Shewanella Loihica PV-4 Research Articles

Using static magnetic field to recover ammonia efficiently by DNRA process

Dissimilatory nitrate reduction to ammonium (DNRA) has garnered attention due to its ability to recover ammonia and reduce greenhouse gas emissions simultaneously. In this study, the potential of using static magnetic field (SMF) to improve DNRA process was explored from the sight of molecular biology. Functional genes, microbial community structure, and metabolism pathways were discussed. SMF of 40 mT shortened the start-up time of DNRA from 75 days to 41 days, while 80 mT SMF delayed it to 103 days. On day 80, DNRA potential rate under 40 mT SMF, reached 174 ± 11 μmol kg−1 h−1, significantly surpassing 0 mT (88 ± 6 μmol kg−1 h−1) and 80 mT SMF (52 ± 4 μmol kg−1 h−1). SMF of 40 mT also accelerated community succession and the enrichment of functional bacteria like Geobacter (from 15.71% to 32.11%). qPCR results suggested that 40 mT SMF promoted the rapid enrichment of DNRA functional gene nrfA and 80 mT SMF promoted the enrichment of nirS gene on day 40. Dynamic responses of Thauera sp. RT1901, Stutzerimonas stutzeri, Shewanella oneidensis MR-1, and Shewanella loihica PV-4 to SMF at transcriptional levels confirmed SMF could improve the nitrogen removal and electron transfer of DNRA and denitrification bacteria. Consequently, this work validated the possibility of using SMF to improve DNRA process for ammonia recovery and investigated the underlying mechanisms, which could promote the application of DNRA in full-scale.

Artificial and Biosynthetic Nanoparticles Boost Bioelectrochemical Reactions via Efficient Bidirectional Electron Transfer of Shewanella loihica.

Bioelectrochemical reactions using whole-cell biocatalysts are promising carbon-neutral approaches because of their easy operation, low cost, and sustainability. Bidirectional (outward or inward) electron transfer via exoelectrogens plays the main role in driving bioelectrochemical reactions. However, the low electron transfer efficiency seriously inhibits bioelectrochemical reaction kinetics. Here, a three dimensional and artificial nanoparticles-constituent inverse opal-indium tin oxide (IO-ITO) electrode is fabricated and employed to connect with exoelectrogens (Shewanella loihica PV-4). The above electrode collected 128-fold higher cell density and exhibited a maximum current output approaching 1.5mAcm-2 within 24h at anode mode. By changing the IO-ITO electrode to cathode mode, the exoelectrogens exhibited the attractive ability of extracellular electron uptake to reduce fumarate and 16 times higher reverse current than the commercial carbon electrode. Notably, Fe-containing oxide nanoparticles are biologically synthesized at both sides of the outer cell membrane and probably contributed to direct electron transfer with the transmembrane c-type cytochromes. Owing to the efficient electron exchange via artificial and biosynthetic nanoparticles, bioelectrochemical CO2 reduction is also realized at the cathode. This work not only explored the possibility of augmenting bidirectional electron transfer but also provided a new strategy to boost bioelectrochemical reactions by introducing biohybrid nanoparticles.

Bioelectricity and CO2-to-butyrate production using photobioelectrochemical cells with bio-hydrogel

Bio-photoelectrochemical cell (BPEC) is an emerging technology that can convert the solar energy into electricity or chemicals. However, traditional BPEC depending on abiotic electrodes is challenging for microbial/enzymatic catalysis because of the inefficient electron exchange. Here, electroactive bacteria (Shewanella loihica PV-4) were used to reduce graphene oxide (rGO) nanosheets and produce co-assembled rGO/Shewanella biohydrogel as a basic electrode. By adsorbing chlorophyll contained thylakoid membrane, this biohydrogel was fabricated as a photoanode that delivered maximum photocurrent 126 μA/cm3 under visible light. Impressively, the biohydrogel could be served as a cathode in BPEC by forming coculture system with genetically edited Clostridium ljungdahlii. Under illumination, the BPEC with above photoanode and cathode yielded ∼ 5.4 mM butyrate from CO2 reduction, 169 % increase compared to dark process. This work provided a new strategy (nanotechnology combined with synthetic biology) to achieve efficient bioelectricity and valuable chemical production in PBEC.

Carbon nanotubes modified reticulated vitreous carbon materials enhancing extracellular electron transfer of Shewanella loihica PV-4

Increasing current output of bioelectrochemical devices is crucial for their real-world application. Electrodes modified with conductive nanoparticles increase the contact area between anodic microorganisms and solid electron acceptors, thus resulting in higher cells concentration and current output. Electron transfer mechanisms in Shewanella loihica PV-4 viable biofilms formed at carbon nanotubes (CNTs)-coated RVC electrode is investigated in potentiostat-controlled electrochemical cells set at oxidative potentials. Chronoamperometry showed a stable biofilm growth with final current output above 150 µA·cm−2 within two weeks, which was 3 times higher than unmodified RVC systems. The current output showed biphasic growth, which could be attributed to outer layer and inner layer biofilm growth, respectively, where aerobic and anaerobic growth occurred, respectively. Cyclic voltammetry and differential pulse voltammetry analysis indicated a contribution from both direct electron transfer (DET) and mediated electron transfer (MET) mechanisms. In the early stage, DET was dominant and contributed to biofilm formation. In the late stage, the contribution from MET to the overall extracellular electron transfer (EET) eventually became dominant. The mediators could be confined into the biofilm matrix and produce continuous current output. Scanning electron microscopy (SEM) showed that the bacteria distributed uniformly on the electrode surface, which enhanced the EET rate. The high surface of the RVC-CNTs electrode was accessible to biofilms in short-term batch experiments, and RVC-CNTs could be a potential anodic material for bioelectrochemical devices.

Cadmium effects on net N2O production by the deep-sea isolate Shewanella loihica PV-4.

Deep-sea mining may lead to the release of high concentrations of metals into the surrounding seabed, which can disturb important ecosystem functions provided by microbial communities. Among these, the production of N2O and its reduction to N2 is of great relevance since N2O is an important greenhouse gas. Metal impacts on net N2O production by deep-sea bacteria are, however, currently unexplored. Here, we evaluated the effects of cadmium (Cd) on net N2O production by a deep-sea isolate, Shewanella loihica PV-4. We performed a series of Cd exposure incubations in oxic conditions and determined N2O fluxes during induced anoxic conditions, as well as the relative expression of the nitrite reductase gene (nirK), preceding N2O production, and N2O reductase gene (nosZ), responsible for N2O reduction. Net N2O production by S. loihica PV-4 exposed to Cd was strongly inhibited when compared to the control treatment (no metal). Both nirK and nosZ gene expression were inhibited in reactors with Cd, but nirK inhibition was stronger, supporting the lower net N2O production observed with Cd. The Cd inhibition of net N2O production observed in this study poses the question whether other deep-sea bacteria would undergo the same effects. Future studies should address this question as well as its applicability to complex communities and other physicochemical conditions, which remain to be evaluated.

Electrochemical Characteristics of Shewanella loihica PV-4 on Reticulated Vitreous Carbon (RVC) with Different Potentials Applied.

The current output of an anodic bioelectrochemical system (BES) depends upon the extracellular electron transfer (EET) rate from electricigens to the electrodes. Thus, investigation of EET mechanisms between electricigens and solid electrodes is essential. Here, reticulated vitreous carbon (RVC) electrodes are used to increase the surface available for biofilm formation of the known electricigen Shewanella loihica PV-4, which is limited in conventional flat electrodes. S. loihica PV-4 utilizes flavin-mediated EET at potential lower than the outer membrane cytochromes (OMC), while at higher potential, both direct electron transfer (DET) and mediated electron transfer (MET) contribute to the current output. Results show that high electrode potential favors cell attachment on RVC, which enhances the current output. DET is the prevailing mechanism in early biofilm, while the contribution of MET to current output increased as the biofilm matured. Electrochemical analysis under starvation shows that the mediators could be confined in the biofilm. The morphology of biofilm shows bacteria distributed on the top layer of honeycomb structures, preferentially on the flat areas. This study provides insights into the EET pathways of S. loihica PV-4 on porous RVC electrodes at different biofilm ages and different set potential, which is important for the design of real-world BES.

A novel real-time TMAO detection method based on microbial electrochemical technology

Trimethylamine N-oxide (TMAO) is considered to be a novel biomarker of cardiovascular diseases. However, the traditional TMAO detection method has failed to meet the requirements of real-time and point-of-care tests. Herein, a novel TMAO detection method based on microbial electrochemical technology is established, which realizes the direct conversion of TMAO concentration into electrical signals. Attached Shewanella loihica PV-4 was first proven to be capable of simultaneous inward extracellular electron transfer and TMAO reduction. The TMAO detection method showed a wide linear range of 0 to 250 μM, a high sensitivity of 23.92 μA/mM, and a low limit of detection of 5.96 μM. In addition, the TMAO detection process was accomplished within 600 s, with an acceptable accuracy of 90% in the real serum, showing high feasibility in clinical applications.

In situ produced hydrogen peroxide by biosynthesized Palladium nanoparticles and natural clay mineral for Highly-efficient Carbamazepine degradation

Fenton reaction is an effective method to remove refractory organics such as carbamazepine (CBZ) from water streams. Nevertheless, its application is greatly compromised by extra hydrogen peroxide (H2O2) addition and iron mud accumulation. Herein, Fenton-like process with in situ produced H2O2 by biosynthesized palladium nanoparticles (bioPd-NPs) and natural iron-bearing clay minerals is proposed for CBZ degradation. The bioPd-NPs prepared by Shewanella loihica PV-4 were in the size range of 5–20 nm, which catalyzed the in situ production of H2O2 from formic acid (FA) and oxygen. Then the in situ generated H2O2 underwent Fenton-like reactions with nontronite for CBZ degradation. With bioPd-NPs and nontronite dosage of 1 g/L and FA concentration of 20 mM, the complete CBZ (10 mg/L) degradation was achieved within 60 min. Oxidative radicals such as HO· and H2O2 generated in our constructed system played key roles in CBZ degradation. Intermediates/products identification and theoretical calculation revealed that hydroxylation was the main CBZ degradation pathway. This work provides a promising Fenton-like technology for elimination of CBZ from environment with prevention of additional H2O2 supplementation and excessive iron mud production.

Structural and Kinetic Characterization of Hyperthermophilic NADH-Dependent Persulfide Reductase from Archaeoglobus fulgidus.

NADH-dependent persulfide reductase (Npsr) has been proposed to facilitate dissimilatory sulfur respiration by reducing persulfide or sulfane sulfur-containing substrates to H2S. The presence of this gene in the sulfate and thiosulfate-reducing Archaeoglobus fulgidus DSM 4304 and other hyperthermophilic Archaeoglobales appears anomalous, as A. fulgidus is unable to respire S0 and grow in the presence of elemental sulfur. To assess the role of Npsr in the sulfur metabolism of A. fulgidus DSM 4304, the Npsr from A. fulgidus was characterized. AfNpsr is specific for persulfide and polysulfide as substrates in the oxidative half-reaction, exhibiting kcat/Km on the order of 104 M−1 s−1, which is similar to the kinetic parameters observed for hyperthermophilic CoA persulfide reductases. In contrast to the bacterial Npsr, AfNpsr exhibits low disulfide reductase activity with DTNB; however, similar to the bacterial enzymes, it does not show detectable activity with CoA-disulfide, oxidized glutathione, or cystine. The 3.1 Å X-ray structure of AfNpsr reveals access to the tightly bound catalytic CoA, and the active site Cys 42 is restricted by a flexible loop (residues 60-66) that is not seen in the bacterial homologs from Shewanella loihica PV-4 and Bacillus anthracis. Unlike the bacterial enzymes, AfNpsr exhibits NADH oxidase activity and also shows no detectable activity with NADPH. Models suggest steric and electrostatic repulsions of the NADPH 2′-phosphate account for the strong preference for NADH. The presence of Npsr in the nonsulfur-reducing A. fulgidus suggests that the enzyme may offer some protection against S0 or serve in another metabolic role that has yet to be identified.

Different mechanisms for riboflavin to improve the outward and inward extracellular electron transfer of Shewanella loihica

Exploring extracellular electron transfer (EET) is important to promote the applications of electrochemically active bacteria (EAB). Shewanella loihica PV-4, which is a model EAB capable of bidirectional EET, has shown promising application prospects in water quality monitoring and nitrogen removal. However, its bidirectional EET mechanism is not clear yet. This study focused on the electrochemical characteristics and electron transfer processes of S. loihica PV-4 bidirectional EET. Results revealed that riboflavin (RF) facilitates both the outward and inward EET, and the enhancement mechanism depends on EET direction. When S. loihica PV-4 performs outward EET, free RF acts as a redox mediator to promote electricity production. When the EET direction is reversed, bound RF is involved in electricity consumption. These results elucidated the effect of RF on S. loihica PV-4 bidirectional EET, providing a basis for the enhancement of S. loihica PV-4 EET.

Dissimilatory Nitrate Reduction to Ammonium (DNRA) and Denitrification Pathways Are Leveraged by Cyclic AMP Receptor Protein (CRP) Paralogues Based on Electron Donor/Acceptor Limitation in Shewanella loihica PV-4.

Under anoxic conditions, many bacteria, including Shewanella loihica strain PV-4, could use nitrate as an electron acceptor for dissimilatory nitrate reduction to ammonium (DNRA) and/or denitrification. Previous and current studies have shown that DNRA is favored under higher ambient carbon-to-nitrogen (C/N) ratios, whereas denitrification is upregulated under lower C/N ratios, which is consistent with our bioenergetics calculations. Interestingly, computational analyses indicate that the common cyclic AMP receptor protein (designated CRP1) and its paralogue CRP2 might both be involved in the regulation of two competing dissimilatory nitrate reduction pathways, DNRA and denitrification, in S. loihica PV-4 and several other denitrifying Shewanella species. To explore the regulatory mechanism underlying the dissimilatory nitrate reduction (DNR) pathways, nitrate reduction of a series of in-frame deletion mutants was analyzed under different C/N ratios. Deletion of crp1 could accelerate the reduction of nitrite to NO under both low and high C/N ratios. CRP1 is not required for denitrification and actually suppresses production of NO and N2O gases. Deletion of either of the NO-forming nitrite reductase genes nirK or crp2 blocked production of NO gas. Furthermore, real-time PCR and electrophoretic mobility shift assays (EMSAs) demonstrated that the transcription levels of DNRA-relevant genes such as nap-β (napDABGH), nrfA, and cymA were upregulated by CRP1, while nirK transcription was dependent on CRP2. There are tradeoffs between the different physiological roles of nitrate/lactate, as nitrogen nutrient/carbon source and electron acceptor/donor and CRPs may leverage dissimilatory nitrate reduction pathways for maximizing energy yield and bacterial survival under ambient environmental conditions.IMPORTANCE Some microbes utilize different dissimilatory nitrate reduction (DNR) pathways, including DNR to ammonia (DNRA) and denitrification pathways, for anaerobic respiration in response to ambient carbon/nitrogen ratio changes. Large-scale industrial nitrogen fixation and fertilizer application raise the concern of emission of N2O, a stable gas with potent global warming potential, as consequence of microbial respiration, thereby aggravating global warming and climate change. However, little is known about the molecular mechanism underlying the choice of two competing DNR pathways. We demonstrate that the global regulator CRP1, which is widely encoded in bacteria, is required for DNRA in S. loihica PV-4 strain, while the CRP2 paralogue is required for transcription of the nitrite reductase gene nirK for denitrification. Sufficient carbon source lead to the predominance of DNRA, while carbon source/electron donor deficiency may result in an incomplete denitrification process, raising the concern of high levels of N2O emission from nitrate-rich and carbon source-poor waters and soils.

Structural and enzymatic analysis of a dimeric cholylglycine hydrolase like acylase active on N-acyl homoserine lactones

The prevalence of substrate cross-reactivity between AHL acylases and β-lactam acylases provides a glimpse of probable links between quorum sensing and antibiotic resistance in bacteria. Both these enzyme classes belong to the N-terminal nucleophile (Ntn)-hydrolase superfamily. Penicillin V acylases alongside bile salt hydrolases constitute the cholylglycine hydrolase (CGH) group of the Ntn-hydrolase superfamily. Here we report the ability of two acylases, Slac1 and Slac2, from the marine bacterium Shewanella loihica-PV4 to hydrolyze AHLs. Three-dimensional structure of Slac1reveals the conservation of the Ntn hydrolase fold and CGH active site, making it a unique CGH exclusively active on AHLs. Slac1homologs phylogenetically cluster separate from reported CGHs and AHL acylases, thereby representing a functionally distinct sub-class of CGH that might have evolved as an adaptation to the marine environment. We hypothesize that Slac1 could provide the structural framework for understanding this subclass, and further our understanding of the evolutionary link between AHL acylases and β-lactam acylases.

Dual detection of biochemical oxygen demand and nitrate in water based on bidirectional Shewanella loihica electron transfer

This study for the first time proposed a method for simultaneously measuring BOD and nitrate in water using electrochemically active bacteria. Firstly, the bidirectional extracellular electron transfer (EET) capability of a model electricigen Shewanella loihica PV-4 was revealed. Then, based on the respective outward and inward EET, S. loihica PV-4 was utilized to detection BOD and nitrate. The results demonstrated a positive correlation between the outward EET and BOD (from 0 mg/L to 435 mg/L) while a negative correlation between the inward EET and nitrate (from 0 mg/L to 7 mg/L); both the relationships were well fitted by the combination of traditional linear model and Michaelis-Menten model (R2>0.96). Finally, a dual detection method for BOD and nitrate measurements was established based on the ano-cathodophilic capability of S. loihica PV-4 biofilm, and exhibited the characteristics of high accuracy (>80%) and fast analysis (<1h), suggesting a promising prospect in water monitoring.

Synergistic improvement of Shewanella loihica PV-4 extracellular electron transfer using a TiO2@TiN nanocomposite

Extracellular electron transfer (EET) allows microorganisms to perform anaerobic respiration using insoluble electron acceptors, including minerals and electrodes. EET-based applications require efficient electron transfer between living and non-living systems. To improve EET efficiency, the TiO2@TiN nanocomposite was used to form hybrid biofilms with Shewanella loihica PV-4 (PV-4). Chronoamperometry showed that peak current was increased 4.6-fold via the addition of the TiO2@TiN nanocomposite. Different biofilms were further tested in a dual-chamber microbial fuel cell. The PV-4 biofilm resulted a maximum power density of 33.4 mW/m2, while the hybrid biofilm of the TiO2@TiN nanocomposite with PV-4 yielded a 92.8% increase of power density. Electrochemical impedance spectroscopy analyses showed a lower electron-transfer resistance in the hybrid biofilm. Biological measurements revealed that both flavin secretion and cytochrome c expression were increased when the TiO2@TiN nanocomposite presented. These results demonstrated that the TiO2@TiN nanocomposite could synergistically enhance the EET of PV-4 through altering its metabolism. Our findings provide a new strategy for optimizing biotic-abiotic interactions in bioelectrochemical systems.

Dissimilatory bioreduction of iron(III) oxides by Shewanella loihica under marine sediment conditions

Shewanella is a genus of marine bacteria capable of dissimilatory iron reduction (DIR). In the context of deep-sea mining activities or submarine mine tailings disposal, dissimilatory iron reducing bacteria may play an important role in biogeochemical reactions concerning iron oxides placed on the sea bed. In this study, batch experiments were performed to evaluate the capacity of Shewanella loihica PV-4 to bioreduce different iron oxides (ferrihydrite, magnetite, goethite and hematite) under conditions similar to those in anaerobic sea sediments. Results showed that bioreduction of structural Fe(III) via oxidation of labile organic matter occurred in all these iron oxides. Based on the aqueous Fe (II) released, derived Fe(II)/acetate ratios and bioreduction coefficients seem to be only up to about 4% of the theoretical ones, considering the ideal stoichiometry of the reaction. A loss of aqueous Fe (II) was caused by adsorption and mineral transformation processes. Scanning electron microscope images showed that Shewanella lohica was attached to the Fe(III)-oxide surfaces during bioreduction. Our findings suggest that DIR of Fe(III) oxides from mine waste placed in marine environments could result in adverse ecological impacts such as liberation of trace metals in the environment.

Differential gene content and gene expression for bacterial evolution and speciation of Shewanella in terms of biosynthesis of heme and heme-requiring proteins

BackgroundMost species of Shewanella harbor two ferrochelatase paralogues for the biosynthesis of c-type cytochromes, which are crucial for their respiratory versatility. In our previous study of the Shewanella loihica PV-4 strain, we found that the disruption of hemH1 but not hemH2 resulted in a significant accumulation of extracellular protoporphyrin IX (PPIX), but it is different in Shewanella oneidensis MR-1. Hence, the function and transcriptional regulation of two ferrochelatase genes, hemH1 and hemH2, are investigated in S. oneidensis MR-1.ResultIn the present study, deletion of either hemH1 or hemH2 in S. oneidensis MR-1 did not lead to overproduction of extracellular protoporphyrin IX (PPIX) as previously described in the hemH1 mutants of S. loihica PV-4. Moreover, supplement of exogenous hemins made it possible to generate the hemH1 and hemH2 double mutant in MR-1, but not in PV-4. Under aerobic condition, exogenous hemins were required for the growth of MR-1ΔhemH1ΔhemH2, which also overproduced extracellular PPIX. These results suggest that heme is essential for aerobic growth of Shewanella species and MR-1 could also uptake hemin for biosynthesis of essential cytochrome(s) and respiration. Besides, the exogenous hemin mediated CymA cytochrome maturation and the cellular KatB catalase activity. Both hemH paralogues were transcribed in wild-type MR-1, and the hemH2 transcription was remarkably up-regulated in MR-1ΔhemH1 mutant to compensate for the loss of hemH1. The periplasmic glutathione peroxidase gene pgpD, located in the same operon with hemH2, and a large gene cluster coding for iron, heme (hemin) uptake systems are absent in the PV-4 genome.ConclusionOur results indicate that the genetic divergence in gene content and gene expression between these Shewanella species, accounting for the phenotypic difference described here, might be due to their speciation and adaptation to the specific habitats (iron-rich deep-sea vent versus iron-poor freshwater) in which they evolved and the generated mutants could potentially be utilized for commercial production of PPIX.

Effect of control mode on the sensitivity of a microbial fuel cell biosensor with Shewanella loihica PV-4 and the underlying bioelectrochemical mechanism

A relatively poor sensitivity is a critical challenge for the application of microbial fuel cell biosensors (MFC-biosensors). This study investigated the effects of two control modes on sensor sensitivity and revealed the underlying bioelectrochemical mechanism. The results demonstrated that the sensitivity of an S. loihica PV-4 MFC-biosensor increased by 6.1 times when the anode was controlled at a constant potential (CP) instead of being operated with a fixed external resistance (ER). This obvious difference in sensor sensitivity was partly attributed to the masking effect of the observable offset current under ER mode and the lower electricity production capacity under CP mode. Moreover, the analysis of metabolic structure showed that under CP mode the anodic biofilm presented lower viability after toxic shock, due to a poorer ability to synthesize and secrete extracellular polymeric substances. Electrochemical measurements further revealed a lower capacitance under CP mode, which favored the permeation of Cd2+ into the biofilm.

Controlled Layer-By-Layer Deposition of Carbon Nanotubes on Electrodes for Microbial Fuel Cells

Carbon nanotubes (CNTs) and polyelectrolyte poly(allylamine hydrochloride) (PAH) composite modified indium tin oxide (ITO) electrodes, by a layer-by-layer (LBL) self-assembly technique, was evaluated as an anode for microbial fuel cells (MFCs). The bioelectrochemistry of Shewanella loihica PV-4 in an electrochemical cell and the electricity generation performance of MFCs with multilayer (CNTs/PAH)n-deposited ITO electrodes as an anode were investigated. Experimental results showed that the current density generated on the multilayer modified electrode increased initially and then decreased as the deposition of the number of layers (n = 12) increased. Chronoamperometric results showed that the highest peak current density of 34.85 ± 2.80 mA/m2 was generated on the multilayer (CNTs/PAH)9-deposited ITO electrode, of which the redox peak current of cyclic voltammetry was also significantly enhanced. Electrochemical impedance spectroscopy analyses showed a well-formed nanostructure porous film on the surface of the multilayer modified electrode. Compared with the plain ITO electrode, the multilayered (CNTs/PAH)9 anodic modification improved the power density of the dual-compartment MFC by 29%, due to the appropriate proportion of CNTs and PAH, as well as the porous nanostructure on the electrodes.

IN SILICO GENOME-SCALE RECONSTRUCTION AND ANALYSIS OF THE SHEWANELLA LOIHICA PV-4 METABOLIC NETWORK

Microbial Fuel Cells (MFCs) are devices that generate electricity directly from organic compounds with microbes (electricigens) serving as anodic catalysts. As a novel environment-friendly energy source, MFCs have extensive practical value. Since the biological features and metabolic mechanism of electricigens have a great effect on the electricity production of MFCs, it is a big deal to screen strains with high electricity productivity for improving the power output of MFC. Reconstructions and simulations of metabolic networks are of significant help in studying the metabolism of microorganisms so as to guide gene engineering and metabolic engineering to improve their power-generating efficiency. Herein, we reconstructed a genome-scale constraint-based metabolic network model of Shewanella loihica PV-4, an important electricigen, based on its genomic functional annotations, reaction databases and published metabolic network models of seven microorganisms. The resulting network model iGX790 consists of 902 reactions (including 71 exchange reactions), 798 metabolites and 790 genes, covering the main pathways such as carbon metabolism, energy metabolism, amino acid metabolism, nucleic acid metabolism and lipid metabolism. Using the model, we simulated the growth rate, the maximal synthetic rate of ATP, the flux variability analysis of metabolic network, gene deletion and so on to examine the metabolism of S. loihica PV-4.

Polyaniline/Carbon Nanotubes Composite Modified Anode via Graft Polymerization and Self-Assembling for Microbial Fuel Cells.

Microbial fuel cells (MFCs) are promising devices for sustainable energy production, wastewater treatment and biosensors. Anode materials directly interact with electricigens and accept electrons between cells, playing an important role in determining the performance of MFCs. In this study, a novel carbon nanotubes (CNTs) and polyaniline (PANI) nanocomposite film modified Indium-tin oxide (ITO) anode was fabricated through graft polymerization of PANI after the modification of γ-aminopropyltriethoxysilane (APTES) on ITO substrate, which was followed by layer-by-layer (LBL) self-assembling of CNTs and PANI alternatively on its surface. (CNTs/PANI)n/APTES/ITO electrode with low charge transfer resistance showed better electrochemical behavior compared to the bare ITO electrode. Twelve layers of CNTs/PANI decorated ITO electrode with an optimal nanoporous network exhibited superior biocatalytic properties with a maximal current density of 6.98 µA/cm2, which is 26-fold higher than that of conventional ITO electrode in Shewanella loihica PV-4 bioelectrochemical system. MFCs with (CNTs/PANI)12/APTES/ITO as the anode harvested a maximum output power density of 34.51 mW/m2, which is 7.5-fold higher than that of the unmodified ITO electrode. These results demonstrate that (CNTs/PANI)12/APTES/ITO electrode has superior electrochemical and electrocatalytic properties compared to the bare ITO electrode, while the cellular toxicity of CNTs has an effect on the performance of MFC with (CNTs/PANI)n/APTES/ITO electrode.