Extracellular Electron Transfer Research Articles

Characterizing Mediated Extracellular Electron Transfer in Lactic Acid Bacteria with a Three-Electrode, Two-Chamber Bioelectrochemical System.

Many bacteria perform extracellular electron transfer (EET), whereby electrons are transferred from the cell to an extracellular terminal electron acceptor. This electron acceptor can be an electrode and electrons can be delivered indirectly via a redox-active mediator molecule. Here, we present a protocol to study mediated EET in Lactiplantibacillus plantarum, a probiotic lactic acid bacterium widely used in the food industry, using a bioelectrochemical system. We detail how to assemble a three-electrode, two-chambered bioelectrochemical system and provide guidance on characterizing EET in the presence of a soluble mediator using chronoamperometry and cyclic voltammetry techniques. We use representative data from 1,4-dihydroxy-2-naphthoic acid (DHNA)-mediated EET experiments with L. plantarum to demonstrate data analysis and interpretation. The techniques described in this protocol can open new opportunities for electro-fermentation and bioelectrocatalysis. Recent applications of this electrochemical technique with L. plantarum demonstrated an acceleration of metabolic flux towards producing fermentation end-products, which are critical flavor components in food fermentation. As such, this system has the potential to be further developed to alter flavors in food production or produce valuable chemicals.

Engineering Shewanella oneidensis-Carbon Felt Biohybrid Electrode Decorated with Bacterial Cellulose Aerogel-Electropolymerized Anthraquinone to Boost Energy and Chemicals Production.

Interfacial electron transfer between electroactive microorganisms (EAMs) and electrodes underlies a wide range of bio-electrochemical systems with diverse applications. However, the electron transfer rate at the biotic-electrode interface remains low due to high transmembrane and cell-electrode interfacial electron transfer resistance. Herein, a modular engineering strategy is adopted to construct a Shewanella oneidensis-carbon felt biohybrid electrode decorated with bacterial cellulose aerogel-electropolymerized anthraquinone to boost cell-electrode interfacial electron transfer. First, a heterologous riboflavin synthesis and secretion pathway is constructed to increase flavin-mediated transmembrane electron transfer. Second, outer membrane c-Cyts OmcF is screened and optimized via protein engineering strategy to accelerate contacted-based transmembrane electron transfer. Third, a S. oneidensis-carbon felt biohybrid electrode decorated with bacterial cellulose aerogel and electropolymerized anthraquinone is constructed to boost the interfacial electron transfer. As a result, the internal resistance decreased to 42 Ω, 480.8-fold lower than that of the wild-type (WT) S. oneidensis MR-1. The maximum power density reached 4286.6 ± 202.1mWm-2, 72.8-fold higher than that of WT. Lastly, the engineered biohybrid electrode exhibited superior abilities for bioelectricity harvest, Cr6+ reduction, and CO2 reduction. This study showed that enhancing transmembrane and cell-electrode interfacial electron transfer is a promising way to increase the extracellular electron transfer of EAMs.

Enhanced denitrification performance and extracellular electron transfer for eletrotrophic bio-cathode mediated by anthraquinone-2, 6-disulfonate (AQDS) at low temperature

The slow electron transfer rate and low temperature are the bottleneck in biological denitrification process, and quinone electron shuttle (e.g. AQDS) has been confirmed as a viable approach to improve the efficiency of biological denitrification by accelerating electron transport. In this study, we employed the eletrotrophic bio-cathode with cold-tolerant to explore the mechanisms of extracellular electron transfer (EET) mediated by AQDS in simultaneous nitrification and denitrification (SND) for the first time. The results indicated that adding 0.5 mmol/L AQDS increased NH4+-N removal rate and SND efficiency by 18.1% and 17.9%, respectively, and the optimal hydraulic retention time (HRT) was shortened from 5 d to 3 d. The potential mechanism was that AQDS resulted in a 1.47-fold increase in the relative abundance of cathodic electroactive microorganisms, and the relative abundance of functional genes such as cytochrome c, flavin, cold acclimation and translocator increased by about 1.15–300.42%, which accelerated the transfer of extracellular electron and significantly improved the energy metabolism efficiency. In addition, AQDS may act as an electron shuttle to accelerate the electronic transition in long-distance electron transfer. In summary, this study expands our understanding of the role of AQDS in the SND process by bio-cathode under low temperature and provides an insight into the EET of eletrotrophic microorganisms with cold-tolerant.

Shortening electron transfer distance to enhance chemicals and electric energy production in Escherichia coli

Intracellular electron abundance affects the efficiency of biosynthetic pathways in microbial cell factories. However, regulating electron transfer efficiency to change intracellular electron abundance is challenging. Herein, we aimed to improve the electron transfer efficiency by shortening electron transfer distance using two systems: an extracellular electron transfer system (EET) and a light-driven intracellular electron regeneration system (ERS). The EET involved reconstructing an efficient electron transfer pathway (AEC4) and using periplasmic and biofilm engineering to improve its efficiency, whereas the ERS involved developing a biologically self-synthetic photosynthetic biohybrid system and using protein cage engineering to enhance its efficiency. As a proof of concept, the EET was implemented using a dynamic AND logic gate to increase the α-ketoglutarate titer to 30.4 g/L and the power density to 561.5 mW/m2 in microbial fuel cells. In a 5 L fermenter, the titer and productivity of α-ketoglutarate reached 67.7 g/L and 1.88 g/L/h, respectively. The ERS was manipulated using a chemo-optogenetic AND logic gate to increase the adipate titer to 43.1 g/L in a 5 L fermenter. Overall, our results provide a potential way to shorten electron transfer distance for the efficient production of valuable chemicals and electric energy.

A single-microbe living bioelectronic sensor for intracellular amperometric analysis

Subcellularly amperometric analysis in situ is crucial for understanding intracellular redox biochemistry and subcellular heterogeneity. Unfortunately, the ultra-small size and complex microenvironment inside the cell pose a great challenge to achieve this goal. To address the challenge, a minimized living microbial sensor has been fabricated in this work for amperometric analysis. Here, by fabricating the dimidiate microelectrode as the working electrode, while fitting a living electroactive bacterium (EAB) as the transducer, outward extracellular electron transfer (EET) of the sensory EAB is correlated with the concentration of lactic acid, which is electrochemically recorded and thus displays an electrical signal output for detection. In specific, the S. oneidensis modified dimidiate microelectrode (S.O.@GNE-NPE) acts as an integrated electroanalytical device to generate the electrical signal in situ. The established microcircuit provides unprecedented precision and sensitivity, contributing to subcellular amperometric measurement. The microbial sensor shows a linear response in the concentration range of 0–60 mM, with a limit of detection (LOD) at 0.3 mM. The microsensor also demonstrates good selectivity against interferences. Additionally, intracellular analysis of lactic acid provides direct evidence of enhanced lactic metabolism in cancer cells as a result of “Warburg Effect”. This work shows an example of nano-, bio- and electric technologies that have been integrated on the EAB-modified dimidiate microelectrode, and achieves intracellular biosensing application through such integration. It may give a new strategy on the combination of micro/nanotechnologies with sensory EAB for the necessary development of bioelectronic devices.

Nanowire Electrode Structures Enhanced Direct Extracellular Electron Transport via Cell-Surface Multi-Heme Cytochromes in Desulfovibrio ferrophilus IS5

Extracellular electron transfer (EET) by sulfate-reducing bacteria (SRB), such as Desulfovibrio ferrophilus IS5, enables bacterial interactions with minerals, which are vital for biogeochemical cycling and environmental chemistry. Here, we explore the direct EET mechanisms through outer-membrane cytochromes (OMCs) using IS5 as a model SRB. We employed nanostructured electrodes arrayed with 0, 50, 200, and 500 nm long nanowires (NWs) coated with indium–tin–doped oxide to examine the impact of electrode morphology on the direct EET efficacy. Compared to flat electrodes, NW electrodes significantly enhanced current production in IS5 with OMCs. However, this enhancement was diminished when OMC expression was reduced. Differential pulse voltammetry revealed that NW electrodes specifically augmented redox peaks associated with OMCs without affecting those related to redox mediators, suggesting that NWs foster direct EET through OMCs. Scanning electron microscopy observations following electrochemical analyses revealed a novel vertical cell attachment and aggregation on NW electrodes, contrasting with the horizontal monolayer cell attachment on flat electrodes. This study presents the first evidence of the critical role of electrode nanoscale topography in modulating SRB cell orientation and aggregation behavior. The findings underscore the significant influence of electrode morphology on the direct EET kinetics, highlighting the potential impact of mineral morphology on mineral reduction and biogeochemical processes.

Biochar accelerates methane production efficiency from Baijiu wastewater: Some viewpoints considering direct interspecies electron transfer

The low pH of Maotai-flavor Baijiu wastewater (MFBW) adversely affects its anaerobic digestion (AD) performance, resulting in low AD efficiency. Here, coconut shell was used to produce biochar. The characteristics of biochar were regulated through acid, alkali, and magnetic modification, respectively. Biochar and modified biochars were applied to assist the AD of MFBW. The results showed that biochar could significantly increase methane yield by 220.8 %–241.7 % with the corresponding soluble chemical oxygen demand (sCOD) degradation increasing by 52.3 %–57.5 % (p < 0.05). Joint modification could significantly enhance the electron donating capacity from 0.0042 to 0.0095 mmol e−1/g (p < 0.05). The combined modification with magnetic alkali had the best stimulating effect on the AD process, which might be related to the conductive particles (Fe3O4) formed during magnetization processes. The modified biochar featured a high degree of surface roughness, a relatively large aperture, and strong electron donating ability, all of which were beneficial to the colonization and microbial growth. Supplementation with biochar resulted in the enrichment of Proteobacteria, Firmicutes, and Actinobacteria, especially for Syntrophomonas (rising from 0.013 % to 6.74 %–10.93 % of relative abundance). These microorganisms are related to the hydrolysis, acidification, and extracellular electron transfer. The enrichment of electroactive microorganism is a prerequisite for improving the direct interspecies electron transfer pathway. This study provides theoretical support for efficient MFBW treatment.

Boosting bioelectricity generation in bioelectrochemical systems with nitrogen-doped three-dimensional graphene aerogel anode

Bioelectricity generation by electrochemically active bacteria has become particularly appealing due to its vast potential in energy production, pollution treatment, and biosynthesis. However, developing high-performance anodes for bioelectricity generation remains a significant challenge. In this study, a highly efficient three-dimensional nitrogen-doped macroporous graphene aerogel anode with a nitrogen content of approximately 4.38 ± 0.50 at% was fabricated using hydrothermal method. The anode was successfully implemented in bioelectrochemical systems inoculated with Shewanella oneidensis MR-1, resulting in a significantly higher anodic current density (1.0 A/m2) compared to the control one. This enhancement was attributed to the greater biocapacity and improved extracellular electron transfer efficiency of the anode. Additionally, the N-doped aerogel anode demonstrated excellent performance in mixed-culture inoculated bioelectrochemical systems, achieving a high power density of 4.2 ± 0.2 W/m², one of the highest reported for three-dimensional carbon-based bioelectrochemical systems to date. Such improvements are likely due to the good biocompatibility of the N-doped aerogel anode, increased extracellular electron transfer efficiency at the bacteria/anode interface, and selectively enrichment of electroactive Geobacter soli within the NGA anode. Furthermore, based on gene-level Picrust2 prediction results, N-doping significantly upregulated the conductive pili-related genes of Geobacter in the three-dimensional anode, increasing the physical connection channels of bacteria, and thus strengthening the extracellular electron transfer process in Geobacter.

High Throughput Screening of Electroactive Bacteria Using a Cross-Bar Architecture in a 3D Design Array

Promoting extracellular electron transfer (EET) in bacteria has many widespread applications including wastewater treatment and environmental remediation. With the development of synthetic biology technologies that can alter microbial electron transfer routes and enhance their electrogenic capacity, there is a need for high throughput systems to identify the responsible genes and the consequent metabolic pathways in an expeditious way. Conventional platforms utilizing single electrodes, or electrochemical cells, and colorimetric detection suffer from low sensitivity and low throughput. Moreover, they require bulky equipment for readout and fluid handling. Moving towards a nuanced miniaturized electrochemical detection, we propose individually addressable microwells used to monitor electron flux from EET-capable bacteria with the ability to screen a large number of electroactive bacteria for various applications. Materials and method : The device is simple and fabricated with cost-effective electrodes. Carbon felt is used as the working electrode to improve bacterial entrapment and assist in capturing maximum electrons donated by the electroactive bacteria. Silver-silver chloride ink (Ag/AgCl) as the reference electrode will help maintain the potential of with respect to the working electrode. Conductive carbon ink as the counter electrode will promote current collection. The proposed device has the elements of a traditional three electrode system arranged in two different planes resulting in a 3D configuration of electrodes. The reference and counter electrode are on the bottom plane with acrylic wells housed right above each electrode pair, while the working electrode is used in the cross-bar architecture from the top plane as shown in Figure 1(a). The fabrication steps are illustrated in Figure 1(b). This system can be scaled to have 10000 microwells on a single platform with individual addressability of each micro-well array. With this layout, connections from the innermost wells can be drawn effortlessly without any signal damping from the surrounding wells. As a proof of concept, we have fabricated a 9 by 9 array (20cm by 20 cm) as shown in Figure 1(a). Five strains of Shewanella oneidensis MR-1, the wild type organism and four modified strains overexpressing variants of MtrA (unmodified MtrA, IV-205, IV-261 and an empty vector), were screened for their current producing capacity [1]. Results:A bacterial load of OD600= 0.065 was sufficient to give a reliable current signal. For each strain, the working electrode was biased at 0.205 V with respect to the reference electrode (Ag/AgCl), and the resulting chronoamperometry signal was recorded as shown in Figure 1(c). The values obtained helped identify the current-producing capacity of each mutant. Among them, the highest peak current was given by mtr A+ (20 μA), and IV – 261 gave a low peak current (2 μA) within 3 minutes, validating the difference in the genetic make-up and EET capabilities. Technical and biological replicates were also conducted for all the 5 strains. The average standard deviation for the technical and biological replicates (n=3, OD600=0.065) was 0.09 μA and 0.56 μA respectively. A similar current trend for the same set of strains was obtained by Ian et al., [1] using a traditional bioelectrochemical system, thus validating our platform’s functionality and integrity. This system allows for parallel screening of wild type and mutant variations of multiple electroactive bacteria as demonstrated. The high throughput feature enabled by this device can also find application to characterize mutants generated by directed evolution workflows. Additionally, mechanical and electrical multiplexing can further improve the electronic instrumentation and connections between the wells for parallel readout and will also minimize human intervention during the current measurements. Chronoamperometry measurements is an important technique to characterize EET. Thus, a low-cost system like ours will help do that in a shorter time frame with minimal bacterial inoculum and for a larger number of electroactive bacterial species. In conclusion, we aim to show that a 3D carbon felt platform is miniaturized, and can be scaled to understand and characterize EET from electroactive bacteria in a high throughput manner. References Ian J. Campbell, Joshua T. Atkinson, Matthew D. Carpenter, Dru Myerscough, Lin Su, Caroline M. Ajo-Franklin, and Jonathan J. Silberg Biochemistry 2022 61 (13), 1337-1350 Figure 1

Bidirectional extracellular electron transfers in Serratia marcescens and Stenotrophomonas sp. correlate to EPS and Cr(VI) removal in single‐chamber bioelectrochemical systems

AbstractBACKGROUNDElectrochemically active bacteria (EAB) capable of bidirectional extracellular electron transfer (EET), either outward or inward EET, largely control the efficiency of interactions and electrical communication between biofilm and electrode and, thus, control the performance of bioelectrochemical systems (BESs) for heavy metals removal. However, the behavior of such metallurgical EAB capable of bidirectional EET has yet to be investigated, and the role of extracellular polymeric substances (EPS) in these switchable EAB with bidirectional EET and in the presence of heavy metals remains unexplored in the single‐chamber BESs treating heavy metal‐based wastewaters that are limited by carbon/electron sources or electron acceptors.RESULTSThe biofilms of the Cr(VI)‐tolerant EAB Stenotrophomonas sp. YS1 and Serratia marcescens Q1 exhibited bidirectional EET metabolizing either organic (acetate) or inorganic (HCO3−) species with simultaneous removal of Cr(VI) in single‐chamber BESs. Q1 inward EET uptake of electrons was more efficient than that of YS1 (165 μA vs. 118 μA); meanwhile, YS1 outward EET was more efficient than Q1 (8.0 μA vs. 4.7–5.2 μA). The adaptive electrochemically‐tunable EPS in both biofilm strains was regulated by the direction of the EET (inward or outward) in the presence of Cr(VI) and circuital current.CONCLUSIONThis study demonstrates the switching properties of EAB, such as Stenotrophomonas sp. or S. marcescens, that are capable of bidirectional EET to or from the electrodes, and it displays the regulation of such responses with the amount and compositional diversity of the biofilms’ EPS, giving a comprehensive appreciation of tunable EPS for Cr(VI)‐wastewater treatment in single‐chamber BESs. © 2024 Society of Chemical Industry (SCI).

Direct transformation of rice straw to electricity and hydrogen by a single yeast strain: Performance and mechanism

Lignocellulosic waste is one of the most abundant renewable energy sources. However, pretreatment or complex microbial consortia are usually required for the biological transformation into energy products. This study demonstrates the direct transformation of raw rice straw biomass into electricity and hydrogen without any pretreatment by employing a single yeast strain, Cystobasidium slooffiae JSUX1, in microbial fuel cells (MFCs). The yeast-MFCs exhibit a maximum power density of 28.56 ± 2.54 mW/m2 with simultaneous production of hydrogen gas (4.9 ± 0.52 L/m3) from raw straw. Further analysis reveals that enzymes secreted by the yeast strain degraded rice straw into sugars or organic acids, serving as fuel for electricity and hydrogen production. In addition, humic acid (HA) and Fe-HA derived from biomass consumption serve as electron mediators for extracellular electron transfer (EET) in MFCs. This study demonstrates the power of the yeast strain for renewable energy recovery from straw, diversifying the toolbox for the lignocellulose industry.

The dmsEFABGH operon encodes an essential and modular electron transfer pathway for extracellular iodate reduction by Shewanella oneidensis MR-1.

Extracellular iodate reduction by Shewanella spp. contributes to iodide generation in the biogeochemical cycling of iodine. However, there is a disagreement on whether Shewanella spp. use different extracellular electron transfer pathways with dependence on electron donors in iodate reduction. In this study, a series of gene deletion mutants of Shewanella oneidensis MR-1 were created to investigate the roles of dmsEFABGH, mtrCAB, and so4357-so4362 operons in iodate reduction. The iodate-reducing activity of the mutants was tested with lactate, formate, and H2 as the sole electron donors, respectively. In the absence of single-dms gene, iodate reduction efficiency of the mutants was only 12.9%-84.0% with lactate at 24 hours, 22.1%-85.9% with formate at 20 hours, and 19.6%-57.7% with H2 at 42 hours in comparison to complete reduction by the wild type. Progressive inhibition of iodate reduction was observed when the dms homolog from the so4357-so4362 operon was deleted in the single-dms gene mutants. This result revealed complementation of dmsEFABGH by so4357-so4362 at the single-gene level, indicating modularity of the extracellular electron transfer pathway encoded by dmsEFABGH operon. Under the conditions of all electron donors, significant inhibition of iodate reduction and accumulation of H2O2 were detected for ΔmtrCAB. Collectively, these results demonstrated that the dmsEFABGH operon encodes an essential and modular iodate-reducing pathway without electron donor dependence in S. oneidensis MR-1. The mtrCAB operon was involved in H2O2 elimination with all electron donors. The findings in this study improved the understanding of molecular mechanisms underlying extracellular iodate reduction.IMPORTANCEIodine is an essential trace element for human and animals. Recent studies revealed the contribution of microbial extracellular reduction of iodate in biogeochemical cycling of iodine. Multiple reduced substances can be utilized by microorganisms as energy source for iodate reduction. However, varied electron transfer pathways were proposed for iodate reduction with different electron donors in the model strain Shewanella oneidensis MR-1. Here, through a series of gene deletion and iodate reduction experiments, we discovered that the dmsEFABGH operon was essential for iodate reduction with at least three electron donors, including lactate, formate, and H2. The so4357-so4362 operon was first demonstrated to be capable of complementing the function of dmsEFABGH at single-gene level.

Solar-driven Methanogenesis through Microbial Ecosystem Engineering on Carbon Nitride.

Semi-biological photosynthesis combines synthetic photosensitizers with microbial catalysts to produce sustainable fuels and chemicals from CO2. However, the inefficient transfer of photoexcited electrons to microbes leads to limited CO2 utilization, restricting the catalytic performance of such biohybrid assemblies. Here, we introduce a biological engineering solution to address the inherently sluggish electron uptake mechanism of a methanogen, Methanosarcina barkeri (M. barkeri), by coculturing it with an electron transport specialist, Geobacter sulfurreducens KN400 (KN400), an adapted strain rich with multiheme c-type cytochromes (c-Cyts) and electrically conductive protein filaments (e-PFs) made of polymerized c-Cyts with enhanced capacity for extracellular electron transfer (EET). Integration of this M. barkeri-KN400 co-culture with a synthetic photosensitizer, carbon nitride, demonstrates that c-Cyts and e-PFs, emanating from live KN400, transport photoexcited electrons efficiently from the carbon nitride to M. barkeri for methanogenesis with remarkable long-term stability and selectivity. The demonstrated cooperative interaction between two microbes via direct interspecies electron transfer (DIET) and the photosensitizer to assemble a semi-biological photocatalyst introduces an ecosystem engineering strategy in photocatalysis to drive sustainable chemical synthesis.

Distribution and response of electroactive microorganisms to freshwater river pollution

Electroactive microorganisms (EAMs) play a vital role in biogeochemical cycles by facilitating extracellular electron transfer. They demonstrate remarkable adaptability to river sediments that are characterized by pollution and poor water quality, significantly contributing to the sustainability of river ecosystems. However, the distribution and diversity of EAMs remain poorly understood. In this study, 16S rRNA gene high-throughput sequencing and real-time fluorescence quantitative PCR were used to assess EAMs in 160 samples collected from eight rivers within the Pearl River Delta of Southern China. The results indicated that specialized EAMs communities in polluted sediments exhibited variations in response to water quality and sediment depth. Compared to clean sediment, polluted sediments showed a 4.5% increase in the relative abundances of EAMs communities (59 genera), with 45- and 17-times higher abundances of Geobacter and cable bacteria. Additionally, the abundance of cable bacteria decreased with increasing sediment depth in polluted sediments, while the abundance of L. varians GY32 exhibited an opposite trend. Finally, the abundances of Geobacter, cable bacteria, and L. varians GY32 were positively correlated with the abundance of filamentous microorganisms (FMs) across all samples, with stronger interactions in polluted sediments. These findings suggest that EAMs demonstrate heightened sensitivity to polluted environments, particularly at the genus (species) level, and exhibit strong adaptability to conditions characterized by high levels of acid volatile sulfide, low dissolved oxygen, and elevated nitrate nitrogen. Therefore, environmental factors could be manipulated to optimize the growth and efficiency of EAMs for environmental engineering and natural restoration applications.

Poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) bioanodes in Co-doped modified microbial fuel cell promote sulfamethoxine degradation with high enrichment of electroactive bacteria and extracellular electron transfer

Microbial fuel cell (MFC) is a new renewable energy technology, which has a wide range of applications. In this paper, the refractory organic matter sulfamethoxazole (SMX) was degraded by MFC technology, and the electricity generation performance was analyzed in detail. Since Ni and Fe ions can promote electron transfer, the presence of S ions can enhance the biocompatibility of the material, in this study, nano-ferro nickel sulfide (NiFe2S4) was prepared by hydrothermal method, and co-doped with ferro oxide (Fe3O4) nanoparticles to modify the poly (3,4-ethylenedioxythiophene): poly (styrenesulfonate) (PEDOT: PSS) hydrogel served as the bioanode of MFC. Through high throughput results, we found the enriched electro producing bacteria Desulfuromonas and Fe-S cluster oxidoreductase, proving that bimetallic sulfides have good synergistic effect with metal oxides, which can provide more active sites and accelerate the EET process. The maximum power density obtained as a bioanode in MFC was 3.67 W/m3. During degradation of SMX, the maximum power density was 2.39 W/m3. At the same time, the degradation efficiency of SMX reached 93.78 %, and the chemical oxygen demand (COD) removal rate was 32.52 %. This provides a valuable reference for the degradation of refractory organic compounds such as antibiotic SMX, and also provides multi-dimensional thinking for renewable energy technologies.

Single-walled Carbon Nanotubes Wrapped with Charged Polysaccharides Enhance Extracellular Electron Transfer.

Microbial electrochemical systems (MESs) rely on the microbes' ability to transfer charges from their anaerobic respiratory processes to electrodes through extracellular electron transfer (EET). To increase the generally low output signal in devices, advanced bioelectrical interfaces tend to augment this problem by attaching conducting nanoparticles, such as positively charged multiwalled carbon nanotubes (CNTs), to the base carbon electrode to electrostatically attract the negatively charged bacterial cell membrane. On the other hand, some reports point to the importance of the magnitude of the surface charge of functionalized single-walled CNTs (SWCNTs) as well as the size of functional groups for interaction with the cell membrane, rather than their polarity. To shed light on these phenomena, in this study, we prepared and characterized well-solubilized aqueous dispersions of SWCNTs functionalized by either positively or negatively charged cellulose-derivative polymers, as well as with positively charged or neutral small molecular surfactants, and tested the electrochemical performance of Shewanella oneidensis MR-1 in MESs in the presence of these functionalized SWCNTs. By simple injection into the MESs, the positively charged polymeric SWCNTs attached to the base carbon felt (CF) electrode, and as fluorescence microscopy revealed, allowed bacteria to attach to these structures. As a result, EET currents continuously increased over several days of monitoring, without bacterial growth in the electrolyte. Negatively charged polymeric SWCNTs also resulted in continuously increasing EET currents and a large number of bacteria on CF, although SWCNTs did not attach to CF. In contrast, SWCNTs functionalized by small-sized surfactants led to a decrease in both currents and the amount of bacteria in the solution, presumably due to the detachment of surfactants from SWCNTs and their detrimental interaction with cells. We expect our results will help researchers in designing materials for smart bioelectrical interfaces for low-scale microbial energy harvesting, sensing, and energy conversion applications.

Evidence for autotrophic growth of purple sulfur bacteria using pyrite as electron and sulfur source.

Microbial utilization of solid-phase substrates constitutes a critical area of focus in environmental microbiology, offering valuable insights into microbial metabolic processes and adaptability. Recent advancements in this field have profoundly deepened our knowledge of microbial physiology pertinent to these scenarios and spurred innovations in biosynthesis and energy production. Furthermore, research into interactions between microbes and solid-phase substrates has directly linked microbial activities to the surrounding mineralogical environments, thereby enhancing our understanding of the relevant biogeochemical cycles. Our study represents a significant step forward in this field by demonstrating, for the first time, the autotrophic growth of purple sulfur bacteria using insoluble pyrite (FeS2) as both the electron and sulfur source. The presented comparative growth profiles, substrate characterizations, and transcriptomic sequencing data shed light on the relationships between electron donor types, photosynthetic reaction center activities, and potential extracellular electron transfer in these organisms capable of anoxygenic photosynthesis. Furthermore, the findings of our study may provide new insights into early-Earth biogeochemical evolutions, offering valuable constraints for understanding the environmental conditions and microbial processes that shaped our planet's history.

Insights into mechanisms of red mud promoting biogas production from waste activated sludge anaerobic digestion

Anaerobic digestion of waste activated sludge is usually limited by the low overall methane yield. The addition of exogenous materials is considered to be a potential method to improve anaerobic methane production. In this study, red mud was added into waste activated sludge anaerobic digester and its effects on anaerobic digestion were investigated. Results showed that methane production was improved with the addition of red mud, and almost reached maximum in 10 g/L red mud added reactor (124.4 mL/g VSS), which was about 24.0 % higher than control. Further investigation indicated that the key genes related to methane metabolism, the activity of the corresponding coenzymes and the rate of extracellular electron transfer were also increased in red mud added reactor. Batch experiments on the effects of red mud in each stage of waste activated sludge anaerobic digestion showed that red mud could improve solubilization, hydrolysis, and acidification processes, but inhibit the methanogenesis process. The potential mechanism behind the enhancement of methane production during whole waste activated sludge anaerobic digestion is that the addition of red mud can intensify the hydrolysis and acidification processes, thereby providing more substrates for methanogens and subsequently enhancing the activity of coenzymes related to methane production.

Light-driven extracellular electron transfer accelerates microbiologically influenced corrosion by Rhodopseudomonas palustris TIE-1

This study investigates the microbiologically influenced corrosion (MIC) of X80 steel accelerated by the phototrophic bacterium Rhodopseudomonas palustris TIE-1. The photorespiration plays a key role in promoting extracellular electron transfer (EET)-induced MIC. In the early corrosion stage, unstable localized corrosion dominated in the dark, while intense diffusion-controlled corrosion occurs in light. Compared to the sterile anaerobic medium, R. palustris TIE-1 accelerated corrosion of X80 steel, with a significantly higher corrosion rate under light conditions, approximately three times that of dark conditions. Inhibition of photosynthetic electron transfer or cessation of photostimulation resulted in pronounced reduction in the corrosion rate.

Multilevel assault dominated by electron-transfer nonradical pathway via electronic metal–support interactions of Bi-C bonds for disinfection

Multilevel assaults on aqueous pathogenic microorganisms were realized by persulfate (PS) activation system with Bi–C based electronic metal-supported interactions (EMSI) of bismuth-embedded carbon chainmail catalyst (Bi@CC) dominated by electron-transfer nonradical pathway. The Bi@CC/PS disinfection system achieved approximately 100 % bacterial inactivation (6.5 log10 cfu/mL) within 35 min. The electron transfer from Bi to the carbon layer via Bi-C bond leading to higher work functions, thus realizing primary assault to the extracellular membrane via bacterial extracellular electron transfer (EET) function. Electron rearrangement of Bi-C is conducive to forming the intermediate complexes (Bi@CC-PS*), leading to higher work functions and enhancing electron transfer pathway (ETP) process to aggravate cell destruction, while radicals (∙OH andSO4∙-) directly attacks the interior cell. The Bi@CC/PS disinfection system possesses excellent environmental adaptability, cycling performance, and safety. This study enriches the heterogeneous mechanism of PS activation and proposes a promising strategy for sewage effluent treatment.