Periodic step polarization accelerates electron recovery by electroactive biofilms (EABs).

Relatively low rate of electron recovery is one of the factors that limit the advancement of bioelectrochemical systems. Here, new periodic polarizations were investigated with electroactive biofilms (EABs) enriched from activated sludge and Geobacter sulfurreducens biofilms. When representative anode potentials (Ea ) were applied, redox centers with midpoint potentials (Emid ) higher than Ea were identified by localized cyclic voltammetry. The electrons held by these redox centers were accessible when Ea was raised to 0.4 V (vs. Ag/AgCl). New periodic polarizations that discharge at 0.4 V recovered electrons faster than normal periodic and fixed-potential polarizations. The best-performing periodic step polarization accelerated electron recovery by 23%-24% and 12%-76% with EABs and G. sulfurreducens biofilms, respectively, compared to the fixed-potential polarization. Quantitative reverse transcription polymerase chain reaction showed an increased abundance of omcZ mRNA transcripts from G. sulfurreducens after periodic step polarization. Therefore, both the rate of energy recovery by EABs and the performance of bioelectrochemical systems can be enhanced by improving the polarization schemes.

Production of polyhydroxyalkanoate (PHA) copolymer from food waste using mixed culture for carboxylate production and Pseudomonas putida for PHA synthesis.

Production of polyhydroxyalkanoates (PHAs) with high concentration of carboxylate, that was accumulated from solid state fermentation (SSF) of food waste (FW), was tested using Pseudomonas putida strain KT2440. Mixed-culture SSF of FW supplied in a high concentration of carboxylate, which caused a high PHA production of 0.56 g PHA/g CDM under nutrients control. Interestingly, this high PHA fraction in CDM was almost constant at 0.55 g PHA/g CDM even under high nutrients concentration (25 mM NH4+), probably due to high reducing power maintained by high carboxylate concentration. PHA characterization indicated that the dominant PHA building block produced was 3-hydroxybutyrate, followed by 3-hydroxy-2-methylvalerate and 3-hydroxyhenxanoate. Carboxylate profiles before and after PHA production suggested that acetate, butyrate, and propionate were the main precursors to PHA via several metabolic pathways. Our result support that mixed culture SSF of FW for high concentration carboxylate and P. putida for PHA production enables sustainable production of PHA in cost-effective manners.

Butyrate production and purification by combining dry fermentation of food waste with a microbial fuel cell.

This study developed and evaluated a high-purity butyrate producing bioprocess from food waste by combining dry fermentation (DF) with a microbial fuel cell (MFC). Acclimatization of a DF reactor with an enrichment culture resulted in high food waste degradation (VS removed, %) and butyrate production. A high VS degradation of 81%, butyrate concentration of up to 24 gCODbutyrate/L and butyrate yields of 497 gCODbutyrate/kg VSadded was obtained in the DF reactor. As a result, butyrate comprised 83% of all short chain fatty acids (SCFA) in the DF broth. Acetate (10%) and propionate (7%) comprised the rest of the SCFA. The butyrate composition was further purified by feeding the DF broth to a multi-electrode MFC enriched with anode respiring bacteria (ARB) such as Geobacter sp. (>55%). The ARB in the MFC removed acetate and propionate while purified butyrate was recovered in the MFC effluent. Butyrate purity in the MFC effluent reached as high as 99% at hydraulic retention time of 72 h. Along with butyrate purification, the MFC produced electric power in a range of 0.1-0.6 Wh/gCODbutyraterecovered (or 0.01-7.85 kWh/ton of food waste), demonstrating that MFCs can be an energy-positive butyrate purification bioprocess.

Kinetics of anaerobic methane oxidation coupled to denitrification in the membrane biofilm reactor.

Anaerobic oxidation of methane coupled to denitrification (AOM-D) in a membrane biofilm reactor (MBfR), a platform used for efficiently coupling gas delivery and biofilm development, has attracted attention in recent years due to the low cost and high availability of methane. However, experimental studies have shown that the nitrate-removal flux in the CH4 -based MBfR (<1.0 g N/m2 -day) is about one order of magnitude smaller than that in the H2 -based MBfR (1.1-6.7 g N/m2 -day). A one-dimensional multispecies biofilm model predicts that the nitrate-removal flux in the CH4 -based MBfR is limited to <1.7 g N/m2 -day, consistent with the experimental studies reported in the literature. The model also determines the two major limiting factors for the nitrate-removal flux: The methane half-maximum-rate concentration (K2 ) and the specific maximum methane utilization rate of the AOM-D syntrophic consortium (kmax2 ), with kmax2 being more important. Model simulations show that increasing kmax2 to >3 g chemical oxygen demand (COD)/g cell-day (from its current 1.8 g COD/g cell-day) and developing a new membrane with doubled methane-delivery capacity (Dm ) could bring the nitrate-removal flux to ≥4.0 g N/m2 -day, which is close to the nitrate-removal flux for the H2 -based MBfR. Further increase of the maximum nitrate-removal flux can be achieved when Dm and kmax2 increase together.

Induction of cathodic voltage reversal and hydrogen peroxide synthesis in a serially stacked microbial fuel cell.

We developed an innovative strategy to address the inhibition of anode-respiring bacteria due to voltage reversal in serially stacked microbial fuel cells by inducing cathodic voltage reversal and H2O2 production. When platinum-coated carbon (Pt/C) cathodes were employed (stacked MFCPt/C) and the MFC was operated with acetate medium, the last unit (MFC 4) caused a voltage reversal of -0.8 V with a substantial anode overpotential of 1.22 V. After replacing the Pt/C cathode with a Pt-free carbon gas diffusion electrode in MFC 4, an electrode overpotential, approximately 0.5 V, was shifted from the anode to the cathode, inducing cathodic voltage reversal. Under cathodic voltage reversal, MFC 4 generated H2O2 at a production rate of 117 mg H2O2/m2-h. Hence, under cathodic voltage reversal induced by Pt-free cathodes, due to less anode polarization, the anode-respiring activity can largely be sustained in a stacked MFC that treats organic wastewater consistently and the quality of treated wastewater may be improved with energy-efficient and on-site generated H2O2.

The Roles of Biofilm Conductivity and Donor Substrate Kinetics in a Mixed-Culture Biofilm Anode.

We experimentally assessed the kinetics and thermodynamics of electron transfer (ET) from the donor substrate (acetate) to the anode for a mixed-culture biofilm anode. We interpreted the results with a modified biofilm-conduction model consisting of three ET steps in series: (1) intracellular ET, (2) non-Ohmic extracellular ET (EET) from an outer membrane protein to an extracellular cofactor (EC), and (3) ET from the EC to the anode by Ohmic-conduction in the biofilm matrix. The steady-state current density was 0.82 ± 0.03 A/m2 in a miniature microbial electrochemical cell operated at fixed anode potential of -0.15 V versus the standard hydrogen electrode. Illumina 16S-rDNA and -rRNA sequences showed that the Geobacter genus was less than 30% of the community of the biofilm anode. Biofilm conductivity was high at 2.44 ± 0.42 mS/cm, indicating that the maximum current density could be as high as 270 A/m2 if only Ohmic-conduction EET was limiting. Due to the high biofilm conductivity, the maximum energy loss for Ohmic-conduction EET was negligible, 0.085 mV. The energy loss in the second ET step also was small, only 20 mV, and the potential for the EC involved in the second ET was -0.15 V, a value documenting that >99% of the EC was in the oxidized state. Monod kinetics for utilization of acetate were relatively slow, and at least 87% of the energy loss was in the intracellular step. Thus, intracellular ET was the main kinetic and thermodynamic bottleneck to ET from donor substrate to the anode for a highly conductive biofilm.

Ohmic resistance affects microbial community and electrochemical kinetics in a multi-anode microbial electrochemical cell.

Multi-anode microbial electrochemical cells (MxCs) are considered as one of the most promising configurations for scale-up of MxCs, but understanding of anode kinetics in multiple anodes is limited in the MxCs. In this study we assessed microbial community and electrochemical kinetic parameters for biofilms on individual anodes in a multi-anode MxC to better comprehend anode fundamentals. Microbial community analysis targeting 16S rRNA Illumina sequencing showed that Geobacter genus was abundant (87%) only on the biofilm anode closest to a reference electrode (low ohmic energy loss) in which current density was the highest among three anodes. In comparison, Geobacter populations were less than 1% for biofilms on other two anodes distant from the reference electrode (high ohmic energy loss), generating small current density. Half-saturation anode potential (EKA) was the lowest at -0.251 to -0.242 V (vs. standard hydrogen electrode) for the closest biofilm anode to the reference electrode, while EKA was as high as -0.134 V for the farthest anode. Our study proves that electric potential of individual anodes changed by ohmic energy loss shifts biofilm communities on individual anodes and consequently influences electron transfer kinetics on each anode in the multi-anode MxC.

Energy-positive food wastewater treatment using an anaerobic membrane bioreactor (AnMBR).

An immersed-membrane anaerobic membrane bioreactor (AnMBR) achieved 88-95% of COD removal for meat-processing wastewater at organic loading rate (OLR) of 0.4-3.2 kgCOD m(-3) d(-1). Membrane flux was stable for low OLR (0.4 and 1.3 kgCOD m(-3) d(-1)), but irrecoverable fouling occurred at high OLR of 3.2 kgCOD m(-3) d(-1). Methane gas yield of 0.13-0.18 LCH4 g(-1)CODremoved was obtained, which accounted for 33-38% of input COD, the most significant electron sink. Dissolved methane was only 3.4-11% of input COD and consistently over-saturated at all OLR conditions. The least accumulation of dissolved methane (25 mg L(-1) and saturation index 1.3) was found for the highest OLR of 3.2 kgCOD m(-3) d(-1) where biogas production rate was the highest. Energy balances showed that AnMBR produced net energy benefit of 0.16-1.82 kWh m(-3), indicating the possibility of energy-positive food wastewater treatment using AnMBRs.

Contribution of Liquid/Gas Mass-Transfer Limitations to Dissolved Methane Oversaturation in Anaerobic Treatment of Dilute Wastewater.

The mechanisms controlling the accumulation of dissolved methane in anaerobic membrane bioreactors (AnMBRs) treating a synthetic dilute wastewater (a glucose medium) were assessed experimentally and theoretically. The AnMBR was maintained at a temperature of 24-26 °C as the organic loading rate increased from 0.39 to 1.1 kg COD/m(3)-d. The measured concentration of dissolved methane was consistently 2.2- to 2.5-fold larger than the concentration of dissolved methane at thermodynamic equilibrium with the measured CH4 partial pressure, and the fraction of dissolved methane was as high as 76% of the total methane produced. The low gas production rate in the AnMBR significantly slowed the mass transport of dissolved methane to the gas phase. Although the production rate of total methane increased linearly with the COD loading rate, the concentration of dissolved methane only slightly increased with an increasing organic loading rate, because the mass-transfer rate increased by almost 5-fold as the COD loading increased from 0.39 to 1.1 kg COD/m(3)-d. Thus, slow mass transport kinetics exacerbated the situation in which dissolved methane accounted for a substantial fraction of the total methane generated from the AnMBR.

Ammonium nitrogen removal from the permeates of anaerobic membrane bioreactors: economic regeneration of exhausted zeolite.

This study revealed that ammonium exchange of natural zeolite could be an economical method of nitrogen removal from the permeates of anaerobic membrane bioreactors (AnMBRs). It was found that the mass ratio of Na+ to Zeolite - NH4+ - N significantly affected regeneration efficiency (RE), not simply NaCI concentration. Batch experiments showed that the mass ratio of 750g Na+/g Zeolite - NH4+ - N was required to achieve RE over 90% in 2h at pH 9. However, the alkaline regeneration at pH 12 significantly decreased the mass ratio down to 4.2 in batch tests. It was confirmed that the alkaline regeneration only needed NaCl 10 g/L (the mass of Na+ to Zeolite - NH4+ - N of 4.2) for RE of 85% in 2 h of reaction time in continuous column tests. Economic analysis showed that this alkaline regeneration decreased chemical costs over 10 times as compared with a conventional regeneration method. A significant bottleneck of zeolite processes would be the requirement of substituting exhausted zeolite with virgin one, due to the reductions of ammonium exchange capacity and RE.

Membranes for bioelectrochemical systems: challenges and research advances.

Increasing energy demand has been a big challenge for current society, as the fossil fuel sources are gradually decreasing. Hence, development of renewable and sustainable energy sources for the future is considered one of the top priorities in national strategic plans. Bioenergy can meet future energy requirements - renewability, sustainability, and even carbon-neutrality. Bioenergy production from wastes and wastewaters is especially attractive because of dual benefits of energy generation and contaminant stabilization. There are several bioenergy technologies using wastes and wastewaters as electron donor, which include anaerobic digestion, dark biohydrogen fermentation, biohydrogen production using photosynthetic microorganisms, and bioelectrochemical systems (BESs). Among them BES seems to be very promising as we can produce a variety of value-added products from wastes and wastewaters, such as electric power, hydrogen gas, hydrogen peroxide, acetate, ethanol etc. Most ofthe traditional BES uses a membrane to separate the anode and cathode chamber, which is essential for improving microbial metabolism on the anode and the recovery of value-added products on the cathode. Performance of BES lacking a membrane can be seriously deteriorated, due to oxygen diffusion or substantial loss of synthesized products. For this reason, usage of a membrane seems essential to facilitate BES performance. However, a membrane can bring several technical challenges to BES application compared to membrane-less BES. These challenges include poor proton permeability, substrate loss, oxygen back diffusion, pH gradient, internal resistance, biofouling, etc. This paper aims to review the major technical barriers associated with membranes and future research directions for their application in BESs.

The effect of solids retention time on dissolved methane concentration in anaerobic membrane bioreactors.

We assessed the effect of solids retention times (SRT) on dissolved methane concentration in a lab-scale anaerobic membrane bioreactor (AnMBR) operated at SRT 20d and 40d at ambient temperature (23 +/- 1 degrees C). Daily methane production was 196 +/- 17 mL/d and 285 +/- 18 mL/d for SRT 20d and 40d, respectively. In comparison, the average concentration of dissolved methane in AnMBR permeates was 9.9 +/- 2.3 mg/L for SRT 20d (close to thermodynamic equilibrium), which was decreased to 4.3 +/- 0.3 mg/L for SRT 40d. We often found oversaturation of dissolved methane at SRT 20d, which means that mass transfer of dissolved methane from liquid to gas phase is dynamic at this short SRT. However, we never detected oversaturation of dissolved methane at SRT 40d, due to slow endogenous decay kinetics. Higher daily methane production at SRT 40d than that at SRT 20d indicates that methane was supplementarily produced from biomass electrons by endogenous decay. This study shows that operation of AnMBRs under long SRT can keep low dissolved methane concentration in AnMBR permeate, along with high methane yield.

Optimization of biofilm conductance measurement with two-electrode microbial electrochemical cells (MECs).

This study was conducted to develop a standardized and consistent method for biofilm conductance measurement for an improved comprehension of extracellular electron transfer. Biofilm conductance (2.12 ± 0.25 × 10-4 S) with and without a fixed anode potential did not show significant difference. The conductance showed a sigmoidal relationship with anode potential. The current-voltage profile of the tested biofilm at applied voltage larger than 100 mV showed deviation from Ohm's law. Up to 69% decrease in biofilm conductance and deviation from Ohm's law were observed in the current-voltage profile when the measurement time increased. By choosing the voltage range (0- 100 mV) and step (25 mV), measurement time (100-s at each voltage step), and anode control mode, these operation settings were found more suitable for consistent and accurate biofilm conductance measurement in the 2-Au MEC system. This represents the first study that comprehensively evaluated the environmental and instrumental parameters for biofilm conductance measurement.

An integrated leachate bed reactor - anaerobic membrane bioreactor system (LBR-AnMBR) for food waste stabilization and biogas recovery.

This study developed an integrated LBR - AnMBR system for efficient stabilization and biogas recovery from food waste (FW) at room temperatures (21-22 °C). First, the leachate recirculation rate (4.4-13.2 L/h) was optimized to maximize hydrolysis and acidification yields. The maximum hydrolysis yield of 551 gSCOD/kg VSadded was achieved at recirculation rate of 13.2 L/h. The VFA concentrations in the FW leachate was as high as 12.5-16.0 g/L, resulting in a high acidification of 468 g CODVFA/kg VS. The solubilized FW was further stabilized by feeding the leachate to AnMBR. Different hydraulic (HRT) and solids retention times (SRT) were tested to achieve high COD removal and methane yields. High COD removal of 86 ± 3% was obtained in the AnMBR at HRT of 13 and SRT of 75 days. High biogas recovery of about 850 kWh per ton FWtreated was achieved along with high quality of AnMBR permeates containing low COD concentration but advantageously high concentration of nutrients (NH4+-N 317-403 mg/L, total phosphate 23-213 mg/L) without any particulates, which can be reused for landscape or liquid fertilizer.

Nitrite reduction using a membrane biofilm reactor (MBfR) in a hypoxic environment with dilute methane under low pressures.

Methane-based membrane biofilm reactors (MBfRs) can be an effective solution for nitrogen control in wastewater, but there is limited information on nitrite reduction for dilute wastewater (e.g., municipal wastewater) in hypoxic MBfRs. This study assessed the impacts of dilute (20 %), low-pressure methane (0.35-2.41 kPa) applied to MBfRs at hydraulic retention times (HRTs) of 2-12 h on nitrite removals, dissolved methane concentrations, and the resulting changes in the microbial community. High nitrite flux along with rapid and virtually complete (>99 %) nitrite removals were observed at methane pressures of 1.03-2.41 kPa at HRTs above 4 h, despite the use of diluted methane gas for the MBfR. The lowest methane pressure (0.35 kPa) was also able to achieve up to 98 % nitrite removals but required HRTs of up to 12 h. All scenarios had low dissolved methane concentrations (<10 mg/L), indicating that dilute methane at low supply pressures can effectively remove nitrite while meeting dissolved methane guidelines in treated effluent. Methylococcus genus was the key bacterium in MBfR biofilm grown at different HRTs and methane pressures, along with Methylocystis and other heterotrophic denitrifiers (Terrimonas and Hyphomicrobium). This study indicates that methane-based denitrification MBfRs can be a valuable tool to meet nitrogen limits for dilute wastewater coupled to partial nitrification, while limiting the release of methane to the environment.

Microbial electrolysis cells for the production of biohydrogen in dark fermentation - A review.

To assess biohydrogen for future green energy, this review revisited dark fermentation and microbial electrolysis cells (MECs). Hydrogen evolution rate in mesophilic dark fermentation is as high as 192 m3 H2/m3-d, however hydrogen yield is limited. MECs are ideal for improving hydrogen yield from carboxylate accumulated from dark fermentation, whereas hydrogen production rate is too slow in MECs. Hence, improving anode kinetic is very important for realizing MEC biohydrogen. Intracellular electron transfer (IET) and extracellular electron transfer (EET) can limit current density in MECs, which is proportional to hydrogen evolution rate. EET does not limit current density once electrically conductive biofilms are formed on anodes, potentially producing 300 A/m2. Hence, IET kinetics mainly govern current density in MECs. Among parameters associated with IET kinetic, population of anode-respiring bacteria in anode biofilms, biofilm density of active microorganisms, biofilm thickness, and alkalinity are critical for current density.

Biohydrogen production and purification: Focusing on bioelectrochemical systems.

Innovative technologies on green hydrogen production become significant as the hydrogen economy has grown globally. Biohydrogen is one of green hydrogen production methods, and microbial electrochemical cells (MECs) can be key to biohydrogen provision. However, MECs are immature for biohydrogen technology due to several limitations including extracellular electron transfer (EET) engineering. Fundamental understanding of EET also needs more works to accelerate MEC commercialization. Interestingly, studies on biohydrogen gas purification are limited although biohydrogen gas mixture requires complex purification for use. To facilitate an MEC-based biohydrogen technology as the green hydrogen supply this review discussed EET kinetics, engineering of EET and direct interspecies electron transfer associated with hydrogen yield and the application of advanced molecular biology for improving EET kinetics. Finally, this article reviewed biohydrogen purification technologies to better understand purification and use appropriate for biohydrogen, focusing on membrane separation.

Acidogenic fermentation of food waste in a leachate bed reactor (LBR) at high volumetric organic Loading: Effect of granular activated carbon (GAC) and sequential enrichment of inoculum.

This study investigated the impact of different granular activated carbon (GAC) loadings and inoculum enrichment on acidogenic fermentation of food waste in a leachate bed reactor (LBR) operated at a high volumetric organic loading of 49 g VS/Lreactor. LBR with a high GAC loading of 0.51 g GAC/g VSfood waste achieved hydrolysis yield of 620 g SCOD/kg VSadded, significantly (P ≤ 0.05) higher to that obtained for LBRs with low or no GAC loading. A high GAC loading also resulted in a higher acidification yield of 507 g CODSCFA/kg VSadded. Butyrate dominated the short-chain fatty acid (SCFA) composition by constituting 57-60 % of total SCFA at high GAC loadings, while the composition of acetate (38-40 %) and butyrate (36-38 %) were similar at lower GAC loadings. Inoculum enrichment further improved the hydrolysis and acidogenesis yields by 10-22 % resulting in the final hydrolysis yield of 683 g SCOD/kg VSadded and acidification yield of 617 g CODSCFA/kg VSadded.

Anaerobic membrane bioreactors for wastewater treatment: Challenges and opportunities.

Anaerobic membrane bioreactors (AnMBRs) have become a new mature technology and entered into the wastewater market, but there are several challenges to be addressed for wide applications. In this review, we discuss challenges and potentials of AnMBRs focusing on wastewater treatment. Nitrogen and dissolved methane control, membrane fouling and its control, and membrane associated cost including energy consumption are main bottlenecks to facilitating AnMBR application in wastewater treatment. Accumulation of dissolved methane in AnMBR permeate decreases the benefit of methane energy and contributes to methane gas emissions to atmosphere. Separate control units for nitrogen and dissolved methane add system complexity and increase capital and operating and maintenance (O & M) costs in AnMBR-centered wastewater treatment. Alternatively, methane-based denitrification can be an ideal nitrogen control process due to simultaneous removal of nitrogen and dissolved methane. Membrane fouling and energy associated with membrane fouling control are major limitations, in addition to membrane cost. More efforts are required to decrease capital and O & M costs associated with the control of dissolved methane nitrogen and membrane fouling to facilitate AnMBRs for wastewater treatment. PRACTITIONER POINTS: AnMBRs can accelerate anaerobic wastewater treatment including dilute wastewater. Nitrogen and dissolved methane control is detrimental for AnMBR application to wastewater treatment. Membrane biofilm reactors using gas-permeable membranes are suitable for simultaneous nitrogen and dissolved methane control. High capital and O & M costs from membranes are a major bottleneck to wide application of AnMBRs. Dynamic membranes could be an option to reduce capital and O & M costs for AnMBRs.

State-of-the-art management technologies of dissolved methane in anaerobically-treated low-strength wastewaters: A review.

The recent advancement in low temperature anaerobic processes shows a great promise for realizing low-energy-cost, sustainable mainstream wastewater treatment. However, the considerable loss of the dissolved methane from anaerobically-treated low-strength wastewater significantly compromises the energy potential of the anaerobic processes and poses an environmental risk. In this review, the promises and challenges of existing and emerging technologies for dissolved methane management are examined: its removal, recovery, and on-site reuse. It begins by describing the working principles of gas-stripping and biological oxidation for methane removal, membrane contactors and vacuum degassers for methane recovery, and on-site biological conversion of dissolved methane into electricity or value-added biochemicals as direct energy sources or energy-compensating substances. A comparative assessment of these technologies in the three categories is presented based on methane treating efficiency, energy-production potential, applicability, and scalability. Finally, current research needs and future perspectives are highlighted to advance the future development of an economically and technically sustainable methane-management technology.