Metal-reducing Bacterium Shewanella Oneidensis Research Articles

Nanoscale Carbonate Ion-Selective Amperometric/Voltammetric Probes Based on Ion-Ionophore Recognition at the Organic/Water Interface: Hidden Pieces of the Puzzle in the Nanoscale Phase.

Here, we report on the successful demonstration and application of carbonate (CO32-) ion-selective amperometric/voltammetric nanoprobes based on facilitated ion transfer (IT) at the nanoscale interface between two immiscible electrolyte solutions. This electrochemical study reveals critical factors to govern CO32--selective nanoprobes using broadly available Simon-type ionophores forming a covalent bond with CO32-, i.e., slow dissolution of lipophilic ionophores in the organic phase, activation of hydrated ionophores, peculiar solubility of a hydrated ion-ionophore complex near the interface, and cleanness at the nanoscale interface. These factors are experimentally confirmed by nanopipet voltammetry, where a facilitated CO32- IT is studied with a nanopipet filled with an organic phase containing the trifluoroacetophenone derivative CO32-ionophore (CO32-ionophore VII) by voltammetrically and amperometrically sensing CO32- in water. Theoretical assessments of reproducible voltammetric data confirm that the dynamics of CO32- ionophore VII-facilitated ITs (FITs) follows the one-step electrochemical (E) mechanism controlled by both water-finger formation/dissociation and ion-ionophore complexation/dissociation during interfacial ITs. The yielded rate constant, k0 = 0.048 cm/s, is very similar to the reported values of other FIT reactions using ionophores forming non-covalent bonds with ions, implying that a weak binding between CO32- ion-ionophore enables us to observe FITs by fast nanopipet voltammetry regardless of the nature of bondings between the ion and ionophore. The analytical utility of CO32--selective amperometric nanoprobes is further demonstrated by measuring the CO32- concentration produced by metal-reducing bacteria Shewanella oneidensis MR-1 as a result of organic fuel oxidation in bacterial growth media in the presence of various interferents such as H2PO4-, Cl-, and SO42-.

Single molecule tracking of bacterial cell surface cytochromes reveals dynamics that impact long-distance electron transport

Using a series of multiheme cytochromes, the metal-reducing bacterium Shewanella oneidensis MR-1 can perform extracellular electron transfer (EET) to respire redox-active surfaces, including minerals and electrodes outside the cell. While the role of multiheme cytochromes in transporting electrons across the cell wall is well established, these cytochromes were also recently found to facilitate long-distance (micrometer-scale) redox conduction along outer membranes and across multiple cells bridging electrodes. Recent studies proposed that long-distance conduction arises from the interplay of electron hopping and cytochrome diffusion, which allows collisions and electron exchange between cytochromes along membranes. However, the diffusive dynamics of the multiheme cytochromes have never been observed or quantified in vivo, making it difficult to assess their hypothesized contribution to the collision-exchange mechanism. Here, we use quantum dot labeling, total internal reflection fluorescence microscopy, and single-particle tracking to quantify the lateral diffusive dynamics of the outer membrane-associated decaheme cytochromes MtrC and OmcA, two key components of EET in S. oneidensis. We observe confined diffusion behavior for both quantum dot-labeled MtrC and OmcA along cell surfaces (diffusion coefficients DMtrC = 0.0192 ± 0.0018 µm2/s, DOmcA = 0.0125 ± 0.0024 µm2/s) and the membrane extensions thought to function as bacterial nanowires. We find that these dynamics can trace a path for electron transport via overlap of cytochrome trajectories, consistent with the long-distance conduction mechanism. The measured dynamics inform kinetic Monte Carlo simulations that combine direct electron hopping and redox molecule diffusion, revealing significant electron transport rates along cells and membrane nanowires.

Electrical properties of outer membrane extensions from Shewanella oneidensis MR-1.

Shewanella oneidensis MR-1 is a metal-reducing bacterium that is able to exchange electrons with solid-phase minerals outside the cell. These bacterial cells can produce outer membrane extensions (OMEs) that are tens of nanometers wide and several microns long. The capability of these OMEs to transport electrons is currently under investigation. Tubular chemically fixed OMEs from S. oneidensis have shown good dc conducting properties when measured in an air environment. However, no direct demonstration of the conductivity of the more common bubble-like OMEs has been provided yet, due to the inherent difficulties in measuring it. In the present work, we measured the electrical properties of bubble-like OMEs in a dry air environment by Scanning Dielectric Microscopy (SDM) in force detection mode. We found that at the frequency of the measurements (∼2 kHz), OMEs show an insulating behavior, with an equivalent homogeneous dielectric constant εOME = 3.7 ± 0.7 and no dephasing between the applied ac voltage and the measured ac electric force. The dielectric constant measured for the OMEs is comparable to that obtained for insulating supramolecular protein structures (εprotein = 3-4), pointing towards a rich protein composition of the OMEs, probably coming from the periplasm. Based on the detection sensitivity of the measuring instrument, the upper limit for the ac longitudinal conductivity of bubble-like OMEs in a dry air environment has been set to σOME,ac < 10-5 S m-1, a value several orders of magnitude smaller than the dc conductivity measured in tubular chemically fixed OMEs. The lack of conductivity of bubble-like OMEs can be attributed to the relatively large separation between cytochromes in these larger OMEs and to the suppression of cytochrome mobility due to the dry environmental conditions.

Effects of flow-interruption on the bacteria transport behavior in porous media

Attachment is an important strategy for microorganisms to survive, grow, and perform biogeochemical functions in soil and groundwater systems. This study investigated the effect of flow-interruption (FI) that is common in subsurface environments on bacterial attachment/detachment using a metal-reducing bacterium Shewanella oneidensis strain (MR1) as an example. Column experiments and one dimensional bacterial transport models were performed by varying flow interruption duration, fluid ionic strength, and solid grain surface electrostatic properties with a goal to understand the effect of FI on bacterial attachment/detachment. The results indicated that MR1 generally behaved as colloidal particles in transport and attachment/detachment under continuous flow conditions. However, the FI events induced a significant release of MR1 in both injection and elution phases, showing a peak MR1 concentration, which was higher than that in the injecting solution, immediately after the FI events. The peak MR1 concentrations were higher in systems with lower adhesive forces between bacteria and solid grains, a lower attachment site capacity in porous media, and a longer FI duration. The diffusive release of the previously attached bacteria during the FI events and the momentum force when flow re-started immediately after the FI events were likely the mechanisms leading to bursting release of bacteria with the release concentrations higher than that in the influent solutions. The results have important implication for understanding bacterial migration and attachment, and their subsequent functions, especially in dynamic hydraulic environments such as in hyporheic zones or in soils subject to dynamic changes in infiltration rates under for example extreme precipitation and drought conditions.

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.

Degradation of 2, 2′, 4, 4′-Tetrabrominated diphenyl ether (BDE-47) via the Fenton reaction driven by the dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1

A microbially facilitated approach was developed to degrade 2, 2′, 4, 4′-tetrabrominated diphenyl ether (BDE-47). This approach consisted of biological production of Fe(II) and H2O2 by the dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1 during the repetitive anoxic/oxic cycles and abiotic production of hydroxyl radical (HO●) with the biologically produced Fe(II) and H2O2 via Fenton reaction. Under the condition tested, BDE-47 did not inhibit the growth of S. oneidensis MR-1. Water soluble Fe(III)-citrate and the solid minerals ferrihydrite [Fe(III)2O3•0.5H2O] and goethite [Fe(III)OOH] were tested in this study. Under anoxic condition, the amounts of Fe(II) produced by S. oneidensis MR-1 varied among the Fe(III)s tested, which decreased in the order of Fe(III)-citrate > ferrihydrite > goethite. Under subsequent oxic condition, H2O2 was produced via O2 reduction by S. oneidensis MR-1. The amounts of H2O2 detected also varied, which decreased in the order of the reactions with Fe(III)-citrate > goethite > ferrihydrite. S. oneidensis MR-1 maintained its ability to produce Fe(II) and H2O2 for up to seven anoxic/oxic cycles. At each end of anoxic/oxic cycle, HO● was detected. The amount of HO● produced decreased in the order of the reactions with ferrihydrite > goethite > Fe(III)-citrate, which was opposite to that of H2O2 detected. Compared to the controls without HO●, the amounts of BDE-47 in the reactions with HO● decreased. The more HO● in the reaction, the less amount of BDE-47 detected. Furthermore, no BDE-47 degradation was observed when HO● was scavenged or ferrihydrite was either omitted or replaced by nitrate. Finally, identification of degradation products, such as hydroxylated BDE-47 and trisBDE, dibromophenol and monobromophenol, suggested that OH-addition and Br-substitution by HO● were the main mechanisms for degrading BDE-47. Collectively, all these results demonstrated for the first time that the Fenton reaction driven by S. oneidensis MR-1 degraded BDE-47 effectively.

Biotic dissolution of autunite under anaerobic conditions: effect of bicarbonates and Shewanella oneidensis MR1 microbial activity.

Uranium is a contaminant of major concern across the US Department of Energy complex that served a leading role in nuclear weapon fabrication for half a century. In an effort to decrease the concentration of soluble uranium, tripolyphosphate injections were identified as a feasible remediation strategy for sequestering uranium in situ in contaminated groundwater at the Hanford Site. The introduction of sodium tripolyphosphate into uranium-bearing porous media results in the formation of uranyl phosphate minerals (autunite) of general formula {X1-2[(UO2)(PO4)]2-1·nH2O}, where X is a monovalent or divalent cation. The stability of the uranyl phosphate minerals is a critical factor that determines the long-term effectiveness of this remediation strategy that can be affected by biogeochemical factors such as the presence of bicarbonates and bacterial activity. The objective of this research was to investigate the effect of bicarbonate ions present in the aqueous phase on Ca-autunite dissolution under anaerobic conditions, as well as the role of metal-reducing facultative bacterium Shewanella oneidensis MR1. The concentration of total uranium determined in the aqueous phase was in direct correlation to the concentration of bicarbonate present in the solution, and the release of Ca, U and P into the aqueous phase was non-stoichiometric. Experiments revealed the absence of an extensive biofilm on autunite surface, while thermodynamic modeling predicted the presence of secondary minerals, which were identified through microscopy. In conclusion, the dissolution of autunite under the conditions studied is susceptible to bicarbonate concentration, as well as microbial presence.

Spin-Dependent Electron Transport through Bacterial Cell Surface Multiheme Electron Conduits.

Multiheme cytochromes, located on the bacterial cell surface, function as long-distance (>10 nm) electron conduits linking intracellular reactions to external surfaces. This extracellular electron transfer process, which allows microorganisms to gain energy by respiring solid redox-active minerals, also facilitates the wiring of cells to electrodes. While recent studies have suggested that a chiral induced spin selectivity effect is linked to efficient electron transmission through biomolecules, this phenomenon has not been investigated in extracellular electron conduits. Using magnetic conductive probe atomic force microscopy, Hall voltage measurements, and spin-dependent electrochemistry of the decaheme cytochromes MtrF and OmcA from the metal-reducing bacterium Shewanella oneidensis MR-1, we show that electron transport through these extracellular conduits is spin-selective. Our study has implications for understanding how spin-dependent interactions and magnetic fields may control electron transport across biotic-abiotic interfaces in both natural and biotechnological systems.

Surface-Induced Formation and Redox-Dependent Staining of Outer Membrane Extensions in Shewanella oneidensis MR-1

The metal-reducing bacterium Shewanella oneidensis MR-1 produces extensions of its outer membrane (OM) and periplasm that contain cytochromes responsible for extracellular electron transfer (EET) to external redox-active surfaces, including minerals and electrodes. While the role of multi-heme cytochromes in transporting electrons across the cell wall is well established, their distribution along S. oneidensis OM extensions is also thought to allow lateral electron transport along these filaments. These proposed bacterial nanowires, which can be several times the cell length, would thereby extend EET to more distant electron acceptors. However, it is still unclear why these extensions form, and to what extent they contribute to respiration in living cells. Here, we investigate physical contributors to their formation using in vivo fluorescence microscopy. While previous studies focused on the display of S. oneidensis outer membrane extensions (OMEs) as a response to oxygen limitation, we find that cell-to-surface contact is sufficient to trigger the production of OMEs, including some that reach >100 µm in length, irrespective of medium composition, agitation, or aeration. To visualize the extent of heme redox centers along OMEs, and help distinguish these structures from other extracellular filaments, we also performed histochemical redox-dependent staining with transmission electron microscopy on wild type and cytochrome-deficient strains. We demonstrate that redox-active components are limited to OMEs and not present on other extracellular appendages, such as pili and flagella. We also observed that the loss of 8 functional periplasmic and outer membrane cytochromes significantly decreased both the frequency and intensity of redox-dependent staining found widespread on OMEs. These results will improve our understanding of the environmental conditions that influence the formation of S. oneidensis OMEs, as well as the distribution and functionality of EET components along extracellular appendages.

Modeling the Structure of Type IV Pili from Shewanella Oneidensis

The metal-reducing bacterium Shewanella oneidensis produces conductive nanowires, which are described as extensions of the outer membrane or type IV pili (T4P), of which the structural details remain unknown. To better understand the structure of S. oneidensis T4P as a nanowire candidate, and the potential mechanism for electron transport along them, theoretical energy-minimized models of these pili were constructed. The models showed a discontinuous geometry consisting of aromatic rings, with large gaps of ∼10 Å, providing a potential pathway for electron hopping, which may account for previously reported conductivity. Moreover, mutations of the pilin subunit based on the models were proposed, which might fill the gaps and possibly increase the conductivity. These models will facilitate further investigation of the nanowires and their applications in bioenergy and bioelectronics.

A mechanistic study of TiO2 nanoparticle toxicity on Shewanella oneidensis MR-1 with UV-containing simulated solar irradiation: Bacterial growth, riboflavin secretion, and gene expression

Toxicity of nanomaterials to ecological systems has recently emerged as an important field of research, and thus, many researchers are exploring the mechanisms of how nanoparticles impact organisms. Herein, we probe the mechanisms of bacteria-nanoparticle interaction by investigating how TiO2 nanoparticles impact a model organism, the metal-reducing bacterium Shewanella oneidensis MR-1. In addition to examining the effect of TiO2 exposure, the effect of synergistic simulated solar irradiation containing UV was explored in this study, as TiO2 nanoparticles are known photocatalysts. The data reveal that TiO2 nanoparticles cause an inhibition of S. oneidensis growth at high dosage without compromising cell viability, yet co-exposure of nanoparticles and illumination does not increase the adverse effects on bacterial growth relative to TiO2 alone. Measurements of intracellular reactive oxygen species and riboflavin secretion, on the same nanoparticle-exposed bacteria, reveal that TiO2 nanoparticles have no effect on these cell functions, but application of UV-containing illumination with TiO2 nanoparticles has an impact on the level of riboflavin outside bacterial cells. Finally, gene expression studies were employed to explore how cells respond to TiO2 nanoparticles and illumination, and these results were correlated with cell growth and cell function assessment. Together these data suggest a minimal impact of TiO2 NPs and simulated solar irradiation containing UV on S. oneidensis MR-1, and the minimal impact could be accounted for by the nutrient-rich medium used in this work. These measurements demonstrate a comprehensive scheme combining various analytical tools to enable a mechanistic understanding of nanoparticle-cell interactions and to evaluate the potential adverse effects of nanoparticles beyond viability/growth considerations.

Microbial reduction of Cr(VI) in the presence of chromate conversion coating constituents

ABSTRACTThe reduction of Cr(VI) by the metal-reducing bacterium Shewanella oneidensis MR-1 was evaluated, to determine the potential for exploiting Cr(VI) bioreduction as a means of treating chromate conversion coating (CCC) waste streams. Inclusion of Cr(VI) at concentrations ≥1 mM inhibited aerobic growth of S. oneidensis, but that organism was able to reduce Cr(VI) at a concentration of up to 1 mM under anaerobic, nongrowth conditions. S. oneidensis reduced Cr(VI) in the presence of common CCC constituents, with the exception of ferricyanide, when these CCC constituents were included at concentrations typical of CCC waste streams. Ferricyanide inhibited neither aerobic growth nor metabolism under aerobic, nitrate- or iron-reducing conditions, suggesting that the ferricyanide-depended inhibition of Cr(VI) reduction is not due to broad metabolic inhibition, but is specific to Cr(VI) reduction. Results indicate that under some conditions, the activities of metal-reducing bacteria, such as S. oneidensis, could be exploited for the removal of Cr(VI) from CCC waste streams under appropriate conditions.

Biochar-Facilitated Microbial Reduction of Hematite.

As an important component of soil organic matter (SOM), the transformation of pyrogenic carbon plays a critical role in the biogeochemical cycles of carbon and other redox-active elements such as iron (Fe). Herein, we studied the influences of wheat straw-derived biochars on the microbial reduction of 100 mM of hematite by the dissimilatory metal reducing bacteria Shewanella oneidensis MR-1 under anoxic conditions. The long-term microbial reduction extent and initial reduction rate of hematite were accelerated by more than 2-fold in the presence of 10 mg L(-1) biochar. Soluble leachate from 10 mg L(-1) biochar enhanced Fe(III) reduction to a similar degree. Microbially prereduced biochar leachate abiotically reduced hematite, consistent with the apparent electron shuttling capacity of biochar leachate. Electron paramagnetic resonance (EPR) analysis suggested that biochar leachate-associated semiquinone functional groups were likely involved in the redox reactions. In addition to electron shuttling effects, biochar particles sorbed 0.5-1.5 mM biogenic Fe(II) and thereby increased the long-term extent of hematite reduction by 1.4-1.7 fold. Our results suggest that Fe redox cycling may be strongly impacted by pyrogenic carbon in soils with relatively high content of indigenous pyrogenic carbon or substantial application of biochar.

Electron Cryo-Tomography of Nanowires in Shewanella Oneidensis MR-1

The dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1 is known to make ‘nanowires,’ conductive cell appendages used in electron transport. Novel live fluorescence imaging techniques recently clarified that the conductive filaments in S. oneidensis are not pili, as previously thought, but rather extensions of the outer membrane. However, their fine structure and mechanism remained unclear. Here, we report the first high-resolution in vivo images of nanowires in frozen-hydrated cells in a near-native state by electron cryo-tomography (ECT). We first developed a method for in vivo production of nanowires in cells grown on EM grids. Using correlative light microscopy and ECT, we then identified and imaged nanowires at high resolution and in three dimensions. Interestingly, we find that nanowires are composed of outer membrane vesicles (OMVs) strung together to form a chain. The size of the vesicles and the length and thickness of the vesicle chain are highly variable. However, even apparently smooth filaments such as those described previously are in fact, at higher resolution, revealed to be chains of OMVs. This work will help us investigate the electron transport mechanism along Shewanella nanowires. Additionally, this work has implications in understanding how OMVs, ubiquitous in Gram-negative bacteria, can assemble into chains.

Multiheme Cytochromes and the Bacterial Nanowires of Shewanella oneidensis MR-1: Regulation, Structure, and Extracellular Electron Transport Mechanisms

Dissimilatory metal-reducing bacteria can extract free energy from their environment by performing electron transfer to solid-phase minerals outside the cell. This extracellular electron transport (EET) has important implications in global elemental cycles as well as renewable energy technologies. Among the pathways for EET, bacterial nanowires have received significant attention in the past decade due to their unique ability to mediate long-range electron transport to electron acceptors microns away from the cell surface. Here we report a comprehensive characterization of the composition, structure, and regulatory network that underlies bacterial nanowires from the metal-reducing bacterium Shewanella oneidensis MR-1. Using fluorescent and atomic force techniques, we find that the Shewanella nanowires are extensions of the outer membrane and periplasm that contain multiple multiheme cytochromes. The localization of decaheme cytochromes MtrC and OmcA supports a multistep redox hopping mechanism, allowing long-range electron transport along a membrane network of heme cofactors that line the nanowires. The electron flux resulting from such a mechanism strongly depends on the cytochrome density and topology. Using correlated electron cryo-tomography and in vivo fluorescent microscopy, we are gaining new insight into the localization patterns of cytochromes along nanowires as well as the morphology and the formation mechanism of these structures. Finally, we report our progress on understanding the underlying regulatory network, by testing targeted mutations and analyzing the transcriptome of Shewanella chemostat cultures as they encounter electron acceptor limitation and form nanowires. The transcriptional response includes an increase in the expression of multiheme cytochromes, heme synthesis enzymes, and cytochrome maturation proteins. Our findings on the regulation, ultrastructure and electron transport mechanism help shape a biophysical understanding of these redox-functionalized membrane and vesicular extensions as a microbial strategy for electron transport and energy distribution.

Metabolite-enabled mutualistic interaction between Shewanella oneidensis and Escherichia coli in a co-culture using an electrode as electron acceptor.

Mutualistic interactions in planktonic microbial communities have been extensively studied. However, our understanding on mutualistic communities consisting of co-existing planktonic cells and biofilms is limited. Here, we report a planktonic cells-biofilm mutualistic system established by the fermentative bacterium Escherichia coli and the dissimilatory metal-reducing bacterium Shewanella oneidensis in a bioelectrochemical device, where planktonic cells in the anode media interact with the biofilms on the electrode. Our results show that the transfer of formate is the key mechanism in this mutualistic system. More importantly, we demonstrate that the relative distribution of E. coli and S. oneidensis in the liquid media and biofilm is likely driven by their metabolic functions towards an optimum communal metabolism in the bioelectrochemical device. RNA sequencing-based transcriptomic analyses of the interacting organisms in the mutualistic system potentially reveal differential expression of genes involved in extracellular electron transfer pathways in both species in the planktonic cultures and biofilms.

High-resolution structure of a type IV pilin from the metal-reducing bacterium Shewanella oneidensis.

BackgroundType IV pili are widely expressed among Gram-negative bacteria, where they are involved in biofilm formation, serve in the transfer of DNA, motility and in the bacterial attachment to various surfaces. Type IV pili in Shewanella oneidensis are also supposed to play an important role in extracellular electron transfer by the attachment to sediments containing electron acceptors and potentially forming conductive nanowires.ResultsThe potential nanowire type IV pilin PilBac1 from S. oneidensis was characterized by a combination of complementary structural methods and the atomic structure was determined at a resolution of 1.67 Å by X-ray crystallography. PilBac1 consists of one long N-terminal α-helix packed against four antiparallel β-strands, thus revealing the core fold of type IV pilins. In the crystal, PilBac1 forms a parallel dimer with a sodium ion bound to one of the monomers. Interestingly, our PilBac1 crystal structure reveals two unusual features compared to other type IVa pilins: an unusual position of the disulfide bridge and a straight α-helical section, which usually exhibits a pronounced kink. This straight helix leads to a distinct packing in a filament model of PilBac1 based on an EM model of a Neisseria pilus.ConclusionsIn this study we have described the first structure of a pilin from Shewanella oneidensis. The structure possesses features of the common type IV pilin core, but also exhibits significant variations in the α-helical part and the D-region.Electronic supplementary materialThe online version of this article (doi:10.1186/s12900-015-0031-7) contains supplementary material, which is available to authorized users.

Photocatalytic properties of zinc sulfide nanocrystals biofabricated by metal-reducing bacterium Shewanella oneidensis MR-1

Accumulation and utilization of heavy metals from wastewater by biological treatment system has aroused great interest. In the present study, a metal-reducing bacterium Shewanella oneidensis MR-1 was used to explore the biofabrication of ZnS nanocrystals from the artificial wastewater. The biogenic H2S produced via the reduction of thiosulfate precipitated the Zn(II) as sulfide extracellularly. Characterization by X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), and field emission scanning electron microscope (FESEM) confirmed the precipitates as ZnS nanocrystals. The biogenic ZnS nanocrystals appeared spherical in shape with an average diameter of 5nm and mainly aggregated in the medium and cell surface of S. oneidensis MR-1. UV–vis DRS spectra showed ZnS nanoparticles appeared a strong absorption below 360nm. Thus, the photocatalytic activity of ZnS was evaluated by the photodegradation of rhodamine B (RhB) under UV irradiation. The biogenic ZnS nanocrystals showed a high level of photodegradation efficiency to RhB coupled with a significant blue-shift of maximum adsorption peak. A detailed analysis indicated the photogenerated holes, rather than hydroxyl radicals, contributed to the photocatalytic decolorization of RhB. This approach of coupling biosynthesis of nanoparticles with heavy metal removal may offer a potential avenue for efficient bioremediation of heavy metal wastewater.

Bacterial Nanowires of Shewanella Oneidensis MR-1 are Outer Membrane and Periplasmic Extensions of the Extracellular Electron Transport Components

Bacterial nanowires offer a pathway for extracellular electron transfer (EET) by linking the respiratory chain of bacteria to external surfaces, including oxidized metals in the environment and engineered electrodes in renewable energy devices. Specifically, nanowires of the model metal-reducing bacterium Shewanella oneidensis MR-1 were previously shown to be conductive under non-physiological conditions. Despite the global, environmental, and technological consequences of bacterial nanowire-mediated EET, the composition, electron transport mechanism, and physiological relevance of these appendages remain unclear. The nanowires of S. oneidensis MR-1 were previously thought, but never shown, to be bacterial pili. In addition, the transport mechanism through bacterial nanowires has been the subject of intense debate, with “metallic-like” band transport and multistep redox hopping between multiheme cytochromes as the two proposed mechanisms. Here we report the first in vivo observations of the formation and respiratory impact of nanowires in S. oneidensis MR-1. Using live fluorescence measurements and quantitative gene expression analysis, we demonstrate that S. oneidensis MR-1 nanowires are extensions of the outer membrane and periplasm, rather than pilin-based structures. We show, through immunolabeling, that multiheme cytochromes localize to nanowires, in turn supporting the multistep redox hopping model as the transport mechanism. Furthermore, these bacterial nanowires are associated with outer membrane vesicles, structures ubiquitous in Gram-negative bacteria, and occasionally appear as membrane vesicle chains that transition to smoother filaments. Redox-functionalized membrane and vesicular extensions may represent a general microbial strategy for electron transport and energy distribution.

Accumulation of amorphous Cr(III)-Te(IV) nanoparticles on the surface of Shewanella oneidensis MR-1 through reduction of Cr(VI).

Industrial effluents constitute a major source of metal pollution of aquatic bodies. Moreover, due to their environmental persistence, toxic metal pollution is of special concern. Microbial reduction is considered a promising strategy for toxic metal removal among the several methods available for metal remediation. Here, we describe the coremediation of toxic Cr(VI) and Te(IV) by the dissimilatory metal reducing bacterium-Shewanella oneidensis MR-1. In the presence of both Cr(VI) and Te(IV), S. oneidensis MR-1 reduced Cr(VI) to the less toxic Cr(III) form, but not Te(IV) to Te(0). The reduced Cr(III) ions complexed rapidly with Te(IV) ions and were precipitated from the cell cultures. Electron microscopic analyses revealed that the Cr-Te complexed nanoparticles localized on the bacterial outer membranes. K-edge X-ray absorption spectrometric analyses demonstrated that Cr(III) produced by S. oneidensis MR-1 was rapidly complexed with Te(IV) ions, followed by formation of amorphous Cr(III)-Te(IV) nanoparticles on the cell surface. Our results could be applied for the simultaneous sequestration and detoxification of both Cr(VI) and Te(IV) as well as for the preparation of nanomaterials through environmental friendly processes.