Continuous production of L-tert-leucine in series of two enzyme membrane reactors

The L-tert-leucine synthesis was performed continuously in series of two enzyme-membrane reactors by reductive amination of trimethylpyruvate with leucine dehydrogenase. The necessary “native” cofactor NADH is regenerated with the aid of a second enzyme, formate dehydrogenase. Considering detailed kinetic studies of initial reaction rates under conditions relevant to the process a kinetic model was developed. The model shows that the overall reaction rate is strongly inhibited by the reaction product. The reactor's models combine the mass balances and proposed kinetic equations. The model adequacy was verified by using it to simulate the experiments and by comparing experimental and computed conversion, space-time yield and enzyme consumption. The calculations for the three reactor's types (batch, single CSTR and a cascade of two CSTRs in series) were compared. The results showed that a single CSTR is no favourable reactor configuration due to the very strong product inhibition. Space-time yield drops from 560 g litre^?1 day^?1 in a batch reactor to 110 g litre^?1 day^?1 in a single CSTR at the highest conversion of 98%. At the conversion of 95% the difference in biocatalyst costs between batch and two CSTR in series is negligible. Therefore the use of two enzyme membrane reactors in series was proposed. The modelling in this work shows that the optimisation of the quantity of the enzyme used results in a minimisation of the biocatalyst costs.     

Mesophilic batch anaerobic co-digestion of pulp and paper sludge and monosodium glutamate waste liquor for methane production in a bench-scale digester

This paper presented results from anaerobic co-digestion of pulp and paper sludge (PPS) and monosodium glutamate waste liquor (MGWL). A bench-scale anaerobic digester, 10 L in volume was developed, to operate under mesophilic (37 ± 2 °C) batch condition. Under versatile and reliable anaerobic conduct, high efficiency for bioconversion of PPS and MGWL were obtained in the system. The accumulative methane yield attained to 200 mL g⁻¹ VS_added and the peak value of methane daily production was 0.5 m³/(m³ d). No inhibitions of volatile fatty acids (VFAs) and ammonia on anaerobic co-digestion were found. pH 6.0-8.0 and alkalinity 1000-4000 mg CaCO₃/L were got without adjustment. This work showed that there was a good potential to the use of PPS and MGWL to anaerobic co-digestion for methane production.     

A continuous process for the production of baculovirus using insect-cell cultures

Summary A continuous culture of insect cells (Spodoptera frugiperda) was used for continuous production of baculovirus (nuclear polyhedrosis virus fromAutographa californica). The system consisted of a cascade of two continuous stirred tank reactors (CSTRs). In CSTR I the insect cells were grown in suspension. This suspension was fed continuously to CSTR II where the virus infection occurred. For a period of about 25 days the average volumetric productivity was about 10⁷ polyhedra (virus particles occluded in protein capsules) and 10⁸ infectious NOVs (non-occluded virus particles) per cm³ effluent. This is equivalent to 25 polyhedra and 250 NOVs per infected cell, respectively. In one case, the percentage of infected cells was 65%, which is close to the theoretical value of 68%. After a run-time of 32 days a decrease of process productivity was observed, probably due to the so-called passage effect, a degeneration of the virus DNA.     

Aerobic granular sludge formation for high strength agro-based wastewater treatment

The present study investigates the formation of aerobic granular sludge in sequencing batch reactor (SBR) fed with palm oil mill effluent (POME). Stable granules were observed in the reactor with diameters between 2.0 and 4.0 mm at a chemical oxygen demand (COD) loading rate of 2.5 kg COD m⁻³ d⁻¹. The biomass concentration was 7600 mg L⁻¹ while the sludge volume index (SVI) was 31.3 mL g SS⁻¹ indicating good biomass accumulation in the reactor and good settling properties of granular sludge, respectively. COD and ammonia removals were achieved at a maximum of 91.1% and 97.6%, respectively while color removal averaged at only 38%. This study provides insights on the development and the capabilities of aerobic granular sludge in POME treatment.     

Biogeochemical processes in intensive zero-effluent marine fish culture with recirculating aerobic and anaerobic biofilters

The biogeochemical processes that drive nutrient transformations and recycling in organic marine sediment-water environments were studied for 17 months in a zero-effluent intensive recirculating culture system. The system consisted of a 10 m³ gilthead seabream (Sparus aurata) tank coupled to aerobic and anaerobic water treatment elements. Nutrients and alkalinity were measured in the system to quantify the main biogeochemical processes. Fractions of the carbon fed in feed were found in fish (18.3%) and in sludge (11%); the missing carbon was respired by fish (45%) and by aerobic (8.4%) and anaerobic (7.7%) microorganisms. Fractions of the nitrogen fed in feed were found in fish (15.4%) and in sludge (14.3%); the missing nitrogen was eliminated by nitrification-denitrification. Most of the phosphorus and ash fed in feed and not found in fish accumulated within the sludge in the system. The rates of nitrification, denitrification and sulphate reduction increased with time, reaching 0.3 g N m^− 2 d^− 1, 53 g N m^− 2 d^− 1 and 145 g S m^− 2 d^− 1, respectively. Nitrification developed more rapidly than denitrification, leading at first to nitrate accumulation (to 20 mmol NO₃ l^− 1 by day 200) and a decrease in alkalinity. Once denitrification surpassed nitrification, nitrate concentrations decreased, eventually being reduced to < 0.3 mmol NO₃ l^− 1 by day 510, and alkalinity stabilized. Toxic hydrogen sulphide, generated within the anaerobic sludge, was oxidized by oxygen and nitrate as it diffused through the anaerobic-aerobic sediment-water interface. When nitrate levels in the water above the sludge dropped below 2 mmol l^− 1, sulphide was also oxidized in the fluidized bed reactor. Denitrification reduced nitrate in the water, respired (jointly with sulphate reduction) carbon in the sludge, oxidized the hydrogen sulphide, and contributed to stabilization of alkalinity and accumulation of polyphosphate in bacteria as a major sink of labile P.     

Energy valorization of industrial biomass: Using a batch frying process for sewage sludge

This paper studies the energy valorization of sewage sludge using a batch fry–drying process. Drying processes was carried out by emerging the cylindrical samples of the sewage sludge in the preheated recycled cooking oil. Experimental frying curves for different conditions were determined. Calorific values for the fried sewage sludge were hence determined to be around 24 MJ kg⁻¹, showing the auto-combustion potential of the fried sludge. A one-dimensional model allowing for the prediction of the water removal during frying was developed. Another water replacement model for oil intake in the fried sewage sludge was also developed. Typical frying curves were obtained and validated against the experimental data.     

Correlation between fouling propensity of soluble extracellular polymeric substances and sludge metabolic activity altered by different starvation conditions

Soluble extracellular polymeric substances (EPSs) cause membrane fouling in membrane bioreactors (MBRs), correlated with MBR sludge characteristics. Effects of F/M ratios on the evolution of soluble EPSs, fouling propensity of supernatants, and sludge metabolic activity were measured in this study in a two-period sequencing batch reactor (SBR). The experimental results show that fouling propensity was directly correlated with soluble-EPS concentration and composition. Sludge that had entirely lost active cells by long-term starvation released 64.4 ± 0.9 mg/L of humic acids, which caused a rapid increase in membrane resistance (40.67 ± 2.24 × 10¹¹ m⁻¹) during fouling tests. During short-term starvation, induced by incubation at a normal to low F/M ratio of 0.05 d⁻¹, sludge can use previously secreted utilization-associated products (UAPs) to maintain endogenous respiration. Therefore, the strategies of accumulating sludge and prolonging sludge retention time in MBRs may create long-term starvation and promote membrane fouling.     

Remediation of chromite ore processing residue by pyrolysis process with sewage sludge

The present work developed a novel technique to treat chromite ore processing residue (COPR). The process involved mixing the COPR with sewage sludge followed by pyrolysis. The gaseous organic fraction generated during pyrolysis of sludge was beneficial to Cr(VI) reduction. Process variables, such as the amount of sludge added to COPR (sludge-to-COPR (S/C) ratio), heating temperature, reaction time and particle size, were systematically varied, and their influences on the Cr(VI) reduction in COPR were investigated. Cr(VI) content had decreased greatly, from 3384 mg kg⁻¹ for untreated COPR to less than 30 mg kg⁻¹ for COPR treated at 600 °C.     

Modeling of two-phase anaerobic process treating traditional Chinese medicine wastewater with the IWA Anaerobic Digestion Model No. 1

The aim of the study was to implement a mathematical model to simulate two-phase anaerobic digestion (TPAD) process which consisted of an anaerobic continuous stirred tank reactor (CSTR) and an upflow anaerobic sludge blanket (UASB) reactor in series treating traditional Chinese medicine (TCM) wastewater. A model was built on the basis of Anaerobic Digestion Model No. 1 (ADM1) while considering complete mixing model for the CSTR, and axial direction discrete model and mixed series connection model for the UASB. The mathematical model was implemented with the simulation software package MATLABTM/Simulinks. System performance, in terms of COD removal, volatile fatty acids (VFA) accumulation and pH fluctuation, was simulated and compared with the measured values. The simulation results indicated that the model built was able to well predict the COD removal rate (−4.8–5.0%) and pH variation (−2.9–1.4%) of the UASB reactor, while failed to simulate the CSTR performance. Comparing to the measured results, the simulated acetic acid concentration of the CSTR effluent was underpredicted with a deviation ratios of 13.8–23.2%, resulting in an underprediction of total VFA and COD concentrations despite good estimation of propionic acid, butyric acid and valeric acid. It is presumed that ethanol present in the raw wastewater was converted into acetic acid during the acidification process, which was not considered by the model. Additionally, due to the underprediction of acetic acid the pH of CSTR effluent was overestimated.     

Production of medium-chain-length polyhydroxyalkanoates by activated sludge enriched under periodic feeding with nonanoic acid

The potential use of activated sludge for the production of medium-chain-length polyhydroxyalkanoates (MCL-PHAs) was investigated. The enrichment of bacterial populations capable of producing MCL-PHAs was achieved by periodic feeding with nonanoic acid in a sequencing batch reactor (SBR). Denaturing gradient gel electrophoresis analysis revealed Pseudomonas aeruginosa strains to be predominant in the bacterial community during the SBR process. The composition of PHA synthesized by the enriched biomass from nonanoic acid consisted of a large concentration (>89 mol%) of MCL monomer units and a small amount of short-chain-length monomer units. Under fed-batch fermentation with continuous feeding of nonanoic acid at a flow rate of 0.225 g/L/h and a C/N ratio of 40, a maximum PHA content of 48.6% dry cell weight and a conversion yield (Y_p/s) of 0.94 g/g were achieved. These results indicate that MCL-PHA production by activated sludge is a promising alternative to typical pure culture approaches.     

Effects of key operational parameters on biohydrogen production via anaerobic fermentation in a sequencing batch reactor

In this study, a series of tests were conducted in a 6 L anaerobic sequencing batch reactor (ASBR) to investigate the effect of pH, hydraulic retention time (HRT) and organic loading rate on biohydrogen production at 28 °C. Sucrose was used as the main substrate to mimic carbohydrate-rich wastewater and inoculum was prepared from anaerobic digested sludge without pretreatment. The reactor was operated initially with nitrogen sparging to form anaerobic condition. Results showed that methanogens were effectively suppressed. The optimum pH value would vary depending on the HRT. Maximum hydrogen production rate and yield of 3.04 L H₂/L reactor d and 2.16 mol H₂/mol hexose respectively were achieved at pH 4.5, HRT 30 h, and OLR 11.0 kg/m³ d. Two relationships involving the propionic acid/acetic acid ratio and ethanol/acetic acid ratio were derived from the analysis of the metabolites of fermentation. Ethanol/acetic acid ratio of 1.25 was found to be a threshold value for higher hydrogen production.     

Achieving nitrite accumulation in a continuous system treating low-strength domestic wastewater: switchover from batch start-up to continuous operation with process control

Although biological nitrogen removal via nitrite is recognized as one of the cost-effective and sustainable biological nitrogen removal processes, nitrite accumulation has proven difficult to achieve in continuous processes treating low-strength nitrogenous wastewater. Partial nitrification to nitrite was achieved and maintained in a lab-scale completely stirred tank reactor (CSTR) treating real domestic wastewater. During the start-up period, sludge with ammonia-oxidizing bacteria (AOB) but no nitrite-oxidizing bacteria (NOB) was obtained by batch operation with aeration time control. The nitrifying sludge with the dominance of AOB was then directly switched into continuous operation. It was demonstrated that partial nitrification to nitrite in the continuous system could be repeatedly and reliably achieved using this start-up strategy. The ratio of dissolved oxygen to ammonium loading rate (DO/ALR) was critical to maintain high ammonium removal efficiency and nitrite accumulation ratio. Over 85% of nitrite accumulation ratio and more than 95% of ammonium removal efficiency were achieved at DO/ALR ratios in an optimal range of 4.0–6.0 mg O₂/g N d, even under the disturbances of ammonium loading rate. Microbial population shift was investigated, and fluorescence in situ hybridization analysis indicated that AOB were the dominant nitrifying bacteria over NOB when stable partial nitrification was established.     

Innovative treatment system for digester liquor using anammox process

This study demonstrated that partial nitritation using nitrifying activated sludge entrapped in a polyethylene glycol (PEG) gel carrier, as a pretreatment to anammox process, could be successfully applied to digester liquor of biogas plant at a nitrogen loading rate of 3.0 kg-N/m³/d. The nitritation process produced an effluent with a NO₂–N/NH₄–N ratio between 1.0 and 1.4, which was found to be suitable for the subsequent anammox process. A high SS concentration (2000–3000 mg/l) in the digester liquor did not affect partial nitritation treatment performances. Effluent from this partial nitritation reactor was successfully treated in the anammox reactor using anammox sludge entrapped in the PEG gel carrier with T-N removal rates of greater than 4.0 kg-N/m³/d. Influent BOD and SS contents did not inhibit anammox activity of the anammox gel carrier. The combination of partial nitritation and anammox reactors using PEG entrapped nitrifying and anammox bacteria was shown to be effective for the removal of high concentration ammonium in the digester liquor of a biogas plant.     

Co-digestion of intermediate landfill leachate and sewage sludge as a method of leachate utilization

This study examines the co-digestion of intermediate landfill leachate and sewage sludge from a municipal wastewater treatment plant. Application of leachate as a co-fermentation component increased the concentrations of soluble organic compounds (expressed as total organic carbon), ammonium nitrogen, and alkalinity in the digester influents.The biogas yield obtained from the co-fermentation of a 20:1 sewage sludge: intermediate leachate mixture was 1.30 m³ per kg of removed volatile solids (VS), while that from a 10:1 mixture was 1.24 m³ per kg of removed VS. These values exceeded the biogas yield for the sludge alone by 13% and 8%, respectively. The leachate addition influenced the proportion of methane to a minor extent. Increased methane yields of 16.9% and 6.2% per kg of removed VS were found for the two sewage sluge:intermediate leachate mixtures, respectively.     

High-solid mesophilic methane fermentation of food waste with an emphasis on Iron, Cobalt, and Nickel requirements

The effect of trace metals on the mesophilic methane fermentation of high-solid food waste was investigated using both batch and continuous experiments. The continuous experiment was conducted by using a CSTR-type reactor with three run. During the first run, the HRT of the reactor was stepwise decreased from 100 days to 30 days. From operation day 50, the reactor efficiency deteriorated due to the lack of trace metals. The batch experiment showed that iron, cobalt, and nickel combinations had a significant effect on food waste. According to the results of the batch experiment, a combination of iron, cobalt, and nickel was added into the CSTR reactor by two different methods at run II, and III. Based on experimental results and theoretical calculations, the most suitable values of Fe/COD, Co/COD, and Ni/COD in the substrate were identified as 200, 6.0, and 5.7 mg/kg COD, respectively.     

Effects of initial lactic acid concentration, HRTs, and OLRs on bio-hydrogen production from lactate-type fermentation

A batch test and continuous operation were performed to identify the effect of lactate on hydrogen production at pH 4.5. When the initial lactic acid concentration was increased from 0 to 8 g/L in the batch test, the hydrogen yield also increased from 1.41 to 1.72 mol-H₂/mol-glucose. The system exhibited a long lag time and an insignificant hydrogen yield with 16 g-lactic acid/L. A continuous stirred tank reactor (CSTR) was operated at different organic loading rates (OLRs: 10, 15, 20 and 40 g/L/day) and hydraulic retention times (HRTs: 6, 12 and 24 h). At an OLR of 20 g-glucose/L/day and 12 h of HRT, the hydrogen yield was 1.2 mol-H₂/mol-glucose. The yield decreased with a 24 h HRT. Even though lactate was one of the major constituents of volatile fatty acids (VFAs), hydrogen production was feasible throughout the operation. Clostridium sp. was the dominant hydrogen-producing bacteria in the system.     

Ethanol production in multistage continuous, single stage continuous, Lactobacillus-contaminated continuous, and batch fermentations

Saccharomyces cerevisiae-based ethanol fermentations were conducted in batch culture, in a single stage continuous stirred tank reactor (CSTR), a multistage CSTR, and in a fermentor contaminated with Lactobacillus that corresponded to the first fermentor of the multistage CSTR system. Using a glucose concentration of 260 g l^–1 in the medium, the highest ethanol concentration reached was in batch (116gl^–1), followed by the multistage CSTR (106gl^–1), and the single stage CSTR continuous production system (60gl^–1). The highest ethanol productivity at this sugar concentration was achieved in the multistage CSTR system where a productivity of 12.7gl^–1h^–1 was seen. The other fermentation systems in comparison did not exceed an ethanol productivity of 3gl^–1h^–1. By performing a continuous ethanol fermentation in multiple stages (having a total equivalent working volume of the tested single stage), a 4-fold higher ethanol productivity was achieved as compared to either the single stage CSTR, or the batch fermentation.     

A two-stage thermophilic anaerobic process for the treatment of source sorted household solid waste

Summary Hydrolysis and acidification of source sorted household solid waste (SSHSW) at 70°C was studied using continuous stirred tank reactor (CSTR). The soluble COD/total initial COD-ratio of the SSHSW increased from 25 to 35% during the CSTR treatment. A thermophilic (55°C) upflow anaerobic sludge blanket (UASB) reactor removed up to 80% of the COD in the liquid fraction of the SSHSW treated at 70°C.     

Microbial community structure and dynamics in a mixotrophic nitrogen removal process using recycled spent caustic under different loading conditions

A laboratory-scale Bardenpho process was established to investigate the proper nitrogen loading rate (NLR) when modified spent caustic (MSC) is applied as electron donor and alkalinity source for denitrification. MSC injection induced autotrophic nitrogen removal with sulfur as electron donor and heterotrophic denitrification. The nitrogen removal rate (NRR) did not increase proportionally to NLR. Based on the total nitrogen concentration in the effluent observed in the trials with MSC, the NLR in the influent should not exceed 0.15 kg N/m³ d in order to satisfy water quality regulations. Microbial communities in the anoxic reactors were characterized by pyrosequencing of 16S rRNA gene sequences amplified by the polymerase chain reaction of DNA extracted from sludge samples. Microbial diversity was lower as MSC dosage was increased, and the injection of MSC caused an increase in SOB belonging to the genus Thiobacillus which is responsible for denitrification using sulfur.     

Enhanced nutrient removal in three types of step feeding process from municipal wastewater

An anoxic/oxic step feeding process was improved to enhance nutrient removal by reconfiguring the process into (1) anaerobic/anoxic/oxic step feeding process or (2) modified University of Capetown (UCT) step feeding process. Enhanced nitrogen and phosphorus removal and optimized organics utilization were obtained simultaneously in the modified UCT type with both internal and sludge recycle ratios of 75% as well as anaerobic/anoxic/oxic volume ratio of 1:3:6. Specifically, the UCT configuration and optimized operational conditions lead to the enrichment of denitrifying phosphorus removal microorganisms and achieved improved anaerobic P-release and anoxic P-uptake activities, which were beneficial to the denitrifying phosphorus removal activities and removal efficiencies. Due to high mixed liquor suspended solid and uneven distributed dissolved oxygen, 35% of total nitrogen was eliminated through simultaneous nitrification and denitrification process in aerobic zones. Moreover, 62 ± 6% of influent chemical oxygen demands was involved in the denitrification or phosphorus release processes.