Potassium channel mediates electroactive biofilm formation via recruiting planktonic Geobacter cells.

Potassium (K+)-channel-based electrical signaling can coordinate microbial actions at a distance that provides an evolutionary advantage to cell communities. Electroactive cells are usually cultured surrounded by an electric field which provided stronger electrical signaling than the K+-mediated electrical signaling. Whether the K+ signaling also plays a role in coordinating the behavior of electroactive microorganisms has not been accurately demonstrated. Thus, we constructed a K+-channel-deficient strain ΔgsuK of Geobacter sulfurreducens to directly investigate roles of K+ signaling in electroactive biofilm formation for the first time. The ΔgsuK strain exhibited significantly inferior biofilm formation (i.e., biomass, thickness and component) and consequently showed weaker electrical performance (i.e., start-up time, current output, electrochemical catalytic behavior and charge transfer resistance) than the wild-type strain. Individual electric generation capacity and the expression of genes involved in biofilm formation and electrical performance in the single cell did not significantly change with the deletion of gsuK, indicating that K+ signaling indeed influenced the recruiting behavior of planktonic cell but not the functioning of the single cell related to biofilm formation or electric generation. This study is intended to provide an in-depth understanding of electroactive biofilm formation and serve as a basis for optimizing its electrical performance via strengthening the recruitment behavior.

Nutrient limitation regulates the properties of extracellular electron transfer and hydraulic shear resistance of electroactive biofilm.

Understanding the roles of nutrient restriction in extracellular electron transfer (EET) and stability of mixed electroactive biofilm is essential in pollutant degradation and bioenergy production. However, the relevant studies are still limited so far. Herein, the effect of nutrient restriction on the EET pathways and stability of mixed electroactive biofilm was explored. It was found that the electroactive Pseudomonas and Geobacter genera were selectively enriched in the biofilms cultured under total nutrient and P-constrained conditions, and two EET pathways including direct and indirect were found, while Rhodopseudomonas genus was enriched in the N-constrained biofilm, which only had the direct EET pathway. Moreover, multiple analyses including 2D confocal Raman spectra revealed that P-constrained biofilm was rich in extracellular polymeric substances (EPS) especially for polysaccharide, presented a dense and uniform layered distribution, and had better stability than N-constrained biofilm with lower EPS and biofilm with heterostructures cultured under total nutrient conditions.

Multistep Signaling in Nature: A Close-Up of Geobacter Chemotaxis Sensing.

Environmental changes trigger the continuous adaptation of bacteria to ensure their survival. This is possible through a variety of signal transduction pathways involving chemoreceptors known as methyl-accepting chemotaxis proteins (MCP) that allow the microorganisms to redirect their mobility towards favorable environments. MCP are two-component regulatory (or signal transduction) systems (TCS) formed by a sensor and a response regulator domain. These domains synchronize transient protein phosphorylation and dephosphorylation events to convert the stimuli into an appropriate cellular response. In this review, the variability of TCS domains and the most common signaling mechanisms are highlighted. This is followed by the description of the overall cellular topology, classification and mechanisms of MCP. Finally, the structural and functional properties of a new family of MCP found in Geobacter sulfurreducens are revisited. This bacterium has a diverse repertoire of chemosensory systems, which represents a striking example of a survival mechanism in challenging environments. Two G. sulfurreducens MCP-GSU0582 and GSU0935-are members of a new family of chemotaxis sensor proteins containing a periplasmic PAS-like sensor domain with a c-type heme. Interestingly, the cellular location of this domain opens new routes to the understanding of the redox potential sensing signaling transduction pathways.

Adaptive Synthesis of a Rough Lipopolysaccharide in Geobacter sulfurreducens for Metal Reduction and Detoxification.

The ability of some metal-reducing bacteria to produce a rough (no O-antigen) lipopolysaccharide (LPS) could facilitate surface interactions with minerals and metal reduction. Consistent with this, the laboratory model metal reducer Geobacter sulfurreducens PCA produced two rough LPS isoforms (with or without a terminal methyl-quinovosamine sugar) when growing with the soluble electron acceptor fumarate but expressed only the shorter and more hydrophilic variant when reducing iron oxides. We reconstructed from genomic data conserved pathways for the synthesis of the rough LPS and generated heptosyltransferase mutants with partial (ΔrfaQ) or complete (ΔrfaC) truncations in the core oligosaccharide. The stepwise removal of the LPS core sugars reduced the hydrophilicity of the cell and increased outer membrane vesiculation. These changes in surface charge and remodeling did not substantially impact planktonic growth but disrupted the developmental stages and structure of electroactive biofilms. Furthermore, the mutants assembled conductive pili for extracellular mineralization of the toxic uranyl cation but were unable to prevent permeation and mineralization of the radionuclide in the cell envelope. Hence, not only does the rough LPS promote cell-cell and cell-mineral interactions critical to biofilm formation and metal respiration but it also functions as a permeability barrier to toxic metal cations. In doing so, the rough LPS maximizes the extracellular reduction of soluble and insoluble metals and preserves cell envelope functions critical to the environmental survival of Geobacter bacteria in metal-rich environments and their performance in bioremediation and bioenergy applications. IMPORTANCE Some metal-reducing bacteria produce an LPS without the repeating sugars (O-antigen) that decorate the surface of most Gram-negative bacteria, but the biological significance of this adaptive feature was not previously investigated. Using the model representative Geobacter sulfurreducens strain PCA and mutants carrying stepwise truncations in the LPS core sugars, we demonstrate the importance of the rough LPS in the control of cell surface chemistry during the respiration of iron minerals and the formation of electroactive biofilms. Importantly, we describe hitherto overlooked roles for the rough LPS in metal sequestration and outer membrane vesiculation that are critical for the extracellular reduction and detoxification of toxic metals and radionuclides. These results are of interest for the optimization of bioremediation schemes and electricity-harvesting platforms using these bacteria.

Evidence of a Streamlined Extracellular Electron Transfer Pathway from Biofilm Structure, Metabolic Stratification, and Long-Range Electron Transfer Parameters.

A strain of Geobacter sulfurreducens, an organism capable of respiring solid extracellular substrates, lacking four of five outer membrane cytochrome complexes (extABCD+ strain) grows faster and produces greater current density than the wild type grown under identical conditions. To understand cellular and biofilm modifications in the extABCD+ strain responsible for this increased performance, biofilms grown using electrodes as terminal electron acceptors were sectioned and imaged using electron microscopy to determine changes in thickness and cell density, while parallel biofilms incubated in the presence of nitrogen and carbon isotopes were analyzed using NanoSIMS (nanoscale secondary ion mass spectrometry) to quantify and localize anabolic activity. Long-distance electron transfer parameters were measured for wild-type and extABCD+ biofilms spanning 5-µm gaps. Our results reveal that extABCD+ biofilms achieved higher current densities through the additive effects of denser cell packing close to the electrode (based on electron microscopy), combined with higher metabolic rates per cell compared to the wild type (based on increased rates of 15N incorporation). We also observed an increased rate of electron transfer through extABCD+ versus wild-type biofilms, suggesting that denser biofilms resulting from the deletion of unnecessary multiheme cytochromes streamline electron transfer to electrodes. The combination of imaging, physiological, and electrochemical data confirms that engineered electrogenic bacteria are capable of producing more current per cell and, in combination with higher biofilm density and electron diffusion rates, can produce a higher final current density than the wild type. IMPORTANCE Current-producing biofilms in microbial electrochemical systems could potentially sustain technologies ranging from wastewater treatment to bioproduction of electricity if the maximum current produced could be increased and current production start-up times after inoculation could be reduced. Enhancing the current output of microbial electrochemical systems has been mostly approached by engineering physical components of reactors and electrodes. Here, we show that biofilms formed by a Geobacter sulfurreducens strain producing ∼1.4× higher current than the wild type results from a combination of denser cell packing and higher anabolic activity, enabled by an increased rate of electron diffusion through the biofilms. Our results confirm that it is possible to engineer electrode-specific G. sulfurreducens strains with both faster growth on electrodes and streamlined electron transfer pathways for enhanced current production.

Engineering Cooperation in an Anaerobic Coculture.

Over the past century, microbiologists have studied organisms in pure culture, yet it is becoming increasingly apparent that the majority of biological processes rely on multispecies cooperation and interaction. While little is known about how such interactions permit cooperation, even less is known about how cooperation arises. To study the emergence of cooperation in the laboratory, we constructed both a commensal community and an obligate mutualism using the previously noninteracting bacteria Shewanella oneidensis and Geobacter sulfurreducens Incorporation of a glycerol utilization plasmid (pGUT2) enabled S. oneidensis to metabolize glycerol and produce acetate as a carbon source for G. sulfurreducens, establishing a cross-feeding, commensal coculture. In the commensal coculture, both species coupled oxidative metabolism to the respiration of fumarate as the terminal electron acceptor. Deletion of the gene encoding fumarate reductase in the S. oneidensis/pGUT2 strain shifted the coculture with G. sulfurreducens to an obligate mutualism where neither species could grow in the absence of the other. A shift in metabolic strategy from glycerol catabolism to malate metabolism was associated with obligate coculture growth. Further targeted deletions in malate uptake and acetate generation pathways in S. oneidensis significantly inhibited coculture growth with G. sulfurreducens The engineered coculture between S. oneidensis and G. sulfurreducens provides a model laboratory system to study the emergence of cooperation in bacterial communities, and the shift in metabolic strategy observed in the obligate coculture highlights the importance of genetic change in shaping microbial interactions in the environment.IMPORTANCE Microbes seldom live alone in the environment, yet this scenario is approximated in the vast majority of pure-culture laboratory experiments. Here, we develop an anaerobic coculture system to begin understanding microbial physiology in a more complex setting but also to determine how anaerobic microbial communities can form. Using synthetic biology, we generated a coculture system where the facultative anaerobe Shewanella oneidensis consumes glycerol and provides acetate to the strict anaerobe Geobacter sulfurreducens In the commensal system, growth of G. sulfurreducens is dependent on the presence of S. oneidensis To generate an obligate coculture, where each organism requires the other, we eliminated the ability of S. oneidensis to respire fumarate. An unexpected shift in metabolic strategy from glycerol catabolism to malate metabolism was observed in the obligate coculture. Our work highlights how metabolic landscapes can be expanded in multispecies communities and provides a system to evaluate the evolution of cooperation under anaerobic conditions.

The electrode potential determines the yield coefficients of early-stage Geobacter sulfurreducens biofilm anodes.

Geobacter sulfurreducens is the model for electroactive microorganisms (EAM). EAM can use solid state terminal electron acceptors (TEA) including anodes via extracellular electron transfer (EET). Yield coefficients relate the produced cell number or biomass to the oxidized substrate or the reduced TEA. These data are not yet sufficiently available for EAM growing at anodes. Thus, this study provides information about kinetics as well as yield coefficients of early-stage G. sulfurreducens biofilms using anodes as TEA at the potentials of -200 mV, 0 mV and +200 mV (vs. Ag/AgCl sat. KCl). The selected microorganism was therefore cultivated in single and double chamber batch reactors on graphite or AuPd anodes. Interestingly, whereas the lag time and maximum current density within 12 days of growth differed, the anode potential does not influence the coulombic efficiency and the formal potential of the EET, which remains constant for all the experiments at ~ -300 to -350 mV. We demonstrated for the first time that the anode potential has a strong influence on single cell yield coefficients which ranged from 2.69 × 1012 cells mole--1 at -200 mV and 1.48 × 1012 cells mole--1 at 0 mV to 2.58 × 1011 cells mole--1 at +200 mV in single chamber reactors and from 1.15 × 1012 cells mole--1 at -200 mV to 8.98× 1011 cells mole--1 at 0 mV in double chamber reactors. This data can be useful for optimization and scaling-up of primary microbial electrochemical technologies.

Electric field stimulates production of highly conductive microbial OmcZ nanowires.

Multifunctional living materials are attractive due to their powerful ability to self-repair and replicate. However, most natural materials lack electronic functionality. Here we show that an electric field, applied to electricity-producing Geobacter sulfurreducens biofilms, stimulates production of cytochrome OmcZ nanowires with 1,000-fold higher conductivity (30 S cm-1) and threefold higher stiffness (1.5 GPa) than the cytochrome OmcS nanowires that are important in natural environments. Using chemical imaging-based multimodal nanospectroscopy, we correlate protein structure with function and observe pH-induced conformational switching to ß-sheets in individual nanowires, which increases their stiffness and conductivity by 100-fold due to enhanced π-stacking of heme groups; this was further confirmed by computational modeling and bulk spectroscopic studies. These nanowires can transduce mechanical and chemical stimuli into electrical signals to perform sensing, synthesis and energy production. These findings of biologically produced, highly conductive protein nanowires may help to guide the development of seamless, bidirectional interfaces between biological and electronic systems.

A New Approach for Evaluating Electron Transfer Dynamics by Using In Situ Resonance Raman Microscopy and Chronoamperometry in Conjunction with a Dynamic Model.

Geobacter sulfurreducens is a good candidate as a chassis organism due to its ability to form thick, conductive biofilms, enabling long-distance extracellular electron transfer (EET). Due to the complexity of EET pathways in G. sulfurreducens, a dynamic approach is required to study genetically modified EET rates in the biofilm. By coupling online resonance Raman microscopy with chronoamperometry, we were able to observe the dynamic discharge response in the biofilm's cytochromes to an increase in anode voltage. Measuring the heme redox state alongside the current allows for the fitting of a dynamic model using the current response and a subsequent validation of the model via the value of a reduced cytochrome c Raman peak. The modeled reduced cytochromes closely fitted the Raman response data from the G. sulfurreducens wild-type strain, showing the oxidation of heme groups in cytochromes until a new steady state was achieved. Furthermore, the use of a dynamic model also allows for the calculation of internal rates, such as acetate and NADH consumption rates. The Raman response of a mutant lacking OmcS showed a higher initial oxidation rate than predicted, followed by an almost linear decrease of the reduced mediators. The increased initial rate could be attributed to an increase in biofilm conductivity, previously observed in biofilms lacking OmcS. One explanation for this is that OmcS acts as a conduit between cytochromes; therefore, deleting the gene restricts the rate of electron transfer to the extracellular matrix. This could, however, be modeled assuming a linear oxidation rate of intercellular mediators.IMPORTANCE Bioelectrochemical systems can fill a vast array of application niches, due to the control of redox reactions that it offers. Although native microorganisms are preferred for applications such as bioremediation, more control is required for applications such as biosensors or biocomputing. The development of a chassis organism, in which the EET is well defined and readily controllable, is therefore essential. The combined approach in this work offers a unique way of monitoring and describing the reaction kinetics of a G. sulfurreducens biofilm, as well as offering a dynamic model that can be used in conjunction with applications such as biosensors.

In Vivo Molecular Insights into Syntrophic Geobacter Aggregates.

Direct interspecies electron transfer (DIET) has been considered as a novel and highly efficient strategy in both natural anaerobic environments and artificial microbial fuel cells. A syntrophic model consisting of Geobacter metallireducens and Geobacter sulfurreducens was studied in this work. We conducted in vivo molecular mapping of the outer surface of the syntrophic community as the interface of nutrients and energy exchange. System for Analysis at the Liquid Vacuum Interface combined with time-of-flight secondary ion mass spectrometry was employed to capture the molecular distribution of syntrophic Geobacter communities in the living and hydrated state. Principal component analysis with selected peaks revealed that syntrophic Geobacter aggregates were well differentiated from other control samples, including syntrophic planktonic cells, pure cultured planktonic cells, and single population biofilms. Our in vivo imaging indicated that a unique molecular surface was formed. Specifically, aromatic amino acids, phosphatidylethanolamine components, and large water clusters were identified as key components that favored the DIET of syntrophic Geobacter aggregates. Moreover, the molecular changes in depths of the Geobacter aggregates were captured using dynamic depth profiling. Our findings shed new light on the interface components supporting electron transfer in syntrophic communities based on in vivo molecular imaging.

Determining incremental coulombic efficiency and physiological parameters of early stage Geobacter spp. enrichment biofilms.

Geobacter spp. enrichment biofilms were cultivated in batch using one-chamber and two-chamber bioelectrochemical reactors. Time-resolved substrate quantification was performed to derive physiological parameters as well as incremental coulombic efficiency (i.e., coulombic efficiency during one batch cycle, here every 6h) during early stage biofilm development. The results of one-chamber reactors revealed an intermediate acetate increase putatively due to the presence of acetogens. Total coulombic efficiencies of two-chamber reactors were considerable lower (19.6±8.3% and 49.3±13.2% for 1st and 2nd batch cycle, respectively) compared to usually reported values of mature Geobacter spp. enrichment biofilms presumably reflecting energetic requirements for biomass production (i.e., cells and extracellular polymeric substances) during early stages of biofilm development. The incremental coulombic efficiency exhibits considerable changes during batch cycles indicating shifts between phases of maximizing metabolic rates and maximizing biomass yield. Analysis based on Michaelis-Menten kinetics yielded maximum substrate uptake rates (vmax,Ac, vmax,I) and half-saturation concentration coefficients (KM,Ac,KM,I) based on acetate uptake or current production, respectively. The latter is usually reported in literature but neglects energy demands for biofilm growth and maintenance as well as acetate and electron storage. From 1st to 2nd batch cycle, vmax,Ac and KM,Ac, decreased from 0.0042-0.0051 mmol Ac- h-1 cm-2 to 0.0031-0.0037 mmol Ac- h-1 cm-2 and 1.02-2.61 mM Ac- to 0.28-0.42 mM Ac-, respectively. Furthermore, differences between KM,Ac/KM,I and vmax,Ac/vmax,I were observed providing insights into the physiology of Geobacter spp. enrichment biofilms. Notably, KM,I considerably scattered while vmax,Ac/vmax,I and KM,Ac remained rather stable indicating that acetate transport within biofilm only marginally affects reaction rates. The observed data variation mandates the requirement of a more detailed analysis with an improved experimental system, e.g., using flow conditions and a comparison with Geobacter spp. pure cultures.

Stratified microbial structure and activity within anode biofilm during electrochemically assisted brewery wastewater treatment.

In a bioelectrochemical system (BES), microbial community of anode biofilm is crucial to BES performance. In this study, the stratified pattern of community structure and activity of an anode-respiring biofilm in a BES fueled with brewery wastewater was investigated over time. The anode biofilm exhibited a superior performance in the removal of ethanol to that of an open-circuit system. The electrical current density reached a high level of 0.55mA/cm2 with a Coulombic efficiency of 71.4%, but decreased to 0.18mA/cm2 in the late stage of operation. A mature biofilm developed a more active outer layer covering a less active inner core, although the activities of the outer and inner layers of biofilm were similar in the early stage. More Geobacter spp., typical exoelectrogens, were enriched in the outer layer than in the inner layer of biofilm in the early stage, while more Geobacter spp. were distributed in the inner layer than in the outer layer in the late stage. The inactive and Geobacter-occupied inner layer of biofilm might be responsible for the decreased electricity generation from wastewater in the late stage of operation. This study provides better understanding of the effect of anode biofilm structure on BES performance.

Disparity of Cytochrome Utilization in Anodic and Cathodic Extracellular Electron Transfer Pathways of Geobacter sulfurreducens Biofilms.

Extracellular electron transfer (EET) in microorganisms is prevalent in nature and has been utilized in functional bioelectrochemical systems. EET of Geobacter sulfurreducens has been extensively studied and has been revealed to be facilitated through c-type cytochromes, which mediate charge between the electrode and G. sulfurreducens in anodic mode. However, the EET pathway of cathodic conversion of fumarate to succinate is still under debate. Here, we apply a variety of analytical methods, including electrochemistry, UV-vis absorption and resonance Raman spectroscopy, quartz crystal microbalance with dissipation, and electron microscopy, to understand the involvement of cytochromes and other possible electron-mediating species in the switching between anodic and cathodic reaction modes. By switching the applied bias for a G. sulfurreducens biofilm coupled to investigating the quantity and function of cytochromes, as well as the emergence of Fe-containing particles on the cell membrane, we provide evidence of a diminished role of cytochromes in cathodic EET. This work sheds light on the mechanisms of G. sulfurreducens biofilm growth and suggests the possible existence of a nonheme, iron-involving EET process in cathodic mode.

Molecular evidence for the adaptive evolution of Geobacter sulfurreducens to perform dissimilatory iron reduction in natural environments.

The electrically conductive pili (e-pili) of Geobacter species enable extracellular electron transfer to insoluble metallic minerals, electrodes and other microbial species, which confers biogeochemical significance and global prevalence on Geobacter in diverse anaerobic environments. E-pili are constructed by truncated PilA which is considered to have evolved from full-length pilin by gene fission under positive evolutionary selection. However, this hypothesis is based on phylogenetic analysis and has not yet been experimentally confirmed. Here, we reconstructed an ancestral strain of G. sulfurreducens (designated COMB) carrying full-length PilA by combining genes GSU1496 and GSU1497. The results demonstrated that strain COMB expressed and assembled the full-length fused PilA and exhibited an outer membrane c-type cytochrome profile similar to the wild-type strain. Surprisingly, the generated COMB-pili were also conductive, indicating the evolution of truncated PilA did not occur for conductivity. Moreover, strain COMB minimally reduced Fe(III) oxides but maintained its ability to respire electrodes, demonstrating the truncation of pilin enables iron respiration. This study provides the first experimental evidence that the truncation of pilin in Geobacter species confers adaption to Fe(III)-mineral-mediated selective pressures, and suggests an evolutionary event during which the separation of the GSU1497 gene helped Geobacter survive and thrive in natural environments.

Flagella act as Geobacter biofilm scaffolds to stabilize biofilm and facilitate extracellular electron transfer.

Flagella are widely expressed in electroactive biofilms; however, their actual role is unknown. To understand the role of flagella, two Geobacter sulfurreducens strains (KN400 and PCA, with and without flagella, respectively) were selected. We restored flagellum expression in trans in strain PCA and prevented flagellum expression in strain KN400. Electrochemical results showed that flagellum restoration in strain PCA promoted current generation, while flagellum deletion in strain KN400 impaired current production. However, the expression of conductive pili and outer surface c-type cytochromes was not affected. Further microscopic analyses demonstrated that flagella promoted the formation of thicker biofilms and served as biofilm matrix scaffolds to accommodate more extracellular cytochromes with an orderly arrangement, which increased the electron diffusion rate within the biofilm. Our findings reveal an unprecedented structural role for flagella in stabilizing electroactive biofilms and highlight the importance of cytochromes in electron transfer across biofilms, which will deepen our understanding of biofilm conductivity.

Environmental-Stress-Induced Increased Softness of Electroactive Biofilms, Determined with a Torsional Quartz Crystal Microbalance.

Electroactive biofilms are intensely studied not only for energy conversion and electrosynthesis, but also as sensing systems. The electrical current produced by the layer is largely proportional to the rate of metabolism and therefore decreases when the biofilm experiences adverse environmental conditions. Acoustic measurements may complement this approach. The layer's softness can be inferred from shifts of resonance frequency and resonance bandwidth of a quartz crystal microbalance (QCM) contacting these layers. The layer's softness responds to the environment. Both negative potentials of the electrode (the equivalent of "suffocation") and lack of nutrient supply (the equivalent of "starvation") were studied. For comprehensive analysis, torsional resonators operating on three different modes of vibration are suited best. Such data can be fitted with a viscoelastic model, leading to a quantitative estimate of the shear modulus. On a more empirical level, one might also use the ratio of the shift in bandwidth to the negative shift in frequency as an indicator of stress. For ease of operation, one might even replace the torsional resonators with thickness-shear resonators.

Peak selection matters in principal component analysis: A case study of syntrophic microbes.

In situ liquid time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a powerful technique to study surface characterization of living biofilms in hydrated conditions. However, ToF-SIMS data analysis is still a great challenge in complicated bacterial biofilms, because many interference peaks from the medium may result in inaccurate interpretation. In this study, two syntrophic Geobacter populations are investigated using in situ liquid ToF-SIMS to reveal the biofilm surface changes between them due to direct interspecies electron transfer. By comparing spectral principal component analysis (PCA) results of all peaks and selected peaks, the authors find that spectral peak overlay is an effective strategy to reduce the matrix effect in handling complex ToF-SIMS data. Additionally, the spectral PCA results of high intensity and high resolution data obtained from liquid ToF-SIMS are compared. Selected peaks, amino acid peaks, and water cluster peaks spectral PCA produce nice separation among samples in both high intensity and high resolution data sets. However, the high resolution data show better separation between coculture planktonic and coculture aggregates, confirming that the higher mass accuracy is useful in the analysis of microbial samples. In conclusion, the results show that peak selection is critical for acquiring effective microbial information and interpretation of syntrophic Geobacter using spectral data from in situ liquid ToF-SIMS.

Building electrode with three-dimensional macroporous interface from biocompatible polypyrrole and conductive graphene nanosheets to achieve highly efficient microbial electrocatalysis.

Bioelectrochemical systems (BESs) possess a great potential for simultaneous wastewater treatment and energy recovery. Rational construction of electrode materials could significantly improve the BESs performance. Three-dimensional macroporous electrode interface with high conductivity is highly desirable but challenging. In this work, we report a hierarchically nanostructured reduced graphene oxide nanosheets-polypyrrole (rGO@PPy) electrode via one-step electrodeposition technique. The prepared electrode was comprehensively studied by scanning/transmission electron microscopy, Raman spectroscopy, X-ray diffraction and electrochemical measurements, which showed that the rGO@PPy possessed a three-dimensional macroporous interconnecting scaffold with superior conductivity. The rGO@PPy electrode was utilized in Geobacter sulfurreducens inoculated BESs, and the maximum current density was 4.10 ±â€¯0.02 mA cm-2, which is 8-fold higher than that of a rGO electrode (0.51 ±â€¯0.03 mA cm-2), and is among the best performance reported for two-dimensional electrodes. The improved performance is ascribed to ultrahigh biomass concentration induced by "best match scale" between rGO@PPy and microbes, excellent extracellular electron transfer, as well as enhanced microbial affinity through the adequate exposure of biocompatible PPy layers. This work demonstrated a synergistic effect between rGO and PPy for the BESs performance improvement, and provided a new insight to design and fabricate a high-performance bioelectrode.

The electrifying physiology of Geobacter bacteria, 30 years on.

The family Geobacteraceae, with its only valid genus Geobacter, comprises deltaproteobacteria ubiquitous in soil, sediments, and subsurface environments where metal reduction is an active process. Research for almost three decades has provided novel insights into environmental processes and biogeochemical reactions not previously known to be carried out by microorganisms. At the heart of the environmental roles played by Geobacter bacteria is their ability to integrate redox pathways and regulatory checkpoints that maximize growth efficiency with electron donors derived from the decomposition of organic matter while respiring metal oxides, particularly the often abundant oxides of ferric iron. This metabolic specialization is complemented by versatile metabolic reactions, respiratory chains, and sensory networks that allow specific members to adaptively respond to environmental cues to integrate organic and inorganic contaminants in their oxidative and reductive metabolism, respectively. Thus, Geobacteraceae are important members of the microbial communities that degrade hydrocarbon contaminants under iron-reducing conditions and that contribute, directly or indirectly, to the reduction of radionuclides, toxic metals, and oxidized species of nitrogen. Their ability to produce conductive pili as nanowires for discharging respiratory electrons to solid-phase electron acceptors and radionuclides, or for wiring cells in current-harvesting biofilms highlights the unique physiological traits that make these organisms attractive biological platforms for bioremediation, bioenergy, and bioelectronics application. Here we review some of the most notable physiological features described in Geobacter species since the first model representatives were recovered in pure culture. We provide a historical account of the environmental research that has set the foundation for numerous physiological studies and the laboratory tools that had provided novel insights into the role of Geobacter in the functioning of microbial communities from pristine and contaminated environments. We pay particular attention to latest research, both basic and applied, that has served to expand the field into new directions and to advance interdisciplinary knowledge. The electrifying physiology of Geobacter, it seems, is alive and well 30 years on.