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.

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.

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.

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.

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.

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.

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.

A modelling study of the spatially heterogeneous mutualism between electroactive biofilm and planktonic bacteria.

Microbial cooperation widely exists in anaerobic reactors degrading complex pollutants, conventionally studied separately inside the biofilm or the planktonic community. Recent experiments discovered the mutualism between the planktonic bacteria and electroactive biofilm treating propionate, an end-product usually accumulated in anaerobic digesters. Here, a one-dimensional multispecies model found the preference on acetate-based pathway over the hydrogen-based in such community, evidenced by the fact that acetate-originated current takes 66% of the total value and acetate-consuming anode-respiring bacteria takes over 80% of the biofilm. Acetate-based anodic respiration most apparently influences biofilm function while propionate fermentation is the dominant planktonic bio-reaction. Additionally, initial planktonic propionate level shows the ability of coordinating the balance between these two extracellular electron transfer pathways. Increasing the propionate concentration from 2 to 50 mM would increase the steady hydrogen-originated current by 210% but decrease the acetate-originated by 26%, suggesting a vital influence of the planktonic microbial process to the metabolic balance in biofilm. Best strategy to promote the biofilm activity is to increase the biomass density and biofilm conductivity simultaneously, which would increase the current density by 875% without thickening the biofilm thickness or prolonging the growth apparently.

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.

Recovery of dissolved methane from anaerobically treated food waste leachate using solvent-based membrane contactor.

The difficulty of dissolved methane recovery remains a major hurdle for mainstream anaerobic wastewater treatment processes. We recently proposed solvent-based membrane contactor (SMC) for high (>90%) methane recovery over a wide temperature range and net-energy production. Here, we investigate the methane recovery efficacy of the SMC process by using an AnMBR effluent from treating food waste leachate. We observed almost identical methane transfer kinetics to the process employing foulant-free methane-saturated feed solutions, with >92% methane recoveries, showing that organic foulants have insignificant impacts on the methane transport in the SMC. We then performed two different membrane contactor experiments: direct-contact membrane-distillation (DCMD, with transmembrane water vapor flow) and SMC (no water vapor flow). From the negligible fouling observed in the SMC experiment, opposite to the DCMD, we elucidate that the absence of water vapor flow renders the SMC process intrinsically robust to membrane fouling. With the low fouling propensity of the SMC process under highly fouling environments, our study highlights the feasibility of SMC processes to enhance the energy production in mainstream anaerobic wastewater treatment processes.

Hydrophilic graphene aerogel anodes enhance the performance of microbial electrochemical systems.

The hydrophilic three-dimensional (3D) structure of graphene materials was produced with reducing agent-ethylene glycol through hydrothermal reduction. Numerous microorganisms with diverse community structure were established in anode surface, as the hydrophilicity of the graphene anode increased; more populations of Proteobacteria and Firmicutes families were identified in a higher hydrophilic anode. In addition, the start-up time of a microbial fuel cell (MFC) equipped with hydrophilic 3D graphene anode was only 43 h, which is much shorter than traditional 3D graphene-based anode systems. The most hydrophilic anode exhibited the maximal power density of 583.8 W m-3, 5 times larger than the least hydrophilic one. The content of oxygen in graphene materials improving hydrophilicity would play an important role in enhancing power density. This study proves that hydrophilic 3D graphene materials as the anode can improve MFC performance and start-up time.

High-rate carboxylate production in dry fermentation of food waste at room temperature.

Dry fermentation of food waste was optimized to achieve the maximum solid loading rate for carboxylate production without clogging events in a dry fermenter run at neutral pH. High inoculum-to-substrate ratio improved food waste solubilization and carboxylate yield, but the ratio 15% completely clogged the dry fermenter. Higher leachate circulate rate tended to enhance food waste fermentation, but partial clogging was observed at 13.2 L/h of leachate circulation rate. The dry fermenter achieved carboxylate yield of 428.5 g/kg food waste and volatile solid reduction of 79% at the solid loading rate 4.82 kg volatile solids/m3-d. This study first tracks chemical oxygen demand (COD) in food waste dry fermentation, showing maximum soluble COD <60% of food waste COD with residual food waste 13.6-16.3%. The operating cost was as low as $1.7/ton FW, implying that food waste treatment will be cost-neutral if recovered carboxylate can create economic benefit over the operating cost.

Mixed dye wastewater treatment in a bioelectrochemical system-centered process.

The feasibility of mixed dye wastewater treatment was evaluated with a novel integrated bioprocess that consisted of a hybrid anaerobic reactor (HAR) with a built-in bioelectrochemical system, an aerobic biofilm reactor (ABFR) and a denitrification reactor (DR). The position of the DR significantly affected chemical oxygen demand (COD) and colority in effluent, and placing the DR after the ABFR improved effluent quality probably due to minimization of the undesired autoxidation of aromatic amine in dye wastewater. The optimal integrated process of HAR + ABFR + DR efficiently treated mixed dye wastewater, and concentrations of COD and TN were decreased down to 75 ± 18 mg/L and 12.91 ± 0.31 mg/L, respectively, along with colority 48 ± 4 times. Total phosphorus reduced to below 0.5 mg/L with coagulation using poly aluminum chloride, and the effluent quality fully met the discharge standard. This comprehensive study suggests the feasibility of the BES based process for practical application to mixed dye wastewater treatment.

Methane-based denitrification kinetics and syntrophy in a membrane biofilm reactor at low methane pressure.

A methane-based membrane biofilm reactor (MBfR) was assessed for a tertiary nitrogen removal process in domestic wastewater treatment. To mitigate effluent dissolved methane concentrations, the MBfR was operated with a 20% methane mixing ratio and a low pressure of 0.003 atm. The nitrate concentration was reduced from 20 to 4 mg/L with a low methane concentration of 3.3 mg/L in the effluent at 4 h hydraulic retention time (HRT). An in situ dissolved oxygen sensor showed a concentration of 0.045 mg/L in the MBfR, demonstrating methane oxidation under hypoxic conditions. Both 16S rRNA gene sequencing and metagenomic analysis identified bacteria capable of oxidation of methane coupled to denitrification (Methylocystis), whereas other bacteria were implicated in either methane oxidation (Methylococcus) or nitrate reduction (Escherichia). Reduced genetic potential for nitrate reduction to nitrite at a shorter HRT coincided with a decreased efficiency of denitrification, suggesting rate limitation by this initial step of denitrification. Genes encoding nitrite reduction to dinitrogen were at similar relative abundance under both HRT conditions. Our results provide mechanistic evidence for microbial syntrophy between aerobic methanotrophs and denitrifiers in methane-fed MBfRs operated under varying HRTs, with important implications for novel biological nitrogen removal to dilute wastewater.

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.