Applying metabolic flux analysis to hydrogen fermentation using a metabolic network constructed for anaerobic mixed cultures.

In this study, a mixed-cultural metabolic network for anaerobic digestion that included the concept of a "universal bacterium" was constructed, and metabolic flux analysis (MFA) applying this network was conducted to evaluate the flow of electrons and materials during H2 fermentation under various conditions. The MFA results from two H2 fermenters feeding glucose with (GP) or without (GA) the addition of peptone suggest that hydraulic retention time (HRT) presents a significant impact on hydrogen production, and the reversed trends could be observed at HRTs below and above 4 h. From the MFA results of lactate/acetate-fed H2 fermenter, the highest flux of H2 production is associated with more significant acetate consumption and the following pathways toward the anaplerotic reactions cycle that produces NADH. The occurrence of acetogenesis in the H2 fermenters using various types of bioethanol-fermented residues (BEFRs) was also identified according to the MFA results. By analyzing the MFA results of all 49 sets of data from H2 fermenters via Pearson's correlation, it was revealed that the flux of H2 production positively correlates to the reduction of ferredoxin with pyruvate oxidation, acetate formation, and acetate emission when lactate was produced in the system. On the contrary, negative relationships were found between the flux of H2 production and these three fluxes. The extended application of MFA provides additional information, including the fluxes between intracellular metabolites, and the information has the potential to be used in decision-making systems during the future operation of anaerobic processes by connecting operational parameters.

Resource recovery from lignocellulosic wastes via biological technologies: Advancements and prospects.

Lignocellulosic wastes were recently considered as biomass resources, however, its conversion to valuable products is still immature although researchers have put lots of effort into this issue. This article reviews the key challenges of the biorefinery utilizing lignocellulosic materials and recent developments to conquer those obstacles. Available biological techniques and processes, from the pretreatments of cellulosic materials to the valorization processes, were emphasized. Biological pretreatments, including hydrolysis using microbial consortia, fungi, enzymes, engineered bacterial/fungal strains, and co-culture systems, could enhance the release of reducing sugar. Resources recovery, including biogases, ethanol, butanol, PHA, etc., from lignocellulosic materials were also discussed, while the influences of composition of lignocellulosic materials and pretreatment options, applications of co-culture system, and integrated treatments with other wastes, were described. In the review, co-culture system and metabolic engineering are emphasized as the promising biological technologies, while perspectives are provided for their future developments.

Biological treatment of volatile organic compounds (VOCs)-containing wastewaters from wet scrubbers in semiconductor industry.

This study investigated biological treatment for two kinds of volatile organic compounds (VOCs)-containing wastewaters collected from wet scrubbers in a semiconductor industry. Batch test results indicated that one wastewater containing highly volatile organic compounds was not suitable for aerated treatment conditions while the other containing much lower volatile organic compounds was suitable for aerobic treatment. Accordingly, two moving bed bioreactors, by adding commercial biocarrier BioNET, were operated under aerobic and anoxic conditions for treating low volatility wastewater (LVW) and high volatility wastewater (HVW), respectively. During 280 days of operation, the aerobic LVW bioreactor attained the highest chemical oxygen demand (COD) removal rate of 98.9 mg-COD/L/h with 81% of COD removal efficiency at hydraulic retention time (HRT) of 1 day. The anoxic HVW bioreactor performed above 80% of COD removal efficiency with the highest COD removal rate of 16.5 mg-COD/L/h at HRT of 2 days after 380 days of operation. The specific COD removal rates at different initial substrate-to-biomass (S0/X0) ratios, using either suspended sludge or microorganisms attached onto BioNET from both bioreactors, followed the Monod-type kinetics, while the half-saturation coefficients were generally higher for the microorganisms onto BioNET due presumably to relatively poor mass transfer efficiency. Based on the results of microbial community analysis using the next generation sequencing technique, the dominant communities of suspended sludge and BioNET, including nitrifiers, denitrifiers, and degraders for polycyclic aromatic hydrocarbons, were similar in the corresponded bioreactors, but microbial community shifts were observed with increased organic loadings.

Effects of copper on biological treatment of NMF- and MDG-containing wastewater from TFT-LCD industry.

This study aimed to evaluate the effects of copper on N-methylformamide (NMF)- and methyl diglycol (MDG)-containing wastewater treatment using batch experiments and a lab-scale anoxic-oxic (A/O) sequencing batch reactor (SBR). Batch experimental results indicated that aerobic degradation of NMF followed Monod-type kinetics. Copper inhibition on nitrification also followed Monod-type inhibition kinetics with copper-to-biomass ratio instead of copper concentration. Specific degradation rates of NMF and MDG under both aerobic and anoxic conditions decreased in the matrix of full-scale wastewater, and high copper dosage would further reduce the degradation rates. In the long-term presence of 0.5 mg/L copper, the A/O SBR could maintain stable and complete degradations of NMF and MDG, 95% of COD removal, and more than 50% of total nitrogen (TN) removal. High concentrations of copper spikes, including 40 mg/L and 110 mg/L, slowed down degradation rates for both NMF and MDG, but did not affect COD and TN removal efficiencies in the full 24 h-cycle operation. The long-term A/O SBR operation revealed that daily dosage of 0.5 mg/L copper was not detrimental to NMF/MDG degradations due to regularly wasting sludge, but 110 mg/L of copper spike obviously reduced NMF/MDG degradation rate although it could be recovered later by regularly wasting sludge and maintaining SRT at 20 days.

Nitrogen removal as nitrous oxide for energy recovery: Increased process stability and high nitrous yields at short hydraulic residence times.

The Coupled Aerobic-anoxic Nitrous Decomposition Operation (CANDO) is a two-stage process for nitrogen removal and resource recovery: in the first, ammonia is oxidized to nitrite in an aerobic bioreactor; in the second, oxidation of polyhydroxyalkanoate (PHA) drives reduction of nitrite to nitrous oxide (N2O) which is stripped for use as a biogas oxidant. Because ammonia oxidation is well-studied, tests of CANDO to date have focused on N2O production in anaerobic/anoxic sequencing batch reactors. In these reactors, nitrogen is provided as nitrite; PHA is produced from acetate or other dissolved COD, and PHA oxidation is coupled to N2O production from nitrite. In a pilot-scale study, N2O recovery was affected by COD/N ratio, total cycle time, and relative time periods for PHA synthesis and N2O production. In follow-up bench-scale studies, different reactor cycle times were used to investigate these operational parameters. Increasing COD/N ratio improved nitrite removal and increased biosolids concentration. Shortening the anaerobic phase prevented fermentation of PHA and improved its utilization. Efficient PHA synthesis and utilization in the anaerobic phase correlated with high N2O production in the anoxic phase. Shortening the anoxic phase prevented reduction of N2O to N2. By shortening both phases, total cycle time was reduced from 24 to 12 h. This optimized operation enabled increased biomass concentrations, increased N2O yields (from 71 to 87%), increased N loading rates (from 0.1 to 0.25 kg N/m3-d), and shorter hydraulic residence times (from 10 to 2 days). Long-term changes in operational performance for the different bioreactor systems tested were generally similar despite significant differences in microbial community structure. Long-term operation at short anaerobic phases selected for a glycogen-accumulating community dominated by a Defluviicoccus-related strain.

Recent advancement on biological technologies and strategies for resource recovery from swine wastewater.

Swine wastewater is categorized as one of the agricultural wastewater with high contents of organics and nutrients including nitrogen and phosphorus, which may lead to eutrophication in the environment. Insufficient technologies to remove those nutrients could lead to environmental problems after discharge. Several physical and chemical methods have been applied to treat the swine wastewater, but biological treatments are considered as the promising methods due to the cost effectiveness and performance efficiency along with the production of valuable products and bioenergies. This review summarizes the characteristics of swine wastewaters in the beginning, and briefly describes the current issues on the treatments of swine wastewaters. Several biological techniques, such as anaerobic digestion, A/O process, microbial fuel cells, and microalgae cultivations, and their future aspects will be addressed. Finally, the potentials to reutilize biomass produced during the treatment processes are also presented under the consideration of circular economy.

Biological treatment of DMSO-containing wastewater from semiconductor industry under aerobic and methanogenic conditions.

This study evaluated biological treatment of dimethyl sulfoxide (DMSO)-containing wastewater from semiconductor industry under aerobic and anaerobic conditions. DMSO concentration as higher as 1.5 g/L did not inhibit DMSO degradation efficiency in aerobic membrane bioreactor (MBR), while specific DMSO degradation rate at different initial DMSO-to-biomass (S0/X0) ratios from batch tests seemed to follow the Haldane-type kinetics. According to the microbial community analysis, Proteobacteria decreased from 88.2% to 26% as influent DMSO concentration increased, while Bacteroidetes, Parcubacteria, Saccharibacteria increased. Within the Bacteroidetes class, Flavobacterium and Laribacter genus significantly increased from less than 0.05%-26.8% and 13.4%, respectively, which might both be related to the DMS degradation. Hyphomicrobium and Thiobacillus, known as aerobic DMSO and DMS degraders, instead, decreased at higher DMSO conditions. Under methanogenic conditions, batch results implied DMSO concentrations higher than 3 g/L could be inhibitory, while DMSO and COD removal achieved 100% and 93%, respectively, using a pilot-scale anaerobic fluidized bed membrane bioreactor (AFMBR) with influent DMSO below 1.5 g/L. Results of terminal restriction fragment length polymorphism (TRFLP) analysis targeting on mcrA functional gene revealed that Methanomethylovorans sp. was dominant in AFMBR after 54 days of operation, indicating its importance on degrading DMS and mathanethiol (MT).

Pilot study of cold-rolling wastewater treatment using single-stage anaerobic fluidized membrane bioreactor.

A pilot-scale single-stage anaerobic fluidized membrane bioreactor (AFMBR) was firstly used in this study to treat cold-rolling emulsion wastewater from steel industry. It was continuously operated for 302 days with influent COD concentration of 860-1120 mg/L. Under a hydraulic retention time of 1.5 d, the average effluent COD concentration of 72 mg/L achieved corresponding 90% of COD removal. The permeate flux was varied between 1.7 and 2.9 L/m2/h during operation which decreased with increased biomass concentration inside AFMBR. The trans-membrane pressure (TMP) was generally around 35-40 kPa, however, it increased up to 60 kPa when volatile suspended solid increased to above 2.5 g/L. Both flux and TMP data reveal the importance of biomass control for AFMBR operation. Results from terminal restriction fragment length polymorphism (T-RFLP) show the genus Methanosaeta was dominant on GAC and it shared dominance with the genera Methanomethylovorans and Methanosarcina in suspended sludge.

Biological acetate production from carbon dioxide by Acetobacterium woodii and Clostridium ljungdahlii: The effect of cell immobilization.

This study investigated the acetate production from gas mixture of hydrogen (H2) and carbon dioxide (CO2) in the ratio of 7:3 using two acetogens: Acetobacterium woodii and Clostridium ljungdahlii. Batch result shows A. woodii performed two-phase degradation with the presence of glucose that lactate was produced from glucose and was reutilized for the production of butyrate and few acetate, while only acetate was detected when providing gas mixture. C. ljungdahlii produced butyrate and ethanol along with acetate when glucose was introduced, while only ethanol and acetate were found by feeding gas mixture. The acetate-to-ethanol (A/E) ratio can be enhanced by cell immobilization, while GAC immobilization produced only acetate and the production rate reached 0.072 mmol/d under fed-batch operation. Acetate production rate increased from 18 to 28 mmol/L/d with GAC immobilization when gas flowrate increased from 100 to 300 mL/min in anaerobic fluidized membrane bioreactor (AFMBR), and a highest A/E ratio of 30 implies the possible application of acetate recovery from H2 and CO2.

Biological hydrogen and methane production from bagasse bioethanol fermentation residues using a two-stage bioprocess.

This study investigated the recovery of H2 and CH4 from bagasse bioethanol fermentation residues (bagasse BEFR) using a two-stage bioprocess. In the hydrogen fermentation bioreactor (HFB), carbohydrate removal efficiency was maintained at 82-93% and the highest hydrogen yield was 8.24mL/gCOD at volumetric loading rate (VLR) of 80kgCOD/m(3)/day. The results indicated a positive correlation between hydrogen yield and butyrate-to-acetate ratio, which might be due to the mechanisms of lactate/acetate utilization for hydrogen production and acetogenesis occurring in the HFB. Remaining volatile fatty acids and alcohols in the HFB effluent were further utilized for methane production in methane fermentation bioreactor (MFB), in which the highest methane yield of 345.2mL/gCOD was attained at VLR of 2.5kgCOD/m(3)/day. Overall, the two-stage bioprocess achieved a maximum COD removal of 81% from bagasse BEFR, and converted 0.3% and 72.8% of COD in the forms of H2 and CH4, respectively.

Enhanced bioenergy recovery from oil-extracted microalgae residues via two-step H2/CH4 or H2/butanol anaerobic fermentation.

Algae-based biodiesel is considered a promising alternative energy; therefore, the treatment of microalgae residues would be necessary. Anaerobic processes can be used for treating oil-extracted microalgae residues (OMR) and at the same time for recovering bioenergy. In this study, anaerobic batch experiments were conducted to evaluate the potential of recovering bioenergy, in the forms of butanol, H2, or CH4, from pretreated OMR. Using pretreated OMR as the only substrate, a butanol yield of 0.086 g/g-carbohydrate was obtained at carbohydrate of 40 g/L. With supplemented butyrate, a highest butanol yield of 0.192 g/g-carbohydrate was achieved at pretreated OMR containing 25 g/L of carbohydrate with 15 g/L of butyrate addition, attaining the highest energy yield of 3.92 kJ/g-OMR and energy generation rate of 0.65 kJ/g-OMR/d. CH4 production from pretreated OMR attained an energy yield of 8.83 kJ/g-OMR, but energy generation rate required further improvement. H2 production alone from pretreated OMR might not be attractive regarding energy yield, but it attained a superb energy generation rate of 0.68 kJ/g-OMR/d by combining H2 production from pretreated OMR and butanol production from pretreated OMR with supplementary butyrate from H2 fermentation supernatant. This study demonstrated an integrated system as an option for treating OMR and recovering bioenergy.

Biological butanol production from microalgae-based biodiesel residues by Clostridium acetobutylicum.

This study conducted batch experiments to evaluate the potential of butanol production from microalgae biodiesel residues by Clostridium acetobutylicum. The results indicated that with 90 g/L of glucose as the sole substrate the highest butanol yield of 0.2 g/g-glucose was found, but the addition of butyrate significantly enhanced the butanol yield. The highest butanol yield of 0.4 g/g-glucose was found with 60 g/L of glucose and 18 g/L of butyrate. Using microalgae biodiesel residues as substrate, C. acetobutylicum produced 3.86 g/L of butanol and achieved butanol yield of 0.13 g/g-carbohydrate via ABE fermentation, but the results indicated that approximately one third of carbohydrate was not utilized by C. acetobutylicum. Biological butanol production from microalgae biodiesel residues can be possible, but further research on fermentation strategies are required to improve production yield.

Metabolic flux network analysis of fermentative hydrogen production: using Clostridium tyrobutyricum as an example.

This study applies metabolic flux network analysis (MFA) to evaluate the metabolic flux of fermentative hydrogen production (FHP) with the use of Clostridium tyrobutyricum fed with either glucose or lactate/acetate as substrates. The MFA results suggest that hydraulic retention time (HRT) presents significant impact on hydrogen production from glucose. At HRT between 4 and 18 h, increase of HRT increased hydrogen production but decreased lactate production, while at HRT below 4 h decrease of HRT increased hydrogen production but decreased lactate production. The flux for lactate, butyrate and acetate seemed to affect H2 production, due presumably to their impacts on the balance of NADH, ferredoxin and ATP. It is suggested that the MFA can be a useful tool to provide valuable information for optimization and design of the fermentative hydrogen production process.

A two-stage bioprocess for hydrogen and methane production from rice straw bioethanol residues.

This study evaluates a two-stage bioprocess for recovering hydrogen and methane while treating organic residues of fermentative bioethanol from rice straw. The obtained results indicate that controlling a proper volumetric loading rate, substrate-to-biomass ratio, or F/M ratio is important to maximizing biohydrogen production from rice straw bioethanol residues. Clostridium tyrobutyricum, the identified major hydrogen-producing bacteria enriched in the hydrogen bioreactor, is likely utilizing lactate and acetate for biohydrogen production. The occurrence of acetogenesis during biohydrogen fermentation may reduce the B/A ratio and lead to a lower hydrogen production. Organic residues remained in the effluent of hydrogen bioreactor can be effectively converted to methane with a rate of 2.8 mmol CH(4)/gVSS/h at VLR of 4.6 kg COD/m(3)/d. Finally, approximately 75% of COD in rice straw bioethanol residues can be removed and among that 1.3% and 66.1% of COD can be recovered in the forms of hydrogen and methane, respectively.

Fermentative biohydrogen production from lactate and acetate.

In this study, a continuous-flow stirred tank reactor (CSTR) fed with lactate and acetate was operated to enrich hydrogen-producing bacteria. By varying the influent substrate concentrations and hydraulic retention times (HRT), the volumetric loading rate (VLR) of 55.64 kg-COD/m(3)/day seemed to be optimum for this enriched culture for fermentative hydrogen production from lactate and acetate. The results of batch experiments confirmed that the enriched culture tended to fulfill the e(-) equiv requirement for cell growth at a lower VLR condition (21.77 kg-COD/m(3)/day), while it could largely distribute the e(-) equiv for hydrogen production at a higher VLR condition. However, a maximum lactate/acetate concentration allowed for enriching this culture existed, especially at a lower HRT condition in which wash-out can be an issue for this enriched culture. Finally, the results of cloning and sequencing indicated that Clostridium tyrobutyricum was considered the major hydrogen-producing bacteria in the CSTR fed with lactate and acetate.

Metabolic and energetic aspects of biohydrogen production of Clostridium tyrobutyricum: The effects of hydraulic retention time and peptone addition.

This study evaluates the microbial metabolism and energy demand in fermentative biohydrogen production using Clostridium tyrobutyricum FYa102 at different hydraulic retention times (HRT) over a period of 1-18 h. The hydrogen yield shows a positive correlation with the butyrate yield, the B/A ratio, and the Y(H2)/2(Y(HAc)+Y(HBu)) ratio, but a negative correlation with the lactate yield. A decrease in HRT, which is accompanied by an increased biomass growth, tends to decrease the B/A ratio, due presumably to a higher energy demand for microbial growth. The production of lactate at a low HRT, however, may involve an unfavorable change in e(-) equiv distribution to result in a reduced hydrogen production. Finally, the relatively high hydrogen yields observed in the bioreactor with the peptone addition may be ascribed to the utilization of peptone as an additional energy and/or amino-acid source, thus reducing the glucose demand for biomass growth during the hydrogen production process.

Evaluation of bioenergy recovery processes treating organic residues from ethanol fermentation process.

This study evaluates a two-stage bioprocess for recovering bioenergy in the forms of hydrogen and methane while treating organic residues of ethanol fermentation from tapioca starch. A maximum hydrogen production rate of 0.77 mmol H(2)/g VSS/h can be achieved at volumetric loading rate (VLR) of 56 kg COD/m(3)/day. Batch results indicate that controlling conditions at S(0)/X(0)=12 with X(0)=4000 mg VSS/L and pH 5.5-6 are important for efficient hydrogen production from fermentation residues. Hydrogen-producing bacteria enriched in the hydrogen bioreactor are likely utilizing lactate and acetate for biohydrogen production from ethanol-fermentation residues. Organic residues remained in the effluent of hydrogen bioreactor can be effectively converted to methane with a rate of 0.37 mmol CH(4)/g VSS/h at VLR of 8 kg COD/m(3)/day. Approximately 90% of COD in ethanol-fermentation residues can be removed and among that 2% and 85.1% of COD can be recovered in the forms of hydrogen and methane, respectively.