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Living bio-nano systems for artificial photosynthesis are of growing interest. Typically, these systems use photoinduced charge transfer to provide electrons for microbial metabolic processes, yielding a biosynthetic solar fuel. Here, we demonstrate an entirely different approach to constructing a living bio-nano system, in which electrogenic bacteria respire semiconductor nanoparticles to support nanoparticle photocatalysis. Semiconductor nanocrystals are highly active and robust photocatalysts for hydrogen (H2) evolution, but their use is hindered by the oxidative side of the reaction. In this system, Shewanella oneidensis MR-1 provides electrons to a CdSe nanocrystalline photocatalyst, enabling visible light-driven H2 production. Unlike microbial electrolysis cells, this system requires no external potential. Illuminating this system at 530 nm yields continuous H2 generation for 168 h, which can be lengthened further by replenishing bacterial nutrients. [ABSTRACT FROM AUTHOR]

Purpose: The extracellular electron transfer (EET) capability enables electroactive microorganisms have a wide range of applications in energy production, wastewater treatment, bioremediation and chemical synthesis. Because the oxygen levels will sharply affect the EET process, we integrated transcriptome changes under different oxygen levels with protein–protein interaction (PPI) network to study the specific changes under these conditions in a typical electroactive microorganism Shewanella oneidensis MR-1 (S. oneidensis MR-1). Methods: First, the mRNA expression data of S. oneidensis MR-1 under different oxygen levels were integrated into its PPI network to construct the active protein networks. Then, we studied the changes of key proteins and their interactions by comparative analysis of multiple pairs of active networks. Results: The analysis of node centrality and its changes in the active networks under high and low oxygen levels shows that most of the key nodes in the network are ribosomes or proteins closely related to ribosomes. The results of our centrality versus centrality change analysis of nodes show that the two proteins FlgB and PetA are the largest changed proteins. Among these proteins, the FlgB protein is the structural component of flagella, while PetA protein is closely related to cytochrome c. Furthermore, we also analyzed the changes in protein–protein interactions in the active networks under different oxygen levels, identified the key interactions in each pair of active networks, and finally screened out the key interaction Tig-RplX that exists in three pairs of active networks. Conclusion: Our results indicate that the translational processes of proteins and the corresponding translation efficiency may play an important role before and after the activation of the EET process of S. oneidensis MR-1. Furthermore, this study can also provide some guidance for identifying key proteins and interactions under different conditions for this species. [ABSTRACT FROM AUTHOR]

Simple Summary: In order to counter the increased deposition of greenhouse gases in the atmosphere, which has resulted in significant climatic changes, the production of alternative fuels to replace conventional fossil fuels has become necessary due to the rapidly diminishing concentration of fossil fuels and the rising global demand for energy. This investigation focused on two different samples that could be used as anolytes to produce energy in single- and double-chamber microbial fuel cells with a graphite electrode. In microbial fuel cells' energy production, the microbes consume organic substrates, use them in their metabolic processes, and produce valuable products that can be used as fuel to produce energy. The highest voltage outputs from the investigated bacterial strains were generated by strains A1 and A2, at 402 mV and 350 mV, respectively. Strain A6 of the ten different bacterial strains produced the least electricity, with a measurement of 35.03 mV. Natural resources are in short supply, and the ecosystem is being damaged as a result of the overuse of fossil fuels. The creation of novel technology is greatly desired for investigating renewable and sustainable energy sources. Microorganisms have received a lot of interest recently for their potential to transform organic waste into sustainable energy and high-value goods. New exoelectrogens that can transmit electrons to electrodes and remove specific wastewater contaminants are expected to be studied. In this study, we examined three distinct samples (as determined by chemical oxygen demand and pH) that can be used as anolytes to generate power in single-chamber and double-chamber microbial fuel cells using graphite electrodes. Wastewater from poultry farms was studied as an exoelectrogenic anolyte for microbial fuel cell power generation. The study examined 10 different bacterial strains, numbered A1 through A10. Due to their highly anticipated capacity to metabolize organic/inorganic chemicals, the diverse range of microorganisms found in poultry wastewater inspired us to investigate the viability of generating electricity using microbial fuel cells. From the investigated bacterial strains, the highest voltage outputs were produced by strains A1 (Lysinibacillus sphaericus) and A2 (Bacillus cereus), respectively, at 402 mV and 350 mV. Among the 10 different bacterial strains, strain A6 generated the least amount of electricity, measuring 35.03 mV. Furthermore, a maximum power density of 16.16 1.02 mW/m2 was achieved by the microbial fuel cell using strain A1, significantly outperforming the microbial fuel cell using a sterile medium. The strain A2 showed significant current and power densities of 35 1.12 mA/m2 and 12.25 1.05 mW/m2, respectively. Moreover, in the two representative strains, chemical oxygen demand removal and Coulombic efficiency were noted. Samples from the effluent anode chamber were taken in order to gauge the effectiveness of chemical oxygen demand removal. Wastewater had an initial chemical oxygen demand content of 350 mg/L on average. Strains A1 and A2 decomposed 94.28% and 91.71%, respectively, of the organic substrate, according to the chemical oxygen demand removal efficiency values after 72 h. Strains A1 and A2 had electron donor oxidation efficiencies for 72 h of 54.1% and 60.67%, respectively. The Coulombic efficiency increased as the chemical oxygen demand decreased, indicating greater microbial electroactivity. With representative strains A1 and A2, Coulombic efficiencies of 10% and 3.5%, respectively, were obtained in the microbial fuel cell. The findings of this study greatly advance the field as a viable source of power technology for alternative energy in the future, which is important given the depletion of natural resources. [ABSTRACT FROM AUTHOR]

Bioreduction of Cr(VI) is cost-effective and environmentally friendly, however, the slow bioreduction rate limits its application. In this study, the potential synergistic enhancement of Cr(VI) bioreduction by shewanella oneidensis MR-1 (S. oneidensis) with goethite and riboflavin (RF) was investigated. The results showed that the S. oneidensis reaction system reduce 29.2% of 20 mg/L Cr(VI) after 42 h reaction, while the S. oneidensis/goethite/RF reaction system increased the Cr(VI) reduction rate to 87.74%. RF as an efficient electron shuttle and Fe(II) from goethite bioreduction were identified as the crucial components in Cr(VI) reduction. XPS analysis showed that the final precipitates of Cr(VI) reduction were Cr(CH3C(O)CHC(O)CH3)3 and Cr2O3 and adhered to the bacterial cell surface. In this process, the microbial surface functional groups such as hydroxyl and carboxyl groups participated in the adsorption and reduction of Cr(VI). Meanwhile, an increase in cytochrome c led to an increase in electron transfer system activity (ETSA), causing a significant enhancement in extracellular electron transfer efficiency. This study provides insight into the mechanism of Cr(VI) reduction in a complex environment where microorganisms, iron minerals and RF coexist, and the synergistic treatment method of Fe(III) minerals and RF has great potential application for Cr(VI) detoxification in aqueous environment. [ABSTRACT FROM AUTHOR]

The terrestrial serpentinite-hosted ecosystem known as “The Cedars” is home to a diverse microbial community persisting under highly alkaline (pH ~ 12) and reducing (Eh magnetite > serpentinite ~ chromite ~ hematite. Genome analysis identified genes for flavin-mediated iron reduction and synthesis of a bacillibactin-like, catechol-type siderophore. Ali-BS5-314 is a Gram-negative, rod-shaped, mesophilic, facultative anaerobic alkaliphile that grows between pH 10–12 and temperatures 10–40°C, with limited growth observed 1–5% NaCl. Nitrate is used as a terminal electron acceptor under anaerobic conditions, which was corroborated by genome analysis. The Ali-BS5-314 genome also includes genes for benzoate-like compound metabolism. Anaero-CMMVII remained difficult to cultivate for physiological studies; however, growth was observed between pH 9–12, with the addition of 0.01–1% yeast extract. Anaero-CMMVII is a probable oxygen-tolerant anaerobic alkaliphile with hydrogenotrophic respiration coupled with nitrate reduction, as determined by genome analysis. Based on single-copy genes, ANI, AAI and dDDH analyses, Paeni-Cedars and Ali-BS5-314 are related to other species (P. glucanolyticus and A. aestuarii, respectively), and Anaero-CMMVII represents a new species. The characterization of these three isolates demonstrate the range of ecophysiological adaptations and metabolisms present in serpentinite-hosted ecosystems, including mineral reduction, alkaliphily, and siderophore production.

Substances inhibiting specific proteins involved in cellular electron transfer chains are used in biochemical research to investigate intracellular electron transfer routes and to redirect the electron fluxes. This also provides an in vivo approach to investigating the extracellular electron transfer (EET) mechanisms within and across biological membranes in bioelectrochemical research. However, the applicability of these specific inhibitors in electrochemical systems remains to be validated, particularly when aiming for (semi-)quantitative evaluation of the contribution of specific redox proteins to the EET. Here we conducted a systematic analysis of commonly used inhibitors and discovered several of them to be electrochemically active and thus capable of interfering with measurements in a bioelectrochemical reactor system in abiotic experiments. We also observed effects in vivo using a biophotovoltaics reactor with Synechocystis sp. PCC 6803 as a model system.

Abstract Direct interspecies electron transfer (DIET) may be most important in methanogenic environments, but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor. To better understand DIET with methanogens, the transcriptome of Geobacter metallireducens during DIET‐based growth with G. sulfurreducens reducing fumarate was compared with G. metallireducens grown in coculture with diverse Methanosarcina. The transcriptome of G. metallireducens cocultured with G. sulfurreducens was significantly different from those with Methanosarcina. Furthermore, the transcriptome of G. metallireducens grown with Methanosarcina barkeri, which lacks outer‐surface c‐type cytochromes, differed from those of G. metallireducens cocultured with M. acetivorans or M. subterranea, which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET. Differences in G. metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable. Cocultures with c‐type cytochrome deletion mutant strains, ∆Gmet_0930, ∆Gmet_0557 and ∆Gmet_2896, never became established with G. sulfurreducens but adapted to grow with all three Methanosarcina. Two porin–cytochrome complexes, PccF and PccG, were important for DIET; however, PccG was more important for growth with Methanosarcina. Unlike cocultures with G. sulfurreducens and M. acetivorans, electrically conductive pili were not needed for growth with M. barkeri. Shewanella oneidensis, another electroactive microbe with abundant outer‐surface c‐type cytochromes, did not grow via DIET. The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.

Abstract Purpose c-Type cytochromes are essential for extracellular electron transfer (EET) in electroactive microorganisms. The expression of appropriate c-type cytochromes is an important feature of these microorganisms in response to different extracellular electron acceptors. However, how these diverse c-type cytochromes are tightly regulated is still poorly understood. Methods In this study, we identified the high co-fitness genes that potentially work with different c-type cytochromes by using genome-wide co-fitness analysis. We also constructed and studied the co-fitness networks that composed of c-type cytochromes and the top 20 high co-fitness genes of them. Results We found that high co-fitness genes of c-type cytochromes were enriched in signal transduction processes in Shewanella oneidensis MR-1 cells. We then checked the top 20 co-fitness proteins for each of the 41 c-type cytochromes and identified the corresponding signal proteins for different c-type cytochromes. In particular, through the analysis of the high co-fitness signal protein for CymA, we further confirmed the cooperation between signal proteins and c-type cytochromes and identified a novel signal protein that is putatively involved in the regulation of CymA. In addition, we showed that these signal proteins form two signal transduction modules. Conclusion Taken together, these findings provide novel insights into the coordinated utilization of different c-type cytochromes under diverse conditions.

ABSTRACT Desulfovibrio vulgaris has been a primary pure culture sulfate reducer for developing microbial corrosion concepts. Multiple mechanisms for how it accepts electrons from Fe0 have been proposed. We investigated Fe0 oxidation with a mutant of D. vulgaris in which hydrogenase genes were deleted. The hydrogenase mutant grew as well as the parental strain with lactate as the electron donor, but unlike the parental strain, it was not able to grow on H2. The parental strain reduced sulfate with Fe0 as the sole electron donor, but the hydrogenase mutant did not. H2 accumulated over time in Fe0 cultures of the hydrogenase mutant and sterile controls but not in parental strain cultures. Sulfide stimulated H2 production in uninoculated controls apparently by both reacting with Fe0 to generate H2 and facilitating electron transfer from Fe0 to H+. Parental strain supernatants did not accelerate H2 production from Fe0, ruling out a role for extracellular hydrogenases. Previously proposed electron transfer between Fe0 and D. vulgaris via soluble electron shuttles was not evident. The hydrogenase mutant did not reduce sulfate in the presence of Fe0 and either riboflavin or anthraquinone-2,6-disulfonate, and these potential electron shuttles did not stimulate parental strain sulfate reduction with Fe0 as the electron donor. The results demonstrate that D. vulgaris primarily accepts electrons from Fe0 via H2 as an intermediary electron carrier. These findings clarify the interpretation of previous D. vulgaris corrosion studies and suggest that H2-mediated electron transfer is an important mechanism for iron corrosion under sulfate-reducing conditions. IMPORTANCE Microbial corrosion of iron in the presence of sulfate-reducing microorganisms is economically significant. There is substantial debate over how microbes accelerate iron corrosion. Tools for genetic manipulation have only been developed for a few Fe(III)-reducing and methanogenic microorganisms known to corrode iron and in each case those microbes were found to accept electrons from Fe0 via direct electron transfer. However, iron corrosion is often most intense in the presence of sulfate-reducing microbes. The finding that Desulfovibrio vulgaris relies on H2 to shuttle electrons between Fe0 and cells revives the concept, developed in some of the earliest studies on microbial corrosion, that sulfate reducers consumption of H2 is a major microbial corrosion mechanism. The results further emphasize that direct Fe0-to-microbe electron transfer has yet to be rigorously demonstrated in sulfate-reducing microbes.

Shewanella oneidensis is able to carry out extracellular electron transfer (EET), although its EET efficiency is largely limited by low flavin concentrations, poor biofilm forming‐ability, and weak biofilm conductivity. After identifying an important role for riboflavin (RF) in EET via in vitro experiments, the synthesis of RF is directed to 837.74 ± 11.42 µm in S. oneidensis. Molecular dynamics simulation reveals RF as a cofactor that binds strongly to the outer membrane cytochrome MtrC, which is correspondingly further overexpressed to enhance EET. Then the cell division inhibitor sulA, which dramatically enhanced the thickness and biomass of biofilm increased by 155% and 77%, respectively, is overexpressed. To reduce reaction overpotential due to biofilm thickness, a spider‐web‐like hybrid biofilm comprising RF, multiwalled carbon nanotubes (MWCNTs), and graphene oxide (GO) with adsorption‐optimized elongated S. oneidensis, achieve a 77.83‐fold increase in power (3736 mW m−2) relative to MR‐1 and dramatically reduce the charge‐transfer resistance and boosted biofilm electroactivity. This work provides an elegant paradigm to boost EET based on a synthetic biology strategy and materials science strategy, opens up further opportunities for other electrogenic bacteria. [ABSTRACT FROM AUTHOR]

Geobacter sulfurreducens is part of a specialized group of microbes with the unique ability to exchange electrons with insoluble materials, such as iron oxides and electrodes. Therefore, G. sulfurreducens plays an essential role in the biogeochemical iron cycle and microbial electrochemical systems. In G. sulfurreducens this ability is primarily dependent on electrically conductive nanowires that link internal electron flow from metabolism to solid electron acceptors in the extracellular environment. Here we show that when carrying conjugative plasmids, which are self-transmissible plasmids that are ubiquitous in environmental bacteria, G. sulfurreducens reduces insoluble iron oxides at much slower rates. This was the case for all three conjugative plasmids tested (pKJK5, RP4 and pB10). Growth with electron acceptors that do not require expression of nanowires was, on the other hand, unaffected. Furthermore, iron oxide reduction was also inhibited in Geobacter chapellei, but not in Shewanella oneidensis where electron export is nanowire-independent. As determined by transcriptomics, presence of pKJK5 reduces transcription of several genes that have been shown to be implicated in extracellular electron transfer in G. sulfurreducens, including pilA and omcE. These results suggest that conjugative plasmids can in fact be very disadvantageous for the bacterial host by imposing specific phenotypic changes, and that these plasmids may contribute to shaping the microbial composition in electrode-respiring biofilms in microbial electrochemical reactors. [ABSTRACT FROM AUTHOR]

Microbial graphene oxide reduction is a developing method that serves to reduce both production costs and environmental impact in the synthesis of graphene. This study demonstrates microbial graphene oxide reduction using Shewanella decolorationis NTOU1 under neutral and mild conditions (pH = 7, 35 °C, and 1 atm). Graphene oxide (GO) prepared via the modified Hummers' method is used as the sole solid electron acceptor, and the characteristics of reduced GO (rGO) are investigated. According to electron microscopic images, the surface structure of GO was clearly changed from smooth to wrinkled after reduction, and whole cells were observed to be wrapped by GO/rGO films. Distinctive appendages on the cells, similar to nanowires or flagella, were also observed. With regard to chemical-bonding changes, after a 24-h reaction of 1 mg mL−1, GO was reduced to rGO, the C/O increased from 1.4 to 3.0, and the oxygen-containing functional groups of rGO were significantly reduced. During the GO reduction process, the number of S. decolorationis NTOU1 cells decreased from 1.65 × 108 to 1.03 × 106 CFU mL−1, indicating the bactericide effects of GO/rGO. In experiments adding consistent concentrations of initial bacteria and lactate, it was shown that with the increase of GO additions (0.5–5.0 mg mL−1), the first-order reaction rate constants (k) of lactate metabolism and acetate production increased accordingly; in experiments adding consistent concentrations of initial bacteria and GO but different lactate levels (1 to 10 mM), the k values of lactate metabolism did not change significantly. The test results of adding different electron transfer mediators showed that riboflavin and potassium ferricyanide were able to boost GO reduction, whereas 2,6-dimethoxy-1,4-benzoquinone and 2,6-dimethyl benzoquinone completely eliminated bacterial activity. [ABSTRACT FROM AUTHOR]

The mechanism behind how flavin mononucleotide (FMN)-producing bacteria attach to a host intestine remains unclear. In order to address this issue, this study isolated the Gram-positive bacteria Lactobacillus plantarum from Mongolian fermented Airag, named L. plantarum MA. These bacteria were further employed as the model microbes, and their electrogenic properties were first identified by their significant expression of type II NADH-quinone oxidoreductase. This study also demonstrated that the electrical activity of L. plantarum MA can be conducted through flavin mononucleotide (FMN)-based extracellular electron transfer, which is highly dependent on the presence of a carbon source in the medium. Our data show that approximately 15 µM of FMN, one of the key electron donors for the generation of electricity, can be produced from L. plantarum MA, as they were cultured in the presence of lactulose for 24 h. We further demonstrated that the electrical activity of L. plantarum MA can promote microbial adhesion and can thus enhance the colonization effectiveness of Caco-2 cells and mouse cecum. Such enhanced adhesiveness was attributed to the increased expression of type I collagens in the intestinal epithelium after treatment with L. plantarum MA. This study reveals the mechanism behind the electrogenic activity of L. plantarum MA and shows how the bacteria utilize electricity to modulate the protein expression of gut tissue for an enhanced adhesion process. [ABSTRACT FROM AUTHOR]

Extracellular electron transfer (EET) is a process that microorganisms use to reduce or oxidize external insoluble electron acceptors or donors. Much of our mechanistic understanding of this process is derived from studies of transmembrane cytochrome complexes and extracellular redox shuttles that mediate outward EET to anodes and external electron acceptors. In contrast, there are knowledge gaps concerning the reverse process of inward EET from external electron donors to cells. Here, we describe a role for soluble iron (exogenous FeCl2) in enhancing EET from cathodes to the model EET bacterium Shewanella oneidensis MR‐1, with fumarate serving as the intracellular electron acceptor. This iron concentration‐dependent electron uptake was eradicated upon addition of an iron chelator and occurred only in the presence of fumarate reductase, confirming an electron pathway from cathodes to this periplasmic enzyme. Moreover, S. oneidensis mutants lacking specific outer membrane and periplasmic cytochromes exhibited significantly decreased current levels relative to wild‐type. These results indicate that soluble iron can function as an electron carrier to the EET machinery of S. oneidensis. [ABSTRACT FROM AUTHOR]

During extracellular electron transfer, Geobacter sulfurreducens constitutively expresses the bc‐cytochrome CbcL, yet cells containing only this menaquinone oxidase fail to respire above −0.1 V vs. SHE. By identifying mutations within cbcL that permit growth at higher potentials, we provide evidence that this cytochrome is regulated by redox potential. Strains expressing only CbcLV205A, CbcLV205G, and CbcLF525Y were capable of growth with high potential electron acceptors including Fe(III) citrate, Mn(IV) oxides, and electrodes poised at +0.1 V vs. SHE. Electrochemical characterization of wild type CbcL revealed oxidative inactivation of electron transfer above −0.1 V, while CbcLV205A, CbcLV205G, and CbcLF525Y remained active. Growth yields of CbcLV205A, CbcLV205G, and CbcLF525Y were only 50 % of WT, consistent with CbcL‐dependent electron transfer conserving less energy. These data support the hypothesis that CbcL has evolved to rapidly shut off in response to redox potential, in order to divert electrons to higher yield oxidases that coexist in the Geobacter membrane. [ABSTRACT FROM AUTHOR]

Carbon source starvation can promote steel corrosion in the presence of a pure culture through extracellular electron transfer (EET). However, the impact of carbon source starvation on corrosion induced by mixed strains is still unknown. This work investigated the impact of carbon source starvation on EH40 steel corrosion in the presence of Desulfovibrio vulgaris and Pseudomonas aeruginosa, typical species of sulfate- and nitrate-reducing bacteria. It was found that the impact of carbon source starvation on corrosion depended on nitrate addition. When nitrate (5 g∙L−1 NaNO3) was not added, the corrosion was promoted by carbon source starvation. However, the corrosion was initially promoted by carbon source starvation, but later inhibited with nitrate addition. The corrosion behaviors in different systems were closely related to different numbers of the strains in biofilms and their metabolic activities, and the mechanisms were revealed. [ABSTRACT FROM AUTHOR]

This review focuses on the development of microbial biofuel cells to demonstrate how similar principles apply to the development of bioelectronic devices. The low specificity of microorganism-based amperometric biosensors can be exploited in designing microbial biofuel cells, enabling them to consume a broader range of chemical fuels. Charge transfer efficiency is among the most challenging and critical issues while developing biofuel cells. Nanomaterials and particular redox mediators are exploited to facilitate charge transfer between biomaterials and biofuel cell electrodes. The application of conductive polymers (CPs) can improve the efficiency of biofuel cells while CPs are well-suitable for the immobilization of enzymes, and in some specific circumstances, CPs can facilitate charge transfer. Moreover, biocompatibility is an important issue during the development of implantable biofuel cells. Therefore, biocompatibility-related aspects of conducting polymers with microorganisms are discussed in this review. Ways to modify cell-wall/membrane and to improve charge transfer efficiency and suitability for biofuel cell design are outlined. [ABSTRACT FROM AUTHOR]

[Display omitted] The low power density originating from poor electroactive bacteria (EAB) adhesion and sluggish extracellular electron transfer (EET) at the anode interface, is a major impediment preventing the practical implementation of microbial fuel cells (MFCs). Tailoring the surface properties of anodes is an effective and powerful strategy for addressing this issue. In this study, we successfully fabricated an efficient anode electrocatalyst, consisting of carbon nanotubes encapsulating iron disulfide (FeS 2 @CNT) micropolyhedrons, using simple hydrothermal and freeze-drying methods, which not only strengthened the anode interaction with EAB but also promoted the EET process at the anode interface. As expected, the MFCs with a FeS 2 @CNT anode yielded an outstanding power density of 1914 mW m−2 at a current density of 4350 mA m−2, which significantly exceeded those of pure CNT (1096.2 mW m−2, 2703.3 mA m−2) and carbon cloth (426.8 mW m−2, 965.6 mA m−2) anodes. The high-power output can be attributed to the synergistic effect between FeS 2 and CNTs, endowing the anode with biocompatibility for biofilm adhesion and colonization, nutrient diffusion, and the presence of abundant Fe and S active sites for EET mediation. Owing to the low cost, facile fabrication process, and excellent electrocatalytic performance toward the redox reactions in biofilms, the synthesized FeS 2 @CNT electrocatalyst is a promising material for high-performance and cost-effective MFCs with commercial applications. [ABSTRACT FROM AUTHOR]

Improving the bacterial loading contents and extracellular electron transfer (EET) efficiency are promising strategies to enhance the overall performance of microbial fuel cells (MFCs). Herein, an aligned carbon nanofiber-bacteria (ACNF-bacteria) hybrid is prepared and used as an efficient anode employing the electrospinning and co-filtration method. The ACNF matrix reveals rich porous structure, large specific surface area, and obviously improved electrocatalytic performance compared to the pristine carbon cloth (CC) anode. Specifically, MFC with the ACNF-bacteria anode achieves a maximum power density of 704 mW m−2, which is 1.7, 2.1 times higher than that of ACNF, CNF and CC anode. Impressively, the coulombic efficiency and chemical oxygen demand (COD) removal rate for ACNF-bacteria anode reach 47.4% and 87.9%, respectively, far exceeding the CC anode (29.3% and 47.6%). Considering all the factors of low cost, facial fabrication, and high-power output, this novel ACNF-bacteria anode provides great potential for high-performance and cost-effective MFCs in a large scale. • Electrospun aligned carbon nanofibers boosted electricity generation of MFC. • Suction filtration method enhanced bacterial loadings. • ACNF-bacteria composite anode possesses rapid EET efficiency. [ABSTRACT FROM AUTHOR]

Microbial fuel cell (MFC) is a promising approach that could utilize microorganisms to oxidize biodegradable pollutants in wastewater and generate electrical power simultaneously. Introducing advanced anode nanomaterials is generally considered as an effective way to enhance MFC performance by increasing bacterial adhesion and facilitating extracellular electron transfer (EET). This review focuses on the key advances of recent anode modification materials, as well as the current understanding of the microbial EET process occurring at the bacteria‐electrode interface. Based on the difference in combination mode of the exoelectrogens and nanomaterials, anode surface modification, hybrid biofilm construction and single‐bacterial surface modification strategies are elucidated exhaustively. The inherent mechanisms may help to break through the performance output bottleneck of MFCs by rational design of EET‐related nanomaterials, and lead to the widespread application of microbial electrochemical systems. [ABSTRACT FROM AUTHOR]

Lactiplantibacillus plantarum is a lactic acid bacterium that is commonly found in the human gut and fermented food products. Despite its overwhelmingly fermentative metabolism, this microbe can perform extracellular electron transfer (EET) when provided with an exogenous quinone, 1,4-dihydroxy-2-naphthoic acid (DHNA), and riboflavin. However, the separate roles of DHNA and riboflavin in EET in L. plantarum have remained unclear. Here, we seek to understand the role of quinones and flavins in EET by monitoring iron and anode reduction in the presence and absence of these small molecules. We found that addition of either DHNA or riboflavin can support robust iron reduction, indicating electron transfer to extracellular iron occurs through both flavin-dependent and DHNA-dependent routes. Using genetic mutants of L. plantarum, we found that flavin-dependent iron reduction requires Ndh2 and EetA, while DHNA-dependent iron reduction largely relies on Ndh2 and PplA. In contrast to iron reduction, DHNA-containing medium supported more robust anode reduction than riboflavin-containing medium, suggesting electron transfer to an anode proceeds most efficiently through the DHNA-dependent pathway. Furthermore, we found that flavin-dependent anode reduction requires EetA, Ndh2, and PplA, while DHNA-dependent anode reduction requires Ndh2 and PplA. Taken together, we identify multiple EET routes utilized by L. plantarum and show that the EET route depends on access to environmental biomolecules and on the electron acceptor. This work expands our molecular-level understanding of EET in Gram-positive microbes and provides additional opportunities to manipulate EET for biotechnology. [ABSTRACT FROM AUTHOR]

The mutual benefits between electroactive bacteria (EAB) and non-EAB promote bioenergy recovery in microbial fuel cell (MFC), while competition between functional and non-functional bacteria on electrodes hinders further performance improvement. Introducing non-electrode conductive carriers into MFC may reduce contact resistance and alleviate interspecies competition. In this study, three-dimensional spherical biochar (BC) granules as independent microbial and charge carriers are introduced into anode chamber of an MFC without direct contact with the electrode (carbon cloth, CC) creating CC-BC-MFC. CC-BC-MFC achieves superior power density of 15 ± 1.4 W m−3 compared to CC-MFC (12 ± 1.4 W m−3) and CC/BC-MFC (10 ± 1.2 W m−3, where BC directly contacts CC). Higher effluent quality highlights the outstanding contribution of adequate biomass in effective containment treatment. Smaller internal resistance and higher coulombic efficiency are obtained in CC-BC-MFC by relieving microbial competition at anode. Syntrophic growth between Geobacter (69.4 %) and Methanosarcina (98 %) is established on BC in CC-BC-MFC, with high expression of Geobacter -associated pili genes and acetotrophic and CO 2 -reducing methanogenic genes associated with methanogens. Therefore, incorporating BC as flexible and independent charge carriers can reduce internal resistance, provide adequate habitat, alleviate microbial competition at anode, and offer a feasible strategy for MFC in wastewater treatment. [Display omitted] • Biochar granules as the independent charge carriers was introduced into MFC. • The composite MFC exhibited higher power generation and COD removal rate. • Introducing independent carriers help to circumvent the extra contact resistance. • Granular charge carriers can alleviate competition for ecological niches at anode. • The syntrophic growth between Geobacter and methanogens was established on biochar. [ABSTRACT FROM AUTHOR]

Tetramethylammonium hydroxide (TMAH), an extensively utilized photoresist developer, is frequently present in ammonium-rich wastewater from semiconductor manufacturing, and its substantial ecotoxicity should not be underestimated. This study systematically investigated the effects of TMAH on the anammox granular sludge (AnGS) system and elucidated its inhibitory mechanisms. The results demonstrated that the median inhibitory concentration of TMAH for anammox was 84.85 mg/L. The nitrogen removal performance of the system was significantly decreased after long-term exposure to TMAH (0–200 mg/L) for 30 days (p < 0.05), but it showed adaptability to certain concentrations (≤50 mg/L). Concurrently, the stability of the granules decreased dramatically, resulting in the breakdown of AnGS. Further investigations indicated that TMAH exposure increased the secretion of extracellular polymeric substances but weakened their defense function. The increase in reactive oxygen species resulted in damage to the cell membrane. Reduced activity of anammox bacteria, impeded electron transfer, and changes in enzyme activity suggested that TMAH affected the metabolic activity. Microbiological analysis revealed that TMAH caused a decrease in the abundance of anammox bacteria and a weakening of symbiotic interactions within the microbial community. These results provide valuable guidance for the AnGS system application in chip wastewater treatment. [Display omitted] • The effects of TMAH on the AnGS system were first studied. • IC 50 of TMAH was detected to be 84.85 mg/L. • EPS secretion is a key strategy of the AnGS system to cope with TMAH stress. • TMAH impaired extracellular electron transport ability of the AnGS system. • The AnGS system showed an adaptive trend when TMAH ≤50 mg/L. [ABSTRACT FROM AUTHOR]

Electrochemically active biofilms (EABs) for nitrite detection have high specificity, rapid response, operational simplicity, and extended lifespan advantages. However, their scale production remains challenging due to time-consuming and uniform preparation. In this study, a novel approach was proposed to fast fabricate an EAB biosensor with a synthetic biofilm electrode for nitrite detection. The biofilm electrode was prepared by coating bioinks with varying conductive materials onto the surface of the graphite sheets, showing short incubation time and good reproducibility. Incorporating conductive materials into the bioinks remarkably enhanced the maximum voltage of the first cycle of bioelectrode incubation, with an increase of up to 633% for carbon nanofibers. The nitrite reduction current was amplified by a factor of 2.97, due to the enhancement of extracellular electron transfer (EET). The developed nitrite biosensor exhibited a detection range of 0.1–15 mg NO 2 --N L−1, with a high sensitivity of 610.8 μA mM−1 cm−2, and a stabilization operation time of at least 280 cycles. This study not only provided valuable insights into conductive materials for synthetic biofilms but also presented a practical approach for the rapid preparation, scale production, and optimization of highly sensitive and stable EAB sensors. [Display omitted] • Bioelectrode was prepared by coating conductive material contained bioink. • Coated bioelectrode showed short preparation time and improved repeatability. • Carbon nanofiber contained biofilm showed a porous and 3D network structure. • 3D network structure enhanced electron transfer and matter transfer. • The bioelectrode showed highly sensitive and stable detection to nitrite. [ABSTRACT FROM AUTHOR]

Radionuclide contamination has become a global environmental concern due to the high mobility and toxicity of certain isotopes. Bioreduction mediated by electrochemically active bacteria (EAB) with unique extracellular electron transfer (EET) capability is recognized as a promising approach for nuclear waste treatment. However, it is difficult to achieve bidirectional regulation of EET pathway through traditional genetic manipulation, limiting the bioremediation application of EAB. Here, we designed and optimized a novel Esa quorum sensing (EQS) system for highly efficient gene expression and interleaved cellular functional output. By promoting dimethyl sulfoxide reductase at low cell density and increasing the synthesis of electron conductive complex and flavins at high cell density, the EQS system dynamically switched the extracellular respiratory pathway of Shewanella oneidensis MR-1 according to cell density. The engineered strain exhibited precise switching and substantial improvement in the extracellular remediation of multiple nuclides, sequentially increasing the reduction of iodine IO 3 - and uranium U(VI) by 2.51- and 2.05-fold compared with the control, respectively. Furthermore, a mobile bacterial biofilm material was fabricated for collecting uranium precipitates coupled with U(VI) reduction. This work clearly demonstrates that EQS system contributes to the bidirectional regulation of EET pathway in EAB, providing an effective and refined strategy for bioremediation of multi-nuclide contamination. [Display omitted] • An EQS system was constructed for efficient gene expression and functional output. • Transmembrane pathway of S. oneidensis was dynamically controlled by EQS system. • The EQS system substantially promoted extracellular reduction of IO 3 - and U(VI). • Uranium precipitate was recovered along with U(VI) reduction by a biofilm material. [ABSTRACT FROM AUTHOR]

Antibiotics often co-exist with other pollutants, posing a significant threat to ecosystems. This review first examines the applications and limitations of microbial treatments for various types of antibiotic-combined pollutants. Then, it explores the mechanisms and application of hybrid systems that integrate advanced oxidation with microbial treatment, categorized into two-stage and intimately hybrid systems. Finally, the review highlights key knowledge gaps in hybrid systems and provides new insight into the removal of combined pollutants. [Display omitted] • Advanced oxidation enhanced the influent's bioavailability to elevate combined pollutant removal in a two-stage hybrid system. • Electron transfer between microorganisms and the catalyst enhances intimately hybrid system performance. • Antibiotic-resistance genes can be reduced through both direct and indirect pathways in hybrid systems. [ABSTRACT FROM AUTHOR]

Enhancing extracellular electron transfer (EET) efficiency is crucial for improving the anaerobic digestion (AD) system's capability to treat recalcitrant wastewater. In this study, a novel S, N co-doped biochar (S-N-BC) was prepared through surface engineering to optimize EET within AD systems. The addition of S-N-BC significantly enhanced the performance of a mesophilic AD system treating Congo red wastewater, increasing the decolorization rate by 78 %, COD degradation rate by 82 %, and methane yield by 87 % compared to the control. Additionally, the shock resistance of anaerobic granular sludge was improved, as evidenced by the formation of the protective extracellular polymeric substances (EPS) barrier and the enhanced activities of the electron transport system. Mechanistic analysis revealed that adding S-N-BC did not alter the Congo red decolorization pathway but significantly enriched various electrochemically active bacteria and established EET pathways between microbial-pollutant and inter-microbial. This significantly accelerated EET efficiency within the AD system, ensuring stable and efficient operation under challenging conditions. This study proposed a novel approach using S-N-BC to simultaneously enhance "dual-pathway EET" between microbial-pollutant and inter-microbial while constructing an EPS protective barrier, addressing the issues of low efficiency and fragile stability of AD systems for treating recalcitrant wastewater. [Display omitted] • Performance of AD system in treating CR was greatly enhanced by S-N-BC. • Microbial-pollutant and inter-microbial electron transfer were enhanced by S-N-BC. • An EPS protective barrier of microorganisms was constructed by S-N-BC. [ABSTRACT FROM AUTHOR]

[Display omitted] • Augmented electroactive microorganisms facilitate extracellular electron transfer. • Extracellular electron transfer enhances organic stabilization and methane output. • Extracellular electron transfer increases resilience to kinetic imbalances. • Augmented electroactive microorganisms ensure stable, high-performance operation. The performance and stability of a bioelectrochemical anaerobic digester (BeAD), continuously augmented with electroactive microorganisms (EAMs), were investigated. The BeAD showcased superior performance, sustaining the high COD removal efficiency and methane production rate of 76.5 % and 0.67 L/(L.d), respectively, in a stable state. Prominently, it exhibited remarkable resilience under hydraulic and organic shock loads, adeptly recuperating from disturbances up to 1000 % of its stable condition. This resilience of up to 300 % shock load was driven by increased levels of electron transport components such as quinones and riboflavins, which act as electron shuttles. However, after extreme shock exposures from 500 % to 1000 %, despite the spike in inhibitory by-products such as humic acids and ammonia, the upregulation of the mtr complex was pivotal in recovering and sustaining methane production. These insights emphasize the BeAD's capability to bolster both performance and stability, thereby providing a potent strategy for practical application of bioelectrochemical systems. [ABSTRACT FROM AUTHOR]

[Display omitted] • Electric wiring between bacteria and an electrode via redox polymer was fabricated. • Electric wiring was established even for non-electroactive bacteria. • Selective measurement of wired bacteria was achieved in a bacterial suspension. Non-electroactive bacteria (n-EAB), constituting the majority of known bacteria to date, have been underutilized in electrochemical conversion technologies due to their lack of direct electron transfer to electrodes. In this study, we established an electric wiring between n-EAB (gram-positive Bacillus subtilis and gram-negative Escherichia coli) and an extracellular electrode via a ferrocene-polyethyleneimine-based redox polymer (Fc-PEI). Chronoamperometry recordings indicated that Fc-PEI can transfer intracellular electrons to the extracellular electrode regardless of the molecular organization of PEI (linear or branched) and the membrane structure of bacteria (gram-positive or -negative). As fluorescence staining suggested, Fc-PEI improves the permeability of the bacterial cell membrane, enabling electron carriers in the cell to react with Fc. In addition, experiments with Fc-immobilized electrodes without PEI suggested the existence of an alternative electron transfer pathway from B. subtilis to the extracellular Fc adsorbed onto the cell membrane. Furthermore, we proposed for the first time that the bacteria/Fc-linear PEI modified structure enables selective measurement of immobilized bacterial activity by physically blocking contact between the electrode surface and planktonic cells co-existing in the surrounding media. Such electrodes can be a powerful analytical tool for elucidating the metabolic activities of specific bacteria wired to the electrode even within complex bacterial communities. [ABSTRACT FROM AUTHOR]

• PDA with ample functional groups and 3D macroporous GA were used to modify anode. • PGA/CB enhanced electrochemical activity, hydrophilicity and microbial adhesion. • PGA/CB enriched exoelectrogens, promoted riboflavin secretion and EET efficiency. • PGA/CB-MFCs achieved superior power output due to the synergy between GA and PDA. The structure and surface physicochemical properties of anode play a crucial role in microbial fuel cells (MFCs). To enhance the enrichment of exoelectrogen and facilitate extracellular electron transfer (EET), a three-dimensional macroporous graphene aerogel with polydopamine coating was successfully introduced to modify carbon brush (PGA/CB). The three-dimensional graphene aerogel (GA) with micrometer pores improved the space utilization efficiency of microorganisms. Polydopamine (PDA) coating enhanced the physicochemical properties of the electrode surface by introducing abundant functional groups and nitrogen-containing active sites. MFCs equipped with PGA/CB anodes (PGA/CB-MFCs) demonstrated superior power generation compared to GA/CB-MFCs and CB-MFCs (MFCs with GA/CB and CB anodes respectively), including a 23.0 % and 30.1 % reduction in start-up time, and an increase in maximum power density by 2.43 and 1.24 times respectively. The higher bioelectrochemical activity exhibited by the biofilm of PGA/CB anode and the promoted riboflavin secretion by PGA modification imply the enhanced EET efficiency. 16S rRNA high-throughput sequence analysis of the biofilms revealed successful enrichment of Geobacter on PGA/CB anodes. These findings not only validate the positive impact of the synergistic effects between GA and PDA in promoting EET and improving MFC performance but also provide valuable insights for electrode design in other bioelectrochemical systems. [ABSTRACT FROM AUTHOR]

Persistent free radicals (PFRs) in biochar are an important electron shuttle for mediating electron transfer, which has significant impact on the biogeochemical redox reactions. Although the influence of biochar on the extracellular electron transfer (EET) for redox cycle has been extensively studied, the molecular mechanism for promoting the EET with PFRs remains poorly understood. This study investigated the oxygen-centered PFRs-mediated Cr(VI) reduction by Shewanella oneidensis MR-1 (MR-1) and exhibited the molecular mechanism of electron transfer between flavin substances and PFRs. Results showed that the Cr(VI) bioreduction rate by MR-1 increased from 31% to 70% with the addition of biochar. Electrochemical results illustrated that biochar increased biocurrent generation in the Cr(VI) bioreduction process. 3D-EEM and LC/MS spectra indicated that MR-1 secreted the flavin mononucleotide (FMN) reductase that relied on the [H] to provide the electrons. Electron paramagnetic resonance spectra illustrated that PFRs in biochar accepted the electrons from FMN reductase and stored those bioelectrons. Because of the oxidation of FMN, the electron transfer from FMN reductase to PFRs would increase the intracellular reactive oxygen species, which further produced the extracellular ·O 2 −. The reduced PFRs released the bioelectrons, accelerating the Cr(VI) reduction by ·O 2 −. Together with the results of the mutant strains experiment, it was found that the EET by c -cytochrome and free radicals contributed to the Cr(VI) bioreduction by 7.1% and 92.9%, respectively. These findings revealed that the PFRs could participate in the EET process and promote the redox reactions, providing a new approach for enhancing the remediation of heavy metal pollution by microorganisms and suggesting the important role of PFRs in the electron transfer process. [Display omitted] • PFRs function as the electron shuttle to enhance the EET by MR-1. • FMN reductase was the predominant substance that sustained the EET. • The PFRs-mediated EET process upregulated the intracellular ROS level. • ·O 2 − generation by MR-1 cells promoted the Cr(VI) bioreduction under anaerobic. [ABSTRACT FROM AUTHOR]

• RR greatly enhanced extracellular electron transfer of Geobacter anode biofilm. • RR specifically interacted with the outer membrane tetraheme cytochrome OmcE. • OmcE-RR interaction favored RR intension and electron transfer within biofilm. • OmcE-RR complex might accept electrons from OmbB-OmaB-OmcB porin. Bioelectrochemical system (BES) is a unique biotechnology for wastewater treatment and energy recovery, and extracellular electron transfer (EET) between microbe and electrode is the key to optimize the performance of BESs. Resazurin is an effective artificial compound that can promote EET in BESs, but the way how it transports electrons is not fully understood. In this study differential pulse voltammetry revealed that the redox potential of resorufin (RR) (intermediate of resazurin reduction, actual electron mediator) within Geobacter sulfurreducens biofilm was positively shifted by 100 mV than that of free RR, and this shift was attenuated by the mutation of outer-membrane cytochrome gene omcE but not by omcS and omcZ mutation, indicating that RR specifically interacted with OmcE. By using heterologously expressed OmcE monomers in Escherichia coli , it was found that RR bonded with OmcE monomers with a moderate intensity (dissociation constant of 720 nM), and their interaction obviously increased the content of α helix in OmcE monomers. Biomolecular analysis indicated that heme II of OmcE monomer might be the binding site for RR (binding energy of -7.01 kJ/mol), which were favorable for electron transfer within OmcE-RR complex. Comparative transcriptomics showed that RZ addition significantly upregulated the expression of omcE , periplasmic cytochrome gene ppcB , and outer-membrane genes omaB, ombB and omcB , thus, it was hypothesized that OmcE-bound RR might serve as potential electron acceptor of OmbB-OmaB-OmcB porin complex which passes electrons across outer membrane. Our work demonstrated a new pathway of artificial electron mediators in facilitating EET in Geobacter species, which may guide the application of electron mediator in improving the performance of BESs. [Display omitted] [ABSTRACT FROM AUTHOR]

• Electrochemical stimulation accelerated and enhanced antibiotics biodegradation. • Microbial metabolic activity was promoted under electrochemical stimulation. • Desulfobulbus and Geobactor were enriched in biocathode and bioanode biofilm. • Electrochemical stimulation facilitated extracellular electron transfer processes. The presence of antibiotics in wastewater poses significant threat to our ecosystems and health. Traditional biological wastewater treatment technologies have several limitations in treating antibiotic-contaminated wastewaters, such as low removal efficiency and poor process resilience. Here, a novel electrochemical-coupled sulfur-mediated biological system was developed for treating wastewater co-contaminated with several antibiotics (e.g., ciprofloxacin (CIP), sulfamethoxazole (SMX), chloramphenicol (CAP)). Superior removal of CIP, SMX, and CAP with efficiencies ranging from 40.6 ± 2.6 % to 98.4 ± 1.6 % was achieved at high concentrations of 1000 μg/L in the electrochemical-coupled sulfur-mediated biological system, whereas the efficiencies ranged from 30.4 ± 2.3 % to 98.2 ± 1.4 % in the control system (without electrochemical stimulation). The biodegradation rates of CIP, SMX, and CAP increased by 1.5∼1.9-folds under electrochemical stimulation compared to the control. The insights into the role of electrochemical stimulation for multiple antibiotics biodegradation enhancement was elucidated through a combination of metagenomic and electrochemical analyses. Results showed that sustained electrochemical stimulation significantly enriched the sulfate-reducing and electroactive bacteria (e.g., Desulfobulbus, Longilinea , and Lentimicrobiumin on biocathode and Geobactor on bioanode), and boosted the secretion of electron transport mediators (e.g., cytochrome c and extracellular polymeric substances), which facilitated the microbial extracellular electron transfer processes and subsequent antibiotics removal in the sulfur-mediated biological system. Furthermore, under electrochemical stimulation, functional genes associated with sulfur and carbon metabolism and electron transfer were more abundant, and the microbial metabolic processes were enhanced, contributing to antibiotics biodegradation. Our study for the first time demonstrated that the synergistic effects of electrochemical-coupled sulfur-mediated biological system was capable of overcoming the limitations of conventional biological treatment processes. This study shed light on the mechanism of enhanced antibiotics biodegradation via electrochemical stimulation, which could be employed in sulfur-mediated bioprocess for treating antibiotic-contaminated wastewaters. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • NCDs (C: N precursor = 1:0.5–1:3) were used to modified the biocathode. • NCDs 1:1 modification achieved highest acetate synthesis of MES (1.9 ± 0.1 mM d−1). • Pyrrolic N positively correlated with Sporomusa abundance and cytochrome expression. • NCDs 1:1 accelerated the EEU of S. ovata by upregulating cytochrome expression. • NCDs 1:1 enhanced the metabolism of S. ovata by increasing 2 H+-pumping step. This study investigated the molecular mechanism behind the highly efficient performance of nitrogen-doped carbon dots (NCDs)-assisted microbial electrosynthesis systems (MESs). The impact of NCDs (C:N precursor = 1:0.5–1:3) on acetogens was examined in the biocathode. The highest electrocatalytic performance was observed with NCDs 1:1. The maximum acetate production rate of 1.9 ± 0.1 mM d−1 was achieved in NCDs 1:1 - modified MESs, which was 26.7–216.7 % higher than other MESs (0.6–1.5 mM d−1). With NCDs 1:1 modified, the biocathode exhibited a 129.3–186.8 % increase in the abundance of Sporomusa , and 38.5–104.6 % increase in cytochrome expression (cydAB , cybH). Transcriptome confirmed that cytochromes played a crucial role in the extracellular electron uptake (EEU) of NCDs 1:1 -modified Sporomusa. NCDs 1:1 enhanced EEU efficiency, thereby increasing the two H+-pumping steps and accelerating microbial CO 2 fixation. These results provide valuable insights into increasing CO 2 fixation by maximizing EEU efficiency in acetogens. [ABSTRACT FROM AUTHOR]

[Display omitted] • Direct transforming raw straw to electricity and H 2 using yeast C. slooffiae JSUX1. • The MFCs delivered a power density of 28.56 mW/m2 and H 2 of 4.9 L/m3. • The enzymes secreted by the yeast JSUX1 consumed raw straw to electricity. • Humic acids derived from raw straw consumption served as electron mediators for MFCs. 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. [ABSTRACT FROM AUTHOR]

The production of reactive oxygen species (ROS) in the rhizosphere is limited by the low extracellular electron transfer capacity of indigenous microorganisms. In the present study, electrical stimulation was used to promote the generation of rhizospheric ROS by accelerating extracellular electron transfer. The result showed that •OH concentrations in the electrically stimulated group (ES group) exceeded the control group by 15.76 %. Accordingly, the removal rate of the target pollutant (i.e., 2,4-dichlorophenol, and sulfamethoxazole) was 20.01 %−24.80 % higher in the ES group than in the control group. The sediment of the ES group had a higher capacity (30.55 %) and a lower electrical resistance (29.15 %) compared to the control group, which subsequently promoted the dissimilatory iron reduction to produce Fe(II) for triggering a Fenton-like process. The increased extracellular respiratory capacity under electrical stimulation could be attributed to the polarization of C-N and C O bonds, which provided more electron storage sites and thus participated in proton-coupled electron transfer. In addition, the concentration of ATP and co-enzymes (NADH/NAD+ and Complex I/Complex III), reflecting electron exchange within respiratory chains, increased distinctly under electrical stimulation. Applying electrical stimulation seemed feasible to increase ROS production and contaminant degradation in the rhizosphere, deepening the understanding of electrical stimulation to promote the production of ROS in the natural system. [Display omitted] • Rhizospheric ROS production was enhanced by 15.76 % under electrical stimulation. • Electrical stimulation promoted sediment extracellular respiratory capacity. • Polarization of C-N and C O bonds enhanced proton-coupled electron transfer. • Co-enzymes reflecting electron exchange in respiratory chains increased distinctly. • The removal rate of the target pollutants was elevated by 20.01 %−24.80 %. [ABSTRACT FROM AUTHOR]

The present study aimed to enhance the co-anaerobic digestion system of mushroom residues and cattle manure by incorporating biosurfactant sophorolipid. Results demonstrated that the addition of 75 mg/L sophorolipid increased cumulative methane production by 33.68%, acetate content by 9–10 times, and the abundance of Methanosarcina by 69.22%. The electroactive microorganisms (Bacteroides , Petrimonas , etc.) were enriched, while the up-regulation of functional genes associated with carbohydrate metabolism and methane metabolism was observed. The metagenomic analysis revealed the significant involvement of inter-microbial communication and extracellular electron transfer in anaerobic digestion. Petrimonas was identified as the predominant host involved in cellular processes and environmental information processing. The supplementation of sophorolipid significantly enhanced its abundance during the late anaerobic digestion period (by 12.30%–64.84%). The results emphasize the crucial function of sophorolipid as biosurfactant in enhancing the efficiency of anaerobic digestion. [Display omitted] • Sophorolipid enhanced the co-anaerobic digestion efficiency. • Sophorolipid promoted the metabolism of functional microorganisms. • Sophorolipid promoted inter-microbial communication and EET. [ABSTRACT FROM AUTHOR]

Bioremediation has garnered considerable interest due to its advantages of economy and no secondary pollution. However, the direct electron transfer rate between microorganisms and Cr(VI) is low. The bioreduction of Cr(VI) by Shewanella oneidensis MR-1 (S. oneidensis) in the presence of formylmethylflavin (FMF) was conducted to understand how FMF mediated the extracellular electron transfer process to improve Cr(VI) removal. In this study, FMF was firstly synthesized by modifing RF to increase its solubility and redox activity. The findings indicate that S. oneidensis /FMF exhibited better Cr(VI) removal performance compared to that of S. oneidensis. Under optimum conditions, 40 mg/L Cr(VI) could be completely removed by S. oneidensis /FMF within 120 h, while only 48.6% of Cr(VI) was removed by S. oneidensis , and the first order rate constant (k) for the Cr(VI) elimination by S. oneidensis /FMF (0.033 h−1) was about 4.1-fold greater than that of S. oneidensis (0.008 h−1). Moreover, the removal of Cr(VI) by S. oneidensis /FMF were dominated by reduction, and supplemented by adsorption and complexation. FMF enhance the extracellular electron transfer rate (EETR) was confirmed by electrochemical cyclic voltammetry (CV) and linear scanning voltammetry (LSV) experiments. This study emphasizes the potential important role of FMF in environmental bioremediation. [Display omitted] • FMF was synthesized by modifing RF to enhance its solubility and redox activity. • 40 mg/L Cr(VI) could be cleared by S. oneidensis /FMF within 120 h. • FMF can act as an electron transfer mediator to greatly enhance the EETR. • Cr(VI) removal was accomplished by reduction, adsorption and complexation. [ABSTRACT FROM AUTHOR]

Bioelectrochemical system (BES) relies on the electrochemical reactions derived from the interaction between microorganisms and solid electrodes to enable processes such as electricity generation and other biotechnological applications. The diversity of these electroactive microorganisms capable of extracellular electron transfer (EET) spread across all three domains of life. The expanding research in this domain focuses on enhancing the EET capabilities of these exoelectrogens and exploring non-exoelectrogens that can support them while reducing waste through various biogeochemical processes. Approaches such as biofilm improvement, genetic modification of electron-conducting proteins, and overexpressing redox mediators are explored to increase EET efficiency. Electrochemically inactive fermentative microorganisms that are non-exoelectrogens often coexist with exoelectrogens. Although their presence has been associated with increased power generation, their excessive proliferation can diminish power output. Therefore, understanding the synergies and intricate balance between exoelectrogens and non-exoelectrogens is necessary. This review discusses the mechanism of EET, strategies to improve EET by engineering microbial communities, the role of non-exoelectrogens involved in the BES, interactions, and synergies within microbial consortia and the factors that affect them, as well as their community structure and dynamics. This review seeks to elucidate the complex interplay within BES and pave the way for future advancements in this field by examining these aspects. • Bioelectrochemical Systems (BES) utilize the exo-electrogenesis to generate electricity. • BES host diverse array of microorganisms including fermentative, methanogensis etc. • Synergistic interactions between exoelectrogens and non-exoelectrogens enhances the BES performance. • Microbial community and its dynamics are affected by internal and external factors. [ABSTRACT FROM AUTHOR]

• A novel 3D N-doped graphene aerogel anode with abundant N was fabricated. • 3D N-doped graphene aerogel anode result in higher bioelectricity generation. • Possible enhanced mechanisms for the N-doped graphene aerogel anode were explored. • The novel anode had good biocompatibility for Geobacter soli enrichment. • N-doping significantly upregulated the conductive pili-related genes of Geobacter. 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. [Display omitted] [ABSTRACT FROM AUTHOR]

Metal oxides rich in oxygen vacancies (O v) possess excellent intrinsic conductivity, variable valence metal center and high capacitance, which were potential candidates as microbial fuel cells anode. However, their high bacterial toxicity greatly hindered the activity of biofilm. To solve this issue, in this work, polydopamine (PDA) covered capacitive O v doped Co 3 O 4 nanosheets were designed and successfully grown on carbon cloth (CC). The introduction of O v significantly promoted the charge storage, the net charge and the accumulated charge were 217.1 and 2680.3 C/m2 , respectively, surpassed Co 3 O 4 anode without O v. In addition, O v also decreased the charge-transfer resistance (R ct), effectively accelerating EET. The R ct of PDA/O v -Co 3 O 4 @CC was 29.51 Ω, lower than CC (382.10 Ω). The cover of PDA prevented the direct contact between metal ions and microorganisms, greatly enhanced the bacterial viability. The power density of MFCs with PDA/O v -Co 3 O 4 @CC anode was 2.77 W/m2, 1.45 times higher than that of bare CC (1.90 W/m2). In addition, PDA/O v -Co 3 O 4 @CC anode displayed excellent coulombic efficiency and daily COD removal amount (23.5 % and 147.6 mg/L/d), higher than the control groups. The community analysis showed the abundance of Geobacter on PDA/O v -Co 3 O 4 @CC anode was 91 %, higher than other anodes. Present study developed a facile and effective PDA coating strategy to simultaneously enhance the biocompatibility and capacitance of O v -rich metal oxides, which will be a promising method for the preparation of high performance MFCs anode. [Display omitted] [ABSTRACT FROM AUTHOR]

• GAC improved CSMYs by 18.8–20.9%, while MNPs decreased CSMYs by 0.8–27.0%. • Redox-active EPSs were over-secreted to protect microbes from MNPs and hinder EET. • MNPs caused DIR by Clostridium sp., which competed with methanogens for substrates. • The medium GAC dose promoted DIET by pili, c-Cyts, and 4 methanogenesis pathways. • Lower MNP doses hindered pili assembly, while the high dose impeded CH 4 conversion. Enhancing direct interspecies electron transfer (DIET) via granular activated carbon (GAC) and magnetite nanoparticles (MNPs) to promote methanation performance in dry anaerobic digestion (AD) systems has been extensively studied in recent years. However, the underlying mechanisms, particularly how GAC and MNPs influence DIET and extracellular polymeric substances (EPSs), remain poorly understood. In this study, the different effects and mechanisms of GAC and MNPs on dry AD fed with pig manure (PM) and corn straw were compared, with additions of 10, 20, and 50% of the volatile solid (VS) of PM. The results indicated that GAC improved the cumulative specific methane yields (CSMYs) by 18.8–20.9%, while a high dose of MNPs (50% of the VS PM) decreased the CSMY and hydrolysis rate constant by 27.0 and 75.4%, respectively. GAC provided microorganism attachment sites and functional groups for electron transfer, improving hydrolysis by enriching fermenters such as Fastidiosipila and Rikenellaceae. Nevertheless, MNPs stimulated intense dissimilatory iron reduction by Clostridium sp. in the early stage, competing with methanogens for substrates. Low and medium GAC doses (10 and 20% of the VS PM) increased the utilization of redox-active substances (e.g., humic substances and proteins) in EPS, while the high MNP dose stimulated their secretion due to microbial self-protection, impairing electron transfer. Metagenomic evidence showed that the medium GAC dose improved four methanogenic pathways and DIET mediated by microbial nanowires, while a high MNP dose impeded the final step of methanogenesis. These results provide deeper insights into the mechanisms of different conductive materials on dry AD. [ABSTRACT FROM AUTHOR]

[Display omitted] • Biochar promoted CH 4 component in biogas to 89.0 ± 6.3 % in the AnMBR. • Biochar increased 0.74 mol e- /g VS /min of electron transfer velocity in AD system. • Biochar was involved in electroactive microbial floravias metabolism via DIET. • The rate for acetate converted into CH 4 was enhanced 3.14 times by biochar. • Ambient temperature anaerobic digestion is a key to achieve net energy output. The anaerobic membrane bioreactor (AnMBR) is an alternative technology with energy neutrality potential when treating municipal wastewater (MWW). However, the poor bio-reaction kinetic conditions caused by low COD concentrations in MWW greatly limits the CH 4 yield. In this study, the average methanogenesis rate was significantly increased from 164.8 to 275.5 mL/g COD with the two times biochar addition of 5 g/L. And following the second time addition of 5 g/L biochar, the CH 4 component significantly increased to a high level of 89.0 ± 6.3 %. In addition, the rate for acetate converted into CH 4 increased from 3.3 to 10.5 mmol/(g VS ·d) with biochar addition of 10 g/L, indicating that acetotrophic methanogenesis route was significantly promoted by biochar. Bio-electrochemistry analysis showed that biochar accelerated 0.74 mol e-/g VS /min of electron transfer velocity in AD system. The addition of biochar enriched organic metabolism bacteria such as Ancinetobacter_lwoffii , Comamonas and Geobacter , and upregulated the expression of functional genes related to organics degradation such as lldD , sucD and fumC, facilitates the direct transfer of electrons and acetates produced by Comamonas and Geobacte r to convert CO 2 into CH 4 through microbial mediation. Thus, the COD flow shifted significantly towards more CH 4 production. The COD mass flow showed that biochar facilitated more bio-catabolism (up about 52.7 %) and less bio-anabolism (down about 92 %), and more COD in wastewater was converted to CH 4 , resulting a significant promoting of energy production. The energy balance analysis indicated that the net energy consumption was reduced by 15 % after two times addition of 5 g/L biochar, and the key to achieve the net energy output during treating MWW is the efficient operation of AnMBR at ambient temperature. [ABSTRACT FROM AUTHOR]

[Display omitted] • Higher mZVI dosages shifted nitrogen distribution from NO 2 –-N to gaseous forms. • Achieved 76.5% total nitrogen removal with the addition of mZVI. • NO 2 –-N was more readily transformed than NO 3 –-N with mZVI. • Addition of mZVI reduced the activation energy for NO 2 –-N transformation by 72%. • Nir activity was twice as high with mZVI (0.31 μg NO 2 –-N min−1 mg−1 protein). This study investigates the denitrification characteristics facilitated by microscale zero-valent iron (mZVI) in conjunction with Klebsiella oxytoca. While Klebsiella oxytoca effectively degrades both nitrate (NO 3 –-N) and nitrite (NO 2 –-N) on its own, the presence of mZVI significantly enhances NO 2 –-N elimination, highlighting mZVI's facilitative role in denitrification. Optimal performance is observed at an mZVI concentration of 1.0 g/L, enabling complete biotic reduction of both NO 3 –-N and NO 2 –-N, particularly at an initial NO 3 –-N concentration of 50 mg/L and a C/N ratio of 16. mZVI promotes the conversion of NO 2 –-N to gaseous nitrogen, with this effect becoming more pronounced at higher mZVI doses. Inhibitory kinetics experiments demonstrate mZVI's ability to mitigate NO 2 –-N inhibition on microorganisms. Thermodynamic and electrochemical analyses reveal that mZVI enhances NO 2 –-N reduction through mechanisms involving Fe(II)/Fe(III) electron pair generation, as well as the secretion of extracellular polymeric substances and flavins. Genetic abundance and enzyme activity analyses further confirm mZVI's pivotal role in enhancing denitrification efficiency and elucidating its underlying mechanisms. [ABSTRACT FROM AUTHOR]

[Display omitted] • PFS can serve as a potential electron acceptor in the process of nitrogen removal. • Fe3+ as an extracellular electron acceptor compensated nitrite deficiency in Anammox. • Feammox-Anammox combined system achieved nitrogen removal of 85 % at NO 2 –-N/NH 4 +-N of 0.5. • Proportion of ammonia loss by extracellular electron transfer increased from 0.3% to 45%. • Ca. Brocadia and Ca.Kuenenia alternately dominated intracellular and extracellular electron transfer. In addressing the issue of the unstable supply of electron acceptors (nitrite) during Anammox, researchers have often overlooked that the electron transfer process in anammox bacteria (AnAOB) is not restricted to the intracellular, extracellular electron transfer can also serve as an alternative pathway. This study utilized Fe3+ as an adjunctive electron acceptor, coupling Anammox with ferric iron reduction (Feammox) by introducing polymeric ferric sulfate (PFS) in the Anammox system. This approach achieved effective nitrogen removal even when nitrite was deficient. At a NO 2 –-N to NH 4 +-N ratio of 0.5, the Feammox-Anammox combined system maintained stable nitrogen removal under the dual electron acceptor, with total nitrogen and NH 4 +-N removal efficiency of 85 % and 91 %, respectively. The contribution of Feammox and Anammox to NH 4 +-N removal was 45 % and 55 %, respectively. TEM-EDS confirmed the presence of iron elements across the periphery, periplasm, and cytoplasm of AnAOB, suggesting the involvement of PFS in the metabolic and electron transfer processes. Notably, the introduction of Fe3+ and the limitation of nitrite availability resulted in increased relative abundances of FeOB (Thermomonas) and FeRB (Ignavibacterium) to 30 % and 3.6 %, respectively, indicating that the electron transfer process with Fe3+ as the electron acceptor was continuously intensified, as well as a cycle of Fe3+ / Fe2+ was established. Due to unique nitrogen metabolism mode and high Fe3+ tolerance, the abundance of Candidatus_Brocadia was enriched up to 7.2 %, which alternately dominated the Anammox process with Candidatus_Kuenenia , providing favorable conditions for AnAOB to carry out the extracellular electron transfer process. These findings provide a novel strategy for improving the stability and efficiency of the Anammox process under deficient nitrite supply. [ABSTRACT FROM AUTHOR]

The influence of dissolved oxygen (DO) and Shewanella algae on the corrosion behavior of pure titanium were systematically studied in this work. The formation of S. algae biofilms on titanium surface was facilitated by the anaerobic environment, which accelerated titanium corrosion. Upon the breakdown of passive film, S. algae acquired electrons from the titanium base substrate through extracellular electron transfer (EET) processes. Various EET-related genes were overexpressed by the low DO condition, leading to enhanced EET kinetics. High concentration DO increased TiO 2 content and the thickness of passive film, which enhanced the protective effect and mitigated microbiologically influenced corrosion. [Display omitted] • Shewanella algae accelerated Ti corrosion while dissolved oxygen reduced MIC effect. • Anaerobic environment facilitated the formation of S. algae biofilms on Ti surface. • Dissolved oxygen increased TiO 2 content and the thickness of passive film. • Various EET-related genes of S. algae were overexpressed in anaerobic condition. [ABSTRACT FROM AUTHOR]