CMR12a, a strain producing considerable amounts of phenazine-1-carboxamide (PCN) and biosurfactants, the electricity generation was improved significantly. Supernatants of Pseudomonas sp. CMR12a_Reg, a regulatory mutant lacking the ability to produce PCN, had no similar improvement effect. Purified PCN, together with rhamnolipids as biosurfactants (1 mg L⁻¹), could clearly improve electricity generation by Brevibacillus sp. PTH1, as well as enable this bacterium to oxidize acetate with concomitant reduction of ferric iron, supplied as goethite (FeOOH). When added alone, PCN had no observable effects on Brevibacillus’ electron transfer. This work demonstrates that metabolites produced by Pseudomonas sp. enable Gram-positive bacteria to achieve extracellular electron transfer. Possibly, this bacterial interaction is a key process in the anodic electron transfer of a MFC, enabling Brevibacillus sp. PTH1 to achieve its dominance. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Characterization of electrochemical activity of a strain ISO2‐3 phylogenetically related to Aeromonas sp. isolated from a glucose‐fed microbial fuel cell

The microbial communities associated with electrodes in closed and open circuit microbial fuel cells (MFCs) fed with glucose were analyzed by 16S rRNA approach and compared. The comparison revealed that bacteria affiliated with the Aeromonas sp. within the Gammaproteobacteria constituted the major population in the closed circuit MFC (harvesting electricity) and considered to play important roles in current generation. We, therefore, attempted to isolate the dominant bacteria from the anode biofilm, successfully isolated a Fe (III)‐reducing bacterium phylogenetically related to Aeromonas sp. and designated as strain ISO2‐3. The isolated strain ISO2‐3 could grow and concomitantly produce current (max. 0.24 A/m²) via oxidation of glucose or hydrogen with an electrode serving as the sole electron acceptor. The strain could ferment glucose, but generate less electrical current. Cyclic voltammetry supported the strain ISO2‐3 was electrically active and likely to transfer electrons to the electrode though membrane‐associated compounds (most likely c‐type cytochrome). This mechanism requires intimate contact with the anode surface. Scanning electron microscopy revealed that the strain ISO2‐3 developed multiplayer biofilms on the anode surface and also produced anchor‐like filamentous appendages (most likely pili) that may promote long‐range electron transport across the thick biofilm. Biotechnol. Bioeng. 2009; 104: 901–910. © 2009 Wiley Periodicals, Inc.     

Characterization of Exoelectrogenic Bacteria Enterobacter Strains Isolated from a Microbial Fuel Cell Exposed to Copper Shock Load

Microorganisms capable of generating electricity in microbial fuel cells (MFCs) have gained increasing interest. Here fourteen exoelectrogenic bacterial strains were isolated from the anodic biofilm in an MFC before and after copper (Cu) shock load by Hungate roll-tube technique with solid ferric (III) oxide as an electron acceptor and acetate as an electron donor. Phylogenetic analysis of the 16S rRNA gene sequences revealed that they were all closely related to Enterobacter ludwigii DSM 16688^T within the Enterobacteriaceae family, although these isolated bacteria showed slightly different morphology before and after Cu shock load. Two representative strains R2B1 (before Cu shock load) and B4B2 (after Cu shock load) were chosen for further analysis. B4B2 is resistant to 200 mg L⁻¹ of Cu(II) while R2B1 is not, which indicated the potential selection of the Cu shock load. Raman analysis revealed that both R2B1 and B4B2 contained c-type cytochromes. Cyclic voltammetry measurements revealed that strain R2B1 had the capacity to transfer electrons to electrodes. The experimental results demonstrated that strain R2B1 was capable of utilizing a wide range of substrates, including Luria-Bertani (LB) broth, cellulose, acetate, citrate, glucose, sucrose, glycerol and lactose to generate electricity, with the highest current density of 440 mA·m⁻² generated from LB-fed MFC. Further experiments indicated that the bacterial cell density had potential correlation with the current density.     

Impedance spectroscopy as a tool for non‐intrusive detection of extracellular mediators in microbial fuel cells

Endogenously produced, diffusible redox mediators can act as electron shuttles for bacterial respiration. Accordingly, the mediators also serve a critical role in microbial fuel cells (MFCs), as they assist extracellular electron transfer from the bacteria to the anode serving as the intermediate electron sink. Electrochemical impedance spectroscopy (EIS) may be a valuable tool for evaluating the role of mediators in an operating MFC. EIS offers distinct advantages over some conventional analytical methods for the investigation of MFC systems because EIS can elucidate the electrochemical properties of various charge transfer processes in the bio‐energetic pathway. Preliminary investigations of Shewanella oneidensis DSP10‐based MFCs revealved that even low quantities of extracellular mediators significantly influence the impedance behavior of MFCs. EIS results also suggested that for the model MFC studied, electron transfer from the mediator to the anode may be up to 15 times faster than the electron transfer from bacteria to the mediator. When a simple carbonate membrane separated the anode and cathode chambers, the extracellular mediators were also detected at the cathode, indicating diffusion from the anode under open circuit conditions. The findings demonstrated that EIS can be used as a tool to indicate presence of extracellular redox mediators produced by microorganisms and their participation in extracellular electron shuttling. Biotechnol. Bioeng. 2009; 104: 882–891. © 2009 Wiley Periodicals, Inc.     

Electricity generation by anaerobic bacteria and anoxic sediments from hypersaline soda lakes

Anaerobic bacteria and anoxic sediments from soda lakes produced electricity in microbial fuel cells (MFCs). No electricity was generated in the absence of bacterial metabolism. Arsenate respiring bacteria isolated from moderately hypersaline Mono Lake (Bacillus selenitireducens), and salt-saturated Searles Lake, CA (strain SLAS-1) oxidized lactate using arsenate as the electron acceptor. However, these cultures grew equally well without added arsenate using the MFC anode as their electron acceptor, and in the process oxidized lactate more efficiently. The decrease in electricity generation by consumption of added alternative electron acceptors (i.e. arsenate) which competed with the anode for available electrons proved to be a useful indicator of microbial activity and hence life in the fuel cells. Shaken sediment slurries from these two lakes also generated electricity, with or without added lactate. Hydrogen added to sediment slurries was consumed but did not stimulate electricity production. Finally, electricity was generated in statically incubated “intact” sediment cores from these lakes. More power was produced in sediment from Mono Lake than from Searles Lake, however microbial fuel cells could detect low levels of metabolism operating under moderate and extreme conditions of salt stress.     

Use of Pseudomonas species producing phenazine-based metabolites in the anodes of microbial fuel cells to improve electricity generation

The rate of anodic electron transfer is one of the factors limiting the performance of microbial fuel cells (MFCs). It is known that phenazine-based metabolites produced by Pseudomonas species can function as electron shuttles for Pseudomonas themselves and also, in a syntrophic association, for Gram-positive bacteria. In this study, we have investigated whether phenazine-based metabolites and their producers could be used to improve the electricity generation of a MFC operated with a mixed culture. Both anodic supernatants obtained from MFCs operated with a Pseudomonas strain (P-PCA) producing phenazine-1-carboxylic acid (PCA) and those from MFCs operated with a strain (P-PCN) producing phenazine-1-carboxamide (PCN) exerted similarly positive effects on the electricity generation of a mixed culture. Replacing supernatants of MFCs operated with a mixed culture with supernatants of MFCs operated with P-PCN could double the currents generated. Purified PCA and purified PCN had similar effects. If the supernatant of an engineered strain overproducing PCN was used, the effect could be maintained over longer time courses, resulting in a 1.5-fold increase in the production of charge. Bioaugmentation of the mixed culture MFCs using slow release tubes containing P-PCN not only doubled the currents but also maintained the effect over longer periods. The results demonstrated the electron-shuttling effect of phenazine-based compounds produced by Pseudomonas species and their capacity to improve the performance of MFCs operated with mixed cultures. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Flexible and Stretchable Biobatteries: Monolithic Integration of Membrane‐Free Microbial Fuel Cells in a Single Textile Layer

The fabrication and performance of a flexible and stretchable microbial fuel cell (MFC) monolithically integrated into a single sheet of textile substrate are reported. The single‐layer textile MFC uses Pseudomonas aeruginosa (PAO1) as a biocatalyst to produce a maximum power of 6.4 µW cm^?2 and current density of 52 µA cm^?2, which are substantially higher than previous textile‐MFCs and are similar to other flexible paper‐based MFCs. The textile MFC demonstrates a stable performance with repeated stretching and twisting cycles. The membrane‐less single‐chamber configuration drastically simplifies the fabrication and improves the performance of the MFC. A conductive and hydrophilic anode in a 3D fabric microchamber maximizes bacterial electricity generation from a liquid environment and a silver oxide/silver solid‐state cathode reduces cathodic overpotential for fast catalytic reaction. A simple batch fabrication approach simultaneously constructs 35 individual devices, which will revolutionize the mass production of textile MFCs. This stretchable and twistable power device printed directly onto a single textile substrate can establish a standardized platform for textile‐based biobatteries and will be potentially integrated into wearable electronics in the future.     

Sustained generation of electricity by the spore-forming,Gram-positive, <Emphasis Type="Italic">Desulfitobacterium hafniense</Emphasis> strain DCB2

Desulfitobacterium hafniense strain DCB2 generates electricity in microbial fuel cells (MFCs) when humic acids or the humate analog anthraquinone-2,6-disulfonate (AQDS) is added as an electron-carrying mediator. When utilizing formate as fuel, the Gram-positive, spore-forming bacterium generated up to 400 mW/m² of cathode surface area in a single-chamber MFC with a platinum-containing air-fed cathode. Hydrogen, lactate, pyruvate, and ethanol supported electricity generation, but acetate, propionate, and butyrate did not. Scanning electron microscopy indicated that strain DCB2 colonized the surface of a current-generating anode but not of an unconnected electrode. The electricity was recovered fully within minutes after the exchange of the medium in the anode chamber and within a week after an exposure of a colonized anode to 90°C for 20 min. Of the six strains of Desulfitobacteria tested, all of which would reduce AQDS, only D. hafniense strain DCB2 continued to reduce AQDS and generate electricity for more than 24 h, indicating that reduction of the humate analog alone is insufficient to sustain electrode reduction.     

Electricity generation from cellulose by rumen microorganisms in microbial fuel cells

In microbial fuel cells (MFCs) bacteria generate electricity by mediating the oxidation of organic compounds and transferring the resulting electrons to an anode electrode. The objective of this study was to test the possibility of generating electricity with rumen microorganisms as biocatalysts and cellulose as the electron donor in two-compartment MFCs. The anode and cathode chambers were separated by a proton exchange membrane and graphite plates were used as electrodes. The medium in the anode chamber was inoculated with rumen microorganisms, and the catholyte in the cathode compartment was ferricyanide solution. Maximum power density reached 55 mW/m(2) (1.5 mA, 313 mV) with cellulose as the electron donor. Cellulose hydrolysis and electrode reduction were shown to support the production of current. The electrical current was sustained for over 2 months with periodic cellulose addition. Clarified rumen fluid and a soluble carbohydrate mixture, serving as the electron donors, could also sustain power output. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified 16S rRNA genes revealed that the microbial communities differed when different substrates were used in the MFCs. The anode-attached and the suspended consortia were shown to be different within the same MFC. Cloning and sequencing analysis of 16S rRNA genes indicated that the most predominant bacteria in the anode-attached consortia were related to Clostridium spp., while Comamonas spp. abounded in the suspended consortia. The results demonstrated that electricity can be generated from cellulose by exploiting rumen microorganisms as biocatalysts, but both technical and biological optimization is needed to maximize power output.     

Electrochemical analysis of Clostridium propionicum and its acrylic acid production in microbial fuel cells

Currently, acrylic acid is produced at a low yield by the resting cells of Clostridium propionicum with the supplement of extra electron acceptors. As an alternative way, acrylic acid production coupled with electricity generation was achieved by C. propionicum‐based microbial fuel cells (MFCs). Electricity was generated in the salt‐bridge MFCs with cysteine and resazurin in the anode chamber as mediators, and K₃Fe(CN)₆ as the cathode electron acceptor. Power generation was 21.78 mW/m² with an internal resistance of 9809 Ω. Cyclic voltammograms indicated the main mechanism of power production was the electron transfer facilitated by mediators in the system. In the salt‐bridge MFC system, 0.694 mM acrylic acid was produced together with electricity generation.     

Effect of enrichment procedures on performance and microbial diversity of microbial fuel cell for Congo red decolorization and electricity generation

The purpose of this study was to determine the effect of enrichment procedure on the performance and microbial diversity of an air-cathode microbial fuel cell (MFC) which was explored for simultaneous azo dye decolorization and electricity generation. Two different enrichment procedures in which glucose and Congo red were added into the MFCs sequentially (EP1) or simultaneously (EP2) were tested by operating parallel MFCs independently for more than 6 months. The power density, electrode potential, Congo red decolorization, biofilm morphology, and bacterial diversity of the MFCs under the two enrichment procedures were compared and investigated. The results showed that the enrichment procedures have a negligible effect on the dye decolorization, but significantly affected the electricity generation. More than 90% decolorization at dye concentration of 300 mg/L was achieved within 170 h for the two tested enrichment procedures. However, the MFC with EP2 achieved a maximum power density of 192 mW/m², which was 75% higher than that of the MFC with EP1 (110 mW/m²). The depressed surfaces of the bacteria in the MFC with EP1 indicated the allergic response caused by the subsequent addition of Congo red. 16S rRNA sequencing analysis demonstrated a phylogenetic diversity in the communities of the anode biofilm and showed clear differences between the anode-attached populations in the MFCs with a different enrichment procedure. This study suggests that the enrichment procedure is important for the MFC explored for simultaneous dye decolorization and electricity generation.     

Electrical energy production from forest detritus in a forested wetland using microbial fuel cells

Microbial fuel cell (MFC) technology has shown great potential for harvesting energy from waste organic materials. Here, we explored the potential of MFC‐based electricity generation from forest detritus, a large untapped biomass pool. Electricity generation from in situ MFCs and relevant environmental parameters (i.e., carbon sources and concentrations, temperature, water depth) in a seasonally flooded freshwater cypress‐tupelo wetland were monitored intensively for two flooding periods. Current outputs ranged from 0 to 1.27 mA (mean of 0.40 mA for flooding period) and were highly sensitive to environmental changes, showing seasonal and diel dependences. Excluding the influence of heavy storms, drought, or wetland icing, current output was highly temperature‐dependent dielly. Seasonally, current output gradually increased in the first 3–4 months (limited by temperature) and decreased slightly during the last 1–2 months (probably limited by carbon and nutrients) of both flooding periods. Litter extract of baldcypress (Taxodium distictum) with lower C/N ratio and aromatic content showed greater stable current outputs (0.57 mA) based on 50 mg l^?1 biological oxygen demand compared to extracts of water tupelo (Nyssa aquatica) and longleaf pine (Pinus palustris), suggesting that the current output of in situ MFCs could depend on the vegetation within a wetland. Our study highlights the potential application of MFC in generating green and sustainable electricity from forest biomass for powering remote sensors in wetland ecosystems.     

Use of acetate for enrichment of electrochemically active microorganisms and their 16S rDNA analyses

A fuel cell-type electrochemical device has been used to enrich microbes oxidizing acetate with concomitant electricity generation without using an electron mediator from activated sludge. The device generated a stable current of around 5 mA with complete oxidation of 5 mM acetate at the hydraulic retention time of 2.5 h after 4 weeks of enrichment. Over 70% of electrons available from acetate oxidation was recovered as current. Carbon monoxide or hydrogen did not influence acetate oxidation or current generation from the microbial fuel cell (MFC). Denaturing gradient gel electrophoresis showed that DNA extracted from the acetate-enriched MFC had different 16S rDNA patterns from those of sludge or glucose+glutamate-enriched MFCs. Nearly complete 16S rDNA sequence analyses showed that diverse bacteria were enriched in the MFC fed with acetate. Electron microscopic observations showed biofilm developed on the electrode, but not microbial clumps observed in MFCs fed with complex fuel such as glucose and wastewater from a corn-processing factory.     

Recent progress in electrodes for microbial fuel cells

The performance and cost of electrodes are the most important aspects in the design of microbial fuel cell (MFC) reactors. A wide range of electrode materials and configurations have been tested and developed in recent years to improve MFC performance and lower material cost. As well, anodic electrode surface modifications have been widely used to improve bacterial adhesion and electron transfer from bacteria to the electrode surface. In this paper, a review of recent advances in electrode material and a configuration of both the anode and cathode in MFCs are provided. The advantages and drawbacks of these electrodes, in terms of their conductivity, surface properties, biocompatibility, and cost are analyzed, and the modification methods for the anodic electrode are summarized. Finally, to achieve improvements and the commercial use of MFCs, the challenges and prospects of future electrode development are briefly discussed.     

Metabolic flux change in <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> L17 by anaerobic respiration in microbial fuel cell

The metabolic flux in microbial fuel cells (MFCs) is significantly different from conventional fermentation because the electrode in MFCs acts as a terminal electron acceptor. In this study, the difference in the carbon metabolism of Klebsiella pnuemoniae L17 (Kp L17) during growth in MFCs and conventional bioreactors was studied using glucose as the sole carbon and energy source. For metabolic flux analysis (MFA), the in silico metabolic flux model of Kp L17 was also constructed. The MFC bioreactor operated in oxidative mode, where electrons are removed by the anode electrode, generated a smaller quantity of reductive metabolites (e.g., lactate, 2,3-butanediol and ethanol) compared to the conventional fermentative bioreactor (non-MFC). Stoichiometric analysis indicated that the cellular metabolism in MFC had partially (or significantly) shifted to anaerobic respiration from fermentation, the former of which was similar to that often observed under micro-aerobic conditions. Electron balance analysis suggested that 30% of the electrons generated from glucose oxidation were extracted from the microbe and transferred to the electrode. These results highlight the potential use of MFCs in regulating the carbon metabolic flux in a bioprocess.     

An iTRAQ characterisation of the role of TolC during electron transfer from Shewanella oneidensis MR‐1

Anodophilic bacteria have the ability to generate electricity in microbial fuel cells (MFCs) by extracellular electron transfer to the anode. We investigated the anode‐specific responses of Shewanella oneidensis MR‐1, an exoelectroactive Gammaproteobacterium, using for the first time iTRAQ and 2D‐LC MS/MS driven membrane proteomics to compare protein abundances in S. oneidensis when generating power in MFCs, and growing in a continuous culture. The regulated dataset produced was enriched in membrane proteins. Proteins shown to be more abundant in anaerobic electroactive anodic cells included efflux pump TolC and an uncharacterised tetratricopeptide repeat (TPR) protein, whilst the TonB2 system and associated uncharacterised proteins such as TtpC2 and DUF3450 were more abundant in microaerobic planktonic cells. In order to validate the iTRAQ data, the functional role for TolC was examined using a δTolC knockout mutant of S. oneidensis. Possible roles for the uncharacterised proteins were identified using comparative bioinformatics. We demonstrate that employing an insoluble extracellular electron acceptor requires multiple proteins involved in cell surface properties. All MS and processed data are available via ProteomeXchange with identifier PXD004090.     

Layer-by-layer construction of graphene-based microbial fuel cell for improved power generation and methyl orange removal

Development of highly efficient anode is critical for enhancing the power output of microbial fuel cells (MFCs). The aim of this work is to investigate whether modification of carbon paper (CP) anode with graphene (GR) via layer-by-layer assembly technique is an effective approach to promote the electricity generation and methyl orange removal in MFCs. Using cyclic voltammetry and electrochemical impedance spectroscopy, the GR/CP electrode exhibited better electrochemical behavior. Scanning electron microscopy results revealed that the surface roughness of GR/CP increased, which was favorable for more bacteria to attach to the anode surface. The MFCs equipped with GR/CP anode achieved a stable maximum power density of 368 mW m^?2 under 1,000 Ω external resistance and a start time for the initial maximum voltage of 180 h, which were, respectively, 51 % higher and 31 % shorter than the corresponding values of the MFCs with blank anode. The anode and cathode polarization curves revealed negligible difference in cathode potentials but obviously difference in anode potentials, indicating that the GR-modified anode other than the cathode was responsible for the performance improvement of MFC. Meanwhile, compared with MFCs with blank anode, 11 % higher decolorization efficiency and 16 % higher the chemical oxygen demand removal rate were achieved in MFC with GR-modified anode during electricity generation. This study might provide an effective way to modify the anode for enhanced electricity generation and efficient removal of azo dye in MFCs.     

Bioelectricity enhancement via overexpression of quorum sensing system in Pseudomonas aeruginosa-inoculated microbial fuel cells

Low electron transfer efficiency from bacteria to electrodes remains one of the major bottlenecks that limit industrial applications of microbial fuel cells (MFCs). Elucidating biological mechanism of the electron transfer processes is of great help in improving the efficiency of MFCs. Here, we reported that Pseudomonas aeruginosa could use different electron shuttles in a MFC under different quorum sensing (QS) expression patterns. An electron shuttle (rather than phenazines) with a high mid-point potential of 0.20 V (vs. Ag/AgCl–KCl saturated electrode) was found to be the dominating shuttle in a wild-type P. aeruginosa strain. Strikingly, upon genetic overexpression of rhl QS system in this wild-type strain, the electron shuttle was substituted by phenazines (pyocyanin and phenazine-1-carboxylate, with a low mid-point potential of −0.17 V and −0.28 V, respectively), which directly resulted in an increase of about 1.6 times of the maximum current of the rhl overexpressed strain over the wild-type strain. Our result implied that manipulating electron transfer pathways to improve MFCs’ efficiency could be achieved by rewiring gene regulatory circuits, thus synthetic biology strategies would be adopted.     

Power production in MFCs inoculated with Shewanella oneidensis MR‐1 or mixed cultures

Power densities and oxidation–reduction potentials (ORPs) of MFCs containing a pure culture of Shewanella oneidensis MR‐1 were compared to mixed cultures (wastewater inoculum) in cube shaped, 1‐, 2‐, and 3‐bottle batch‐fed MFC reactor configurations. The reactor architecture influenced the relative power produced by the different inocula, with the mixed culture generating 68–480% more power than MR‐1 in each MFC configuration. The mixed culture produced the maximum power density of 858 ± 9 mW m^?2 in the cubic MFC, while MR‐1 produced 148 ± 20 mW m^?2. The higher power by the mixed culture was primarily a result of lower internal resistances than those produced by the pure culture. Power was a direct function of ohmic resistance for the mixed culture, but not for strain MR‐1. ORP of the anode compartment varied with reactor configuration and inoculum, and it was always negative during maximum power production but it did not vary in proportion to power output. The ORP varied primarily at the end of the cycle when substrate was depleted, with a change from a reductive environment during maximum power production (approximately ?175 mV for mixed and approximately ?210 mV for MR‐1 in cubic MFCs), to an oxidative environment at the end of the batch cycle (～250 mV for mixed and ～300 mV for MR‐1). Mixed cultures produced more power than MR‐1 MFCs even though their redox potential was less negative. These results demonstrate that differences between power densities produced by pure and mixed cultures depend on the MFC architecture. Biotechnol. Bioeng. 2010; 105: 489–498. © 2009 Wiley Periodicals, Inc.