Effects of the substrate and cell concentration on bio-hydrogen production from ground wheat by combined dark and photo-fermentation

Effects of the substrate and cell concentration on bio-hydrogen production from ground wheat solution were investigated in combined dark-light fermentations. The ratio of the dark to light bacteria concentration (D/L) was kept constant at 1/10 while the wheat powder (WP) concentration was changed between 2.5 and 20 g L⁻¹ with a total cell concentration of 0.41 g L⁻¹ in the first set of experiments. Cell concentration was changed between 0.5 and 5 g L⁻¹ in the second set of experiments while the wheat powder concentration was constant at 5 g L⁻¹ with a D/L ratio of 1/7. The highest cumulative hydrogen (135 ml) and formation rate (3.44 ml H₂ h⁻¹) were obtained with the 20 g L⁻¹ wheat powder concentration. However, the highest yield (63.9 ml g⁻¹ starch) was obtained with the 2.5 g L⁻¹ wheat powder. In variable cell concentration experiments, the highest cumulative hydrogen (118 ml) and yield (156.8 ml H₂ g⁻¹ starch) were obtained with 1.1 g L⁻¹ cell concentration yielding an optimal biomass/substrate ratio of 0.22 g cells/g WP.     

Optimization of fermentative hydrogen production process by response surface methodology

The effect of temperature, initial pH and glucose concentration on fermentative hydrogen production by mixed cultures was investigated in batch tests, and the optimization of fermentative hydrogen production process was conducted by response surface methodology with a central composite design. Experimental results showed that temperatures, initial pH and glucose concentrations had impact on fermentative hydrogen production individually and interactively. The maximum hydrogen yield of 289.8 mL/g glucose was estimated at the temperature of 38.6 °C, the initial pH of 7.2 and the glucose concentration of 23.9 g/L. The maximum hydrogen production rate of 28.2 mL/h was estimated at the temperature of 37.8 °C, the initial pH of 7.2 and the glucose concentration of 27.6 g/L. The maximum substrate degradation efficiency of 96.9% was estimated at the temperature of 39.3 °C, the initial pH of 7.0 and the glucose concentration of 26.8 g/L. Response surface methodology was a better method to optimize the fermentative hydrogen production process. Modified logistic model could describe the progress of cumulative hydrogen production in the batch tests of this study successfully.     

Effect of sulfate on hydrogen production from the organic fraction of municipal solid waste using co-culture of E. coli and Enterobacter aerogenes

In the present study, the effect of sulfate on the hydrogen production from the organic fraction of municipal solid (OFMSW) waste using co-culture of Enterobacter aerogenes and E. coli has been studied under varying pH conditions. The presence of sulfate in the feedstock declines hydrogen production efficiency. To evaluate the effect of sulfate on hydrogen production from OFMSW, COD/sulfate ratio of 17.5, 15.0, 12.5, 10.0, 7.5, 5.0 and 2.5 were applied at different pH conditions (i.e. pH 5.5, 6.0 and 6.5). The hydrogen production continuously declined with the decreasing COD/sulfate ratio and increase in pH. The cumulative hydrogen production decreased from 220.8 ± 10.5 mL in control to a minimum of 98.3 ± 10.5 mL, 74.4 ± 10.4 mL, and 44.6 ± 2.6 mL at pH 5.5, 6.0 and 6.5 respectively. The major content of gaseous composition included hydrogen and CO₂ at higher COD/sulfate ratio and low pH, while H₂S formation started with the decrease in COD/sulfate ratio and increase in the pH. Similarly, sulfate removal efficiency was found to be influenced by COD/sulfate ratio and pH condition. Soluble metabolite analysis revealed that total volatile fatty acid concentration was not affected by sulfate addition. Thus, Sulfate removal is essential prior to fermentation in order to improve hydrogen yield.     

Effect of substrate concentration on fermentative hydrogen production from sweet sorghum extract

The aim of the present study was to assess the influence of substrate concentration on the fermentative hydrogen production from sweet sorghum extract, in a continuous stirred tank bioreactor. The reactor was operated at a Hydraulic Retention Time (HRT) of 12 h and carbohydrate concentrations ranging from 9.89 to 20.99 g/L, in glucose equivalents. The maximum hydrogen production rate and yield were obtained at the concentration of 17.50 g carbohydrates/L and were 2.93 ± 0.09 L H₂/L reactor/d and 0.74 ± 0.02 mol H₂/mol glucose consumed, corresponding to 8.81 ± 0.02 L H₂/kg sweet sorghum, respectively. The main metabolic product at all steady states was butyric acid, while ethanol production was high at high substrate concentrations. The experiments showed that hydrogen productivity depends significantly on the initial carbohydrate concentration, which also influences the distribution of the metabolic products.     

Electrohydrolysis of landfill leachate organics for hydrogen gas production and COD removal

Hydrogen gas production with simultaneous COD removal was realized by application of DC voltages (0.5-5.0 V) to landfill leachate. The rate and the yield of hydrogen gas production were investigated at different DC voltages by using aluminum electrodes and DC power supply. The highest cumulative hydrogen production (5000 mL), hydrogen yield (2400 mL H₂ g⁻¹ COD), daily hydrogen gas formation (1277 mL d⁻¹), and percent hydrogen (99%) in the gas phase were obtained with 4 V DC voltage. Energy conversion efficiency (H₂ energy/electrical energy) reached the highest level (80.6%) with 1 V DC voltage. Hydrogen gas production was mainly realized by electrohydrolysis of leachate organics due to negligible H₂ gas production in water and leachate control experiments. The highest COD removal (77%) was also obtained with 4 V DC voltage. Electrohydrolysis of landfill leachate was proven to be an effective method for hydrogen gas production with simultaneous COD removal.     

Simultaneous hydrogen gas formation and COD removal from cheese whey wastewater by electrohydrolysis

Cheese whey (CW) was subjected to DC voltages between 0.5 and 5 V for hydrogen gas production with simultaneous COD removal by electrohydrolysis of CW organics. Hydrogen gas formation and COD removal were investigated at different DC voltages using aluminum electrodes. The highest cumulative hydrogen production (5551 mL), hydrogen yield (1709 mL H₂ g⁻¹ COD), hydrogen gas formation rate (913 ml d⁻¹), and percent hydrogen (99%) in the gas phase were obtained with 5 V DC voltage within 158 h. Energy conversion efficiency reached the highest level (80.7%) at 3 V DC voltage with cumulative hydrogen production of 4808 mL and hydrogen yield of 1366 mL H₂ g⁻¹ COD. Hydrogen gas was mainly produced by electrohydrolysis of CW organics due to low H₂ gas production in water and CW control experiments. The highest COD removal (22%) was also obtained with 3 V DC voltage. Major COD removal mechanism was anaerobic degradation of carbohydrates producing volatile fatty acids (VFA) and CO₂. Hydrogen gas was produced by reaction of protons released from VFAs and electrons provided by DC current. Hydrogen gas production by electrohydrolysis of CW solution was proven to be an effective method with simultaneous COD removal.     

Hydrogen gas production from vinegar fermentation wastewater by electro-hydrolysis: Effects of initial COD content

Vinegar fermentation wastewater with different initial COD contents (9.66–48.6 g L⁻¹) were used for hydrogen gas production with simultaneous COD removal by electro-hydrolysis. The applied DC voltage was constant at 4 V. The highest cumulative hydrogen production (3197 ml), hydrogen yield (2766 ml H₂ g⁻¹ COD), hydrogen formation rate (799 ml d⁻¹), and percent hydrogen (99.5%) in the gas phase were obtained with the highest initial COD of 48.6 g COD L⁻¹. The highest energy efficiency (48%) was obtained with the lowest COD content of 9.66 g L⁻¹. Hydrogen gas production by water electrolysis was less than 250 ml and wastewater control resulted in less than 25 ml H₂ in 96 h. The highest (12%) percent COD removal was obtained with the lowest COD content. Hydrogen gas was produced by reaction of (H⁺) ions present in raw WW ( pH = 3.0) and protons released from acetic acid with electrons provided by electrical current. Electro-hydrolysis of vinegar wastewater was proven to be an effective method of H₂ gas production with some COD removal.     

Prevention of substrate and product inhibitions by using a dilution strategy during dark fermentative hydrogen production from molasses

Substrate and product inhibitions have a significant effect on dark fermentative hydrogen gas production. Particularly, rapid formation of volatile fatty acids leads to fast pH decreases shifting the metabolic pathway. Therefore, controlling volatile fatty acid accumulation has great importance in maintaining effective hydrogen production. In this context, a dilution strategy was applied to regulate volatile fatty acids levels within the desired concentration range. A three-factor Box-Behnken statistical experiment design was established to assess the effects of dilution time, dilution percentage and initial COD concentration on hydrogen formation yield and rate. Highest hydrogen yield (7.7 mL H₂/mL_reactor) and rate (21. 47 mL H₂/h or 9.38 mmol/L_reactor.h) were achieved when 85 gCOD/L containing fermentation media was diluted with a percentage of 130 of the initial working volume at the 3rd hour of the fermentation period. Moreover, this strategy enabled to start fermentation with 55 g glucose/L.     

Optimizing hydrogen production from organic wastewater treatment in batch reactors through experimental and kinetic analysis

Anaerobic hydrogen production from organic wastewater, an emerging biotechnology to generate clean energy resources from wastewater treatment, is critical for environmental and energy sustainability. In this study, hydrogen production, biomass growth and organic substrate degradation were comprehensively examined at different levels of two critical parameters (chemical oxygen demand (COD) and pH). Hydrogen yields had a reverse correlation with COD concentrations. The highest specific hydrogen yield (SHY) of 2.1 mole H₂/mole glucose was achieved at the lowest COD of 1 g/L and decreased to 0.7 mole H₂/mole glucose at the highest COD of 20 g/L. The pH of 5.5–6.0 was optimal for hydrogen production with the SHY of 1.6 mole H₂/mole glucose, whereas the acidic pH (4.5) and neutral pH (6.0–7.0) lowered the hydrogen yields. Under all operational conditions, acetate and butyrate were the main components in the liquid fermentation products. Additionally, a comprehensive kinetic analysis of biomass growth, substrate degradation and hydrogen production was performed. The maximum rates of microbial growth (μ_m) and substrate utilization (R_su) were 0.03 g biomass/g biomass/day and 0.25 g glucose/g biomass/day, respectively. The optimum pH for the rate of hydrogen production (RH₂$R_{{\text{H}}_2}$) and SHY were 5.89 and 5.74 respectively. Based on the kinetic analysis, the highest RH₂$R_{{\text{H}}_2}$ and SHY for batch-mode anaerobic hydrogen production systems were projected to be 13.7 mL/h and 2.32 mole H₂/mole glucose.     

Effects of temperature and substrate concentration on biological hydrogen production from starch

This study investigates the effects of temperature and substrate concentration on biological hydrogen production from starch using mixed cultures. In this work, although hydrogen was successfully produced under the thermophilic condition, stable hydrogen production was not observed under the mesophilic condition. In the thermophilic reactor, the maximum hydrogen yield was 2.8 mol H₂/mol glucose at 20 g/l-starch; however, hydrogen yield decreased drastically with the change of by-product distribution when substrate concentration was over 30 g/l-starch. A negative correlation was observed between the hydrogen yield and the total concentration of undissociated acids.     

Hydrogen gas production from waste anaerobic sludge by electrohydrolysis: Effects of applied DC voltage

Waste anaerobic sludge was subjected to different DC voltages (0.5-5 V) for hydrogen gas production by using aluminum electrodes and a DC power supply. Effects of applied DC voltage on the rate and extent of hydrogen gas production were investigated. The highest cumulative hydrogen production (2775 ml), daily hydrogen gas formation (686.7 ml d⁻¹), hydrogen yield (96 ml H₂ g⁻¹ COD) and percent hydrogen (94.3%) in the gas phase were obtained with 2 V DC voltage. Energy conversion efficiency (H₂ energy/electrical energy) also reached the highest level (74%) with 2 V DC voltage application. Control experiments with no voltage application to the sludge yielded almost the same level of COD removal, but no hydrogen gas production. Voltage application to water resulted in much lower hydrogen gas production as compared to sludge indicating negligible electrolysis of water. The results indicated that the sludge was naturally decomposed by the active cells removing COD and releasing hydrogen ions to the medium which reacted with the electrons provided by DC current to produce hydrogen gas. Hydrogen gas production from electrohydrolysis of waste sludge was found to be a fast and effective method with high energy efficiency.     

Electro-hydrolysis of cheese whey solution for hydrogen gas production and chemical oxygen demand (COD) removal using photo-voltaic cells (PVC)

Diluted cheese whey (CW) solution was used for hydrogen gas production by electro-hydrolysis using photo-voltaic cells (PVC) as source of electricity. Effects of initial chemical oxygen demand (COD) concentration on the rate and yield of hydrogen gas production were investigated using a completely mixed and sealed reactor with aluminum electrodes. Cumulative hydrogen gas formation (CHF) increased with increasing initial COD concentration. The highest cumulative hydrogen gas volume (26472 mL), hydrogen gas production rate (4553 mL d⁻¹), hydrogen yield (7004 mL H₂ g⁻¹ COD), and percent COD removal (21.5%) were obtained with initial COD of 35172 mg L⁻¹. H₂ gas formation from water control was only 5365 mL. pH of the CW solution increased with decreasing conductivities during the course of experiments. Gas phase contained more than 99% H₂ at the end of experiments. The highest energy efficiency (20.4%) was also obtained with the highest COD content. Nearly pure hydrogen gas formation by electro-hydrolysis of cheese whey using PVC panels was proven to be an effective method.     

Experimental kinetics and dynamics of hydrogen production on glucose by hydrogen forming bacteria (HFB) culture

The kinetic study was performed using a modified “initial rate-method” and the dynamic ones by the relaxation time methodology. The approach was tested on glucose as sole carbon source while the hydrogen forming bacteria HFB were obtained by acid treatment of anaerobic sludge. A large spectrum of substrate concentration from 5 g/l to 90 g/l was experimentally tested. During the test biogas evolution, gas composition, glucose concentration as well as pH and Red-Ox Potential (ROP) were monitored. At the end of the tests ethanol and VFA were measured to evaluate a reference molar H₂ yield (Y*). The biogas composition ranged in (40–60%) for H₂ and rest CO₂, no CH₄ was observed. A first order kinetic equation for glucose with a kinetic constant of 0.0041 h⁻¹ and an inhibited kinetic equation for biogas evolution with a maximum production rate of 100 ml/l h were set-up. The dynamic study evidences the strong role of the pH in the regulation of activity of the Ferrodoxin and Hydrogenase pools. Lastly a test with a bioreactor of 2 l with pH adjustments validated the dynamics of the system showing an increase of 2.8 times of efficiency of glucose conversion into H₂ compared with tests without pH adjustments and agitation.     

Efficient hydrogen gas production from cassava and food waste by a two-step process of dark fermentation and photo-fermentation

A two-step process of sequential anaerobic (dark) and photo-heterotrophic fermentation was employed to produce hydrogen from cassava and food waste. In dark fermentation, the average yield of hydrogen was approximately 199 ml H₂ g⁻¹ cassava and 220 ml H₂ g⁻¹ food waste. In subsequent photo-fermentation, the average yield of hydrogen from the effluent of dark fermentation was approximately 611 ml H₂ g⁻¹ cassava and 451 ml H₂ g⁻¹ food waste. The total hydrogen yield in the two-step process was estimated as 810 ml H₂ g⁻¹ cassava and 671 ml H₂ g⁻¹ food waste. Meanwhile, the COD decreased greatly with a removal efficiency of 84.3% in cassava batch and 80.2% in food waste batch. These results demonstrate that cassava and food waste could be ideal substrates for bio-hydrogen production. And a two-step process combining dark fermentation and photo-fermentation was highly improving both bio-hydrogen production and removal of substrates and fatty acids.     

Effect of different substrate concentrations and salinity on hydrogen production from mariculture organic waste (MOW)

In this paper, the influence of substrate concentrations and salinity on hydrogen production from mariculture organic waste (MOW) at mesophilic condition in batch reactors was determined. It was found that the hydrogen yield and hydrogen content were influenced by the initial substrate concentrations and salinity. The optimum concentration for hydrogen production was 20 g/L. The salinity could produce inhibit effect for hydrogen production. The low hydrogen content was detected at high salinity condition. The nutrients changing and metabolites composition could also be significantly influenced by the salinity. The releasing of carbohydrate from MOW could be easily used for hydrogen production, and protein could be accumulated and assimilated during the hydrogen fermentation. The enhancing of salinity was disadvantage for total metabolites accumulation.     

Two stage biodiesel and hydrogen production from molasses by oleaginous fungi and Clostridium acetobutylicum ATCC 824

In the present study biodiesel was produced by various fungal species isolated from Egypt using sugarcane molasses as substrate. In the first stage 6 oleaginous fungi, namely, Alternaria alternata, Cladosporium cladosporioides, Epicoccum nigrum, Fusarium oxysporum, Aspergillus parasiticus and Emericella nidulans var. lata were used for lipid production. Subsequent to fungal cultivation on sugarcane molasses the cultures were filtered and biodiesel was prepared by direct esterification of dry fungal biomass. Methyl esters of palmitic, stearic, linoleic and elaidic represented the major components while palmitoleic represented a minor component of biodiesel produced from tested oleaginous fungi. In the second stage, the spent medium of fungal culture was used as the fermentation medium for hydrogen production by Clostridium acetobutylicum ATCC 824. The maximum total H₂ yield was obtained with the spent medium of E. nigrum and A. alternata. The results presented in this study suggest a possibility of interlinking the biodiesel production technology by fungi with hydrogen production by C. acetobutylicum ATCC 824 to exploit the residual sugars in the spent media and therefore increase the economic feasibility of the biofuel production from molasses.     

Enhanced polyhydroxybutyrate production through incorporation of a hydrogen fuel cell and electro-fermentation system

The integration of a hydrogen fuel cell with an electro-fermentation system represents a novel approach for improving polyhydroxybutyrate (PHB) accumulation in Ralstonia eutropha H16, using a sustainable energy source. In this study, electro-fermentation noticeably affected cell growth, biomass production, substrate consumption, and PHB accumulation. Final residual biomass concentrations and maximum specific growth rates were enhanced by supplying a 10-mA electric current. Furthermore, a remarkable enhancement in PHB content (30% higher than control) was achieved by redox-mediated electro-fermentation with a 10 mA electric current, upon the addition of a redox mediator. Two-stage cultivation limited the growth suppression caused by redox-mediated electro-fermentation, and also increased the maximum PHB productivity of the system. The additional electrons supplied upon supplementation of the redox mediator accelerated the glycolytic pathway and redox cycling of NADH/NAD⁺, led to a spontaneous boost for adenosine triphosphate (ATP) generation, and further facilitated the biosynthesis of PHB.     

Efficiency assessment of a photo electrochemical chloralkali process for hydrogen and sodium hydroxide production

In present study, a new reactor configuration is developed which integrates photochemical hydrogen production with an electrochemical chloralkali process. The effects of different parameters on rate of hydrogen, chlorine and sodium hydroxide production are experimentally examined and discussed. The parameters include applied voltage, varied from 4 V to 5 V, amount of catalyst, varied from 1 g/425 mL to 4 g/425 mL, and light intensity, varied from 20 W/m² to 55 W/m². Factorial design of experiments is applied and an analysis of variance (ANOVA) is used to analyze the experimental results. Energy and exergy efficiencies are also examined. An optimization study is performed to find the optimal catalyst concentration. An optimized catalyst concentration in salty water is used to examine its effect on the rate of hydrogen production.