Dissimilatory Metal-reducing Bacteria Research Articles

Microbial corrosion on underwater pottery relics with typical biological condensation disease

Underwater pottery relics salvaged from shipwrecks are vulnerable to physical, chemical and biological corrosion. In this study, the microscopic morphology and structural composition of glazed pottery fragments excavated from the South China Sea as well as purple clay teapot samples from the Yangtze River Estuary II shipwreck were analyzed by means of optical microscopy, scanning electron microscope equipped with energy dispersive spectrometer, X-ray diffraction, X-ray photoelectron spectroscopy, Fourier infrared transform spectroscopy and photoluminescence spectroscopy. According to the research results, the corrosion products deposited on the surface of the glazed pottery fragment from the South China Sea mainly included FeOOH and MnO2, whereas those on the surface of the purple clay teapot fragment from the Yangtze River Estuary II shipwreck primarily incorporated FeS2 and FeS. According to the corrosion phenomenon, it was speculated that the underwater pottery relics were mainly corroded by different microorganisms in the underwater environment, including manganese-oxidizing bacteria, dissimilatory metal-reducing bacteria and sulfate-reducing bacteria. As the corrosion process of these cultural artifacts was described and related models were established, this study provides valuable reference for the disease removal and protection of pottery relics.

Ammonium loss microbiologically mediated by Fe(III) and Mn(IV) reduction along a coastal lagoon system

Anaerobic ammonium oxidation, associated with both iron (Feammox) and manganese (Mnammox) reduction, is a microbial nitrogen (N) removal mechanism recently identified in natural ecosystems. Nevertheless, the spatial distributions of these non-canonical Anammox (NC-Anammox) pathways and their environmental drivers in subtidal coastal sediments are still unknown. Here, we determined the potential NC-Anammox rates and abundance of dissimilatory metal-reducing bacteria (Acidomicrobiaceae A6 and Geobacteraceae) at different horizons (0–20 cm at 5 cm intervals) of subtidal coastal sediments using the 15N isotope-tracing technique and molecular analyses. Sediments were collected across three sectors (inlet, transition, and inner) in a coastal lagoon system (Bahia de San Quintin, Mexico) dominated by seagrass meadows. The positive relationship between 30N2 production rates and dissimilatory Fe and Mn reduction provided evidence for Feammox’s and Mnammox’s co-occurrence. N loss through NC-Anammox was detected in subtidal sediments, with potential rates of 0.07–0.62 μg N g−1 day−1. NC-Anammox process in vegetated sediments tended to be higher than those in adjacent unvegetated ones. NC-Anammox rates showed a subsurface peak (between 5 and 15 cm) in the vegetated sediments but decreased consistently with depth in the adjacent bare bottoms. Thus, the presence/absence of seagrasses and sediment characteristics, particularly the availability of organic carbon and microbiologically reducible Fe(III) and Mn(IV), affected the abundance of dissimilatory metal-reducing bacteria, which mediated NC-Anammox activity and the associated N removal. An annual loss of 32.31 ± 3.57 t N was estimated to be associated with Feammox and Mnammox within the investigated area, accounting for 2.8–4.7% of the gross total import of reactive N from the ocean into the Bahia de San Quintin. Taken as a whole, this study reveals the distribution patterns and controlling factors of the NC-Anammox pathways along a coastal lagoon system. It improves our understanding of the coupling between N and trace metal cycles in coastal environments.

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.

Growth and genome-based insights of Fe(III) reduction of the high-temperature and NaCl-tolerant Shewanella xiamenensis from Changqing oilfield of China.

A facultative anaerobe bacterium Shewanella xiamenensis CQ-Y1 was isolated from the wastewater of Changqing oilfield in Shaanxi Province of China. Shewanella is the important dissimilatory metal-reducing bacteria. It exhibited a well potential application in biodegradation and bioremediation. Genome sequencing, assembling and functional annotation were conducted to explore the genome information of CQ-Y1. The effect of temperatures and NaCl concentrations on the CQ-Y1 growth and Fe(III) reduction were investigated by UV visible spectrophotometry, SEM and XRD. Genomic analysis revealed its complete genome was a circular chromosome of 4,710,887 bp with a GC content of 46.50% and 4,110 CDSs genes, 86 tRNAs and 26 rRNAs. It contains genes encoding for Na+/H+ antiporter, K+/Cl- transporter, heat shock protein associated with NaCl and high-temperature resistance. The presence of genes related to flavin, Cytochrome c, siderophore, and other related proteins supported Fe(III) reduction. In addition, CQ-Y1 could survive at 10% NaCl (w/v) and 45°C, and temperature showed more pronounced effects than NaCl concentration on the bacterial growth. The maximum Fe(III) reduction ratio of CQ-Y1 reached 70.1% at 30°C without NaCl, and the reduction reaction remained active at 40°C with 3% NaCl (w/v). NaCl concentration was more effective than temperature on microbial Fe(III) reduction. And the reduction products under high temperature and high NaCl conditions were characterized as Fe3(PO4)2, FeCl2 and Fe(OH)2. Accordingly, a Fe(III) reduction mechanism of CQ-Y1 mediated by Cytochrome c and flavin was hypothesised. These findings could provide information for a better understanding of the origin and evolution of genomic and metabolic diversity of S. xiamenensis.

Single Strain-Triggered Biogeochemical Cycle of Arsenic.

The microbial metabolism of arsenic plays a prominent role in governing the biogeochemical cycle of arsenic. Although diverse microbes are known to be involved in the redox transformation of inorganic arsenic, the underlying mechanisms about the arsenic redox cycle mediated by a single microbial strain remain unclear yet. Herein, we discover that Shewanella putrefaciens CN32, a well-known arsenate-respiring and dissimilatory metal-reducing bacterium, could mediate the reversible arsenic redox transformation under aerobic conditions. Genetic analysis shows that S. putrefaciens CN32 contains both ars and arr operon but lacks an As(III) oxidase encoding gene. Arsenic(V) reduction tests demonstrate that the ars operon is advantageous but not essential for As(V) respiration in S. putrefaciens CN32. The Arr complex encoded by the arr operon not only plays a crucial role in arsenate respiration under anaerobic conditions but also participates in the sequential process of As(V) reduction and As(III) oxidation under aerobic conditions. The Arr enzyme also contributes to the microbial As(III) resistance. The expression and catalysis directionality of Arr in S. putrefaciens CN32 are regulated by the carbon source types. Our results highlight the complexity of arsenic redox biotransformation in environments and provide new insights into the important contribution of Arr to the As biogeochemical cycle in nature.

Unravelling key factors controlling vivianite formation during anaerobic digestion of waste activated sludge

As a product of phosphorous recovery from anaerobic digestion (AD) of waste activated sludge (WAS), vivianite has received increasing attention. However, key factors controlling vivianite formation have not yet been fully addressed. Thus, this study was initiated to ascertain key factors controlling vivianite formation. A simulation of chemical equilibriums indicates that interfering ions such as metallic ions and inorganic compounds may affect vivianite formation, especially at a PO43- concentration lower than 3 mM. The experiments demonstrated that the rate of ferric bio-reduction conducted by dissimilatory metal-reducing bacteria (DMRB) and the competition of methane-producing bacteria (MPB) with DMRB for VFAs (acetate) were not the key factors controlling vivianite formation, and that ferric bio-reduction of DMRB can proceed when a sufficient amount of Fe3+ exists in WAS. The determined affinity constants (Ks) of both DMRB and MPB on acetate revealed that the KHAc constant (4.2 mmol/g VSS) of DMRB was almost 4 times lower than that of MPB (15.67 mmol/g VSS) and thus MPB could not seriously compete for VFAs (acetate) with DMRB. As a result, vivianite formation was controlled mainly by the amount of Fe3+ in WAS. In practice, a Fe/P molar ratio of 2:1 should be enough for vivianite formation in AD of WAS. Otherwise, exogenously dosing Fe3+ or Fe2+ into AD must be applied in AD.

Biochar facilitated bacterial reduction of Cr(VI) by Shewanella Putrefaciens CN32: Pathways and surface characteristics

Biochar can facilitate the microbial reduction of various pollutants in soil and groundwater environments, but its impact on Cr(VI) reduction by dissimilatory metal reducing bacteria (DMRB) remains to be systematically investigated. In this study, we prepared biochars at 500 °C and 700 °C from wheat straw and grass, and investigated the impact of these biochars on Cr(VI) reduction by a model DMRB, Shewanella Putrefaciens CN32 (CN32). Pristine biochars abiotically reduced Cr(VI), which decreased the concentration and toxicity of chromium to CN32 cells, and brought about higher overall Cr(VI) removal extent after CN32 were added sequentially; on the other hand, no enhancement effect were observed when biochars and CN32 were added simultaneously. Further tests between biologically reduced biochars and Cr(VI) revealed that the reaction rates between bioreduced biochars and Cr(VI) are relatively sluggish compared to that of direct Cr(VI) reduction by CN32, which prohibited biochars from directly accelerating the Cr(VI) reduction by CN32 in simultaneous-addition scenario. The relative importance of biochars’ surface functional groups and surface areas on their reactivities towards Cr(VI) reduction were also investigated. This study deepened our understanding towards the role of biochar played during bacterial Cr(VI) reduction and could potentially contribute to optimizing the biochar-based Cr(VI) bioremediation strategies.

Enhancement effects of dissolved organic matter leached from sewage sludge on microbial reduction and immobilization of Cr(VI) by Geobacter sulfurreducens

Sewage sludge has a high concentration of dissolved organic matter (DOM) which contains compounds that can serve as electron donors or shuttles for metal reduction by dissimilatory metal reducing bacteria (DMRB). In this study, Cr(VI) removal by G. sulfurreducens, a common DMRB present in anaerobic soils, was examined in the presence or absence of sludge DOM. Two different types of sludge DOM were tested; composted sludge DOM (C-DOM) and anaerobically digested sludge DOM (A-DOM). Both sludge DOMs enhanced Cr(VI) reduction by G. sulfurreducens, but C-DOM was more effective likely because it had higher concentrations of humic substances that served as electron shuttles. Transcriptomic studies indicated that G. sulfurreducens utilizes several different mechanisms to tolerate chromium including extracellular Cr(VI) reduction and immobilization by outer membrane c-type cytochromes and electrically conductive pili, intracellular Cr(VI) reduction by triheme cytochromes and NAD(P)H FMN reductase proteins, and chromium efflux by several P-type ATPase and RND transporter proteins. Microscopy experiments also showed that Cr(III) crystals formed on the surface of the cells, indicating that extracellular Cr(VI) reduction and adsorption was involved in the chromium removal process. These results help provide insight into the potential use of sewage sludge as an additive to enhance the bioremediation of chromium contaminated soils.

Size-dependent visible-light-enhanced Cr(VI) bioreduction by hematite nanoparticles

Light irradiation would affect the electron transfer between dissimilatory metal-reducing bacteria (DMRB) and semiconducting minerals, which may impose a great influence on the biogeochemistry cycle of heavy metals. However, the size effect of semiconducting minerals on the its electron transfer with DMRB and microbial Cr(VI) reduction under visible light irradiation is little known. Herein, the Cr(VI) reduction by Shewanella oneidensis MR-1 (MR-1) was investigated in the presence of hematite nanoparticles with average diameters of 10 nm and 50 nm in dark and under visible light irradiation. It is found that hematite nanoparticles adhered onto MR-1 cells to form the composites, leading to the decrease in surface sites and Zeta potential. Hematite mediated-Cr(VI) bioreduction rate under visible light irradiation was 0.342 h−1, which is 3.4 folds enhancement compared with that in dark and 4.4 folds compared with the MR-1 alone under visible light irradiation. Decreasing nanoparticle size of hematite from 50 nm to 10 nm promoted the Cr(VI) reduction under visible light irradiation but impeded it in dark. It was deduced that the bioelectrons from MR-1 could promote the separation of photoelectron-hole pairs of light-irradiated hematite, which consequently enhanced the Cr(VI) bioreduction by MR-1-hematite composites. Moreover, mutant strains experiments demonstrated the vital role of c-cytochrome for the conducting network actively established by MR-1 with hematite nanoparticles. Those findings expand the understanding of the electron transfer pathway for enhancing Cr(VI) reduction by hematite-MR-1 composites, and the impact of particle size on the interaction between semiconducting mineral and electroactive bacteria under light irradiation.

The transformation of U(VI) and V(V) in carnotite group minerals during dissimilatory respiration by a metal reducing bacterium

Recent results from laboratory and field studies support that dissimilatory metal reducing (DMR) bacteria influence the fate and transport of uranium in anaerobic subsurface environments. To date, most research efforts have focused on the reduction of soluble U(VI) by DMR bacteria to form insoluble uraninite (UO2). Subsurface environments harbor, however, large reservoirs of U(VI) in solid or mineral form. Uranium that is structure-bound in minerals is expected to be more refractory to microbial reduction than soluble U, based on analogy with Fe respiration. The reducibility of U(VI) could impact the fate of U(IV) by controlling mineral precipitation reactions, which has implications for the long-term immobilization of U in subsurface environments. We studied anoxic cultures of Shewanella putrefaciens CN32 incubated with natural carnotite-group minerals by X-ray diffraction, electron microscopy, scanning transmission X-ray microscopy (STXM). Near-edge X-ray absorption fine structure (NEXAFS) spectroscopy measurements at U-N4,5, V-L2,3, and O-K edges on cultures incubated up to 10 months show that V(V) was reduced to V(IV), whereas U was not reduced. In contrast, V(V) and U(VI) in solution were both completely reduced to lower oxidation states by CN32, as precipitates within the exopolymer surrounding the bacteria. Assays for the toxicity of U and V to CN32 showed that biofilm formation was stimulated at 0.001 M U(VI), and growth was inhibited at concentrations of U(VI) greater than 0.001 M. Vanadium did not inhibit growth or stimulate biofilm formation at any concentration tested. Investigations of the bacteria-mineral and bacteria-metal interface at the nanometer and molecular scales provide new insights into the co-respiration of V and U that help explain their biogeochemical cycling and have implications for subsurface bioremediation of these elements.

New Mobilization Pathway of Antimonite: Thiolation and Oxidation by Dissimilatory Metal-Reducing Bacteria via Elemental Sulfur Respiration.

Antimony (Sb) mobilization is widely explored with dissimilatory metal-reducing bacteria (DMRB) via microbial iron(III)-reduction. Here, our study found a previously unknown pathway whereby DMRB release adsorbed antimonite (SbIII-O) from goethite via elemental sulfur (S0) respiratory reduction under mild alkaline conditions. We incubated SbIII-O-loaded goethite with Shewanella oneidensis MR-1 in the presence of S0 at pH 8.5. The incubation results showed that MR-1 reduced S0 instead of goethite, and biogenic sulfide induced the formation of thioantimonite (SbIII-S). SbIII-S was then oxidized by S0 to mobile thioantimonate (SbV-S), resulting in over fourfold greater Sb release to water compared with the abiotic control. SbIV-S was identified as the intermediate during the oxidation process by Fourier transform ion cyclotron resonance mass spectrometry and electron spin resonance analysis. The existence of SbIV-S reveals that the oxidation of SbIII-S to SbV-S follows a two-step consecutive one-electron transfer from Sb to S atoms. SbV-S then links with SbIII-S by sharing S atoms and inhibits SbIII-S polymerization and SbIII2S3 precipitation like a "capping agent". This study clarifies the thiolation and oxidation pathway of SbIII-O to SbV-S by S0 respiration and expands the role of DMRB in the fate of Sb.

Biogenic iron sulfide functioning as electron-mediating interface to accelerate dissimilatory ferrihydrite reduction by Shewanella oneidensis MR-1

Microbially driven iron and sulfur geochemical cycles co-exist ubiquitously in subsurface environments and are of environmental relevance. Shewanella species (dissimilatory metal-reducing bacteria) are capable of reducing Fe(III)-(oxyhydr)oxide minerals and diverse sulfur sources using corresponding metabolic pathways and producing FeS secondary minerals. In spite of the ability in promoting bacterial extracellular electron transfer (EET), the specific role of FeS in mediating EET between microbe/mineral interface is still unclear. In this work, the electron-mediating function of biogenic FeS on promoting the reduction of ferrihydrite by S. oneidensis MR-1 using thiosulfate as sulfur source was investigated in terms of Fe(III) reduction percentage, X-ray diffraction and scanning electron microscopy. The results showed that the microbial ferrihydrite reduction was pH-dependent and positively correlated with the addition of thiosulfate. In the presence of thiosulfate, biogenic FeS in nano-scale were formed and deposited on the surfaces of S. oneidensis MR-1 and ferrihydrite to build an interfacial electron transfer bridge between them. The addition of either thiosulfate and in-vitro FeS could rescue the entirely inactivated ability of the mutant (△omcA/mtrC) in ferrihydrite reduction to some extent, but which was obviously inferior to the wild-type strain. Meanwhile, the effect of the biogenic FeS in-situ coating on the surfaces of S. oneidensis MR-1 cells on promoting microbial ferrihydrite reduction was significantly superior to the in-vitro ones. Thus, the in-situ formed biogenic FeS secondary minerals were demonstrated to mediate and accelerate interfacial electron transfer from S. oneidensis MR-1 cells to ferrihydrite through interfacing with the bacterial EET routes, especially Mtr pathway. This work provides an insight into the secondary minerals-mediating interfacial electron transfer between microbes and minerals in the presence of biological S (-II), which has important biogeochemical and environmental implications.

Enhanced Bioreduction of Radionuclides by Driving Microbial Extracellular Electron Pumping with an Engineered CRISPR Platform.

Dissimilatory metal-reducing bacteria (DMRB) with extracellular electron transfer (EET) capability show great potential in bioremediating the subsurface environments contaminated by uranium through bioreduction and precipitation of hexavalent uranium [U(VI)]. However, the low EET efficiency of DMRB remains a bottleneck for their applications. Herein, we develop an engineered CRISPR platform to drive the extracellular electron pumping of Shewanella oneidensis, a representative DMRB species widely present in aquatic environments. The CRISPR platform allows for highly efficient and multiplex genome editing and rapid platform elimination post-editing in S. oneidensis. Enabled by such a platform, a genomic promoter engineering strategy (GPS) for genome-widely engineering the EET-encoding gene network was established. The production of electron conductive Mtr complex, synthesis of electron shuttle flavin, and generation of NADH as intracellular electron carrier are globally optimized and promoted, leading to a significantly enhanced EET ability. Applied to U(VI) bioreduction, the edited strains achieve up to 3.62-fold higher reduction capacity over the control. Our work endows DMRB with an enhanced ability to remediate the radionuclides-contaminated environments and provides a gene editing approach to handle the growing environmental challenges of radionuclide contaminations.

Iron Cycle Tuned by Outer-Membrane Cytochromes of Dissimilatory Metal-Reducing Bacteria: Interfacial Dynamics and Mechanisms In Vitro.

The biogeochemical cycle of iron is of great importance to living organisms on Earth, and dissimilatory metal-reducing bacteria (DMRB) with the capability of reducing hematite (α-Fe2O3) by outer-membrane (OM) cytochromes play a great role in the iron cycle. However, the dynamic binding of cytochromes to α-Fe2O3 at the molecular level and the resulting impact on the photon-to-electron conversion of α-Fe2O3 for the iron cycle are not fully understood. To address these issues, two-dimensional IR correlation analysis coupled with molecular dynamics (MD) simulations was conducted for an OmcA-Fe2O3 system as OmcA bonds stronger with hematite in a typical DMRB,Shewanella. The photoelectric response of α-Fe2O3 with the OmcA coating was evaluated at three different potentials. Specifically, the binding groups from OmcA to α-Fe2O3 were in the sequence of carboxyl groups, amide II, and amide I. Further MD analysis reveals that both electrostatic interactions and hydrogen bonds played essential roles in the binding process, leading to the structural changes of OmcA to facilitate iron reduction. Moreover, the OmcA coating could store the photogenerated electrons from α-Fe2O3 like a capacitor and utilize the stored electrons for α-Fe2O3 reduction in dark and anoxic environments, further driving the biogeochemical cycle of iron. These investigations give the dynamic information on the OM protein/hematite interaction and provide fundamental insights into the biogeochemical cycle of iron by taking the photon-induced redox chemistry of iron oxide into consideration.

Sunlight-triggered synergy of hematite and Shewanella oneidensis MR-1 in Cr(VI) removal

Hematite is usually considered as a terminal electron acceptor for dissimilatory metal-reducing bacteria (DMRB), such as Shewanella oneidensis MR-1. However, hematite is also a semiconductor with visible light response. How the photocatalytic activity of hematite affects its electrical interplay with DMRB, as well as its role in relevant biogeochemical processes under sunlight, is still unclear. In this study, we investigated the effect of hematite on Cr(VI) removal by S. oneidensis MR-1 in the dark versus under stimulated sunlight using both batch experiments and photoelectrochemical analysis in a solar-assisted microbial photoelectrochemical system with a hematite photoanode covered by S. oneidensis MR-1. Under the dark conditions, hematite at low mineral-to-cell ratios can promote Cr(VI) removal through adsorbing both Cr(VI) and bacteria on/near hematite surface, which facilitates Cr(VI) bio-reduction and also alleviates self-poisoning processes of cells with time. However, as mineral-to-cell ratios reach a high level, hematite particles may cover cell surface and impact Cr(VI) bio-reduction, leading to the decreased Cr(VI) removal with increasing hematite particles. Under simulated sunlight, S. oneidensis MR-1 generates electrons from lactate metabolism and utilizes them to fill photoexcited holes in hematite, generating photoexcited electrons to reduce Cr(VI). Thus, in addition to directly enzymatic reduction of Cr(VI), the new light-triggered electron transfer pathway: lactate → S. oneidensis MR-1 → hematite → Cr(VI) further increases Cr(VI) removal and lactate metabolism. Also, the time-dependent cell survival is increased by the presence of hematite under the simulated sunlight, probably owing to the promoted Cr(VI) reduction and accumulation of Cr(III)-products on hematite surface. Moreover, organic hole scavenger, such as Ethylenediaminetetraacetic acid (EDTA), can further enhance Cr(VI) removal by hematite and S. oneidensis MR-1 under light irradiation. The photoelectrochemical results confirm that the light-triggered electron transfer pathway can be promptly and repeatedly produced upon illumination, and the rapid decrease of photocurrents after spiking Cr(VI) indicates Cr(VI) reduction by the photogenerated electrons from hematite. These findings suggest that semiconducting minerals, like hematite, can harvest solar energy to boost microbial metabolism and contaminant transformation by non-phototrophic, electroactive bacteria, which in turn increases bacterial tolerance toward toxic compounds in surrounding environments.

Ferrihydrite Reduction by Photosynthetic Synechocystis sp. PCC 6803 and Its Correlation With Electricity Generation.

Microbial extracellular electron transfer (EET) to solid-state electron acceptors such as anodes and metal oxides, which was originally identified in dissimilatory metal-reducing bacteria, is a key process in microbial electricity generation and the biogeochemical cycling of metals. Although it is now known that photosynthetic microorganisms can also generate (photo)currents via EET, which has attracted much interest in the field of biophotovoltaics, little is known about the reduction of metal (hydr)oxides via photosynthetic microbial EET. The present work quantitatively assessed the reduction of ferrihydrite in conjunction with the EET of the photosynthetic microbe Synechocystis sp. PCC 6803. Microbial reduction of ferrihydrite was found to be initiated in response to light but proceeded at higher rates when exogenous glucose was added, even under dark conditions. These results indicate that current generation from Synechocystis cells does not always need light irradiation. The qualitative trends exhibited by the ferrihydrite reduction rates under various conditions showed significant correlation with those of the microbial currents. Notably, the maximum concentration of Fe(II) generated by the cyanobacterial cells under dark conditions in the presence of glucose was comparable to the levels observed in the photic layers of Fe-rich microbial mats.

Shewanella oneidensis MR‐1 in situ biosynthesis of Ag nanoparticles on TiO 2 nanotubes with enhanced photocatalytic performance

Biogenetic nanocomposites research provides valuable methods for the green synthesis of nanomaterials. As a dissimilatory metal-reducing bacterium, Shewanella oneidensis MR-1 is used to reduce Ag+ to Ag nanoparticles (Ag NPs) (diameter about 10 nm) under anaerobic conditions, resulting in the in-situ formation of Ag NPs immobilised on TiO2 nanotubes (TNTs) (Ag/TNTs nanocomposites). The loading amount of Ag nanocrystals on the TNT surface can be controlled easily through adjusting the AgNO3 concentration and further influence visible-light absorption efficiency of Ag/TNTs nanocomposites by decreasing the Ag loading amount. Ag/TNTs nanocomposites show superior photocatalytic efficiency under simulated sunlight than single TiO2 nanomaterials. Moreover, the photocatalytic capacity of Ag/TNTs nanocomposites synergistic by S. oneidensis MR-1 is further enhanced and the degradation ratio of methylene blue reaches 92.3% within 30 min which attributed to a synergistic effect.

Extracellular electron transfer through visible light induced excited-state outer membrane C-type cytochromes of Geobacter sulfurreducens

Dissimilatory metal-reducing bacteria (DMRB) have a variety of c-type cytochromes (OM c-cyts) intercalated in their outer membrane, and this structure serves as the physiological basis for DMRB to carry out the extracellular electron transfer processes. Using Geobacter sulfurreducens as a model DMRB, we demonstrated that visible-light illumination could alter the electronic state of OM c-cyts from the ground state to the excited state in vivo. The existence of excited-state OM c-cyts in vivo was confirmed by spectroscopy. More importantly, excited-state OM c-cyts had a more negative potential compared to their ground-state counterparts, conferring DMRB with an extra pathway to transfer electrons to semi-conductive electron acceptors. To demonstrate this, using a TiO2-coated electrode as an electron acceptor, we showed that G. sulfurreducens could directly utilise the conduction band of TiO2 as an electron acceptor under visible-light illumination (λ > 420 nm) without causing TiO2 charge separation. When G. sulfurreducens was subject to visible-light illumination, the rate of extracellular electron transfer (EET) to TiO2 accelerated by over 8-fold compared to that observed under dark conditions. Results of additional electrochemical tests provided complementary evidence to support that G. sulfurreducens utilised excited-state OM c-cyts to enhance EET to TiO2.

Effects of biofilm transfer and electron mediators transfer on Klebsiella quasipneumoniae sp. 203 electricity generation performance in MFCs

BackgroundExtracellular electron transfer (EET) is essential in improving the power generation performance of electrochemically active bacteria (EAB) in microbial fuel cells (MFCs). Currently, the EET mechanisms of dissimilatory metal-reducing (DMR) model bacteria Shewanella oneidensis and Geobacter sulfurreducens have been thoroughly studied. Klebsiella has also been proved to be an EAB capable of EET, but the EET mechanism has not been perfected. This study investigated the effects of biofilm transfer and electron mediators transfer on Klebsiella quasipneumoniae sp. 203 electricity generation performance in MFCs.ResultsHerein, we covered the anode of MFC with a layer of microfiltration membrane to block the effect of the biofilm mechanism, and then explore the EET of the electron mediator mechanism of K. quasipneumoniae sp. 203 and electricity generation performance. In the absence of short-range electron transfer, we found that K. quasipneumoniae sp. 203 can still produce a certain power generation performance, and coated-MFC reached 40.26 mW/m2 at a current density of 770.9 mA/m2, whereas the uncoated-MFC reached 90.69 mW/m2 at a current density of 1224.49 mA/m2. The difference in the electricity generation performance between coated-MFC and uncoated-MFC was probably due to the microfiltration membrane covered in anode, which inhibited the growth of EAB on the anode. Therefore, we speculated that K. quasipneumoniae sp. 203 can also perform EET through the biofilm mechanism. The protein content, the integrity of biofilm and the biofilm activity all proved that the difference in the electricity generation performance between coated-MFC and uncoated-MFC was due to the extremely little biomass of the anode biofilm. To further verify the effect of electron mediators on electricity generation performance of MFCs, 10 µM 2,6-DTBBQ, 2,6-DTBHQ and DHNA were added to coated-MFC and uncoated-MFC. Combining the time–voltage curve and CV curve, we found that 2,6-DTBBQ and 2,6-DTBHQ had high electrocatalytic activity toward the redox reaction of K. quasipneumoniae sp. 203-inoculated MFCs. It was also speculated that K. quasipneumoniae sp. 203 produced 2,6-DTBHQ and 2,6-DTBBQ.ConclusionsTo the best of our knowledge, the three modes of EET did not exist separately. K. quasipneumoniae sp.203 will adopt the corresponding electron transfer mode or multiple ways to realize EET according to the living environment to improve electricity generation performance.

Biomineralization of Cu2S Nanoparticles by Geobacter sulfurreducens.

Biomineralization of Cu has been shown to control contaminant dynamics and transport in soils. However, very little is known about the role that subsurface microorganisms may play in the biogeochemical cycling of Cu. In this study, we investigate the bioreduction of Cu(II) by the subsurface metal-reducing bacterium Geobacter sulfurreducens Rapid removal of Cu from solution was observed in cell suspensions of G. sulfurreducens when Cu(II) was supplied, while transmission electron microscopy (TEM) analyses showed the formation of electron-dense nanoparticles associated with the cell surface. Energy-dispersive X-ray spectroscopy (EDX) point analysis and EDX spectrum image maps revealed that the nanoparticles are rich in both Cu and S. This finding was confirmed by X-ray absorption near-edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) analyses, which identified the nanoparticles as Cu2S. Biomineralization of CuxS nanoparticles in soils has been reported to enhance the colloidal transport of a number of contaminants, including Pb, Cd, and Hg. However, formation of these CuxS nanoparticles has only been observed under sulfate-reducing conditions and could not be repeated using isolates of implicated organisms. As G. sulfurreducens is unable to respire sulfate, and no reducible sulfur was supplied to the cells, these data suggest a novel mechanism for the biomineralization of Cu2S under anoxic conditions. The implications of these findings for the biogeochemical cycling of Cu and other metals as well as the green production of Cu catalysts are discussed.IMPORTANCE Dissimilatory metal-reducing bacteria are ubiquitous in soils and aquifers and are known to utilize a wide range of metals as terminal electron acceptors. These transformations play an important role in the biogeochemical cycling of metals in pristine and contaminated environments and can be harnessed for bioremediation and metal bioprocessing purposes. However, relatively little is known about their interactions with Cu. As a trace element that becomes toxic in excess, Cu can adversely affect soil biota and fertility. In addition, biomineralization of Cu nanoparticles has been reported to enhance the mobilization of other toxic metals. Here, we demonstrate that when supplied with acetate under anoxic conditions, the model metal-reducing bacterium Geobacter sulfurreducens can transform soluble Cu(II) to Cu2S nanoparticles. This study provides new insights into Cu biomineralization by microorganisms and suggests that contaminant mobilization enhanced by Cu biomineralization could be facilitated by Geobacter species and related organisms.