Microbial Extracellular Electron Transfer Research Articles

Activated Carbon Application Simultaneously Alleviates Paddy Soil Arsenic Mobilization and Carbon Emission by Decreasing Porewater Dissolved Organic Matter.

Flooding of paddy fields during the rice growing season enhances arsenic (As) mobilization and greenhouse gas (e.g., methane) emissions. In this study, an adsorbent for dissolved organic matter (DOM), namely, activated carbon (AC), was applied to an arsenic-contaminated paddy soil. The capacity for simultaneously alleviating soil carbon emissions and As accumulation in rice grains was explored. Soil microcosm incubations and 2-year pot experimental results indicated that AC amendment significantly decreased porewater DOM, Fe(III) reduction/Fe2+ release, and As release. More importantly, soil carbon dioxide and methane emissions were mitigated in anoxic microcosm incubations. Porewater DOM of pot experiments mainly consisted of humic-like fluorophores with a molecular structure of lignins and tannins, which could mediate microbial reduction of Fe(III) (oxyhydr)oxides. Soil microcosm incubation experiments cospiking with a carbon source and AC further consolidated that DOM electron shuttling and microbial carbon source functions were crucial for soil Fe(III) reduction, thus driving paddy soil As release and carbon emission. Additionally, the application of AC alleviated rice grain dimethylarsenate accumulation over 2 years. Our results highlight the importance of microbial extracellular electron transfer in driving paddy soil anaerobic respiration and decreasing porewater DOM in simultaneously remediating As contamination and mitigating methane emission in paddy fields.

Enhancing Extracellular Electron Transfer of a 3D-Printed Shewanella Bioanode with Riboflavin-Modified Carbon Black Bioink.

3D printing of a living bioanode holds the potential for the rapid and efficient production of bioelectrochemistry systems. However, the ink (such as sodium alginate, SA) that formed the matrix of the 3D-printed bioanode may hinder extracellular electron transfer (EET) between the microorganism and conductive materials. Here, we proposed a biomimetic design of a 3D-printed Shewanella bioanode, wherein riboflavin (RF) was modified on carbon black (CB) to serve as a redox substance for microbial EET. By introducing the medicated EET pathways, the 3D-printed bioanode obtained a maximum power density of 252 ± 12 mW/m2, which was 1.7 and 60.5 times higher than those of SA-CB (92 ± 10 mW/m2) and a bare carbon cloth anode (3.8 ± 0.4 mW/m2). Adding RF reduced the charge-transfer resistance of a 3D-printed bioanode by 75% (189.5 ± 18.7 vs 47.3 ± 7.8 Ω), indicating a significant acceleration in the EET efficiency within the bioanode. This work provided a fundamental and instrumental concept for constructing a 3D-printed bioanode.

Biomineralized manganese oxide mediated nitrogen-contained wastewater treatment

In recent years, manganese (Mn) has emerged as an accelerator for nitrogen metabolism. However, the bioactivity of manganese is limited by the restricted contact between microbes and manganese minerals in the solid phase and by the toxicity of manganese to microbes. To enhance the bioactivity of solid-phase manganese, biomineralized manganese oxide (MnOx) modified by Lactobacillus was introduced. Nitrogen removal performance have confirmed the effective role of biomineralized MnOx in accelerating the removal of total inorganic nitrogen (TIN). Metagenomic analysis has confirmed the enhancement of the nitrogen metabolic pathway and microbial extracellular electron transfer (MEET) in biomineralized MnOx treatment group (BIOA group). Additionally, the enrichment of manganese oxidation and denitrification genus indicates a coupling between nitrogen metabolism and manganese metabolism. One point of views is that biomineralized MnOx-mediated nitrogen transformation processes could serve as a substitute for traditional nitrogen removal processes.

Lack of physiological evidence for cytochrome filaments functioning as conduits for extracellular electron transfer.

Unraveling 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.

All roads lead to Rome: Cyclic di-GMP differentially regulates extracellular electron transfer in <i>Geobacter</i> biofilms

<p>Microbial extracellular electron transfer (EET) in dissimilatory metal-reducing microorganisms (DMRMs) is a widespread biological process and is involved in biogeochemical cycling of a variety of elements on the planet of Earth. However, the regulatory networks controlling such important process have been under-investigated. Here, we reported that the intracellular messenger bis-(3��-5��) cyclic dimeric guanosine monophosphate (c-di-GMP) signaling network controls EET in <i>Geobacter sulfurreducens</i>. The low and high levels of c-di-GMP both improved EET in <i>G. sulfurreducens</i> electrode-respiring biofilms by differentially regulating the expression of EET-associated genes. In particular, we found that a low c-di-GMP level reduced the formation of the anode biofilm but enhanced EET by upregulating the transcription of all known nanowire genes (i.e., <i>pilA</i>, <i>omcS</i>, <i>omcZ</i> and <i>omcE</i>). Upregulated <i>omcZ</i> transcription was further determined to play a decisive role in improving EET. Given that c-di-GMP is present in diverse DMRMs, this study substantially expands our understanding of the regulatory role of c-di-GMP signaling and the varied strategies for efficient EET employed by DMRMs. In addition to be fundamentally significant to understand microbe-mineral and microbe-microbe interactions driven by EET, it is also instructive to develop effective engineered microbial systems for practical applications.</p>

Combined electrical-electrochemical phenotypic profiling of antibiotic susceptibility of in vitro biofilm models.

More than 65% of bacterial infections are caused by biofilms. However, standard biofilm susceptibility tests are not available for clinical use. All conventional biofilm models suffer from a long formation time and fail to mimic in vivo microbial biofilm conditions. Moreover, biofilms make it difficult to monitor the effectiveness of antibiotics. This work creates a powerful yet simple method to form a target biofilm and develops an innovative approach to monitoring the antibiotic's efficacy against a biofilm-associated infection. A paper-based culture platform can provide a new strategy for rapid microbial biofilm formation through capillary action. A combined electrical-electrochemical technique monitors bacterial metabolism rapidly and reliably by measuring microbial extracellular electron transfer (EET) and using electrochemical impedance spectroscopy (EIS) across a microbe-electrode interface. Three representative pathogens, Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus, form their biofilms controllably within an hour. Within another hour their susceptibilities to three frontline antibiotics with different action modes (gentamicin, ciprofloxacin, and ceftazidime) are examined. Our antibiotic susceptibility testing (AST) technique provides a quantifiable minimum inhibitory concentration (MIC) of those antibiotics against the in vitro biofilm models and characterizes their action mechanisms. The results will have an important positive effect because they provide immediately actionable healthcare information at a reduced cost, revolutionizing public healthcare.

Enhancing microbial extracellular electron transfer efficiency through increased configuration entropy of bioanodes

Microbial fuel cells (MFCs) have garnered significant attention in the field of bioelectrochemistry as a clean energy source, with a focus on increasing their power density. This study reports the successful application of a spinel-type high-entropy oxide (Mg0.2Co0.2Ni0.2Cu0.2Zn0.2)Fe2O4 as an anode material for microbial fuel cells. The samples were prepared through ball milling and sintering at 900 °C. Electrochemical tests demonstrated that the high-entropy oxide significantly enhanced the electrochemical performance and biocompatibility of the anode. The MFCs loaded with (Mg0.2Co0.2Ni0.2Cu0.2Zn0.2)Fe2O4 exhibited a power density of up to 3.43 W/m2, which is significantly higher than that of a simple mixture of six metals (2.61 W/m2) and pure Fe2O3 (2.64 W/m2). This outstanding performance results from the disorder induced by the high-entropy composition. This disorder facilitates improved coordination among the primary constituent oxides, maximizing their advantages. Simultaneously, it minimizes the reduction in active sites caused by uneven distribution. High-throughput sequencing results revealed an enrichment of Geobacter in the anodic biofilm, reaching a content as high as 52.51% after domestication. This study elucidates the contribution of entropy to biocatalysis from an entropy perspective and provides a new strategy for developing high-performance materials for microbial fuel cells.

Predicting microbial extracellular electron transfer activity in paddy soils with soil physicochemical properties using machine learning

Predicting microbial extracellular electron transfer activity in paddy soils with soil physicochemical properties using machine learning

Bi-directional regulation of electroactive microbial community using Au/antimicrobial peptide nanocomposite

The power generation and pollutants removal efficiencies of mixed electroactive microbial community rely on the effective electron transfer between outer membrane proteins of electroactive electricigens. However, the poor conductivity of protein and various non-electrogenic bacteria greatly hinder the electron transfer and effective contact between electricigens. Herein, we report a novel “bi-directional” microbial community regulation method that can selectively inhibit Gram-positive non-electrogenic bacteria and enhances the conductivity and electron transfer between electrogenic bacteria and the electrode in MFC by taking the advantages of the excellent conductive and bacteriostatic properties of Au/Nisin nanocomposite. The Au/Nisin stimulation endows microbial fuel cell (MFC) a high-efficiency functional community, which increases the power density and decolorization rate by 103 % and 33 %, respectively. The high conductivity, and selectivity and antimicrobial activity of the nanocomposite improve the electrochemical activity of the bioanode, hereby promoting the microbial extracellular electron transfer and increasing the proportion of electrogenic bacteria such as Proteobacteria, Bacteroidetes and Desulfovibrio in the biofilm. Our work successfully demonstrates that the construction of multifunctional metallic peptide nanocomposite can effectively regulate microbial community to improve the electrochemical performance of MFC.

Enforcing energy consumption promotes microbial extracellular respiration for xenobiotic bioconversion.

Extracellular electron transfer (EET) empowers electrogens to catalyse the bioconversion of a wide range of xenobiotics in the environment. Synthetic bioengineering has proven effective in promoting EET output. However, conventional strategies mainly focus on modifications of EET-related genes or pathways, which leads to a bottleneck due to the intricate nature of electrogenic metabolic properties and intricate pathway regulation that remain unelucidated. Herein, we propose a novel EET pathway-independent approach, from an energy manipulation perspective, to enhance microbial EET output. The Controlled Hydrolyzation of ATP to Enhance Extracellular Respiration (CHEER) strategy promotes energy utilization and persistently reduces the intracellular ATP level in Shewanella oneidensis, a representative electrogenic microbe. This approach leads to the accelerated consumption of carbon substrate, increased biomass accumulation and an expanded intracellular NADH pool. Both microbial electrolysis cell and microbial fuel cell tests exhibit that the CHEER strain substantially enhances EET capability. Analysis of transcriptome profiles reveals that the CHEER strain considerably bolsters biomass synthesis and metabolic activity. When applied to the bioconversion of model xenobiotics including methyl orange, Cr(VI) and U(VI), the CHEER strain consistently exhibits enhanced removal efficiencies. This work provides a new perspective and a feasible strategy to enhance microbial EET for efficient xenobiotic conversion.

Interactions between arsenic migration and CH4 emission in a soil bioelectrochemical system under the effect of zero-valent iron

Dissimilatory soil arsenic (As) reduction and release are driven by microbial extracellular electron transfer (EET), while reverse EET mediates soil methane (CH4) emission. Nevertheless, the detailed biogeochemical mechanisms underlying the tight links between soil As migration and methanogenesis are unclear. This study used a bioelectrochemical-based system (BES) to explore the potential effects of zero-valent iron (ZVI) addition on “As migration–CH4 emission” interactions from chemical and microbiological perspectives. Voltage and ZVI amendment experiments showed that dissolved As was efficiently immobilized with increased CH4 production in the soil BES, As release and CH4 production exhibited a high negative exponential correlation, and reductive As dissolution could be entirely inhibited in the methanogenic stage. Gene quantification and bacterial community analysis showed that in contrast to applied voltage, ZVI changed the spatial heterogeneity of the distribution of electroactive microorganisms in the BES, significantly decreasing the relative abundance of arrA and dissimilatory As/Fe-reducing bacteria (e.g., Geobacter) while increasing the abundance of aceticlastic methanogens (Methanosaeta), which then dominated CH4 production and As immobilization after ZVI incorporation. In addition to biogeochemical activities, coprecipitation with ferric (iron) contributed 77–93% dissolved As removal under ZVI addition. This study will enhance our knowledge of the processes and microorganisms controlling soil As migration and CH4 emission.

Evidence for the feasibility of transmembrane proton gradient regulating oxytetracycline extracellular biodegradation mediated by biosynthesized palladium nanoparticles

Extracellular biodegradation is a promising technology for removing antibiotics and repressing the spread of resistance genes, but the strategy is limited by the low extracellular electron transfer (EET) efficiency of microorganisms. In this work, biogenic Pd0 nanoparticles (bio-Pd0) were introduced in cells in situ to enhance oxytetracycline (OTC) extracellular degradation and the effects of transmembrane proton gradient (TPG) on EET and energy metabolism mediated by bio-Pd0 were investigated. The results indicated that the intracellular OTC concentration gradually decreased with increase in pH due to the simultaneous decreases of OTC adsorption and TPG-dependent OTC uptake. On the contrary, the efficiency of OTC biodegradation mediated by bio-Pd0@B. megaterium showed a pH-dependent increase. The negligible intracellular OTC degradation, the high dependence of OTC biodegradation on respiration chain and the results on enzyme activity and respiratory chain inhibition experiments showed that NADH-dependent (rather than FADH2-dependent) EET process mediated by substrate-level phosphorylation modulated OTC biodegradation due to high energy storage and proton translocation capacity. Moreover, the results showed that altering TPG is an efficient approach to improve EET efficiency, which can be attributed to the increased NADH generation by the TCA cycle, enhanced transmembrane electron output efficiency (as evidenced by increased intracellular electron transfer system (IETS) activity, the negative shift of onset potential, and enhanced one-electron transfer through bound flavin) and stimulation of substrate-level phosphorylation energy metabolism catalyzed by succinic thiokinase (STH) under low TPG conditions. The results of structural equation model that OTC biodegradation was directly and positively modulated by the net outward proton flux as well as STH activity, and indirectly regulated by TPG through NADH level and IETS activity confirmed the previous findings. This study provides a new perspective for engineering microbial EET and application of bioelectrochemistry processes in bioremediation.

Promoting vivianite recovery: Crucial role of tightly-bound extracellular polymeric substances

Vivianite is an important bio-induced product of dissimilated iron reduction (DIR), which exists widely in activated sludge and digested sludge of wastewater treatment plants (WWTPs). Extracellular polymeric substances (EPS) surrounding dissimilatory iron-reducing bacteria (DIRB) were reported to be transient media for microbial extracellular electron transfer (EET) and DIR. However, what happens when a layer of EPS envelops microbial cells during vivianite recovery? This study tried to assess the relationship between iron reduction and electron transfer by stripping EPS layer-by-layer, then clarified the delamination of different EPS layers on the performance of vivianite recovery. After stripping total EPS (PC-EPS batches) of DIRB, the Fe (III) reduction efficiencies (RFe), Fe (III) reduction rate (VFe), and vivianite formation efficiency (RV) decreased by 33%, 37%, and 30%, respectively. Tightly bound-EPS (TB-EPS) was confirmed to play a crucial role in vivianite formation, which contributed 28–51% to vivianite recovery and 34–55% to iron reduction. Electronic shuttle substances were contained in TB-EPS, which enhanced the EET rate and efficiency between cells and Fe (III). The polysaccharide content of TB-EPS was positively related to adhesiveness between cells and mineral, as well as the iron reduction efficiency. Furthermore, redox proteins (PpcA, OmcB) were embedded in EPS, which shortened the distance of electron hopping to across the EPS layer and further accelerated vivianite recovery from wastewater. Long-term operation of anaerobic reactors confirmed that TB-EPS played critical roles in bio-induced vivianite formation.

Multiple roles of released c-type cytochromes in tuning electron transport and physiological status of Geobacter sulfurreducens.

Dissimilatory metal-reducing bacteria (DMRB) can transfer electrons to extracellular insoluble electron acceptors and play important roles in geochemical cycling, biocorrosion, environmental remediation, and bioenergy generation. c-type cytochromes (c-Cyts) are synthesized by DMRB and usually transported to the cell surface to form modularized electron transport conduits through protein assembly, while some of them are released as extracellularly free-moving electron carriers in growth to promote electron transport. However, the type of these released c-Cyts, the timing of their release, and the functions they perform have not been unrevealed yet. In this work, after characterizing the types of c-Cyts released by Geobacter sulfurreducens under a variety of cultivation conditions, we found that these c-Cyts accumulated up to micromolar concentrations in the surrounding medium and conserved their chemical activities. Further studies demonstrated that the presence of c-Cyts accelerated the process of microbial extracellular electron transfer and mediated long-distance electron transfer. In particular, the presence of c-Cyts promoted the microbial respiration and affected the physiological state of the microbial community. In addition, c-Cyts were observed to be adsorbed on the surface of insoluble electron acceptors and modify electron acceptors. These results reveal the overlooked multiple roles of the released c-Cyts in acting as public goods, delivering electrons, modifying electron acceptors, and even regulating bacterial community structure in natural and artificial environments.

Electro-polarization of protein-like substances accelerates trans-cell-wall electron transfer in microbial extracellular respiration

Electrical stimulation has been used to strengthen microbial extracellular electron transfer (EET), however, the deep-seated reasons remain unclear. Here we reported that Bacillus subtilis, a typical gram-positive bacterium capable of extracellular respiration, obtained a higher EET capacity after the electrical domestication. After the electrical domestication, the current generated by the EET of B.subtilis was 23.4-fold that of the control group without pre-domestication. Multiple lines of evidence in bacterial cells of B.subtilis, their cell walls, and a model tripeptide indicated that the polarization of amide groups after the electrical stimulation forwarded the H-bonds recombination and radical generation of protein-like substances to develop extracellular electron transfer via the proton-coupled pattern. The improved electrochemical properties of protein-like substances benefited the trans-cell-wall electron transfer and strengthen extracellular respiration. This study was the first exploration to promote microbial extracellular respiration by improving the electrochemical properties of protein-like substances in cell envelopes.

Inhibition of microbial extracellular electron transfer corrosion of marine structural steel with multiple alloy elements

The microbial corrosion of marine structural steels (09CrCuSb low alloy steel (LAS) and Q235 carbon steel (CS)) in Desulfovibrio vulgaris medium and Pseudomonas aeruginosa medium based on seawater was investigated. In the D. vulgaris medium, the weight loss and maximum pit depth of 09CrCuSb LAS were 0.59 and 0.56 times as much as those of Q235 CS, respectively. Meanwhile, in the P. aeruginosa medium, the values were 0.53 and 0.67 times, respectively. Compared to Q235 CS, 09CrCuSb LAS contains more alloy elements (Cr, Ni, Cu, Al and Sb), which led to obvious inhibition of sessile bacteria growth but had no effect on planktonic bacteria. The number of live sessile cells on the 09CrCuSb LAS surface was 23.4 % and 26.9 % of that on the Q235 CS surface in the D. vulgaris medium and P. aeruginosa medium, respectively. Fewer sessile cells on the steel surface led to a lower extracellular electron transfer (EET) rate so that less corrosion occurred. In addition, the combined effect of alloying elements on grain refinement and passive film formation also improved the anti-corrosion property of the steels.

Electroactive bacteria as whole-cell biosensor for sensitive detection of hydrogen peroxide

Hydrogen peroxide (H2O2) is a widely used strong oxidizer and has harmful effects on human health with excessive exposure. Herein, we proposed a simple and effective method for H2O2 detection by utilizing electroactive bacteria (EAB) as a whole-cell biosensor. The proposed method was achieved by utilizing the multiheme c-type cytochromes (central components of microbial extracellular electron transfer) of model EAB Shewanella oneidensis MR-1 to catalyze the oxidation of tetramethylbenzidine in the presence of H2O2. Owing to the excellent peroxidase-like activity of the multiheme c-cyts, this H2O2 detection method has a wide dynamic range between 0.1 and 1000 μM and a low detection limit of 0.17 μM. The proposed method, when coupled with glucose oxidase, displayed a high sensitivity (0.57 μM) and broad linear range (1 μM - 10 mM) for glucose. Moreover, this method has high specificity and good recovery for detecting H2O2-spiked real samples. Overall, this study has developed a novel biosensing platform that provides an alternative approach to H2O2 detection.

Label-Free Optical Imaging of the Electron Transfer in Single Live Microbial Cells.

Measurement of electron transfer at the single-particle or -cell level is crucial to the in situ study of basic chemical and biological processes. However, it remains challenging to directly probe the microbial extracellular electron transfer process due to the weakness of signals and the lack of techniques. Here, we present a label-free and noninvasive imaging method that is able to measure the electron transfer in microbial cells. We measured the extracellular electron transfer processes by imaging the redox reaction of c-type outer membrane cytochromes in microbial cells using a plasmonic imaging technique, and obtained the electrochemical activity parameters (formal potential and number of electrons transferred) of multiple individual microbial cells, allowing for unveiling ample heterogeneities in electron transfer at the single-cell level. We anticipate that this method will contribute to the study of electron transfer in various biological and chemical processes.

Microbial electrosynthesis with Clostridium ljungdahlii benefits from hydrogen electron mediation and permits a greater variety of products.

Microbial electrosynthesis (MES) is a very promising technology addressing the challenge of carbon dioxide recycling into organic compounds, which might serve as building blocks for the (bio)chemical industry. However, poor process control and understanding of fundamental aspects such as the microbial extracellular electron transfer (EET) currently limit further developments. In the model acetogen Clostridium ljungdahlii, both direct and indirect electron consumption via hydrogen have been proposed. However, without clarification neither targeted development of the microbial catalyst nor process engineering of MES are possible. In this study, cathodic hydrogen is demonstrated to be the dominating electron source for C. ljungdahlii at electroautotrophic MES allowing for superior growth and biosynthesis, compared to previously reported MES using pure cultures. Hydrogen availability distinctly controlled an either planktonic- or biofilm-dominated lifestyle of C. ljungdahlii. The most robust operation yielded higher planktonic cell densities in a hydrogen mediated process, which demonstrated the uncoupling of growth and biofilm formation. This coincided with an increase of metabolic activity, acetate titers, and production rates (up to 6.06 g L-1 at 0.11 g L-1 d-1). For the first time, MES using C. ljungdahlii was also revealed to deliver other products than acetate in significant amounts: here up to 0.39 g L-1 glycine or 0.14 g L-1 ethanolamine. Hence, a deeper comprehension of the electrophysiology of C. ljungdahlii was shown to be key for designing and improving bioprocess strategies in MES research.

In Vivo Voltammetric Imaging of Metal Nanoparticle-Catalyzed Single-Cell Electron Transfer by Fermi Level-Responsive Graphene.

Metal nanomaterials can facilitate microbial extracellular electron transfer (EET) in the electrochemically active biofilm. However, the role of nanomaterials/bacteria interaction in this process is still unclear. Here, we reported the single-cell voltammetric imaging of Shewanella oneidensis MR-1 at the single-cell level to elucidate the metal-enhanced EET mechanism in vivo by the Fermi level-responsive graphene electrode. Quantified oxidation currents of ~20 fA were observed from single native cells and gold nanoparticle (AuNP)-coated cells in linear sweep voltammetry analysis. On the contrary, the oxidation potential was reduced by up to 100 mV after AuNP modification. It revealed the mechanism of AuNP-catalyzed direct EET decreasing the oxidation barrier between the outer membrane cytochromes and the electrode. Our method offered a promising strategy to understand the nanomaterials/bacteria interaction and guide the rational construction of EET-related microbial fuel cells.