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A subset of microorganisms that perform respiration can endogenously utilize insoluble electron donors, such as Fe(II) or a cathode, in a process called extracellular electron transfer (EET). However, it is unknown whether similar endogenous EET can be performed by primarily fermentative species like lactic acid bacteria. We report for the first time electron uptake from a cathode by Lactiplantibacillus plantarum, a primarily fermentative bacteria found in the gut of mammals and in fermented foods. L. plantarum consumed electrons from a cathode and coupled this oxidation to the reduction of both an endogenous organic (pyruvate) and an exogenous inorganic electron acceptor (nitrate). This electron uptake from a cathode reroutes glucose fermentation toward lactate degradation and provides cells with a higher viability upon sugar exhaustion. Moreover, the associated genes and cofactors indicate that this activity is mechanistically different from that one employed by lactic acid bacteria to reduce an anode and to perform respiration. Our results expand our knowledge of the diversity of electroactive species and of the metabolic and bioenergetic strategies used by lactic acid bacteria.

Shewanella oneidensis MR-1 (S. oneidensis MR-1) has been shown to benefit from microbial electrosynthesis (MES) due to its exceptional electron transfer efficiency. In this study, genes involved in both extracellular electron uptake (EEU) and intracellular CO 2 conversion processes were examined and regulated to enhance MES performance. The key genes identified for MES in the EEU process were mtrB, mtrC, mtrD, mtrE, omcA and cctA. Overexpression of these genes resulted in 1.5–2.1 times higher formate productivity than that of the wild-type strains (0.63 mmol/(L·μg protein)), as 0.94–1.61 mmol/(L·μg protein). In the intracellular CO 2 conversion process, overexpression of the nadE, nadD, nadR, nadV, pncC and petC genes increased formate productivity 1.3-fold–3.4-fold. Moreover, overexpression of the formate dehydrogenase genes fdhA1, fdhB1 and fdhX1 in modified strains led to a 2.3-fold–3.1-fold increase in formate productivity compared to wild-type strains. The co-overexpression of cctA, fdhA1 and nadV in the mutant strain resulted in 5.59 times (3.50 mmol/(L·μg protein)) higher formate productivity than that of the wild-type strains. These findings revealed that electrons of MES derived from the electrode were utilized in the energy module for synthesizing ATP and NADH, followed by the synthesis of formate in formate dehydrogenase by the combinatorial effects of ATP, NADH, electrons and CO 2. The results provide new insights into the mechanism of MES in S. oneidensis MR-1 and pave the way for genetic improvements that could facilitate the further application of MES. • Regulating genes related to EEU and intracellular CO 2 conversion processes improved formate productivity in the MES of S. oneidensis MR-1. • The co-overexpression of cctA , fhdA1 , and nadV in S. oneidensis MR-1 increased the productivity of formate by 5.59 times higher than that of the WT strain. • Distinct functional genes between S. oneidensis MR-1 EEU and EET were discovered. • Further information about the MES process of S. oneidensis MR-1 was revealed. [ABSTRACT FROM AUTHOR]

A subset of microorganisms that perform respiration can endogenously utilize insoluble electron donors, such as Fe(II) or a cathode, in a process called extracellular electron transfer (EET). However, it is unknown whether similar endogenous EET can be performed by primarily fermentative species like lactic acid bacteria. We report for the first time electron uptake from a cathode by Lactiplantibacillus plantarum, a primarily fermentative bacteria found in the gut of mammals and in fermented foods. L. plantarum consumed electrons from a cathode and coupled this oxidation to the reduction of both an endogenous organic (pyruvate) and an exogenous inorganic electron acceptor (nitrate). This electron uptake from a cathode reroutes glucose fermentation toward lactate degradation and provides cells with a higher viability upon sugar exhaustion. Moreover, the associated genes and cofactors indicate that this activity is mechanistically different from that one employed by lactic acid bacteria to reduce an anode and to perform respiration. Our results expand our knowledge of the diversity of electroactive species and of the metabolic and bioenergetic strategies used by lactic acid bacteria.

Harvesting electricity from microbial electron transfer is believed as a promising way of renewable energy generation. However, a major challenge lies in the still-unknown mechanisms of extracellular electron transfer, especially how microbes consume electrons from the cathode to catalyze oxygen reduction. Here we report a previously undescribed yet significant extracellular electron uptake pathway mediated by inevitably produced H 2 O 2 , contributing up to 45% of the total biocurrent. This new H 2 O 2 -based bioelectrochemical respiration depends on the continuous supply of electrons from the electrode and the presence of the catalase katG . Selective enhancement of two-electron oxygen reduction on the cathode results in a 2.4-fold increase in biocurrent, and both autotrophic biosynthesis and energy production pathways are upregulated to sustain the H 2 O 2 -based respiration. Our results highlight the importance of two-electron oxygen reduction in bioelectron uptake at the cathode and provide a basis for the design of bioelectricity production systems.

Microbial electrosynthesis (MES) is a rapidly growing technology at the forefront of sustainable chemistry, leveraging the ability of microorganisms to catalyze electrochemical reactions to synthesize valuable compounds from renewable energy sources. The reduction of CO 2 is a major target application for MES, but research in this area has been stifled, especially with the use of direct electron transfer (DET)-based microbial systems. The major fundamental hurdle that needs to be overcome is the low efficiency of CO 2 reduction largely attributed to minimal microbial access to CO 2 owing to its low solubility in the electrolyte. With their tunable physical properties, ionic liquids present a potential solution to this challenge and have previously shown promise in facilitating efficient CO 2 electroreduction by increasing the CO 2 solubility. However, the use of ionic liquids in MES remains unexplored. In this study, we investigated the role of 1-ethyl-3-methylimidazolium acetate ([EMIM][Ac]) using Shewanella oneidensis MR-1 as a model DET strain. Electrochemical investigations demonstrated the ability of S. oneidensis MR-1 biocathodes to directly convert CO 2 to formate with a faradaic efficiency of 34.5 ± 26.1%. The addition of [EMIM][Ac] to the system significantly increased cathodic current density and enhanced the faradaic efficiency to 94.5 ± 4.3% while concurrently amplifying the product yield from 34 ± 23 μM to 366 ± 34 μM. These findings demonstrate that ionic liquids can serve as efficient, biocompatible cosolvents for microbial electrochemical reduction of CO 2 to value-added products, holding promise for more robust applications of MES.

Electromethanogenesis has emerged as a biological branch of Power-to-X technologies that implements methanogenic microorganisms, as an alternative to chemical Power-to-X, to convert electrical power from renewable sources, and CO 2 into methane. Unlike biomethanation processes where CO 2 is converted via exogenously added hydrogen, electromethanogenesis occurs in a bioelectrochemical set-up that combines electrodes and microorganisms. Thereby, mixed, or pure methanogenic cultures catalyze the reduction of CO 2 to methane via reducing equivalents supplied by a cathode. Recent advances in electromethanogenesis have been driven by interdisciplinary research at the intersection of microbiology, electrochemistry, and engineering. Integrating the knowledge acquired from these areas is essential to address the specific challenges presented by this relatively young biotechnology, which include electron transfer limitations, low energy and product efficiencies, and reactor design to enable upscaling. This review approaches electromethanogenesis from a multidisciplinary perspective, putting emphasis on the extracellular electron uptake mechanisms that methanogens use to obtain energy from cathodes, since understanding these mechanisms is key to optimize the electrochemical conditions for the development of these systems. This work summarizes the direct and indirect extracellular electron uptake mechanisms that have been elucidated to date in methanogens, along with the ones that remain unsolved. As the study of microbial corrosion, a similar bioelectrochemical process with Fe0 as electron source, has contributed to elucidate different mechanisms on how methanogens use solid electron donors, insights from both fields, biocorrosion and electromethanogenesis, are combined. Based on the repertoire of mechanisms and their potential to convert CO 2 to methane, we conclude that for future applications, electromethanogenesis should focus on the indirect mechanism with H 2 as intermediary. By summarizing and linking the general aspects and challenges of this process, we hope that this review serves as a guide for researchers working on electromethanogenesis in different areas of expertise to overcome the current limitations and continue with the optimization of this promising interdisciplinary technology. • Electromethanogenesis is a promising technology being optimized at the lab scale. • Interdisciplinary work is required to overcome the system limitations for upscaling. • New mechanisms on electron uptake have been elucidated in methanogens. • Electrochemical parameters must suit the methanogenic electron uptake mechanisms. • Non-electrotrophic methanogens are key biocatalysts for this technology. [ABSTRACT FROM AUTHOR]

Stutzerimonas stutzeri strain FeN3W is an iron-oxidizing bacterium isolated from marine sediment. FeN3W's 5.9 Mb genome encodes complete pathways for glycolysis, gluconeogenesis, TCA cycle, pentose phosphate pathway, and aerobic and anaerobic (nitrate) respiration. The genome contains 32 putative heme-binding proteins predicted to localize to the cell envelope., Competing Interests: The authors declare no conflict of interest.

Microbes synthesize cell‐associated nanoparticles (NPs) and utilize their physicochemical properties to produce energy under unfavorable metabolic conditions. Iron sulfide (FeS) NPs are ubiquitous and are predominantly biosynthesized by sulfate‐reducing bacteria (SRB). However, the biological role of FeS NPs in SRB remains understudied. Now, conductive FeS NPs function is demonstrated as an electron conduit enabling Desulfovibrio vulgaris Hildenborough, an SRB strain, to utilize solid‐state electron donors via direct electron uptake. After forming FeS NPs on the cell surface, D. vulgaris initiated current generation coupled with sulfate reduction on electrodes poised at −0.4 V versus standard hydrogen electrode. Single‐cell activity analysis showed that the electron uptake and metabolic rate via FeS NPs in D. vulgaris were about sevenfold higher than those via native cell‐surface proteins in other SRB. [ABSTRACT FROM AUTHOR]

• A lithographic strategy was used to induce biofilm formation in a modified Shewanella oneidensis cdrAB strain to investigate the relationship between biofilm thickness and cathodic activity. • Differences in cell density at the electrode interface (controlled by blue light exposure time) resulted in measurable differences in biologically driven cathodic current density. • Electrochemical Impedance Spectroscopy (EIS) demonstrated that differences in impedance corresponded to biofilm thickness and activity. • An equivalent circuit model, that showed less than 5 % error on fitted parameters, was used to calculate the biofilm charge transfer resistance. • A linear relationship between biological cathodic current and biofilm charge transfer resistance was observed with correlation score r = 0.87 (r2 = 0.773). A range of biotechnological applications for converting electricity to fixed metabolic substrates is fuelling the study of cathodic bioelectrochemical systems. Shewanella oneidensis MR-1 has emerged as an important model system for extracellular electron uptake on cathodes, as many of the proteins involved in this process overlap with the organism's previously characterized extracellular electron transport machinery. However, there are still many questions surrounding the mechanics of electron uptake in Shewanella stemming largely from the challenge of quantifying biomass on electrode surfaces. This limits our understanding of the physiologic and kinetic constraints of electron uptake, as well as our ability to make meaningful comparisons across systems. To investigate the relationship between cathodic activity and biomass, we used a Shewanella oneidensis strain genetically modified with cell aggregation protein CdrAB behind a blue light-controlled promotor. Using blue light exposure to control cell deposition, we then investigated the relationship between cathodic activity and biomass. Electrochemical impedance spectroscopy (EIS) confirmed a decrease in biofilm impedance over a range of blue light exposures (i.e., 2 to 8 h). Consistent with previous results, after this timepoint, a drop-in electrochemical activity was observed, and impedances increased. For biofilms within the 2–8 h light exposure range, we observed a trend towards increased biological current consumption by quantifying the difference between pre and post kill currents. Comparing EIS data between pre and post kill experiments supported an increase in impedance post addition of killing agents and a trend towards the lowest biofilm impedances observed in the longest blue light exposed systems. Using an equivalent circuit model to extrapolate specific biofilm parameters we quantified the charger transfer resistance within the biofilm that corresponds to varying biofilm thicknesses and matches previously observed activities. For example, electrochemical activity was highest for the 8 h blue light exposed biofilm condition, with a maximum cathodic biologic current of -5.49 ± 0.85 µA, and a biofilm charge transfer resistance measured at 6909.5 ± 2136.5 Ω. On an individual reactor basis, we correlate this biofilm charge transfer resistance with biologic cathodic current. We observed a linear trend with a correlation score of 0.87 (r2 = 0.773). To the best of our knowledge, this is the first investigation of biofilm physiology on Shewanella cathodes using EIS. Continued efforts in this direction will further our understanding of biofilm-electrode interface during extracellular electron uptake with the goal of enhancing applications to bioelectrochemical systems. [Display omitted] [ABSTRACT FROM AUTHOR]

Methanogens capable of accepting electrons from Fe0 cause severe corrosion in anoxic conditions. In previous studies, all iron-corrosive methanogenic isolates were obtained from marine environments. However, the presence of methanogens with corrosion ability using Fe0 as an electron donor and their contribution to corrosion in freshwater systems is unknown. Therefore, to understand the role of methanogens in corrosion under anoxic conditions in a freshwater environment, we investigated the corrosion activities of methanogens in samples collected from groundwater and rivers. We enriched microorganisms that can grow with CO2/NaHCO3 and Fe0 as the sole carbon source and electron donor, respectively, in ground freshwater. Methanobacterium sp. TO1, which induces iron corrosion, was isolated from freshwater. Electrochemical analysis revealed that strain TO1 can uptake electrons from the cathode at lower than −0.61 V vs SHE and has a redox-active component with electrochemical potential different from those of other previously reported methanogens with extracellular electron transfer ability. This study indicated the corrosion risk by methanogens capable of taking up electrons from Fe0 in anoxic freshwater environments and the necessity of understanding the corrosion mechanism to contribute to risk diagnosis.

Direct electron uptake by prokaryotes is a recently described mechanism with a potential application for energy and CO2 storage into value added chemicals. Members of Methanosarcinales, an environmentally and biotechnologically relevant group of methanogens, were previously shown to retrieve electrons from an extracellular electrogenic partner performing Direct Interspecies Electron Transfer (DIET) and were therefore proposed to be electroactive. However, their intrinsic electroactivity has never been examined. In this study, we tested two methanogens belonging to the genus Methanosarcina, M. barkeri, and M. horonobensis, regarding their ability to accept electrons directly from insoluble electron donors like other cells, conductive particles and electrodes. Both methanogens were able to retrieve electrons from Geobacter metallireducens via DIET. Furthermore, DIET was also stimulated upon addition of electrically conductive granular activated carbon (GAC) when each was co-cultured with G. metallireducens. However, when provided with a cathode poised at −400 mV (vs. SHE), only M. barkeri could perform electromethanogenesis. In contrast, the strict hydrogenotrophic methanogen, Methanobacterium formicicum, did not produce methane regardless of the type of insoluble electron donor provided (Geobacter cells, GAC or electrodes). A comparison of functional gene categories between the two Methanosarcina showed differences regarding energy metabolism, which could explain dissimilarities concerning electromethanogenesis at fixed potentials. We suggest that these dissimilarities are minimized in the presence of an electrogenic DIET partner (e.g., Geobacter), which can modulate its surface redox potentials by adjusting the expression of electroactive surface proteins.

• Dual-electric centers catalysts cooperate with microbes to effectively remove CIP. • Delocalized electrons of absorbed CIP captured by microbes at the electron-rich area. • The catalysts synergize extracellular electron transfer for the cleavage of CIP. • The accumulation of antibiotic resistance genes was reduced by the catalysts. Antibiotics and antibiotic resistance genes (ARGs) are still a problem in biological treatment. Herein, we propose a synergetic strategy between microbes and dual-electric centers catalysts (CCN/Cu-Al 2 O 3 /ceramsite) for Ciprofloxacin (CIP)-contained (5 mg/L) water treatment in an up-flow biological filter. CIP was cleaved into small molecules by the catalyst, bringing a 57.6% removal and reducing 10.5% ARG. The characterization results verified that a Cu-π electrostatic force occurs on the catalyst surface, forming electron-rich areas around Cu and electron-poor areas at the carbon-doped g -C 3 N 4 (CCN) aromatic ring. Thus, the electrons of adsorbed CIP were delocalized and then captured by the adsorbed extracellular polymeric substance at the electron-rich areas. Therefore, the synergetic process weakened the stress of CIP on bacteria and reduced ARG accumulation. It also enriched more electro-active bacteria on the surface of CCN/Cu-Al 2 O 3 /ceramsite, promoting the expression of extracellular electron transfer-related genes and reconstructing the energy metabolism mode. This result provides an opportunity for refractory antibiotic treatment in the biological process. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • In-situ biogas upgrading was achieved in a AD/PSB-coupled MEC. • Photosynthetic CO 2 fixation under electrical stimulation increased by 83.3%. • Electrically-stimulated PSB fixed CO 2 more efficiently even under power off. • Electrical stimulation enhanced the electroactivity and photoactivity of PSB. • Electrical stimulation changed the structural of photosynthetic reaction center. Biogas from anaerobic digestion usually contains 30%–50% of CO 2 , which needs to be upgraded before industrial utilization. However, current upgrading techniques typically consume large amounts of chemicals and energy. This study designed a green and low-energy-consumption microbial electrolysis cell (MEC)-based anaerobic digester for in-situ biogas upgrading, in which the cathode with photosynthetic bacteria (PSB) was used to fix CO 2. Results showed that charged MEC-AD increased photosynthetic CO 2 fixation by 83.3% and CH 4 production by 62.8% compared with uncharged open-circuit MEC-AD. Adding an extracellular electron uptake inhibitor to the cathode decreased the CO 2 fixation, accompanied by the decline of current between the two electrodes, which suggested that the cathode electron uptake was the main reason for the facilitated CO 2 fixation of PSB. The electrically-stimulated PSB fixed CO 2 more efficiently than unstimulated PSB even under power off, likely related to their enhanced electroactivity and photoactivity that could acquire more extracellular electrons and light energy for CO 2 fixation. The carbonyl-related hydrogen bonds in the LH1 complex of photosynthetic reaction centers increased under electrical stimulation, resulting in a red shift of absorbance spectra to improve the energy utilization of PSB. This upgrading technique consumed about 0.37 kWh/Nm3 CO 2removed of electric energy, much lower than other MEC-based techniques, providing a green and feasible strategy for in-situ biogas upgrading. [ABSTRACT FROM AUTHOR]

Microbial biofilms are ubiquitous. In marine and freshwater ecosystems, microbe-mineral interactions sustain biogeochemical cycles, while biofilms found on plants and animals can range from pathogens to commensals. Moreover, biofouling and biocorrosion represent significant challenges to industry. Bioprocessing is an opportunity to take advantage of biofilms and harness their utility as a chassis for biocommodity production. Electrochemical bioreactors have numerous potential applications, including wastewater treatment and commodity production. The literature examining these applications has demonstrated that the cell-surface interface is vital to facilitating these processes. Therefore, it is necessary to understand the state of knowledge regarding biofilms' role in bioprocessing. This mini-review discusses bacterial biofilm formation, cell-surface redox interactions, and the role of microbial electron transfer in bioprocesses. It also highlights some current goals and challenges with respect to microbe-mediated bioprocessing and future perspectives., (© The Author(s) 2022. Published by Oxford University Press on behalf of Society of Industrial Microbiology and Biotechnology.)

Methanogens capable of accepting electrons from Fe0 cause severe corrosion in anoxic conditions. In previous studies, all iron-corrosive methanogenic isolates were obtained from marine environments. However, the presence of methanogens with corrosion ability using Fe0 as an electron donor and their contribution to corrosion in freshwater systems is unknown. Therefore, to understand the role of methanogens in corrosion under anoxic conditions in a freshwater environment, we investigated the corrosion activities of methanogens in samples collected from groundwater and rivers. We enriched microorganisms that can grow with CO2/NaHCO3 and Fe0 as the sole carbon source and electron donor, respectively, in ground freshwater. Methanobacterium sp. TO1, which induces iron corrosion, was isolated from freshwater. Electrochemical analysis revealed that strain TO1 can uptake electrons from the cathode at lower than −0.61 V vs SHE and has a redox-active component with electrochemical potential different from those of other previously reported methanogens with extracellular electron transfer ability. This study indicated the corrosion risk by methanogens capable of taking up electrons from Fe0 in anoxic freshwater environments and the necessity of understanding the corrosion mechanism to contribute to risk diagnosis. [ABSTRACT FROM AUTHOR]