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Examples of how metalloproteins feature in electron transfer processes in biological systems are reviewed. Attention is focused on the electron transport chains of cellular respiration and photosynthesis, and on metalloproteins that directly couple electron transfer to a chemical reaction. Brief mention is also made of extracellular electron transport. While covering highlights of the recent and the current literature, this review is aimed primarily at introducing the senior undergraduate and the novice postgraduate student to this important aspect of bioinorganic chemistry. [ABSTRACT FROM AUTHOR]

Microbial fuel cell (MFC) has been regarded as a promising green and clean electricity generation device. The anode plays a crucial role in the electricity generation process within the MFC. This study employed a hydrothermal method for the doping of sulfur onto MXene nanosheets to alleviate the stacking, followed by an in situ polymerization process to fabricate sufur-doped MXene/polypyrrole (S–MXene/PPy) composite hydrogel on carbon felt (CF). The MFC constructed with this anode exhibited the highest power density of 8.05 W m−3 and a maximum output voltage of 646 mV, higher than that of MXene/PPy (623 mV, 6.03 W m−3), PPy (594 mV, 4.91 W m−3), and unmodified CF (574 mV, 4.05 W m−3). This improved performance can be attributed to the reduction of charge transfer resistance in the material while improving the bacteria affinity, leading to enhanced extracellular electron transfer (EET) efficiency. High-throughput sequencing indicates that the excellent biocompatibility significantly promotes the attachment and growth of electroactive microorganisms (Geobacter, 45.34%) on the S–MXene/PPy anode. This study demonstrates that S–MXene/PPy hydrogel has good potential as an anode from the perspective of MFC electricity generation and microbial analysis. [ABSTRACT FROM AUTHOR]

The contribution of Fe(II)-oxidizing bacteria to iron cycling in freshwater, groundwater, and marine environments has been widely recognized in recent years. These organisms perform extracellular electron transfer (EET), which constitutes the foundations of bioelectrochemical systems for the production of biofuels and bioenergy. It was proposed that the Gram-negative bacterium Sideroxydans lithotrophicus ES-1 oxidizes soluble ferrous Fe(II) at the surface of the cell and performs EET through the Mto redox pathway. This pathway is composed by the periplasmic monoheme cytochrome MtoD that is proposed to bridge electron transfer between the cell exterior and the cytoplasm. This makes its functional and structural characterization, as well as evaluating the interaction process with its physiological partners, essential for understanding the mechanisms underlying EET. Here, we report the complete assignment of the heme proton and carbon signals together with a near-complete assignment of 1H, 13C and 15N backbone and side chain resonances for the reduced, diamagnetic form of the protein. These data pave the way to identify and structurally map the molecular interaction regions between the cytochrome MtoD and its physiological redox partners, to explore the EET processes of S. lithotrophicus ES-1. [ABSTRACT FROM AUTHOR]

Mechanistic studies of life's lower metabolic limits have been limited due to a paucity of tractable experimental systems. Here, we show that redox-cycling of phenazine-1-carboxamide (PCN) by Pseudomonas aeruginosa supports cellular maintenance in the absence of growth with a low mass-specific metabolic rate of 8.7 × 10−4 W (g C)−1 at 25°C. Leveraging a high-throughput electrochemical culturing device, we find that non-growing cells cycling PCN tolerate conventional antibiotics but are susceptible to those that target membrane components. Under these conditions, cells conserve energy via a noncanonical, facilitated fermentation that is dependent on acetate kinase and NADH dehydrogenases. Across PCN concentrations that limit cell survival, the cell-specific metabolic rate is constant, indicating the cells are operating near their bioenergetic limit. This quantitative platform opens the door to further mechanistic investigations of maintenance, a physiological state that underpins microbial survival in nature and disease. [Display omitted] • Anaerobic PCN cycling supports a low metabolic rate (∼103 ATP s−1 cell−1 at 25°C) • During PCN cycling, single cells maintain metabolic activity but do not grow • A high-throughput electrochemical platform enables mechanistic studies of maintenance • Disrupting membrane integrity or bioenergetic components impairs survival in this state Anaerobic phenazine cycling by an opportunistic bacterial pathogen supports a low-power, non-growth metabolism that lends itself to quantitative and mechanistic study of maintenance physiology. [ABSTRACT FROM AUTHOR]

In recent years, exploring interactions between sunlight, semiconducting minerals and microorganisms in nature has attracted great attention. However, relatively little research has been conducted on the interaction between minerals and microorganisms in the plain areas of the Yellow River Basin under sunlight. In this study, the mineral composition and microbial community of red soil from the Ningxia Plain, China were analyzed. X–ray diffraction (XRD) results showed that red soil contained semiconducting minerals such as birnessite, hematite and goethite. 16S rRNA analysis found that electroactive microorganisms such as Proteobacteria, Firmicutes and Actinobacteriota were enriched in in–situ microbial community of red soil. Afterwards, electrochemical tests such as linear sweep voltammetry (LSV) and I–t curves, indicated that red soil exhibited good semiconducting properties under light irradiation. Finally, a dual chamber system and electrochemical techniques were used to explore the electron transfer relationship between red soil and microorganisms. The open circuit voltage (350 mV) and maximum power density (1242.72 mW/m2) of red soil cathode system were significantly improved compared to blank graphite electrode, indicating that red soil could serve as electron acceptors for microbial extracellular respiration. Red soil electrode in live bacteria exhibited the highest photocurrent density (0.29 μA/cm2) and the lowest charge transfer resistance (Rct) (223 Ω) under light conditions, indicating that red soil accelerated the rapid transfer of electrons, reduced polarization loss, and enhanced extracellular electron transfer (EET) process under light conditions. This study demonstrated that the participation of semiconducting minerals in microbial EET processes under sunlight in the Ningxia Plain, China. [ABSTRACT FROM AUTHOR]

Oxic microbial electrosynthesis (oMES) allows the utilization of renewable electricity and industrial gas streams containing CO2 and O2 for biomass production by cultivating aerobic, autotrophic, hydrogen‐oxidizing bacteria, commonly known as Knallgas bacteria. oMES is likely not a direct competitor to conventional anoxic microbial electrosynthesis as harnessing aerobic hydrogen‐oxidizing bacteria depends on energetically inefficient assimilatory CO2 reduction pathways. However, it might be a complementary approach to classical biomass production from the perspective of limited land use and the availability of cheap renewable energy. The best characterized Knallgas bacterium is Cupriavidus necator. Extensively studied as lithoautotrophic production host, C. necator already offers a broad arsenal of genetic tools. In contrast, mechanistical knowledge about the recently discovered Kyrpidia spormannii is limited, but this species shows remarkable growth when cultivated as cathodic biofilm in bioelectrochemical systems. In addition, first experiments indicate a low energy demand for biomass production, which is in the order of magnitude of gas fermentation with C. necator or heterotrophic and methanotrophic technologies. Still, many aspects of the electrochemical cultivation of K. spormannii need to be better understood and rigorously improved to be a competitive technology in the making, including electron transfer and microbial kinetics, cultivation conditions, mass and energy balances, and reactor design. [ABSTRACT FROM AUTHOR]

Microbial fuel cells (MFCs) are a promising technology for obtaining energy in wastewater. Effective extracellular electron transfer is one of the key factors for its practical application. In this work, carbon dots (CDs) enriched with oxygen-containing groups on the surface were synthesized as an efficient anode modifier using a simple hydrothermal method and common reactants. The experimental findings indicated that anodes modified with CDs exhibited increased electrical conductivity and greater hydrophilicity. These modifications facilitated increased microorganism loading and contributed to enhancing electrochemical processes within the anode biofilm. The CD-modified MFCs exhibited higher maximum power density (661.1 ± 42.6 mW·m−2) and open-circuit voltage (534.50 ± 6.4 mV), which were significantly better than those of the blank group MFCs (484.1 ± 14.1 mW·m−2 and 447.50 ± 12.1 mV). The use of simple carbon materials to improve the microbial loading on the MFCs anode and the electron transfer between the microbial-electrode may provide a new idea for the design of efficient MFCs. [ABSTRACT FROM AUTHOR]

Dissimilatory iron reduction (DIR) is an important form of microbial respirations and a key part of iron biogeochemical cycle. A wide range of both bacteria and archaea that can conserve energy through Fe(III) reduction are called dissimilatory iron-reducing microorganisms (DIRMs). They have been increasingly recognized as important for coupling organic carbon oxidation in diverse anaerobic environments, such as soil, sediments, freshwater, marine water as well as extreme environments. In parallel with their phylogenetic diversity, DIRMs possess metabolic versatility, including multiple extracellular electron transfer (EET) pathways and various electron donors as well as acceptors. In this review, phylogenetic, environmental distribution of DIRMs was demonstrated comprehensively by summarizing 51 isolated DIRMs belonging to 27 genera in previous literature. EET mechanisms were further elaborated on based on four DIRMs representatives: Geobacter, Shewanella, Gram-positive bacteria and archaea. Various electron donors, acceptors, and novel metabolisms revealed recently prompt the development of DIRMs biotechnological applications, including bioleaching, bioremediation, biosynthesis, anaerobic fermentation, and production of bioelectricity. Although past decades have witnessed a great increase of the publications in DIRMs, further investigation are required for deep understanding and practical applications, such as their roles in natural environments, EET mechanisms in different DIRMs, cooperation with other microbes, and mechanisms of improved bioproduction by adding iron-oxides. DIRMs in real world. (1) they can be used for bioleaching; (2) they can be used for bioremediation for heavy metals and organic pollutants; (3) in aquatic environments, they can conduct iron reduction and form iron precipitate; (4) they can be used to produce current; (5) they can synthesize bioproducts; (6) they can form stable network with other soil microorganisms; (7) in various environments, they can form microbial iron cycle with iron-oxidizing microorganisms and influence many processes. [ABSTRACT FROM AUTHOR]

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 NH 4 +-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. [Display omitted] • AQDS enhanced the removal rate of NH 4 +-N and SND efficiency of electrotrophic bio-cathode at low temperature. • AQDS promoted the energy metabolism of eletrotrophic microorganisms with cold-tolerant. • AQDS increased the abundance of functional genes, ehancing electorn transfer. • AQDS immerged in EPS accelarated electorn hopping by act as an electron shuttle. [ABSTRACT FROM AUTHOR]

The combination of microbial CO 2 reduction process and photocatalysis can meet the economic goal of sustainable carbon neutrality. Solar-energy as driven force can be achieved, by mimicking natural photosynthesis through the interaction between photo-responsive semiconductor nano-materials and electroautotrophic bacteria (EAB). Therefore, we reviewed the engineering strategies developed in recent years for photocatalytic microbial fixation of CO 2 to improve extracellular electron transfer (EET) rate and enhance the conversion efficiency of solar-to-chemical energy, including (1) the application principles of semiconductor nanomaterials in microbial CO 2 reduction systems; (2) photocathode; (3) photoanode; (4) dual photoelectrodes and (5) inorganic-microbe hybrid system; (6) perspectives on future challenges and opportunities for photocatalytic microbial fixation of CO 2 for sustainable energy and chemical production. This review will provide a theoretical basis for the future development of photocatalytic microbial fixation of CO 2 in energy harvesting, CO 2 fixation, and chemical synthesis. [ABSTRACT FROM AUTHOR]

Some sulfate-reducing bacteria (SRB), mainly belonging to the Desulfovibrionaceae family, have evolved the capability to conserve energy through microbial extracellular electron transfer (EET), suggesting that this process may be more widespread than previously believed. While previous evidence has shown that mobile genetic elements drive the plasticity and evolution of SRB and iron-reducing bacteria (FeRB), few have investigated the shared molecular mechanisms related to EET. To address this, we analyzed the prevalence and abundance of EET elements and how they contributed to their differentiation among 42 members of the Desulfovibrionaceae family and 23 and 59 members of Geobacteraceae and Shewanellaceae, respectively. Proteins involved in EET, such as the cytochromes PpcA and CymA, the outer membrane protein OmpJ, and the iron–sulfur cluster-binding CbcT, exhibited widespread distribution within Desulfovibrionaceae. Some of these showed modular diversification. Additional evidence revealed that horizontal gene transfer was involved in the acquiring and losing of critical genes, increasing the diversification and plasticity between the three families. The results suggest that specific EET genes were widely disseminated through horizontal transfer, where some changes reflected environmental adaptations. These findings enhance our comprehension of the evolution and distribution of proteins involved in EET processes, shedding light on their role in iron and sulfur biogeochemical cycling. [ABSTRACT FROM AUTHOR]

Chemical‐to‐bioelectricity by using different biocatalysts was considered as a next‐generation green power source. However, bioelectricity production using macromolecular substrate usually encountered low Coulombic efficiency (CE) and power density due to inefficient electron releasing and sluggish electron collection. Here, a rationally engineered biocascade (including depolymerization module, fermentation module, and electro‐respiration module) embedded in highly conductive 3D graphene hydrogel (electron collection module) was designed and fabricated as a modular platform to simultaneously improve the substrate degradation, enhance the electron releasing and reinforce the electron collection. As a result, this modular platform enabled a ~15‐fold improvement on power density and reached the highest CE (46.3%) and power density (780 mW/m2) ever reported for bioelectricity production from starch (a model macromolecular substrate). This work demonstrated a promising approach for rationally harvesting bioelectricity with complicated substrates, which would open up a new avenue for practical applications. [ABSTRACT FROM AUTHOR]

Cable bacteria are filamentous bacteria that couple the oxidation of sulfide in sediments to the reduction of oxygen via long-distance electron transport over centimeter distances through periplasmic wires. However, the capability of cable bacteria to perform extracellular electron transfer to acceptors, such as electrodes, has remained elusive. In this study, we demonstrate that living cable bacteria actively move toward electrodes in different bioelectrochemical systems. Carbon felt and carbon fiber electrodes poised at +200 mV attracted live cable bacteria from the sediment. When the applied potential was switched off, cable bacteria retracted from the electrode. qPCR and scanning electron microscopy corroborated this finding and revealed cable bacteria in higher abundance present on the electrode surface compared with unpoised controls. These experiments raise new possibilities to study metabolism of cable bacteria and cultivate them in bioelectrochemical devices for bioelectronic applications, such as biosensing and bioremediation. [ABSTRACT FROM AUTHOR]

Microbial electrochemical systems that integrate the advantages of inorganic electrocatalysis and microbial catalysis are expected to provide sustainable solutions to the increasing energy shortages, resource depletion, and climate degradation. However, sluggish extracellular electron transfer (EET) at the interface between electroactive microorganisms and inorganic electrode materials is a critical bottleneck that limits the performance of systems. Electrocatalytic nanomaterials are highly competitive in overcoming this obstacle due to their effective association with microbial catalysis. Therefore, this review focuses on the cutting-edge applications and enhancement mechanisms of nanomaterials with electrocatalytic activity in promoting microbial EET. First, the EET mechanism of microbial electrocatalysis in both microbial anodes and cathodes is briefly introduced, and then recent applications of various electrocatalytic nanomaterials in diverse microbial electrochemical systems are summarized, including heteroatom-doped carbons and precious metal, as well as transition metal oxides, sulfides, carbides, and nitrides. The synergistic effects of nanomaterial electrocatalysis and microbial catalysis on enhancing interfacial EET are analyzed. Finally, the challenges and perspectives of realizing high-performance microbial electrochemical systems are also discussed in order to offer some reference for further research. [ABSTRACT FROM AUTHOR]

Studies have used anaerobic-digester sludge and/or effluent as inocula for bioelectrochemical systems (BESs), such as microbial fuel cells (MFCs), for power generation, while limited studies have isolated and characterized electrochemically active bacteria (EAB) that inhabit anaerobic digesters. In the present work, single-chamber MFCs were operated using the anaerobic-digester effluent as the sole source of organics and microbes, and attempts were made to isolate EAB from anode biofilms in MFCs by repeated anaerobic cultivations on agar plates. Red colonies were selected from those grown on the agar plates, resulting in the isolation of three phylogenetically diverse strains affiliated with the phyla Bacillota, Campylobacterota and Deferribacterota. All these strains are capable of current generation in pure-culture BESs, while they exhibit different electrochemical properties as assessed by cyclic voltammetry. The analyses of their cell-free extracts show that cytochromes are abundantly present in their cells, suggesting their involvement in current generation. The results suggest that anaerobic digesters harbor diverse EAB, and it would be of interest to examine their ecological niches in anaerobic digestion. [ABSTRACT FROM AUTHOR]

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Abstract Oxic microbial electrosynthesis (oMES) allows the utilization of renewable electricity and industrial gas streams containing CO2 and O2 for biomass production by cultivating aerobic, autotrophic, hydrogen‐oxidizing bacteria, commonly known as Knallgas bacteria. oMES is likely not a direct competitor to conventional anoxic microbial electrosynthesis as harnessing aerobic hydrogen‐oxidizing bacteria depends on energetically inefficient assimilatory CO2 reduction pathways. However, it might be a complementary approach to classical biomass production from the perspective of limited land use and the availability of cheap renewable energy. The best characterized Knallgas bacterium is Cupriavidus necator. Extensively studied as lithoautotrophic production host, C. necator already offers a broad arsenal of genetic tools. In contrast, mechanistical knowledge about the recently discovered Kyrpidia spormannii is limited, but this species shows remarkable growth when cultivated as cathodic biofilm in bioelectrochemical systems. In addition, first experiments indicate a low energy demand for biomass production, which is in the order of magnitude of gas fermentation with C. necator or heterotrophic and methanotrophic technologies. Still, many aspects of the electrochemical cultivation of K. spormannii need to be better understood and rigorously improved to be a competitive technology in the making, including electron transfer and microbial kinetics, cultivation conditions, mass and energy balances, and reactor design.

316L stainless steel (SS) is widely applied as microimplant anchorage (MIA) due to its excellent mechanical properties. However, the risk that the oral microorganisms can corrode 316L SS is fully neglected. Microbiologically influenced corrosion (MIC) of 316L SS is essential to the health and safety of all patients because the accelerated corrosion caused by the oral microbiota can trigger the release of Cr and Ni ions. This study investigated the corrosion behavior and mechanism of subgingival microbiota on 316L SS by 16S rRNA and metagenome sequencing, electrochemical measurements, and surface characterization techniques. Multispecies biofilms were formed by the oral subgingival microbiota in the simulated oral anaerobic environment on 316L SS surfaces, significantly accelerating the corrosion in the form of pitting. The microbiota samples collected from the subjects differed in biofilm compositions, corrosion behaviors, and mechanisms. The oral subgingival microbiota contributed to the accelerated corrosion of 316L SS via acidic metabolites and extracellular electron transfer. Our findings provide a new insight into the underlying mechanisms of oral microbial corrosion and guide the design of oral microbial corrosion-resistant materials.

A variety of electron transfer mechanisms link bacterial cytosolic electron pools with functionally diverse redox activities in the cell envelope and extracellular space. In Listeria monocytogenes, the ApbE-like enzyme FmnB catalyzes extracytosolic protein flavinylation, covalently linking a flavin cofactor to proteins that transfer electrons to extracellular acceptors. L. monocytogenes uses an energy-coupling factor (ECF) transporter complex that contains distinct substrate-binding, transmembrane, ATPase A, and ATPase A' subunits (RibU, EcfT, EcfA, and EcfA') to import environmental flavins, but the basis of extracytosolic flavin trafficking for FmnB flavinylation remains poorly defined. In this study, we show that the EetB and FmnA proteins are related to ECF transporter substrate-binding and transmembrane subunits, respectively, and are essential for exporting flavins from the cytosol for flavinylation. Comparisons of the flavin import versus export capabilities of L. monocytogenes strains lacking different ECF transporter subunits demonstrate a strict directionality of substrate-binding subunit transport but partial functional redundancy of transmembrane and ATPase subunits. Based on these results, we propose that ECF transporter complexes with different subunit compositions execute directional flavin import/export through a broadly conserved mechanism. Finally, we present genomic context analyses that show that related ECF exporter genes are distributed across members of the phylum Firmicutes and frequently colocalize with genes encoding flavinylated extracytosolic proteins. These findings clarify the basis of ECF transporter export and extracytosolic flavin cofactor trafficking in Firmicutes. IMPORTANCE Bacteria import vitamins and other essential compounds from their surroundings but also traffic related compounds from the cytosol to the cell envelope where they serve various functions. Studying the foodborne pathogen Listeria monocytogenes, we find that the modular use of subunits from a prominent class of bacterial transporters enables the import of environmental vitamin B2 cofactors and the extracytosolic trafficking of a vitamin B2-derived cofactor that facilitates redox reactions in the cell envelope. These studies clarify the basis of bidirectional small-molecule transport across the cytoplasmic membrane and the assembly of redox-active proteins within the cell envelope and extracellular space.

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.

Microbial fuel cells (MFCs) have the potential to directly convert the chemical energy in organic matter into electrical energy, making them a promising technology for achieving sustainable energy production alongside wastewater treatment. However, the low extracellular electron transfer (EET) rates and limited bacteria loading capacity of MFCs anode materials present challenges in achieving high power output. In this study, three-dimensionally heteroatom-doped carbonized grape (CG) monoliths with a macroporous structure were successfully fabricated using a facile and low-cost route and employed as independent anodes in MFCs for treating brewery wastewater. The CG obtained at 900 °C (CG-900) exhibited excellent biocompatibility. When integrated into MFCs, these units initiated electricity generation a mere 1.8 days after inoculation and swiftly reached a peak output voltage of 658 mV, demonstrating an exceptional areal power density of 3.71 W m−2. The porous structure of the CG-900 anode facilitated efficient ion transport and microbial community succession, ensuring sustained operational excellence. Remarkably, even when nutrition was interrupted for 30 days, the voltage swiftly returned to its original level. Moreover, the CG-900 anode exhibited a superior capacity for accommodating electricigens, boasting a notably higher abundance of Geobacter spp. (87.1%) compared to carbon cloth (CC, 63.0%). Most notably, when treating brewery wastewater, the CG-900 anode achieved a maximum power density of 3.52 W m−2, accompanied by remarkable treatment efficiency, with a COD removal rate of 85.5%. This study provides a facile and low-cost synthesis technique for fabricating high-performance MFC anodes for use in microbial energy harvesting. [ABSTRACT FROM AUTHOR]

[Display omitted] Although lots of nanomaterials modified anodes have been reported to improve the bacterial attachment and extracellular electron transfer (EET) in microbial fuel cells (MFCs), the lack of a three dimensional (3D) conductive and capacitive network severely limited MFCs performance. In this work, 3D conductive networks derived from mucor mycelia were grown on carbon cloth (CC), and capacitive FeMn phosphides/oxides were further anchored on these 3D networks by electrochemical deposition (denoted as FeMn/CMM@CC) to simultaneously address the above challenges. As a result, the multivalent metal active sites were evenly distributed on 3D conductive network, which favored the enrichment of exoelectrogens, mass transport and EET. Consequently, the as-prepared FeMn/CMM@CC anode displayed accumulated charge of 131.4C/m2, higher than bare CC. Meanwhile, FeMn/CMM@CC anode substantially promoted flavin excretion and the amounts of nano conduits. The abundance of Geobacter was 63 % on bare CC, and greatly increased to 83 % on FeMn/CMM@CC. MFCs equipped by FeMn/CMM@CC anode presented the power density of 3.06 W/m2 and coulombic efficiency (29.9 %), evidently higher than bare CC (1.29 W/m2, 7.3 %), and the daily chemical oxygen demand (COD) removal amount also increased to 92.6 mg/L/d. This work developed a facile method to optimize the abiotic-biotic interface by introducing 3D conductive and capacitive network, which was proved to be a promising strategy to modify macro-porous electrodes. [ABSTRACT FROM AUTHOR]

The physiological role of Geobacter sulfurreducens extracellular cytochrome filaments is a matter of debate and the development of proposed electronic device applications of cytochrome filaments awaits methods for large‐scale cytochrome nanowire production. Functional studies in G. sulfurreducens are stymied by the broad diversity of redox‐active proteins on the outer cell surface and the redundancy and plasticity of extracellular electron transport routes. G. sulfurreducens is a poor chassis for producing cytochrome nanowires for electronics because of its slow, low‐yield, anaerobic growth. Here we report that filaments of the G. sulfurreducens cytochrome OmcS can be heterologously expressed in Shewanella oneidensis. Multiple lines of evidence demonstrated that a strain of S. oneidensis, expressing the G. sulfurreducens OmcS gene on a plasmid, localized OmcS on the outer cell surface. Atomic force microscopy revealed filaments with the unique morphology of OmcS filaments emanating from cells. Electron transfer to OmcS appeared to require a functional outer‐membrane porin‐cytochrome conduit. The results suggest that S. oneidensis, which grows rapidly to high culture densities under aerobic conditions, may be suitable for the development of a chassis for producing cytochrome nanowires for electronics applications and may also be a good model microbe for elucidating cytochrome filament function in anaerobic extracellular electron transfer. [ABSTRACT FROM AUTHOR]

Dark fermentation is a promising technology for biohydrogen (bioH 2) production. Augmenting the dark fermentation process with conductive materials has drawn significant research interests improving the bioH 2 yield. This study evaluated the feasibility of using carbon fiber brush (CB) as an additive to increase the efficiency of mesophilic dark fermentation using glucose as substrate and heat-shock pretreated anaerobic sludge as inoculum. Effect of electrode packing density on bioH 2 yield, soluble metabolite product formation, and substrate degradation was investigated. Maximal bioH 2 yield of 334.6 mL/g-glucose was obtained using medium-sized CB with 72.5% higher than the control (194 mL/g-glucose) and butyrate-type fermentation was the dominant metabolic pathway. Further, enhanced concentrations of proteins and polysaccharides on CB biofilm suggested the critical role of extracellular polymeric substances in microbial electron transfer. This was supported via electron transport activity in CB biofilm. Finally, microbial community analysis revealed a selective enrichment of the potentially electroactive Clostridium_sensu_stricto_1 on the CB (56.6%) compared to 40.9% abundance in control. Insights gained from this study demonstrated that CB aided dark fermentation can be an economical and environmentally benign strategy for improving bioH 2 yield. [Display omitted] • Carbon brush (CB) feasibility in dark fermentation was investigated. • Electrode packing density influenced bioH 2 yield and soluble metabolite products. • Highest bioH 2 yield of 334.6 mL/g-glucose was obtained with CB-medium. • CB enhanced extracellular polymeric secretion and electron transport activity. • Clostridium_sensu_stricto_1 was enriched on the CB surface. [ABSTRACT FROM AUTHOR]

Lactococcus lactis, a lactic acid bacterium used in food fermentations and commonly found in the human gut, is known to possess a fermentative metabolism. L. lactis, however, has been demonstrated to transfer metabolically generated electrons to external electron acceptors, a process termed extracellular electron transfer (EET). Here, we investigated an L. lactis mutant with an unusually high capacity for EET that was obtained in an adaptive laboratory evolution (ALE) experiment. First, we investigated how global gene expression had changed, and found that amino acid metabolism and nucleotide metabolism had been affected significantly. One of the most significantly upregulated genes encoded the NADH dehydrogenase NoxB. We found that this upregulation was due to a mutation in the promoter region of NoxB, which abolished carbon catabolite repression. A unique role of NoxB in EET could be attributed and it was directly verified, for the first time, that NoxB could support respiration in L. lactis. NoxB, was shown to be a novel type-II NADH dehydrogenase that is widely distributed among gut microorganisms. This work expands our understanding of EET in Gram-positive electroactive microorganisms and the special significance of a novel type-II NADH dehydrogenase in EET. [ABSTRACT FROM AUTHOR]

Microbial electrosynthesis (MES) is a promising technology that mainly utilizes microbial cells to convert CO2 into value-added chemicals using electrons provided by the cathode. However, the low electron transfer rate is a solid bottleneck hindering the further application of MES. Thus, as an effective strategy, genetic tools play a key role in MES for enhancing the electron transfer rate and diversity of production. We describe a set of genetic strategies based on fundamental characteristics and current successes and discuss their functional mechanisms in driving microbial electrocatalytic reactions to fully comprehend the roles and uses of genetic tools in MES. This paper also analyzes the process of nanomaterial application in extracellular electron transfer (EET). It provides a technique that combines nanomaterials and genetic tools to increase MES efficiency, because nanoparticles have a role in the production of functional genes in EET although genetic tools can subvert MES, it still has issues with difficult transformation and low expression levels. Genetic tools remain one of the most promising future strategies for advancing the MES process despite these challenges. [ABSTRACT FROM AUTHOR]

In this study, a constructed wetland was coupled with a microbial fuel cell to establish a coupled system known as the constructed wetland–microbial fuel cell (CW–MFC), utilized for the treatment of X-3B azo dye wastewater at varying concentrations. Experimental results indicated that the anodic region made the primary contributions to the discoloration of azo dyes and COD removal, with a contribution rate of 60.9–75.8% for COD removal and 57.8–83.0% for the effectiveness of discoloration. Additionally, the role of plants in the constructed wetland area could achieve the removal of small molecular substances and further discoloration. In comparison to open-circuit conditions, under closed-circuit conditions the CW–MFC effectively degraded X-3B azo dye wastewater. Under an external resistance of 2000 Ω, a maximum COD removal rate of 60.0% and a maximum discoloration rate of 85.8% were achieved for X-3B azo dye at a concentration of 100 mg/L. Improvements in the treatment efficiency of X-3B dye wastewater were achieved by altering the external resistance. Under an external resistance of 100 Ω and an influent concentration of X-3B of 800 mg/L, the COD removal rate reached 78.6%, and the decolorization rate reached 85.2%. At this point, the CW–MFC exhibited a maximum power density of 0.024 W/m3 and an internal resistance of 99.5 Ω. Spectral analysis and GC–MS results demonstrated the effective degradation of azo dyes within the system, indicating azo bond cleavage and the generation of numerous small molecular substances. Microbial analysis revealed the enrichment of electrogenic microorganisms under low external resistance conditions, where Geobacter and Trichococcus were dominant bacterial genera under an external resistance of 100 Ω, playing crucial roles in power generation and azo dye degradation within the system. [ABSTRACT FROM AUTHOR]

Electron shuttles are pivotal in enhancing the conversion efficiency at the biotic‐abiotic interface within microbial photoelectrochemical systems. Nevertheless, the potentially toxic effects of electron shuttles on microbial cells and their instability when exposed to light are often overlooked. This concept highlights the attention paid to electron shuttles in microbial photoelectrochemical systems, such as the cytotoxicity, the photoexcited state, and the photostability. Furthermore, it offers theoretical guidance for selecting electron shuttles to optimize sustainable energy production and environmental remediation. [ABSTRACT FROM AUTHOR]

• rGO was biosynthesized via the reduction of GO by Geobacter sulfurreducens PCA. • rGO-PCA combining system achieved complete nitrobenzene (NB) removal within one day. • rGO as electron shuttle accelerated the extracellular electron transfer (EET) process of Geobacter. • EET at microbial cell/rGO interface promoted the bioreduction of NB. • The underlying mechanism of NB biodegradation mediated by graphite material was proposed. In order to explore an efficient and green method to deal with nitrobenzene (NB) pollutant, reduced graphene oxide (rGO) as an electron shuttle was applied to enhance the extracellular electron transfer (EET) process of Geobacter sulfurreducens , which was a typical electrochemically active bacteria (EAB). In this study, rGO biosynthesis was achieved via the reduction of graphene oxide (GO) by G. sulfurreducens PCA within 3 days. Also, the rGO-PCA combining system completely reduced 50-200 µmol/L of NB to aniline as end product within one day. SEM characterization revealed that PCA cells were partly wrapped by rGO, and therefore the distance of electron transfer between strain PCA and rGO material was reduced. Beside, the I D /I G of GO, rGO, and rGO-PCA combining system were 0.990, 1.293 and 1.31, respectively. Moreover, highest currents were observed in rGO-PCA-NB as 12.950 µA/-12.560 µA at -408 mV/156 mV, attributing to the faster electron transfer efficiency in EET process. Therefore, the NB reduction was mainly due to: (I) direct EET process from G. sulfurreducens PCA to NB; (II) rGO served as electron shuttle and accelerated electron transfer to NB, which was the main degradation pathway. Overall, the biosynthesis of rGO via GO reduction by Geobacter promoted the NB removal process, which provided a facile strategy to alleviate the problematic nitroaromatic pollution in the environment. [Display omitted] [ABSTRACT FROM AUTHOR]

The microbial hybrid system modified by magnetic nanomaterials can enhance the interfacial electron transfer and energy conversion under the stimulation of a magnetic field. However, the bioelectrocatalytic performance of a hybrid system still needs to be improved, and the mechanism of magnetic field-induced bioelectrocatalytic enhancements is still unclear. In this work, γ-Fe2O3 magnetic nanoparticles were coated on a Shewanella putrefaciens CN32 cell surface and followed by placing in an electromagnetic field. The results showed that the electromagnetic field can greatly boost the extracellular electron transfer, and the oxidation peak current of CN32@γ-Fe2O3 increased to 2.24 times under an electromagnetic field. The enhancement mechanism is mainly due to the fact that the surface modified microorganism provides an elevated contact area for the high microbial catalytic activity of the outer cell membrane's cytochrome, while the magnetic nanoparticles provide a networked interface between the cytoplasm and the outer membrane for boosting the fast multidimensional electron transport path in the magnetic field. This work sheds fresh scientific light on the rational design of magnetic-field-coupled electroactive microorganisms and the fundamentals of an optimal interfacial structure for a fast electron transfer process toward an efficient bioenergy conversion. [ABSTRACT FROM AUTHOR]

Objectives: Evaluate electrochemically active biofilms as high energy density rechargeable microbial batteries toward providing persistent power in applications where traditional battery technology is limiting (, remote monitoring applications). Results: Here we demonstrated that an electrochemically active biofilm was able to store and release electrical charge for alternating charge/discharge cycles of up to 24 h periodicity (50% duty cycle) with no significant decrease in average current density (0.16 ± 0.04 A/m2) for over 600 days. However, operation at 24 h periodicity for > 50 days resulted in a sharp decrease in the current to nearly zero. This current crash was recoverable by decreasing the periodicity. Overall, the coulombic efficiency remained near unity within experimental error (102 ± 3%) for all of the tested cycling periods. Electrochemical characterization here suggests that electron transfer occurs through multiple routes, likely a mixture of direct and mediated mechanisms. Conclusions: These results indicate that bidirectional electrogenic/electrotrophic biofilms are capable of efficient charge storage/release over a wide range of cycling frequency and may eventually enable development of sustainable, high energy density rechargeable batteries. [ABSTRACT FROM AUTHOR]

Magnetotactic bacteria reside in sediments and stratified water columns. They are named after their ability to synthesize internal magnetic particles that allow them to align and swim along the Earth's magnetic field lines. Here, we show that two magnetotactic species, Magnetospirillum magneticum strain AMB-1 and Magnetospirillum gryphiswaldense strain MSR-1, are electroactive. Both M. magneticum and M. gryphiswaldense were able to generate current in microbial fuel cells with maximum power densities of 27 and 11 µW/m2, respectively. In the presence of the electron shuttle resazurin both species were able to reduce the crystalline iron oxide hematite (Fe2O3). In addition, M. magneticum could reduce poorly crystalline iron oxide (FeOOH). Our study adds M. magneticum and M. gryphiswaldense to the growing list of known electroactive bacteria, and implies that electroactivity might be common for bacteria within the Magnetospirillum genus. [ABSTRACT FROM AUTHOR]

ABSTRACT 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.

High salinity inhibits microbial activity in the bioremediation of saline wastewater. To alleviate osmotic stress, glycine betaine (GB), an osmoprotectant, is added to enhance the secretion of extracellular polymeric substances (EPS). These EPS are pivotal in withstanding environmental stressors, yet the intricate interplay between GB supplementation and microbial responses through EPS modifications—encompassing composition, molecular architecture, and electrochemical features—remains elusive in hypersaline conditions. Here we show microbial strategies for salinity endurance by investigating the impact of GB on the dynamic alterations of EPS properties. Our findings reveal that GB supplementation at 3.5% salinity elevates the total EPS (T-EPS) content from 12.50 ± 0.05 to 24.58 ± 0.96 mg per g dry cell weight. The observed shift in zeta potential from −28.95 to −6.25 mV at 0% and 3.5% salinity, respectively, with GB treatment, indicates a reduction in electrostatic repulsion and compaction. Notably, the EPS protein secondary structure transition from β-sheet to α-helix, with GB addition, signifies a more compact protein configuration, less susceptible to salinity fluctuations. Electrochemical analyses, including cyclic voltammetry (CV) and differential pulse voltammetry (DPV), reveal GB's role in promoting the release of exogenous electron shuttles, such as flavins and c-type cytochromes (c-Cyts). The enhancement in DPV peak areas (QDPV) with GB addition implies an increase in available extracellular electron transfer sites. This investigation advances our comprehension of microbial adaptation mechanisms to salinity through EPS modifications facilitated by GB in saline habitats.

Dark fermentation is considered a promising strategy for clean energy production, where hydrogen can directly be produced from organic residues in the role of functional microbes. However, the low efficiency and poor stability of dark fermentation systems inhibit them from moving to industrial-scale applications. Biochar, the carbonaceous additive, has demonstrated great potential in improving the fermentation performance of the systems resulting from its outstanding physicochemical properties. This review critically summarizes the recent advances in adding biochar to improve the performance of dark fermentation systems. The production methods and physicochemical properties of biochar, as well as their influencing factors during pyrolysis, are first discussed. Then, the promoting effect of biochar on hydrogen production and its underlying mechanisms are explained in detail. Finally, the concluding remarks and perspectives are proposed. This review will deepen the understanding on the specific mechanisms by which biochar enhances hydrogen production in dark fermentation systems. It can also provide a reference for future study to build better biochar-added systems with higher efficiency and more stable environment. •Biochar exhibits great application potential in enhancing biohydrogen production. • The achievements and challenges of biochar in dark fermentation systems are reviewed. • The correlation between hydrogen promotion and biochar properties is analyzed. • The underlying mechanisms of biochar in hydrogen promotion are elucidated. • The impact of biochar on EET in dark fermentation systems is intensively discussed. [ABSTRACT FROM AUTHOR]

In the context of the anaerobic ammonium oxidation process (anammox), great scientific advances have been made over the past two decades, making anammox a consolidated technology widely used worldwide for nitrogen removal from wastewaters. This review provides a detailed and comprehensive description of the anammox process, the microorganisms involved and their metabolism. In addition, recent research on the application of the anammox process with alternative electron acceptors is described, highlighting the biochemical reactions involved, its advantages and potential applications for specific wastewaters. An updated description is also given of studies reporting the ability of microorganisms to couple the anammox process to extracellular electron transfer to insoluble electron acceptors; particularly iron, carbon-based materials and electrodes in bioelectrochemical systems (BES). The latter, also referred to as anodic anammox, is a promising strategy to combine the ammonium removal from wastewater with bioelectricity production, which is discussed here in terms of its efficiency, economic feasibility, and energetic aspects. Therefore, the information provided in this review is relevant for future applications. [ABSTRACT FROM AUTHOR]

Two-dimensional nanostructured materials have emerged as very promising electrode materials for bioelectrochemical systems, enabling high bioelectrical output. We constructed air-cathode microbial fuel cells (MFCs) with electrical energy storage material POP M-TFP -rGO modified anodes for methyl orange (MO) decolorization and investigated the MFC performance. The kinetics of these processes were further analyzed. The results showed that MO decolorization efficiency in the POP M-TFP -rGO material-modified anode MFC reached 69.5% at 10 h. Compared with the unmodified electrode MFC, the efficiency and the kinetic constant of MO decolorization increased by 26.4% and 219%, respectively. Compared with control (40.5 mA m-2, 4.16 mW m-2), the stable current density and maximum power density (162.5 mA m-2, 48.84 mW m-2) were enhanced by 301% and 1070%, respectively. Further analysis of extracellular electron transfer kinetics showed that after modification of POP M-TFP -rGO, the number of electrons n and the charge transfer coefficient α (4, 0.56) increased by 74% and 65%, respectively, compared with control (2.3, 0.34). Therefore, POP M-TFP -rGO could be considered as a suitable and effective option for azo dye wastewater treatment by MFCs. [Display omitted] • Compared with CP only, POP M-TFP -rGO increased MO decolorization efficiency by 69.5%. • POP M-TFP -rGO improved the MO degradation kinetic constant by 219% than CP only. • POP M-TFP -rGO enhanced the maximum power density by 1070% than CP only. • POP M-TFP -rGO increased the kinetic constant of electron transfer process n and α. [ABSTRACT FROM AUTHOR]

Microbial fuel cell (MFC) performance is affected by the metabolic activity of bacteria and the extracellular electron transfer (EET) process. The deficiency of nanostructures on macroporous anode obstructs the enrichment of exoelectrogens and the EET. Herein, a N-doped carbon nanowire-modified macroporous carbon foam was prepared and served as an anode in MFCs. The anode has a hierarchical porous structure, which can solve the problem of biofilm blockage, ensure mass transport, favor exoelectrogen enrichment, and enhance the metabolic activity of bacteria. The microscopic morphology, spectroscopy, and electrochemical characterization of the anode confirm that carbon nanowires can penetrate biofilm, decrease charge resistance, and enhance long-distance electron transfer efficiency. In addition, pyrrolic N can effectively reduce the binding energy and electron transfer distance of bacterial outer membrane hemin. With this hierarchical anode, a maximum power density of 5.32 W/m3 was obtained, about 2.5-fold that of bare carbon cloth. The one-dimensional nanomaterial-modified macroporous anodes in this study are a promising strategy to improve the exoelectrogen enrichment and EET for MFCs. [ABSTRACT FROM AUTHOR]

The focus of this study is to develop a high-performance anode material for microbial fuel cells (MFCs). PEDOT:PSS and nitrogen-modified MXene were combined to create a hydrogel composite material called PPNM, which was drop-cast onto carbon felt (CF) as the MFCs anode. The PPNM exhibited a higher peak power density of 4.78 W m−2, an increase of 332% compared to the CF anode. It is worth noting that the PPNM Hydrogel maintains its rough and porous structure, providing favorable sites for bacterial colonization. The introduction of N-MXene has improved the electrochemical performance of the hydrogel, particularly impacting the mediated electron transfer process. Microbial community analysis revealed the presence of more electrochemically active species on the PPNM anode. These findings highlight the potential of PPNM hydrogel and pave the way for similar strategies in achieving high-performance anodes in MFCs. [ABSTRACT FROM AUTHOR]

Soil extracellular electron transfer (EET) is a pivotal biological process within the realm of soil. Unfortunately, EET suffers from a lack of predictive models. Herein, an intricately crafted machine learning model has been developed for the purpose of predicting soil EET by using the physicochemical properties of soil as independent input variables and the EET capabilities in terms of current density (jmax) and Coulombic charge (Cout) as dependent output variables. An autoencoder ensemble stacking (AES) model was developed to address the aforementioned issue by integrating support vector machine, multilayer perceptron, extreme gradient boosting, and light gradient boosting machine algorithms as the stacking algorithms. With 10-fold cross-validation, the AES model exhibited notable improvements in predicting jmax and Cout, with average test R2 values of 0.83 and 0.84, respectively, surpassing those of single machine learning (ML) models and the basic ensemble model. By utilizing partial correlation plots (PDPs), Shapley Additive explanations (SHAP) values, and SHAP decision plots, we quantitatively explained the impact and contribution of the input molecules on the AES model's predictions of jmax and Cout. In the context of the SHAP method for the AES model, total carbon (TC) was identified as the most correlated descriptor for jmax, while total organic carbon (TOC) stood out as the most relevant descriptor for Cout. In the prediction tasks of jmax and Cout within the AES model, employing a multitask ML approach allowed the model to benefit from the shared information of input variables, thereby enhancing its overall generalizability. This study provides a feasible tool for the prediction of soil EET from soil physiochemical properties and an advanced understanding of the relationship between soil physiochemical properties and EET capability. [ABSTRACT FROM AUTHOR]

Microbial fuel cells (MFCs) employing Pseudomonas putida B6-2 (ATCC BAA-2545) as an exoelectrogen have been developed to harness energy from various conventional substrates, such as acetate, lactate, glucose, and fructose. Owing to its metabolic versatility, P. putida B6-2 demonstrates adaptable growth rates on diverse, cost-effective carbon sources within MFCs, exhibiting distinct energy production characteristics. Notably, the anode chamber's pH rises with carboxylates' (acetate and lactate) consumption and decreases with carbohydrates' (glucose and fructose) utilization. The MFC utilizing fructose as a substrate achieved the highest power density at 411 mW m−2. Initial analysis revealed that P. putida B6-2 forms biofilms covered with nanowires, contributing to bioelectricity generation. These microbial nanowires are likely key players in direct extracellular electron transport through physical contact. This study established a robust foundation for producing valuable compounds and bioenergy from common substrates in bioelectrochemical systems (BESs) utilizing P. putida as an exoelectrogen.

The biogenic synthesis of silver nanoparticles (AgNPs) by microorganisms has been a subject of increasing attention. Despite extensive studies on this biosynthetic pathway, the mechanisms underlying the involvement of proteins and enzymes in AgNPs production have not been fully explored. Herein, we reported that Burkholderia contaminans ZCC was able to reduce Ag+ to AgNPs with a diameter of (10±5) nm inside the cell. Exposure of B. contaminans ZCC to Ag+ ions led to significant changes in the functional groups of cellular proteins, with approximately 5.72% of the (C-OH) bonds being converted to (C-C/C-H) (3.61%) and CO (2.11%) bonds, and 4.52% of the CO (carbonyl) bonds being converted to (C-OH) bonds. Furthermore, the presence of Ag+ and AgNPs induced the ability of extracellular electron transfer for ZCC cells via specific membrane proteins, but this did not occur in the absence of Ag+ ions. Proteomic analysis of the proteins and enzymes involved in heavy metal efflux systems, protein secretion system, oxidative phosphorylation, intracellular electron transfer chain, and glutathione metabolism suggests that glutathione S-transferase and ubiquinol-cytochrome c reductase iron-sulfur subunit play importance roles in the biosynthesis of AgNPs. These findings contribute to a deeper understanding of the functions exerted by glutathione S-transferase and ferredoxin-thioredoxin reductase iron-sulfur subunits in the biogenesis of AgNPs, thereby hold immense potential for optimizing biotechnological techniques aimed at enhancing the yield and purity of biosynthetic AgNPs.

ABSTRACTExtracellular cytochrome filaments are proposed to serve as conduits for long-range extracellular electron transfer. The primary functional physiological evidence has been the reported inhibition of Geobacter sulfurreducens Fe(III) oxide reduction when the gene for the filament-forming cytochrome OmcS is deleted. Here we report that the OmcS-deficient strain from that original report reduces Fe(III) oxide as well as the wild-type, as does a triple mutant in which the genes for the other known filament-forming cytochromes were also deleted. The triple cytochrome mutant displayed filaments with the same 3 nm diameter morphology and conductance as those produced by Escherichia coli heterologously expressing the G. sulfurreducens PilA pilin gene. Fe(III) oxide reduction was inhibited when the pilin gene in cytochrome-deficient mutants was modified to yield poorly conductive 3 nm diameter filaments. The results are consistent with the concept that 3 nm diameter electrically conductive pili (e-pili) are required for G. sulfurreducens long-range extracellular electron transfer. In contrast, rigorous physiological functional evidence is lacking for cytochrome filaments serving as conduits for long-range electron transport.IMPORTANCEUnraveling microbial extracellular electron transfer mechanisms has profound implications for environmental processes and advancing biological applications. This study on Geobacter sulfurreducens challenges prevailing beliefs on cytochrome filaments as crucial components thought to facilitate long-range electron transport. The discovery of an OmcS-deficient strain’s unexpected effectiveness in Fe(III) oxide reduction prompted a reevaluation of the key conduits for extracellular electron transfer. By exploring the impact of genetic modifications on G. sulfurreducens’ performance, this research sheds light on the importance of 3-nm diameter electrically conductive pili in Fe(III) oxide reduction. Reassessing these mechanisms is essential for uncovering the true drivers of extracellular electron transfer in microbial systems, offering insights that could revolutionize applications across diverse fields.