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Microbial conversion of carbon monoxide (CO) to acetate is a promising upcycling strategy for carbon sequestration. Herein, we demonstrate that CO conversion and acetate production rates of Eubacterium limosum KIST612 strain can be improved by in silico prediction and in vivo assessment. The mimicked CO metabolic model of KIST612 predicted that overexpressing the CO dehydrogenase (CODH) increases CO conversion and acetate production rates. To validate the prediction, we constructed mutant strains overexpressing CODH gene cluster and measured their CO conversion and acetate production rates. A mutant strain (ELM031) co-overexpressing CODH, coenzyme CooC2 and ACS showed a 3.1 × increased specific CO oxidation rate as well as 1.4 × increased specific acetate production rate, compared to the wild type strain. The transcriptional and translational data with redox balance analysis showed that ELM031 has enhanced reducing potential from up-regulation of ferredoxin and related metabolism directly linked to energy conservation.

Voltage reversal (VR) in series connection of multiple membrane electrode assembly installed microbial fuel cells (mMEA-MFC) is eliminated by manipulating the resistor control. Discharge test results collected from two mMEA-MFCs initially operated (designated as P1 and P2) confirm that the performance of P2 exceeds that of P1. Thus, driving P1 and P2 as serially stacked MFCs generate the VR in P1. Controlling the inserted resistor adjust the current production of P2 to maintain balance with P1, and the VR in P1 is eliminated in the operation of stacking mode. Thus, manipulating the internal resistance provide an applicable approach to suppress VR in the stacking of mMEA-MFCs system.

In this work, three multi-electrode-embedded microbial fuel cells (MFCs) were connected sequentially and operated in series and parallel modes, fed by effluent of an anaerobic digester continuously operated using swine wastewater. The anaerobic digester achieved ~0.75 CH4 L d−1 while removing 71.2% of COD and 0.8% of ammonia, which was comparable to the literatures reported. The MFCs removed additional COD from the anaerobic digester effluent, achieving the lowest concentration in the last unit, leading to a voltage reversal in the serially-connected unit. The MFCs connected and operated in parallel mode showed the highest power density of ~25 W m−3, which is 18% higher compared to the one operated in series mode. These results definitively show that differences in substrate concentrations among MFC units are inevitable with sequential flow. Further, a parallel connection mode of operation is necessary to achieve stable, long-term power generation from MFC units, without any electrical malfunction.

This study analyzed the effect of methanol on the metabolism of syngas components (i.e., H2 and CO) by the syngas fermenting acetogenic strain E. limosum KIST612. The culture characteristics and relevant proteomic expressions (as fold changes) were carefully analyzed under CO/CO2 and H2/CO2 conditions with and without methanol addition, as well as, under methanol/CO2 conditions. The culture characteristics (specific growth rate and H2 consumption rate) under H2/CO2 conditions were greatly enhanced in the presence of methanol, by 4.0 and 2.7 times, respectively. However, the promoting effect of methanol was not significant under CO/CO2 conditions. Proteomic fold changes in most enzyme expression levels in the Wood-Ljungdahl pathway and chemiosmotic energy conservation also exhibited high correspondence between methanol and H2/CO2 but not between methanol and CO/CO2. These findings suggest the advantages of methanol addition to H2/CO2 for biomass enhancement and faster consumption of gaseous substrates during syngas fermentation.

The aim of this work is to study for concurrent harvesting bioelectricity and struvite mineral from mineral rich wastewater containing with nitrogen (N) and phosphorous (P) contents using MFCs and a chemical precipitation system. Whole reaction was constructed to sequentially run hybrid reactor (consisting of MFCs and struvite precipitation), gravitational sedimentation, nitrogen purging and MFCs. The MFCs generated around 6.439 ± 0.481 mA and 2.084 ± 0.310 mW as Imax and Pmax, respectively under 2g/l of COD. More than 70% of C source, and around 95% of P and N sources have been removed. Struvite mineral was precipitated in the hybrid reactor after the injection of Mg2+ and collected in sedimentation tank. Economic feasibility and beneficial concerns were carefully investigated, and it is proposed for applications in the “decentralised treatment process” of agriculture and livestock wastewater in order to realise circular and strong economy in agriculture by creating virtuous cycles.

The gas–liquid mass transfer coefficient (kLa) of O2 was investigated in a bubble column reactor (BCR) using a sintered gas filter (SF), ceramic membrane module (CMM), and hollow fiber membrane module (HFM), which have different ranges of gas supply areas. kLa was enhanced by increasing flow rate in all of the spargers. Different responses when changing the gas supply area were obtained depending on the sparger type. Average values of kLa that were 52 and 258% higher were obtained using a CMM-integrated BCR compared to SFs and HFMs. CO-water kLa was investigated using CMMs for application to gas fermentation. The CO-water kLa ranged from 28.3 to 113.7/h under the experimental conditions. Based on the experimental data from CO and O2, a model to predict kLa was constructed for CMM-integrated BCRs. A dimensionless number indicating a gas supply area of the sparger was newly defined and included in the developed model.

This study investigated the effects of electrolytes (CaCl2, K2HPO4, MgSO4, NaCl, and NH4Cl) on CO mass transfer and ethanol production in a HFMBR. The hollow fiber membranes (HFM) were found to generate tiny gas bubbles; the bubble coalescence was significantly suppressed in electrolyte solution. The volumetric gas-liquid mass transfer coefficients (kLa) increased up to 414% compared to the control. Saturated CO (C∗) decreased as electrolyte concentrations increased. Overall, the maximum mass transfer rate (Rmax) in electrolyte solution ranged from 106% to 339% of the value obtained in water. The electrolyte toxicity on cell growth was tested using Clostridium autoethanogenum. Most electrolytes, except for MgSO4, inhibited cell growth. The HFMBR operation using a medium containing 1% MgSO4 achieved 119% ethanol production compared to that without electrolytes. Finally, a kinetic simulation using the parameters got from the 1% MgSO4 medium predicted a higher ethanol production compared to the control.

Microbial fuel cells (MFCs) were investigated for use in removing total ammonia nitrogen (TAN) and residual COD from effluent digested in an anaerobic digester (AD) fed with actual swine wastewater for 32 days in batch mode. Cumulative COD removal in the AD was as high as 59,647±2096 mg/L (80.5% removed), whereas TAN removal in the AD was negligible at 296±116 mg-N/L (5.8% removed), causing a decrease in the COD/TAN ratio from 14.5 to 3.0. In a subsequent MFC system, 77.5% of TAN was removed at 36 days, leading to an increase in COD/TAN ratio from 4.6 to 8.1. As a result, the COD in the anode was further reduced from 19,319±417 mg/L to 7519±554 mg/L (61.1% removed). From these results, removing the TAN in MFCs was found to increase the COD/TAN ratio, with the COD being further degraded.

Microbial conversion of syngas to energy-dense biofuels and valuable chemicals is a potential technology for the efficient utilization of fossils (e.g., coal) and renewable resources (e.g., lignocellulosic biomass) in an environmentally friendly manner. However, gas-liquid mass transfer and kinetic limitations are still major constraints that limit the widespread adoption and successful commercialization of the technology. This review paper provides rationales for syngas bioconversion and summarizes the reaction limited conditions along with the possible strategies to overcome these challenges. Mass transfer and economic performances of various reactor configurations are compared, and an ideal case for optimum bioreactor operation is presented. Overall, the challenges with the bioprocessing steps are highlighted, and potential solutions are suggested. Future research directions are provided and a conceptual design for a membrane-based syngas biorefinery is proposed.

In this study, the electrochemical properties of a Shewanella oneidensis MR-1 biofilm were investigated using a mini-microbial electrochemical system. The performance of the biofilm was shown, using discharge test and cyclic voltammetry investigations, to improve over time. Differential pulse voltammograms were also acquired to determine the type of extracellular electron transfer that took place and to characterize the structure of the microbial biofilm formed on the electrode of the electrochemical system. These results indicated that extracellular electron transfer via a flavin-like mediator chemical predominated as the biofilm grew. The results, combined with a comparison of the measured current density with the calculated value of a seamless single-layered biofilm, also suggested that S. oneidensis MR-1 formed a multi-layered biofilm on the electrode.

In order to assess the effects of biofilm capacitance on self-recovering voltage reversals, the restored current is determined and compared with the measured biofilm capacitance by analyzing the results of electrochemical impedance spectroscopy. This comparison demonstrates that self-recovering voltage reversals are caused by temporary damage to, and the recovery of, biofilm capacitance which arises due to the ability of redox enzymes in the electron transfer system to temporarily store electrons. Thus, the development of procedures for voltage reversal control and for the maintenance of serially connected microbial fuel cells (MFCs) should take into account such temporary voltage reversal phenomenon. This discovery and characterization of self-recovering voltage reversals is expected to be practically useful to enhance the reliability of MFCs to be scaled up and implemented in practical systems.

The acetate-assisted cultivation of Eubacterium limosum KIST612 was found to provide a way for enhancing cell mass, the carbon monoxide (CO) consumption rate, and butyrate production using CO as an electron and energy source. Cell growth (146%), μmax (121%), and CO consumption rates (151%) increased significantly upon the addition of 30mM acetate to microbial cultures. The main product of CO fermentation by E. limosum KIST612 shifted from acetate to butyrate in the presence of acetate, and 5.72mM butyrate was produced at the end of the reaction. The resting cell experimental conditions indicated acetate uptake and an increase in the butyrate concentration. Three routes to acetate assimilation and energy conservation were suggested based on given experimental results and previously genome sequencing data. Acetate assimilation via propionate CoA-transferase (PCT) was expected to produce 1.5mol ATP/mol butyrate, and was thus anticipated to be the most preferred route.

This study employed a simultaneous gas feeding and cell-recycled reaction (SGCR) system to ferment CO using Eubacterium limosum KIST612. A bubble column reactor was equipped with an ex-situ hollow fiber membrane module to enable cell recycling. The internal gas circulation rate was adjusted by controlling the pump speed to provide sufficient gas supplement to the microorganism. Gas feedings were conducted by either the use of a gas-tight bag (Batch), a pressurized gas cylinder (Continuous), or a sequential combination of the two (Mixed feeding). Mixed feeding mode achieved higher biomass (9.7 g/L) and acetate (9.8 g/L) concentrations than Batch mode (3.2 g/L biomass and 7.0 g/L acetate) or Continuous mode (5.0 g/L biomass and 8.1 g/L acetate). The high acetate titer in Mixed feeding mode was achieved due to the high concentration of cells secured in a short time at the initial operation stage and maintaining a high specific growth rate.

CO dehydrogenase (CODH) employed in a dissolved CO biosensor development study harbors a solvent-exposed cofactor capable of DET to electrode. Here, CODH was immobilized on arrays of AuNPs of various dimensions to determine the effect of the size and shape of the electrode surface on the direct electrical connection between CODH and electrode surface. The results showed the degree of proximity between the CODH cofactor and electrode surface, which varied with AuNP size and caused significant changes to the electrical connection at the interface as well as to the substrate accessibility. Consequently, a high-density nanoscale SRS was fabricated on electrode to further facilitate direct electrical connection as well as to enable distribution of CODH into monolayer or near-monolayer for lowering the barrier of CO diffusion toward enzyme. The findings show the feasibility of controlling the direct electrical connection between CODH and the electrode as well as controlling the substrate accessibility.

This study proposed a submerged hollow fibre membrane bioreactor (HFMBR) system capable of achieving high carbon monoxide (CO) mass transfer for applications in microbial synthesis gas conversion systems. Hydrophobic polyvinylidene fluoride (PVDF) membrane fibres were used to fabricate a membrane module, which was used for pressurising CO in water phase. Pressure through the hollow fibre lumen ( P ) and membrane surface area per unit working volume of the liquid ( A S / V L ) were used as controllable parameters to determine gas–liquid volumetric mass transfer coefficient ( k L a ) values. We found a k L a of 135.72 h −1 when P was 93.76 kPa and A S / V L was fixed at 27.5 m −1 . A higher k L a of 155.16 h −1 was achieved by increasing A S / V L to 62.5 m −1 at a lower P of 37.23 kPa. Practicality of HFMBR to support microbial growth and organic product formation was assessed by CO/CO 2 fermentation using Eubacterium limosum KIST612.

The effects of azide on electron transport of exoelectrogens were investigated using air-cathode MFCs. These MFCs enriched with azide at the concentration higher than 0.5mM generated lower current and coulomb efficiency (CE) than the control reactors, but at the concentration lower than 0.2mM MFCs generated higher current and CE. Power density curves showed overshoot at higher azide concentrations, with power and current density decreasing simultaneously. Electrochemical impedance spectroscopy (EIS) showed that azide at high concentration increased the charge transfer resistance. These analyses might reflect that a part of electrons were consumed by the anode microbial population rather than transferred to the anode. Bacterial population analyses showed azide-enriched anodes were dominated by Deltaproteobacteria compared with the controls. Based on these results it is hypothesized that azide can eliminate the growth of aerobic respiratory bacteria, and at the same time is used as an electron acceptor/sink.

Comparative genome analysis of Shewanella strains predicted that the strains metabolize preferably two- and three-carbon carbohydrates as carbon/electron source because many Shewanella genomes are deficient of the key enzymes in glycolysis (e.g., glucokinase). In addition, all Shewanella genomes are known to have only one set of genes associated with the phosphotransferase system required to uptake sugars. To engineer Shewanella strains that can utilize five- and six-carbon carbohydrates, we constructed glucose-utilizing Shewanella oneidensis MR-1 by introducing the glucose facilitator (glf; ZMO0366) and glucokinase (glk; ZMO0369) genes of Zymomonas mobilis. The engineered MR-1 strain was able to grow on glucose as a sole carbon/electron source under anaerobic conditions. The glucose affinity (Ks) and glucokinase activity in the engineered MR-1 strain were 299.46 mM and 0.259 ± 0.034 U/g proteins. The engineered strain was successfully applied to a microbial fuel cell system and exhibited current generation using glucose as the electron source.

Five rigid graphite plates were embedded in evenly divided sections of sediment, ranging from 2 cm (A1) to 10 cm (A5) below the top sediment layer. The maximum power and current of the MFCs increased in depth order; however, despite the increase in the internal resistance, the power and current density of the A5 MFC were 2.2 and 3.5 times higher, respectively, than those of the A1 MFC. In addition, the anode open circuit potentials (OCPs) of the sediment microbial fuel cells (SMFCs) became more negative with sediment depth. Based on these results, it could be then concluded that as the anode-embedding depth increases, that the anode environment is thermodynamically and kinetically favorable to anodophiles or electrophiles. Therefore, the anode-embedding depth should be considered an important parameter that determines the performance of SMFCs, and we posit that the anode potential could be one indicator for selecting the anode-embedding depth.

A microbiological process was established to harvest electricity from the carbon monoxide (CO). A CO fermenter was enriched with CO as the sole carbon source. The DGGE/DNA sequencing results showed that Acetobacterium spp. were enriched from the anaerobic digester fluid. After the fermenter was operated under continuous mode, the products were then continuously fed to the microbial fuel cell (MFC) to generate electricity. Even though the conversion yield was quite low, this study proved that synthesis gas (syn-gas) can be converted to electricity with the aid of microbes that do not possess the drawbacks of metal catalysts of conventional methods.

In this work, ammonia removal paths in microbial fuel cells (MFCs) under different initial pH conditions (pH 7.0, 8.0, and 8.6) were investigated. At a neutral pH condition (pH 7.0), MFC used an electrical energy of 27.4% and removed 23.3% of total ammonia by electrochemical pathway for 192h. At the identical pH condition, 36.1% of the total ammonia was also removed by the biological path suspected to be biological ammonia oxidation process (e.g., Anammox). With the initial pH increased, the electrochemical removal efficiency decreased to less than 5.0%, while the biological removal efficiency highly increased to 61.8%. In this study, a neutral pH should be maintained in the anode to utilize MFCs for ammonia recovery via electrochemical pathways from wastewater stream.

To prevent the occurrence of the organic crossover in membraneless microbial fuel cells (ML-MFCs), dual-anode MFC (DA-MFC) was designed from multi-anode concept to ensure anode zone. The anode zone addressed increase the utilization of organic matter in ML-MFCs, as the result, the organic crossover was prevented and performance of MFCs were enhanced. The maximum power of the DA-MFC was 0.46mW, which is about 1.56 times higher than the ML-MFC (0.29mW). Furthermore, the DA-MFC had advantage in correlation of organic substance concentration and dissolved oxygen concentration, and even electric over-potential. In addition, in terms of cathode fouling, the DA-MFC showed clearer surface. Hence, the anode zone should be considered in the advanced ML-MFC for practically use in wastewater treatment process, and also for scale-up of MFCs.

This study determines and compares the intrinsic kinetic parameters (Ks and Ki) of selected Thermococcus onnurineus NA1 strains (wild-type (WT), and mutants MC01, MC02, and WTC156T) using the substrate inhibition model. Ks and Ki values were used to find the optimum dissolved CO (CL) conditions inside the reactor. The results showed that in terms of the maximum specific CO consumption rates (qCO(max)) of WT, MC01, MC02, and WTC156T the optimum activities can be achieved by maintaining the CL levels at 0.56mM, 0.52mM, 0.58mM, and 0.75mM, respectively. The qCO(max) value of WTC156T at 0.75mM was found to be 1.5-fold higher than for the WT strain, confirming its superiority. Kinetic modeling was then used to predict the conditions required to maintain the optimum CL levels and high cell concentrations in the reactor, based on the kinetic parameters of the WTC156T strain.

A microbial fuel cell (MFC) was optimized in terms of MFC design factors and operational parameters for continuous electricity production using artificial wastewater (AW). The performance of MFC was analyzed through the polarization curve method under different conditions using a mediator-less MFC. The highest power density of 0.56 W/m2 was achieved with AW of 300 mg/l fed at the rate of 0.53 ml/min at 35 degrees C. The power per unit cell working volume was 102 mW/l, which was over 60 times higher than those reported in the previous mediator-less MFCs which did not use a cathode or an anode mediator. The power could be stably generated over 2 years.

The cathode reaction is one of the most important limiting factors in bioelectrochemical systems even with precious metal catalysts. Since aerobic bacteria have a much higher affinity for oxygen than any known abiotic cathode catalysts, the performance of a microbial fuel cell can be improved through the use of electrochemically-active oxygen-reducing bacteria acting as the cathode catalyst. These consume electrons available from the electrode to reduce the electron acceptors present, probably conserving energy for growth. Anaerobic bacteria reduce protons to hydrogen in microbial electrolysis cells (MECs). These aerobic and anaerobic bacterial activities resemble those catalyzing microbially-influenced corrosion (MIC). Sulfate-reducing bacteria and homoacetogens have been identified in MEC biocathodes. For sustainable operation, microbes in a biocathode should conserve energy during such electron-consuming reactions probably by similar mechanisms as those occurring in MIC. A novel hypothesis is proposed here which explains how energy can be conserved by microbes in MEC biocathodes.

The effect of two different anode-embedding orientations, lengthwise- and widthwise-embedded anodes was explored, on the performance of sediment microbial fuel cells (SMFCs) using a chessboard anode. The maximum current densities and power densities in SMFCs having lengthwise-embedded anodes (SLA1-SLA10) varied from 38.2mA/m(2) to 121mA/m(2) and from 5.5mW/m(2) to 20mW/m(2). In comparison, the maximum current densities and maximum power densities in SMFCs having anodes widthwise-embedded between 0cm to 8cm (SWA2-SWA5) increased from 82mA/m(2) to 140mA/m(2) and from 14.7mW/m(2) to 31.1mW/m(2) as the anode depth became deeper. Although there was a difference in the performance among SWA5-SWA10, it was considered negligible. Hence, it is concluded that it is important to embed anodes widthwise at the specific anode depths, in order to improve of SMFC performance. Chessboard anode used in this work could be a good option for the determination of optimal anode depths.

A microbial fuel cell (MFC) was successfully developed as a toxicity biomonitoring system, giving a quick response to Cu(II) toxic events. The objective was to increase MFC sensitivity to Cu(II) toxicity by evaluating the impact of shear rate caused by mixing and intermittent nitrogen sparging on the biofilm structure. Low shear rate – achieved by continuously feeding the wastewater into the MFC at a low flow rate of 1.3 mL min−1 during the enrichment period – resulted in low biomass density (124 g VSS L−1 of biofilm), high porosity and reduced levels of extracellular polymeric substances (EPS). Consequently, the sensitivity was improved. Scattered nitrogen sparging also increased the sensitivity by reducing the EPS level. It suggested that MFC enriched under low flow rate with intermittent nitrogen sparging could produce an anodic biofilm that was less dense, more porous, contained less EPS and ultimately displayed higher sensitivity to Cu(II) toxicity.

The external resistance is perhaps the easiest way to influence the operation of a microbial fuel cell (MFC). In this paper, three enrichment strategies, whereby the external resistance was fixed at: (1) a high value in order to maximize the cell voltage (U strategy); (2) a low value in order to maximize the current (I strategy); and (3) a value equal to the internal resistance of the MFC to maximize the power output (P strategy), were investigated. The I strategy resulted in increased maximum power generation and the likely reason is that electron transfer was facilitated under low external resistance, which in turn, favored the development of an electrochemically active biofilm. This experiment was conducted in a single-chamber MFC system equipped with a membrane electrode assembly, and a comparison of the performance achieved by five different membranes is also provided. Selemion was found to be a suitable alternative to Nafion.

The present study emphasizes the importance of overcoming proton limitation in a microbial fuel cell operated on domestic wastewater. When the anode-treated effluent was allowed to trickle into the cathodic compartment (full-loop operation), high COD and suspended solids removal efficiencies over 75% and 84%, respectively, were achieved while ensuring substantial and sustainable power generation. Lower removal efficiencies resulted in decreased cell electromotive force caused by excess substrate crossover. By decreasing the pH in the cathodic compartment to values below 2, we were able to further increase the maximum power generation by 180% in batch mode and 380% in continuous mode as compared to a negative control (tap water of pH 7.6). Under the optimized conditions, the internal resistance and electromotive force were 11 Ω and 0.6 V, respectively, which correspond to the state of the art.

The aim of this study was to evaluate limiting factors affecting electricity output from sediment microbial fuel cells (sediment MFCs). In laboratory tests, various factors likely to be encountered in field application were divided into controllable and uncontrollable ones. Based on the findings, it could be suggested that the sediment MFCs can be operated with an anode to cathode area ratio of at least 5:1 and at high external loads (1000 ohms) when the cathode is closely placed to the anode, though DO concentration at the cathode must be kept above 3 mg/l. Furthermore, no significant effect on current production over a prolonged period was observed within the sediment temperature range of 20-35 degrees C, but was negatively affected by lower temperatures (10 degrees C). These observations provide important factors with respect to the construction and operation of sediment MFCs at field sites, which will aid in maximizing electricity output.