Bioproduction of hydrogen from food waste by pilot-scale combined hydrogen/methane fermentation

A pilot-scale two-phase hydrogen/methane fermentation system generated 3.9 L biogas per unit time and reactor volume from food waste, of which the fraction of H₂ was approximately 60% at a hydraulic retention time (HRT) of 21 h. As substrate, 90% of the carbohydrates in the organic compounds were consumed, based on COD removal efficiency, and the hydrogen yield was approximately 1.82 (H₂-mol/glucose-mol). The maximum decomposition rate coefficient of hydrogen fermentation was observed at an HRT of 21 h, indicating that reducing HRTs improves hydrogen production. Over 80% of the methane was produced in the methane fermentation tank and the predominant fraction of organic acids after methane fermentation comprised acetic acid. Based on our economic evaluation, two-phase hydrogen/methane fermentation has greater potential for recovering energy than methane-only fermentation.     

Production of hydrogen and methane by one and two stage fermentation of food waste

Anaerobic digestion is an attractive process for generation of hydrogen and methane, which involves complex microbial processes on decomposition of organic wastes and subsequent conversion of metabolic intermediates to hydrogen and methane. Comparative performance of a sequential hydrogen and methane fermentation in two stage process and methane fermentation in one stage process were tested in batch reactor at varying ratios of feedstock to microbial inoculum (F/M) under mesophilic incubation. F/M ratios influence biogas yield, production rate, and potential. The highest H₂ and CH₄ yields of 55 and 94 mL g⁻¹ VS were achieved at F/M of 7.5 in two stage process, while the highest CH₄ yield of 82 mL g⁻¹ VS in one stage process was observed at the same F/M. Acetic and butyric acids are the main volatile fatty acids (VFAs) produced in the hydrogen fermentation stage with the concentration range 10–25 mmol L⁻¹. Little concentrations of VFAs were accumulated in methane fermentation in both stage processes. Total energy recovery in two stage process is higher than that in one stage by 18%. This work demonstrated two stage fermentation achieved a better performance than one stage process.     

Bioaugmentation on hydrogen production from food waste

The aim of this work was to evaluate the effect of two hydrolytic (Paenibacillus polymyxa and Bacillus subtilis) and two fermentative (Clostridium saccharobutylicum and Clostridium beijerinckii) strains on hydrogen (H₂) production in dark fermentation by batch testing. Food waste was used as a substrate, pretreated anaerobic sludge was used as the inoculum, and different concentrations of the evaluated microorganisms were used. Bioaugmentation with 3.5 × 10⁹ CFU/mL/L_reactor B. subtilis showed the best performance, obtaining a production of 84.5 mL H₂/g SV and a reduction in the lag phase (from 7.9 h in control to 3.5 h). Bioaugmentation with B. subtilis in an anaerobic sequencing batch reactor exhibited a significant effect on volumetric productivity, reaching a maximal increase of 344% of H₂ production in comparison with that obtained without the addition of the strain. The increase in H₂ was observed in a short period of time (4 cycles), after which H₂ production returned to the original H₂ production baseline. During all reactor operations, the main volatile fatty acids produced were acetic acid and butyric acid. Microbial community analysis when bioaugmentation was applied showed an importance of lactic acid bacteria abundance, such as that of Bifidobacterium and Lactobacillus, whose metabolic activity was crucial in reactor performance. The added concentration of microorganisms is a critical parameter for the bioaugmentation process.     

Viability of ultrasonication of food waste for hydrogen production

Batch anaerobic studies were conducted to study the effect of ultrasonication as a pre-treatment method for pulp waste prior to anaerobic hydrogen production. Pre-treatment was conducted by sonicating a 100 mL of pulp waste at different sonication times varying from 0.5 min to 30 min. The ultimate hydrogen production increased with increasing sonication time. The highest ultimate hydrogen production was achieved at a sonication time of 30 min and reflected an 88% increase over the unsonicated food waste, of 80 mL/g VS_added. The highest final VFAs concentration after fermentation (corresponding to 70% increase over the unsonicated food waste) was also achieved at a sonication time of 30 min. There were no significant differences between the acetate-to-butyrate ratios (HAc/HBu) for the all sonication times. The maximum hydrogen production rate at sonication time of 30 min was about 145% higher than that the unsonicated food waste.     

Effect of inoculum pre-treatment on mesophilic hydrogen and methane production from food waste using two-stage anaerobic digestion

Two-stage anaerobic digestion of food waste was performed using four different inoculum pre-treatment methods to enrich hydrogen (H₂) producing bacteria from sludge. The pretreatments used in this study included heat shock, alkaline treatment, aeration, and a novel pretreatment using waste frying oil (WFO). Alkaline pretreatment and aeration did not completely inhibit methanogens in the first stage while no methane (CH₄) was detected in the reactors cultivated either with heat shock or WFO-pretreated inocula. The highest H₂ and CH₄ yields (76.1 and 598.2 mL/gVS, respectively) were obtained using the inoculum pretreated with WFO. The highest total energy yield (21.96 kJ/gVS) and total organic carbon (TOC) removal efficiencies (95.77%) were obtained using inoculum pretreatment with WFO. The total energy yield trend obtained using the different pretreatments was as follows: WFO > alkaline > heat > aeration > control.     

Effect of biochar in enhancing hydrogen production by mesophilic anaerobic digestion of food wastes: The role of minerals

The role of minerals in biochar in promoting hydrogen (H₂) production by anaerobic digestion of food waste was investigated. The cultures with the addition of biochar, leached biochar, metal sulphate solution and leached biochar combined with metal sulphate solution, respectively, were placed in bench-scale reactors and incubated at incubator at 32 °C. Daily H₂ production and volatile fatty acids (VFAs) were measured and the cumulative H₂ yield (Y_H) and maximum H₂ production rate (R_H) were calculated. The microbial analysis was performed using Illumina MiSeq sequencing. Biochar addition significantly increased the maximum Y_H by 107% and R_H by 54%. However, the addition of leached biochar only increased the maximum Y_H by 39% and R_H by 45% than control. The primary elements in biochars that contribute to H₂ production (Fe, K and Ca) were shown to increase the acetic acid, butyric acid and prevalence of the H₂ producing bacteria Clostridium butyricum.     

Thermophilic hydrogen production from starch wastewater using two-phase sequencing batch fermentation coupled with UASB methanogenic effluent recycling

The aim of this study was to evaluate the performance of thermophilic hydrogenesis coupled with mesophilic methanogenesis in which the effluent was recycled to the hydrogen reactor for starch wastewater treatment. With this system, the hydrogen production rate and yield were 3.45 ± 0.25 L H₂/(L·d) and 5.79 ± 0.41 mmol H₂/g COD_added respectively, and thus higher than the values of the control group without methanogenic effluent recycling. In addition, relatively higher contents of acetate and butyrate were obtained in the hydrogen reactor with recirculation. The methane reactors were operated with the effluent from the hydrogen reactor, and methane yield was stabilized at 0.21–0.23 L/g COD_removal in both. Analysis of the microbial communities further showed that methanogenic effluent recirculation enriched microbial communities in the hydrogen reactor. Two species of bacteria effective in hydrogenesis, Thermoanaerobacterium thermosaccharolyticum and Clostridium thermosaccharolyticum, dominated during hydrogen production, whereas archaea belonging to Euryarchaeota were detected and cultured in the methane reactor. The recycled effluent supplied alkaline substrates for the hydrogen producing bacteria. Alkali balance calculations showed that the amount of added alkali was reduced by 88%. This amount, required for hydrogen production from starch wastewater, was contributed by alkali in the methanogenic effluent, (2225 ± 140 mg CaCO₃/L), resulting in lower operational costs.     

Automatic process control for stable bio-hythane production in two-phase thermophilic anaerobic digestion of food waste

The paper reports the results of a long term (310 days) pilot-scale trial where food waste as sole substrate was treated in a two-phase thermophilic anaerobic digestion process. This was optimized for concurrent hydrogen and methane production. First phase's optimization for hydrogen production was obtained recirculating the effluent coming from the methanogenic phase and without the addition of external chemicals. A drawback of such approach is the recirculation of ammonia into the first phase reactor for hydrogen production with possibility of consequent inhibition.     

Natural inducement of hydrogen from food waste by temperature control

An easy and simple method of producing H₂ from food waste was devised. Although there was no inoculum addition or pretreatment, food waste was naturally decomposed and converted to H₂ when cultivated at 50-60 °C in anaerobic state. Both the highest H₂ yield of 1.79 mol H₂/mol hexose_added and a production rate of 369.1 ml H₂/L/h were observed at 50 °C. While butyrate was the main by-product of the food waste cultivated at 50 °C, lactate whose producing-reaction is non-hydrogenic was dominant at 35 °C where the worst performance was observed. The degradation efficiency of volatile solids and carbohydrate was similar to 50% and 90%, respectively, at both temperatures. Polymerase chain reaction-denaturing gradient gel electrophoresis analysis clearly revealed that the role of temperature control was the microbial selection. At high temperature, the activity of indigenous lactic acid bacteria was suppressed while H₂-producing bacteria, such as Clostridium sp., Acetanaerobacterium elongatum, and Caloramater indicus, were predominantly cultivated.     

Lime mud from paper-making process addition to food waste synergistically enhances hydrogen fermentation performance

The effect of lime mud from paper-making process (LMP) addition on the H₂ fermentation of food waste (FW) was investigated. It was found that a slight addition of LMP (1.0–4.0 g in 200 g FW) significantly enhanced the H₂ fermentation performance, not only increasing the total amount of H₂ produced but also accelerating the whole reaction, shortening the lag period, and increasing the H₂ production rate. Fermentation stability and microbial germination were also facilitated by LMP addition. This was attributed to the existence of Ca, Fe, Mn and alkaline substances such as CaCO₃ and NaOH. The batch process treating a mixture of FW and LMP was showed that the highest hydrogen production of 137.6 mL H₂/g VS was achieved at final pH 5.0, adding 3 g LMP (in 200 g FW) to the fermentation process, which lag-phase time was about 2.5 h.     

Kinetic study of biological hydrogen production by anaerobic fermentation

The growth kinetics of hydrogen producing bacteria using three different substrates, namely sucrose, non-fat dry milk (NFDM), and food waste were investigated in dark fermentation through a series of batch experiments. The results showed that hydrogen production potential and hydrogen production rate increased with an increasing substrate concentration. The maximum hydrogen yields from sucrose, NFDM, and food waste were 234, 119, and 101 mL/g COD, respectively. The low pH (pH<4)$(\mathrm{pH}<4)$ inhibited hydrogen production and resulted in lower carbohydrate fermentation at high substrate concentration. Michaelis–Menten equation was employed to model the hydrogen production rate at different substrate concentrations. The equation gave a good approximation of the maximum hydrogen production rate and the half saturation constant (_Ks)$(K_{\mathrm{s}})$ with correlation coefficient (R²)$(R^2)$ over 0.85. The _Ks$K_{\mathrm{s}}$ values of sucrose, NFDM, and food waste were 1.4, 6.6, and 8.7 g COD/L, respectively. Based on _Ks$K_{\mathrm{s}}$ values, the substrate affinity of the enriched hydrogen producing culture was found to depend on carbohydrate content of the substrate. The substrate containing high carbohydrate showed a lower _Ks$K_{\mathrm{s}}$ value. The maximum hydrogen production rate was governed by the complexity of carbohydrates in the substrate.     

Effect of acid-pretreatment on hydrogen fermentation of food waste: Microbial community analysis by next generation sequencing

This work presents the effect of acid-pretreatment on H₂ fermentation of food waste with detailed microbial information by next generation sequencing. The pretreated food waste at pH 1.0–4.0 was cultivated under mesophilic conditions without external inoculum addition. From the food waste acid-pretreated at pH 1–3, H₂ yields in the range of 1.37–1.74 mol H₂/mol hexose_added were achieved, attaining the highest value at pH 2. Clostridium sp. such as Clostridium acetobutylicum ATCC 824 and Clostridium perfringens occupied more than 70% of total number of sequences at pH 1–3. On the other hand, in the control (no pretreatment) and at pH 4, lactic acid bacteria such as Lactobacillus and Streptococcus were found to be the dominant genus (>90% of total number of sequences), resulting in a low H₂ yield. In addition, the effect of substrate concentration on H₂ fermentation was investigated, and the maximum H₂ productivity was estimated to be 27.2 L H₂/L/d by Andrew's model.     

Effect of organic loading rate on continuous hydrogen production from food waste in submerged anaerobic membrane bioreactor

The characteristics of hydrogen fermentation in a membrane bioreactor (HF-MBR) fed with food waste were investigated at thermophilic condition. The HF-MBR was operated at three different organic loading rates (OLRs) of 70.2, 89.4 and 125.4 kg-COD/m³/day. Biogas production rate increased from 22.4 to 32.8 and 62.5 l/day with OLR. The maximum Hydrogen yield and production rate were 111.1 mL-H₂/g-VS _added and 10.7 l-H₂/L/day at an OLR of 125.4 kg-COD/m³/day. The total carbohydrate degradation was better than 96% throughout the experimental runs. Continuous H₂ production from food waste with CH₄-free biogas was successfully sustained in the HF-MBR for 90 days. The microbial community was dominated by Clostridium sp. strain Z6. The H₂ production was significantly improved by shortening the retention time and increasing the OLRs. The HF-MBR showed an H₂ production capacity at the high OLRs due to its higher cell retention.     

Evaluation of hydrogen producing cultures using pretreated food waste

In order to enhance bio-hydrogen production from food waste, pretreatment methods are widely used. The influence of the initial pH and autoclaving were investigated in batch experiments. Fermentative studies showed that pure cultures like Clostridium beijerinckii could directly utilize raw food waste to produce hydrogen, while other cultures (Clostridium butyricum and Clostridium pasteurianum) could produce hydrogen only after pH adjustment. In this case, the optimal starting pH of the culture was found to be 7. Autoclaving could further enhance hydrogen yields due to increased hydrolysis of food waste. The maximum hydrogen yield was achieved by C. butyricum (38.9 mL-H₂/g-VS_added) after autoclaving food waste with pH adjustment at 7. In addition, the ratio acetic to butyric acid was decreased by autoclaving pretreatment, because butyrate metabolic pathway was favored in the fermentation process. However, suitable pH for bacteria growth and the low ammonia production could be achieved from autoclaving food waste.     

Optimization of key factors affecting hydrogen production from food waste by anaerobic mixed cultures

Key factors (inoculums concentration, substrate concentration and citrate buffer concentration) affecting hydrogen yield (HY) and specific hydrogen production rate (SHPR) from food waste in batch fermentation by anaerobic mixed cultures were optimized using Response Surface Methodology with Central Composite Design. The experiments were conducted in 120 ml serum bottles with a working volume of 70 mL. Under the optimal condition of 2.30 g-VSS/L of inoculums concentration, 2.54 g-VS/L of substrate concentration, and 0.11 M of citrate buffer concentration, the predicted maximum HY and SHPR of 104.79 mL H₂/g-VS_added and 16.90 mL H₂/g-VSS.h, respectively, were obtained. Concentrations of inoculums, substrate and citrate buffer all had an individual effect on HY and SHPR (P < 0.05). The substrate concentration and citrate buffer concentration had the greatest interactive effect on SHPR (P = 0.0075) while their effects on HY (P = 0.0131) were profound. These results were reproduced in confirmation experiments under optimal conditions and generated an HY of 104.58 mL H₂/g-VS_added and an SHPR of 16.86 mL H₂/g-VSS.h. This was only 0.20% and 0.24%, respectively, different from the predicted values. Microbial community analysis by PCR-DGGE indicated that Clostridium was the pre-dominant hydrogen producer at the optimum and worst conditions. The presence of Lactobacillus sp. and Enterococcus sp. might be responsible for the low HY and SHPR at the worst condition.     

Effect of food to microorganism ratio on biohydrogen production from food waste via anaerobic fermentation

The effect of different food to microorganism ratios (F/M) (1–10) on the hydrogen production from the anaerobic batch fermentation of mixed food waste was studied at two temperatures, 35 ± 2 °C and 50 ± 2 °C. Anaerobic sludge taken from anaerobic reactors was used as inoculum. It was found that hydrogen was produced mainly during the first 44 h of fermentation. The F/M between 7 and 10 was found to be appropriate for hydrogen production via thermophilic fermentation with the highest yield of 57 ml-H₂/g VS at an F/M of 7. Under mesophilic conditions, hydrogen was produced at a lower level and in a narrower range of F/Ms, with the highest yield of 39 ml-H₂/g VS at the F/M of 6. A modified Gompertz equation adequately (R² > 0.946) described the cumulative hydrogen production yields. This study provides a novel strategy for controlling the conditions for production of hydrogen from food waste via anaerobic fermentation.     

Effect of ammonia on biohydrogen production from food waste via anaerobic fermentation

The effect of different additive ammonia (0–10 g/l as nitrogen) on hydrogen production from the anaerobic batch mesophilic fermentation of food waste was studied at two feed-to-microorganism ratios (F/M), 3.9 and 8.0. Anaerobic sludge taken from an anaerobic digester was used as inoculum. The hydrogen yield at F/M 3.9 and 8.0 without additive ammonia was 77.2 and 51.0 ml-H₂/gVS, respectively. At F/M 3.9, the hydrogen production was enhanced by adding additive ammonia in the system when the total ammonia nitrogen (TAN) concentration was no higher than 6.0 g/l. A maximum hydrogen yield of 121.4 ml-H₂/gVS was obtained at a TAN concentration of 3.5 g/l. At F/M 8.0, the enhancement of hydrogen production was found in a narrower range of additive TAN concentrations, with a highest yield of 60.9 ml-H₂/gVS at the TAN of 1.5 g/l. Hydrogen production was inhibited at higher additive TAN concentrations for both F/M ratios. This study provides a novel strategy for controlling ammonia for production of hydrogen from food waste via anaerobic fermentation.     

Alkali-treated sewage sludge as a seeding source for hydrogen fermentation of food waste leachate

In the present work, alkali-treated sewage sludge (SS) was used as a seeding source for H₂ fermentation of food waste leachate (FWL). The role of alkaline treatment of SS was to suppress the activity of non-H₂-producers in SS and also to enhance the solubility of SS. The effect of pretreatment pH and FWL:SS ratio on H₂ production was crucial, by changing the pH conditions and selecting the dominant species. High pretreatment pH and high SS content resulted in high initial pH conditions. The highest H₂ yield of 2.1 mol H₂/mol hexose_added was achieved at pretreatment pH 10 and a mixing ratio of FWL:SS = 3:5. At these conditions, the initial pH was 7.9, and cultivation pH was maintained within the reported optimum range of 5.5–6.5 during fermentation. It was found that pretreatment pH 9 was not strong enough to suppress the activity of non-H₂-producers in SS, in particular, lactic acid bacteria (LAB). Microbial analysis results confirmed that LAB such as Lactobacillus sp. and Enterococcus sp. were the dominant species at pretreatment pH 9 while Clostridium sp., the main anaerobic H₂-producers, were dominant at pretreatment pH 10.     

Bio-hythane production from food waste by dark fermentation coupled with anaerobic digestion process: A long-term pilot scale experience

In this paper are presented the results of the investigation on optimal process operational conditions of thermophilic dark fermentation and anaerobic digestion of food waste, testing a long-term run, applying an organic loading rate of 16.3 kgTVS/m³d in the first phase and 4.8 kgTVS/m³d in the second phase. The hydraulic retention times (HRTs) were maintained at 3.3 days and 12.6 days, respectively, for the first and second phase. Recirculation of anaerobic digested sludge, after a mild solid separation, was applied to the dark fermentation reactor in order to control the pH in the optimal hydrogen production range of 5–6. It was confirmed the possibility to obtain a stable hydrogen production, without using external chemicals for pH control, in a long-term test, with a specific hydrogen production of 66.7 l per kg of total volatile solid (TVS) fed and a specific biogas production in the second phase of 0.72 m³ per kgTVS fed; the produced biogas presented a typical composition with a stable presence of hydrogen and methane in the biogas mixture around 6 and 58%, respectively, carbon dioxide being the rest.     

Feasibility of biohydrogen production by anaerobic co-digestion of food waste and sewage sludge

Anaerobic co-digestion of food waste and sewage sludge for hydrogen production was performed in serum bottles under various volatile solids (VS) concentrations (0.5–5.0%) and mixing ratios of two substrates (0:100–100:0, VS basis). Through response surface methodology, empirical equations for hydrogen evolution were obtained. The specific hydrogen production potential of food waste was higher than that of sewage sludge. However, hydrogen production potential increased as sewage sludge composition increased up to 13–19% at all the VS concentrations. The maximum specific hydrogen production potential of 122.9 ml/g carbohydrate-COD was found at the waste composition of 87:13 (food waste:sewage sludge) and the VS concentration of 3.0%. The relationship between carbohydrate concentration, protein concentration, and hydrogen production potential indicated that enriched protein by adding sewage sludge might enhance hydrogen production potential. The maximum specific hydrogen production rate was 111.2 ml H₂/g VSS/h. Food waste and sewage sludge were, therefore, considered as a suitable main substrate and a useful auxiliary substrate, respectively, for hydrogen production. The metabolic results indicated that the fermentation of organic matters was successfully achieved and the characteristics of the heat-treated seed sludge were similar to those of anaerobic spore-forming bacteria, Clostridium sp.