Protective effect of a reverse membrane bioreactor against toluene and naphthalene in anaerobic digestion.

Raw syngas contains tar contaminants including toluene and naphthalene, which inhibit its conversion to methane. Cell encasement in a hydrophilic reverse membrane bioreactor (RMBR) could protect the cells from hydrophobic contaminants. This study aimed to investigate the inhibition of toluene and naphthalene and the effect of using RMBR. In this work, toluene and naphthalene were added at concentrations of 0.5-1.0 and 0.1-0.2 g/L in batch operation. In continuous operation, concentration of 0-6.44 g/L for toluene and 0-1.28 g/L for naphthalene were studied. The results showed that no inhibition was observed in batch operation for toluene and naphthalene at concentrations up to 1 and 0.2 g/L, respectively. In continuous operation of free cell bioreactors (FCBRs), inhibition of toluene and naphthalene started at 2.05 and 0.63 g/L, respectively. When they were present simultaneously, inhibition of toluene and naphthalene occurred at concentrations of 3.14 and 0.63 g/L, respectively. In continuous RMBRs, no inhibition for toluene and less inhibition for naphthalene were observed, resulting in higher methane production from RMBR than that of FCBR. These results indicated that RMBR system gave a better protection effect against inhibitors compared with FCBR.

Cultivation of edible filamentous fungus Aspergillus oryzae on volatile fatty acids derived from anaerobic digestion of food waste and cow manure.

In a circular economy approach, edible filamentous fungi (single cell protein) can be cultivated on volatile fatty acids (VFAs) derived from anaerobic digestion (AD) of organic-rich waste streams. In this study, the effect of pH, concentration/distribution of VFAs, nutrient supplementation, and type of waste on Aspergillus oryzae cultivation on synthetic VFAs, and actual VFAs derived from AD of food waste and cow manure were investigated. The optimal pH for A. oryzae growth on VFAs were 6 and 7 with maximum acetic acid consumption rates of 0.09 g/L.h. The fungus could thrive on high concentrations of acetic (up to 9 g/L) yielding 0.29 g dry biomass/gVFAsfed. In mixed VFAs cultures, A. oryzae primarily consumed caproic and acetic acids reaching a biomass yield of 0.26 g dry biomass/gVFAsfed (containing up to 41% protein). For waste-derived VFAs at pH 6, the fungus successfully consumed 81-100% of caproic, acetic, and butyric acids.

Utilization of food waste-derived volatile fatty acids for production of edible Rhizopus oligosporus fungal biomass.

Rhizopus oligosporus is an edible filamentous fungus that can contribute to meet the growing demand for single-cell protein. Volatile fatty acids (VFAs) are favorable potential substrates for producing R. oligosporus biomass due to their capacity to be synthesized from a wide range of low-value organic solid wastes via anaerobic digestion. The goal of this work was to cultivate R. oligosporus using food waste-derived VFAs as the sole carbon source. To maintain the requisite low substrate concentrations, the fed-batch cultivation technique was applied. This resulted in a four-fold improvement in biomass production relative to standard batch cultivation. Maximum biomass yield of 0.21 ± 0.01 g dry biomass/g VFAs COD eq. consumed, containing 39.28 ± 1.54% crude protein, was obtained. In the bubble-column bioreactors, the complete uptake of acetic acid was observed, while the consumptions of caproic and butyric acids reached up to 97.64% and 26.13%, respectively.

Effect of Effluent Recirculation on Biogas Production Using Two-stage Anaerobic Digestion of Citrus Waste.

Citrus waste is a promising potential feedstock for anaerobic digestion, yet the presence of inhibitors such as d-limonene is known to limit the process. Effluent recirculation has been proven to increase methane yield in a semi-continuous process for recalcitrant material, but it has never been applied to toxic materials. This study was aimed to investigate the effect of recirculation on biogas production from citrus waste as toxic feedstock in two-stage anaerobic digestion. The first digestion was carried out in a stirred tank reactor (STR). The effluent from the first-stage was filtered using a rotary drum filter to separate the solid and the liquid phase. The solid phase, rich in hydrophobic D-limonene, was discarded, and the liquid phase containing less D-limonene was fed into the second digester in an up-flow anaerobic sludge bed (UASB) reactor. A high organic loading rate (OLR 5 g VS/(L·day)) of citrus waste was fed into the first-stage reactor every day. The effluent of the first-stage was then fed into the second-stage reactor. This experiment was run for 120 days. A reactor configuration without recirculation was used as control. The result shows that the reactor with effluent recirculation produced a higher methane yield (160⁻203 NmL/g·VS) compared to that without recirculation (66⁻113 NmL/g·VS). More stable performance was also observed in the reactor with recirculation as shown by the pH of 5⁻6, while without recirculation the pH dropped to the range of 3.7⁻4.7. The VS reduction for the reactor with recirculation was 33⁻35% higher than that of the control without recirculation. Recirculation might affect the hydrolysis-acidogenesis process by regulating pH in the first-stage and removing most of the D-limonene content from the substrate through filtration.

Semi-Continuous Reverse Membrane Bioreactor in Two-Stage Anaerobic Digestion of Citrus Waste.

The presence of an antimicrobial compound called D-Limonene in citrus waste inhibits methane production from such waste in anaerobic digestion. In this work, a two-stage anaerobic digestion method is developed using reverse membrane bioreactors (rMBRs) containing cells encased in hydrophilic membranes. The purpose of encasement is to retain a high cell concentration inside the bioreactor. The effectiveness of rMBRs in reducing cell washout is evaluated. Three different system configurations, comprising rMBRs, freely suspended cells (FCs), and a combination of both (abbreviated to rMBR⁻FCs), are incubated at three different organic loading rates (OLRs) each, namely 0.6, 1.2, and 3.6 g COD/(L cycle). Incubation lasts for eight feeding cycles at 55 °C. Methane yield and biogas composition results show that rMBRs perform better than rMBR⁻FCs and FCs at all three OLRs. Volatile fatty acid profiles and H2 production show that the reactors are working properly and no upset occurs. Additionally, a short digestion time of 4 days can be achieved using the rMBR configuration in this study.

Erratum to "Inhibitory Effect of Long-Chain Fatty Acids on Biogas Production and the Protective Effect of Membrane Bioreactor".

[This corrects the article DOI: 10.1155/2016/7263974.].

Inhibitory Effect of Long-Chain Fatty Acids on Biogas Production and the Protective Effect of Membrane Bioreactor.

Anaerobic digestion of lipid-containing wastes for biogas production is often hampered by the inhibitory effect of long-chain fatty acids (LCFAs). In this study, the inhibitory effects of LCFAs (palmitic, stearic, and oleic acid) on biogas production as well as the protective effect of a membrane bioreactor (MBR) against LCFAs were examined in thermophilic batch digesters. The results showed that palmitic and oleic acid with concentrations of 3.0 and 4.5 g/L resulted in >50% inhibition on the biogas production, while stearic acid had an even stronger inhibitory effect. The encased cells in the MBR system were able to perform better in the presence of LCFAs. This system exhibited a significantly lower percentage of inhibition than the free cell system, not reaching over 50% at any LCFA concentration tested.

Improvement of biogas production from orange peel waste by leaching of limonene.

Limonene is present in orange peel wastes and is known as an antimicrobial agent, which impedes biogas production when digesting the peels. In this work, pretreatment of the peels to remove limonene under mild condition was proposed by leaching of limonene using hexane as solvent. The pretreatments were carried out with homogenized or chopped orange peel at 20-40°C with orange peel waste and hexane ratio (w/v) ranging from 1 : 2 to 1 : 12 for 10 to 300 min. The pretreated peels were then digested in batch reactors for 33 days. The highest biogas production was achieved by treating chopped orange peel waste and hexane ratio of 12 : 1 at 20°C for 10 min corresponding to more than threefold increase of biogas production from 0.061 to 0.217 m(3) methane/kg VS. The solvent recovery was 90% using vacuum filtration and needs further separation using evaporation. The hexane residue in the peel had a negative impact on biogas production as shown by 28.6% reduction of methane and lower methane production of pretreated orange peel waste in semicontinuous digestion system compared to that of untreated peel.

Effects of lactone, ketone, and phenolic compounds on methane production and metabolic intermediates during anaerobic digestion.

Fruit waste is a potential feedstock for biogas production. However, the presence of fruit flavors that have antimicrobial activity is a challenge for biogas production. Lactones, ketones, and phenolic compounds are among the several groups of fruit flavors that are present in many fruits. This work aimed to investigate the effects of two lactones, i.e., γ-hexalactone and γ-decalactone; two ketones, i.e., furaneol and mesifurane; and two phenolic compounds, i.e., quercetin and epicatechin on anaerobic digestion with a focus on methane production, biogas composition, and metabolic intermediates. Anaerobic digestion was performed in a batch glass digester incubated at 55 °C for 30 days. The flavor compounds were added at concentrations of 0.05, 0.5, and 5 g/L. The results show that the addition of γ-decalactone, quercetin, and epicathechin in the range of 0.5-5 g/L reduced the methane production by 50 % (MIC50). Methane content was reduced by 90 % with the addition of 5 g/L of γ-decalactone, quercetin, and epicathechin. Accumulation of acetic acid, together with an increase in carbon dioxide production, was observed. On the contrary, γ-hexalactone, furaneol, and mesifurane increased the methane production by 83-132 % at a concentration of 5 g/L.

Biogas production from lignocelluloses by N-methylmorpholine-N-oxide (NMMO) pretreatment: effects of recovery and reuse of NMMO.

The effects of N-methylmorpholine-N-oxide (NMMO) pretreatment on barley straw and forest residues were investigated for biogas production. The pretreatments were performed at 90°C with 85% NMMO for 3-30h. The best pretreatment conditions resulted in 100% improvement in methane yield during the subsequent digestion compared to that of the untreated lignocelluloses. Methane yields of 0.23 and 0.15Nm(3) CH4/kg VS were obtained from barley straw and forest residues, respectively, corresponding to 88% and 83% of the theoretical yields. In addition, the effects of the pretreatment with recovered and reused NMMO was also studied over the course of five cycles. Pretreatment with recycled NMMO showed the same performance as the fresh NMMO on barley straw. However, pretreatment of forest residues with recycled NMMO resulted in 55% reduction in methane yield.

Pretreatment of oil palm empty fruit bunch (OPEFB) by N-methylmorpholine-N-oxide (NMMO) for biogas production: structural changes and digestion improvement.

Pretreatment of OPEFB (oil palm empty fruit bunch) by NMMO (N-methylmorpholine-N-oxide) on its subsequent digestions was investigated. The pretreatments were carried out at 90 and 120 °C for 1, 3, and 5h in three different modes of dissolution (by 85% NMMO solution), ballooning (79% NMMO solution), and swelling (73% NMMO solution). The total solid recovery after the pretreatment was 89-94%. The pretreatment process did not have a major impact on the composition of OPEFB, other than a reduction of ash from 5.4% up to 1.3%. The best improvement in biogas production was achieved by a dissolution mode pretreatment of OPEFB, using conditions of 85% NMMO, 3h, and 120 °C. It resulted in 0.408 Nm(3)/kg VS methane yield and 0.032 Nm(3)CH(4)/kg VS/day initial methane production rate, which correspond in improving by 48% and 167% compared to the untreated OPEFB, respectively.

Structural changes of oil palm empty fruit bunch (OPEFB) after fungal and phosphoric acid pretreatment.

Oil palm empty fruit bunch (OPEFB) was pretreated using white-rot fungus Pleurotus floridanus, phosphoric acid or their combination, and the results were evaluated based on the biomass components, and its structural and morphological changes. The carbohydrate losses after fungal, phosphoric acid, and fungal followed by phosphoric acid pretreatments were 7.89%, 35.65%, and 33.77%, respectively. The pretreatments changed the hydrogen bonds of cellulose and linkages between lignin and carbohydrate, which is associated with crystallinity of cellulose of OPEFB. Lateral Order Index (LOI) of OPEFB with no pretreatment, with fungal, phosphoric acid, and fungal followed by phosphoric acid pretreatments were 2.77, 1.42, 0.67, and 0.60, respectively. Phosphoric acid pretreatment showed morphological changes of OPEFB, indicated by the damage of fibre structure into smaller particle size. The fungal-, phosphoric acid-, and fungal followed by phosphoric acid pretreatments have improved the digestibility of OPEFB's cellulose by 4, 6.3, and 7.4 folds, respectively.

A pilot study on lignocelluloses to ethanol and fish feed using NMMO pretreatment and cultivation with zygomycetes in an air-lift reactor.

A complete process for the production of bioethanol and fungal biomass from spruce and birch was investigated. The process included milling, pretreatment with N-methylmorpholine-N-oxide (NMMO), washing of the pretreated wood, enzymatic hydrolysis, and cultivation of the zygomycetes fungi Mucor indicus. Investigated factors included wood chip size (0.5-16 mm), pretreatment time (1-5h), and scale of the process from bench-scale to 2m high air-lift reactor. Best hydrolysis yields were achieved from wood chips below 2mm after 5h of pretreatment. Ethanol yields (mg/g wood) of 195 and 128 for spruce, and 175 and 136 for birch were achieved from bench-scale and airlift, respectively. Fungal biomass yields (mg/g wood) of 103 and 70 for spruce, and 86 and 66 for birch from bench scale and airlift respectively were simultaneously achieved. NMMO pretreatment and cultivation with M. indicus appear to be a good alternative for ethanol production from birch and spruce.

Temperature shifts for extraction and purification of zygomycetes chitosan with dilute sulfuric acid.

The temperature-dependent hydrolysis and solubility of chitosan in sulfuric acid solutions offer the possibility for chitosan extraction from zygomycetes mycelia and separation from other cellular ingredients with high purity and high recovery. In this study, Rhizomucor pusillus biomass was initially extracted with 0.5 M NaOH at 120 °C for 20 min, leaving an alkali insoluble material (AIM) rich in chitosan. Then, the AIM was subjected to two steps treatment with 72 mM sulfuric acid at (i) room temperature for 10 min followed by (ii) 120 °C for 45 min. During the first step, phosphate of the AIM was released into the acid solution and separated from the chitosan-rich residue by centrifugation. In the second step, the residual AIM was re-suspended in fresh 72 mM sulfuric acid, heated at 120 °C and hot filtered, whereby chitosan was extracted and separated from the hot alkali and acid insoluble material (HAAIM). The chitosan was recovered from the acid solution by precipitation at lowered temperature and raised pH to 8-10. The treatment resulted in 0.34 g chitosan and 0.16 g HAAIM from each gram AIM. At the start, the AIM contained at least 17% phosphate, whereas after the purification, the corresponding phosphate content of the obtained chitosan was just 1%. The purity of this chitosan was higher than 83%. The AIM subjected directly to the treatment with hot sulfuric acid (at 120 °C for 45 min) resulted in a chitosan with a phosphate impurity of 18.5%.

A novel process for ethanol or biogas production from cellulose in blended-fibers waste textiles.

A novel process has been developed for separation of the cellulose, i.e. cotton and viscose, from blended-fibers waste textiles. An environmentally friendly cellulose solvent, N-methylmorpholine-N-oxide (NMMO) was used in this process for separation and pretreatment of the cellulose. This solvent was mixed with blended-fibers textiles at 120 °C and atmospheric pressure to dissolve the cellulose and separate it from the undissolved non-cellulosic fibers. Water was then added to the solution in order to precipitate the cellulose, while both water and NMMO were reused after separation by evaporation. The cellulose was then either hydrolyzed by cellulase enzymes followed by fermentation to ethanol, or digested directly to produce biogas. The process was verified by testing 50/50 polyester/cotton and 40/60 polyester/viscose-blended textiles. The polyesters were purified as fibers after the NMMO treatments, and up to 95% of the cellulose fibers were regenerated and collected on a filter. A 2-day enzymatic hydrolysis and 1-day fermentation of the regenerated cotton and viscose resulted in 48 and 50 g ethanol/g regenerated cellulose, which were 85% and 89% of the theoretical yields, respectively. This process also resulted in a significant increase of the biogas production rate. While untreated cotton and viscose fibers were converted to methane by respectively, 0.02% and 1.91% of their theoretical yields in 3 days of digestion, the identical NMMO-treated fibers resulted into about 30% of yield at the same period of time.

Process design and economic analysis of a citrus waste biorefinery with biofuels and limonene as products.

Process design and economic analysis of a biorefinery for the treatment of citrus wastes (CW) at different capacities was carried out. The CW is hydrolyzed using dilute sulfuric acid and then further processed to produce limonene, ethanol and biogas. The total cost of ethanol for base case process with 100,000 tons/year CW capacity was calculated as 0.91 USD/L, assuming 10 USD/ton handling and transportation cost of CW to the plant. However, this price is sensitive to the plant capacity. With constant price of methane and limonene, changing the plant capacity from 25,000 to 400,000 tons CW per year results in reducing ethanol costs from 2.55 to 0.46 USD/L in an economically feasible process. In addition, the ethanol production cost is sensitive to the transportation cost of CW. Increasing this cost from 10 to 30 USD/ton for the base case results in increasing the ethanol costs from 0.91 to 1.42 USD/L.

Production of biofuels, limonene and pectin from citrus wastes.

Production of ethanol, biogas, pectin and limonene from citrus wastes (CWs) by an integrated process was investigated. CWs were hydrolyzed by dilute-acid process in a pilot plant reactor equipped with an explosive drainage. Hydrolysis variables including temperature and residence time were optimized by applying a central composite rotatable experimental design (CCRD). The best sugar yield (0.41g/g of the total dry CWs) was obtained by dilute-acid hydrolysis at 150 degrees C and 6min residence time. At this condition, high solubilization of pectin present in the CWs was obtained, and 77.6% of total pectin content of CWs could be recovered by solvent recovery. Degree of esterification and ash content of produced pectin were 63.7% and 4.23%, respectively. In addition, the limonene of the CWs was effectively removed through flashing of the hydrolyzates into an expansion tank. The sugars present in the hydrolyzates were converted to ethanol using baker's yeast, while an ethanol yield of 0.43g/g of the fermentable sugars was obtained. Then, the stillage and the remaining solid materials of the hydrolyzed CWs were anaerobically digested to obtain biogas. In summary, one ton of CWs with 20% dry weight resulted in 39.64l ethanol, 45m(3) methane, 8.9l limonene, and 38.8kg pectin.

Determination of glucosamine and N-acetyl glucosamine in fungal cell walls.

A new method was developed to determine glucosamine (GlcN) and N-acetyl glucosamine (GlcNAc) in materials containing chitin and chitosan, such as fungal cell walls. It is based on two steps of hydrolysis with (i) concentrated sulfuric acid at low temperature and (ii) dilute sulfuric acid at high temperature, followed by one-step degradation with nitrous acid. In this process, chitin and chitosan are converted into anhydromannose and acetic acid. Anhydromannose represents the sum of GlcN and GlcNAc, whereas acetic acid is a marker for GlcNAc only. The method showed recovery of 90.1% of chitin and 85.7-92.4% of chitosan from commercial preparations. Furthermore, alkali insoluble material (AIM) from biomass of three strains of zygomycetes, Rhizopus oryzae, Mucor indicus, and Rhizomucor pusillus, was analyzed by this method. The glucosamine contents of AIM from R. oryzae and M. indicus were almost constant (41.7 +/- 2.2% and 42.0 +/- 1.7%, respectively), while in R. pusillus, it decreased from 40.0 to 30.0% during cultivation from 1 to 6 days. The GlcNAc content of AIM from R. oryzae and R. pusillus increased from 24.9 to 31.0% and from 36.3 to 50.8%, respectively, in 6 days, while it remained almost constant during the cultivation of M. indicus (23.5 +/- 0.8%).

Ethanol production from glucose and dilute-acid hydrolyzates by encapsulated S. cerevisiae.

The performance of encapsulated Saccharomyces cerevisiae CBS 8066 in anaerobic cultivation of glucose, in the presence and absence of furfural as well as in dilute-acid hydrolyzates, was investigated. The cultivation of encapsulated cells in 10 sequential batches in synthetic media resulted in linear increase of biomass up to 106 g/L of capsule volume, while the ethanol productivity remained constant at 5.15 (+/-0.17) g/L x h (for batches 6-10). The cells had average ethanol and glycerol yields of 0.464 and 0.056 g/g in these 10 batches. Addition of 5 g/L furfural decreased the ethanol productivity to a value of 1.31 (+/-0.10) g/L x h with the encapsulated cells, but it was stable in this range for five consecutive batches. On the other hand, the furfural decreased the ethanol yield to 0.41-0.42 g/g and increased the yield of acetic acid drastically up to 0.068 g/g. No significant lag phase was observed in any of these experiments. The encapsulated cells were also used to cultivate two different types of dilute-acid hydrolyzates. While the free cells were not able to ferment the hydrolyzates within at least 24 h, the encapsulated yeast successfully converted glucose and mannose in both of the hydrolyzates in less than 10 h with no significant lag phase. However, since the hydrolyzates were too toxic, the encapsulated cells lost their activity gradually in sequential batches.

Kinetic study of detoxification of dilute-acid hydrolyzates by Ca(OH)2.

Detoxification of dilute-acid hydrolyzates by addition of Ca(OH)(2) (overliming) and cultivation of the detoxified hydrolyzates by Saccharomyces cerevisiae were examined. The examined overliming involves increasing the pH of the hydrolyzates to 9, 10, 11 or 12, keeping up to 90 min at different temperatures of 30, 45 and 60 degrees C, followed by readjustment of the pH to 5. Increasing the pH, time and/or temperature resulted in more effective degradation of furans and resulted in better fermentability for both of the tested hydrolyzates, but higher loss of the sugars was observed as well. Overliming of glucose and furfural solution at pH 12 showed a rapid decrease in concentration of these chemicals followed by a slow degradation process. Therefore, a kinetic model was proposed for the detoxification, where the sugars or furans make transient complexes with calcium ions and this complex will then be converted to the degradation product. The ANOVA analysis of the model resulted in an average R(2) of 0.99 for the model fitted to all the experimental data points.