Hydrogen production by combined dark and light fermentation of ground wheat solution

Ground waste wheat was subjected to combined dark and light batch fermentation for hydrogen production. The dark to light biomass ratio (D/L) was changed between 1/2 and 1/10 in order to determine the optimum D/L ratio yielding the highest hydrogen formation rate and the yield. Hydrogen production by only dark and light fermentation bacteria was also realized along with the combined fermentations. The highest cumulative hydrogen formation (CHF = 76 ml), hydrogen yield (176 ml H₂ g⁻¹ starch) and formation rate (12.2 ml H₂ g⁻¹ biomass h⁻¹) were obtained with the D/L ratio of 1/7 while the lowest CHF was obtained with the D/L ratio of 1/2. Dark–light combined fermentation with D/L ratio of 1/7 was faster as compared to the dark and light fermentations alone yielding high hydrogen productivity and reduced fermentation time. Dark and light fermentations alone also yielded considerable cumulative hydrogen, but slower than the combined fermentation.     

Photo-fermentative hydrogen gas production from dark fermentation effluent of ground wheat solution: Effects of light source and light intensity

Dark fermentation effluent of wheat powder solution was subjected to light fermentation for bio-hydrogen production using different light sources and intensities. Tungsten, fluorescent, infrared (IR), halogen lamps were used as light sources with a light intensity of 270 Wm⁻² along with sunlight. Pure culture of Rhodobacter sphaeroides-RV was used in batch light fermentation experiments. Halogen lamp was found to be the most suitable light source yielding the highest cumulative hydrogen formation (CHF, 252 ml) and yield (781 ml H₂ g⁻¹ TVFA). In the second set of experiments, light fermentations were performed at different light intensities (1–10 klux) using halogen lamp. The optimum light intensity was found to be 5 klux (approx. 176 Wm⁻²) resulting in the highest CHF (88 ml) and hydrogen yield (1037 ml H₂ g⁻¹TVFA). Hydrogen formation was limited by the availability of light at low light intensities below 5 klux and was inhibited by the excess light above 5 klux.     

Bio-hydrogen production from acid hydrolyzed wheat starch by photo-fermentation using different Rhodobacter sp

Hydrogen gas production from sugar solution derived from acid hydrolysis of ground wheat starch by photo-fermentation was investigated. Three different pure strains of Rhodobacter sphaeroides (RV, NRLL and DSZM) were used in batch experiments to select the most suitable strain. The ground wheat was hydrolyzed in acid solution at pH = 3 and 90 °C in an autoclave for 15 min. The resulting sugar solution was used for hydrogen production by photo-fermentation after neutralization and nutrient addition. R. sphaeroides RV resulted in the highest cumulative hydrogen gas formation (178 ml), hydrogen yield (1.23 mol H₂ mol⁻¹ glucose) and specific hydrogen production rate (46 ml H₂ g⁻¹ biomass h⁻¹) at 5 g l⁻¹ initial total sugar concentration among the other pure cultures. Effects of initial sugar concentration on photo-fermentation performance were investigated by varying sugar concentration between 2.2 and 13 g l⁻¹ using the pure culture of R. sphaeroides RV. Cumulative hydrogen volume increased from 30 to 232 ml when total sugar concentration was increased from 2.2 to 8.5 g l⁻¹. Further increases in initial sugar concentration resulted in decreases in cumulative hydrogen formation. The highest hydrogen formation rate (3.69 ml h⁻¹) and yield (1.23 mol H₂ mol⁻¹ glucose) were obtained at a sugar concentration of 5 g l⁻¹.     

Bio-hydrogen production by photo-fermentation of dark fermentation effluent with intermittent feeding and effluent removal

Pure culture of Rhodobacter sphaeroides (NRRL- B1727) was used for continuous photo-fermentation of volatile fatty acids (VFA) present in the dark fermentation effluent of ground wheat starch. The feed contained 1950 ± 50 mg L⁻¹ total VFA with some nutrient supplementation. Hydraulic residence time (HRT) was varied between 24 and 120 hours. The highest steady-state daily hydrogen production (55 ml d⁻¹) and hydrogen yield (185 ml H₂ g⁻¹ VFA) were obtained at HRT = 72 hours (3 days). Biomass concentration increased with increasing HRT. Volumetric and specific hydrogen formation rates were also maximum at HRT = 72 h. High extent of TVFA fermentation at HRT = 72 h resulted in high hydrogen gas production.     

Optimization of media composition for hydrogen gas production from hydrolyzed wheat starch by dark fermentation

Effects of N/C, P/C and Fe(II)/C ratios in fermentation medium on biohydrogen production by dark fermentation of acid-hydrolyzed wheat starch was investigated. The powdered wheat was autoclaved at pH = 3 and 90 °C for 15 min and the resulting sugar solution was fermented after external addition of N, P and Fe(II) to overcome nutrient limitations. Box–Wilson statistical experiment design was used by considering the N/C (0–0.05, w w⁻¹), P/C (0–0.02) and Fe(II)/C (0–0.03) ratios as the independent variables while the hydrogen yield and specific hydrogen production rate (SHPR) were the objective functions to be optimized. A quadratic response function was used to correlate the response functions with the independent variables. Low levels of the variables (N/C < 0.02, P/C < 0.01, Fe(II)/C < 0.01) resulted in low hydrogen yield and SHPR due to nutrient limitations and high levels of nutrients caused inhibitions. The optimum conditions yielding the maximum hydrogen yield (Y = 2.84 mol H₂ mol⁻¹ glucose) were N/C = 0.02, P/C = 0.008 and Fe(II)/C = 0.015. The maximum SHPR (96 mL H₂ g⁻¹ biomass h⁻¹) was obtained at N/C = 0.025, P/C = 0.008 and Fe(II)/C = 0.015 (w w⁻¹).     

Photo-fermentative hydrogen gas production from dark fermentation effluent of acid hydrolyzed wheat starch with periodic feeding

Hydrogen gas production by photo-fermentation of dark fermentation effluent of acid hydrolyzed wheat starch was investigated at different hydraulic residence times (HRT = 1-10 days). Pure Rhodobacter sphaeroides (NRRL B-1727) culture was used in continuous photo-fermentation by periodic feeding and effluent removal. The highest daily hydrogen gas production (85 ml d⁻¹) was obtained at HRT = 4 days (96 h) while the highest hydrogen yield (1200 ml H₂ g⁻¹ TVFA) was realized at HRT = 196 h. Specific and volumetric hydrogen formation rates were also the highest at HRT = 96 h. Steady-state biomass concentrations and biomass yields increased with increasing HRT. TVFA loading rates of 0.32 g L⁻¹ d⁻¹ and 0.51 g L⁻¹ d⁻¹ resulted in the highest hydrogen yield and formation rate, respectively. Hydrogen gas yield obtained in this study compares favorably with the relevant literature reports probably due to operation by periodic feeding and effluent removal.     

Biohydrogen production from cornstalk wastes by anaerobic fermentation with activated sludge

An anaerobic fermentation process to produce hydrogen from cornstalk wastes was systematically investigated in this work. Batch experiments numbered series I, II and III were designed to investigate the effects of acid pretreatment, enzymatic hydrolysis (enzymatic temperature, enzymatic time and enzymatic pH) on hydrogen production by using the natural sludge as inoculant. A maximum cumulative H₂ yield of 126.22 ml g⁻¹-CS (Cornstalk, or 146.94 ml g⁻¹-TS, Total Solid) and an average H₂ production rate of 9.58 ml g⁻¹-CS h⁻¹ were obtained from fermentation cornstalk with a concentration of 20 g/L and an initial pH of 7.0 at 36 °C through an optimal pretreatment process. The optimal process was that the substrate was soaked with an HCl concentration of 0.6 wt% at 90 °C for 2 h, and subsequently enzymatic hydrolysis for 72 h at 50 °C and pH 4.8 before fermentation. The biogas consisted of only H₂ and CO₂. In addition, the fermentation system was the typical ethanol-type fermentation according to ethanol and acetate as the main liquid by-products.     

Hydrogen gas production by electrohydrolysis of volatile fatty acid (VFA) containing dark fermentation effluent

Dark fermentation effluents of wheat powder (WP) solution containing different concentrations of volatile fatty acids (VFAs) were subjected to low voltage (1–3 V) DC current to produce hydrogen gas. Graphite and copper electrodes were tested and the copper electrode was found to be more effective due to higher electrical conductivity. The effects of solution pH (2–7), applied voltage (1–3 V) and the total VFA (TVFA) concentration (1–5 g L⁻¹) on hydrogen gas production were investigated. Hydrogen production increased with decreasing pH and became maximum at pH = 2. Increases in applied voltage and the TVFA concentration also increased the cumulative hydrogen formation. The most suitable conditions for the highest cumulative hydrogen production was pH = 2, with 3 V applied voltage and 5 g TVFA L⁻¹. Up to 110 ml hydrogen gas was obtained with 5 g L⁻¹ TVFA at pH = 5.8 and 2 V applied voltage within 37.5 h. The highest energy efficiency (56%) was obtained with the 2 V applied voltage and 10.85 g L⁻¹ TVFA. Hydrogen production by electrolysis of water in control experiments was negligible for pH > 4. Hydrogen production by electrohydrolysis of VFA containing anaerobic treatment effluents was found to be an effective method with high energy efficiency.     

Microbial culture selection for bio-hydrogen production from waste ground wheat by dark fermentation

Hydrogen formation performances of different anaerobic bacteria were investigated in batch dark fermentation of waste wheat powder solution (WPS). Serum bottles containing wheat powder were inoculated with pure cultures of Clostridium acetobutylicum (CAB), Clostridium butyricum (CB), Enterobacter aerogenes (EA), heat-treated anaerobic sludge (ANS) and a mixture of those cultures (MIX). Cumulative hydrogen formation (CHF), hydrogen yield (HY) and specific hydrogen production rate (SHPR) were determined for every culture. The heat-treated anaerobic sludge was found to be the most effective culture with a cumulative hydrogen formation of 560 ml, hydrogen yield of 223 ml H₂ g⁻¹ starch and a specific hydrogen production rate of 32.1 ml H₂ g⁻¹ h⁻¹.     

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.     

Effects of sludge pre-treatment method on bio-hydrogen production by dark fermentation of waste ground wheat

Three different pre-treatment methods were applied on two different anaerobic sludge cultures and their mixtures in order to investigate the effects of pre-treatment methods on bio-hydrogen production from dark fermentation of waste ground wheat solution. Repeated heat, chloroform and combinations of heat and chloroform pre-treatment methods were applied to anaerobic sludges from different sources. Repeated heat treatment (2 × 5 h) was found to be more effective in selecting hydrogen producing bacteria compared to the other treatment methods tested on the basis of cumulative hydrogen production. The highest hydrogen formation (652 ml) and specific hydrogen production rate (SHPR = 25.7 ml H₂ g⁻¹ cells h⁻¹) were obtained with the anaerobic sludge pre-treated by repeated boiling. Both the type of anaerobic sludge and the pre-treatment method had considerable effects on bio-hydrogen production from wheat powder solution (WPS) by dark fermentation.     

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.     

Dark fermentation of ground wheat starch for bio-hydrogen production by fed-batch operation

Ground wheat solution was used for bio-hydrogen production by dark fermentation using heat-treated anaerobic sludge in a completely mixed fermenter operating in fed-batch mode. The feed wheat powder (WP) solution was fed to the anaerobic fermenter with a constant flow rate of 8.33 mL h⁻¹ (200 mL d⁻¹). Cumulative hydrogen production, starch utilization and hydrogen yields were determined at three different WP loading rates corresponding to the feed WP concentrations of 10, 20 and 30 g L⁻¹. The residual starch (substrate) concentration in the fermenter decreased with operation time while starch consumption was increasing. The highest cumulative hydrogen production (3600 mL), hydrogen yield (465 mL H₂ g⁻¹ starch or 3.1 mol H₂ mol⁻¹ glucose) and hydrogen production rate (864 mL H₂ d⁻¹) were obtained after 4 days of fed-batch operation with the 20 g L⁻¹ feed WP concentration corresponding to a WP loading rate of 4 g WP d⁻¹. Low feed WP concentrations (10 g L⁻¹) resulted in low hydrogen yields and rates due to substrate limitations. High feed WP concentrations (30 g L⁻¹) resulted in the formation of volatile fatty acids (VFAs) in high concentrations causing inhibition on the rate and yield of hydrogen production.     

Improving hydrogen production from cassava starch by combination of dark and photo fermentation

The combination of dark and photo fermentation was studied with cassava starch as the substrate to increase the hydrogen yield and alleviate the environmental pollution. The different raw cassava starch concentrations of 10–25 g/l give different hydrogen yields in the dark fermentation inoculated with the mixed hydrogen-producing bacteria derived from the preheated activated sludge. The maximum hydrogen yield (HY) of 240.4 ml H₂/g starch is obtained at the starch concentration of 10 g/l and the maximum hydrogen production rate (HPR) of 84.4 ml H₂/l/h is obtained at the starch concentration of 25 g/l. When the cassava starch, which is gelatinized by heating or hydrolyzed with α-amylase and glucoamylase, is used as the substrate to produce hydrogen, the maximum HY respectively increases to 258.5 and 276.1 ml H₂/g starch, and the maximum HPR respectively increases to 172 and 262.4 ml H₂/l/h. Meanwhile, the lag time (λ) for hydrogen production decreases from 11 h to 8 h and 5 h respectively, and the fermentation duration decreases from 75–110 h to 44–68 h. The metabolite byproducts in the dark fermentation, which are mainly acetate and butyrate, are reused as the substrates in the photo fermentation inoculated with the Rhodopseudomonas palustris bacteria. The maximum HY and HPR are respectively 131.9 ml H₂/g starch and 16.4 ml H₂/l/h in the photo fermentation, and the highest utilization ratios of acetate and butyrate are respectively 89.3% and 98.5%. The maximum HY dramatically increases from 240.4 ml H₂/g starch only in the dark fermentation to 402.3 ml H₂/g starch in the combined dark and photo fermentation, while the energy conversion efficiency increases from 17.5–18.6% to 26.4–27.1% if only the heat value of cassava starch is considered as the input energy. When the input light energy in the photo fermentation is also taken into account, the whole energy conversion efficiency is 4.46–6.04%.     

Effects of light source,intensity and lighting regime on bio-hydrogen production from ground wheat starch by combined dark and photo-fermentations

Combined dark and photo-fermentation of ground wheat starch was carried out by using different light sources, intensities and lighting regime. A mixture of heat treated anaerobic sludge and Rhodobacter sphaeroides-RV with a certain light/dark bacteria ratio was used in batch experiments. Tungsten, fluorescent, infrared (IR), tungsten + infrared, halogen lamps were used as light sources with a light intensity of 270 Wm⁻² along with sunlight. Halogen lamp was found to be the most suitable light source yielding the highest cumulative hydrogen formation (178 ml) and yield (218 ml g⁻¹ starch). Combined fermentations were performed at different light intensities (1–10 klux) using the halogen lamp in the second set of experiments. The optimum light intensity was found to be 10 klux (approx. 352 Wm⁻²) resulting in the highest cumulative hydrogen (111 ml) and hydrogen yield (139 ml H₂ g⁻¹ starch). Hydrogen formation was limited by the availability of light at low light intensities below 10 klux. Durations of dark/light cycles were changed to determine the most suitable lighting regime. Hydrogen gas formation increased with increasing cycle time and continuous lighting resulted in the highest cumulative hydrogen formation and hydrogen yield.     

Effects of pH value and substrate concentration on hydrogen production from the anaerobic fermentation of glucose

A series of batch experiments were conducted to investigate the effects of pH and glucose concentrations on biological hydrogen production by using the natural sludge obtained from the bed of a local river as inoculant. Batch experiments numbered series I and II were designed at an initial and constant pH of 5.0–7.0 with 1.0 increment and four different glucose concentrations (5.0, 7.5, 10 and 20 g glucose/L). The results showed that the optimal condition for anaerobic fermentative hydrogen production is 7.5 g glucose/L and constant pH 6.0 with a maximum H₂ production rate of 0.22 mol H₂ mol⁻¹ glucose h⁻¹, a cumulative H₂ yield of 1.83 mol H₂ mol⁻¹ glucose and a H₂ percentage of 63 in biogas.     

Hydrogen gas production from acid hydrolyzed wheat starch by combined dark and photo-fermentation with periodic feeding

Hydrogen gas production from acid hydrolyzed waste wheat starch by combined dark and photo-fermentation was investigated in continuous mode with periodic feeding and effluent removal. A mixture of heat treated anaerobic sludge and Rhodobacter sphaeroides (NRRL-B 1727) were used as the seed culture for dark and light fermentations, respectively with biomass ratio of Rhodobacter/sludge = 3. Hydraulic residence time (HRT) was changed between 1 and 8 days by adjusting the feeding periods. Ground waste wheat was acid hydrolyzed at pH = 3 and 121 °C for 30 min using an autoclave and the resulting sugar solution was used as the substrate for combined fermentation after pH adjustment and nutrient addition. The highest daily hydrogen gas production (41 ml d⁻¹), hydrogen yield (470 ml g⁻¹ total sugar = 3.4 mol H₂ mol⁻¹glucose), volumetric and specific hydrogen production rates were obtained at the HRT of 8 days. The highest biomass and the lowest total volatile fatty acids (TVFA) concentrations were also realized at HRT = 8 days indicating VFA fermentation by Rhodobacter sp. at high HRTs. The lowest total sugar loading rate of 0.625 g L⁻¹ d⁻¹ resulted in the highest hydrogen yield and formation rate. Hydrogen gas production by combined fermentation with periodic feeding was proven to be an effective method resulting in high hydrogen yields at long HRTs.     

Thermophilic dark fermentation of acid hydrolyzed waste ground wheat for hydrogen gas production

Batch dark fermentation experiments were performed to investigate the effects of biomass and substrate concentration on bio-hydrogen production from acid hydrolyzed ground wheat at 55 °C. In the first set of experiments, the substrate concentration was constant at 20 g total sugar L⁻¹ and biomass concentration was varied between 0.52 and 2.58 g L⁻¹. Total sugar concentration was varied between 4.2 and 23.7 g L⁻¹ in the second set of experiments with a 1.5 g L⁻¹ constant biomass concentration. The highest cumulative hydrogen formation (582 mL, 30 °C, 1 atm), formation rate (5.43 mL h⁻¹) and final total volatile fatty acid (TVFA) concentration (6.54 g L⁻¹) were obtained with 1.32 g L⁻¹ biomass concentration. In variable substrate concentration experiments, the highest cumulative hydrogen (365 mL) and TVFA concentration (4.8 g L⁻¹) were obtained with 19.25 g L⁻¹ initial total sugar concentration while hydrogen gas formation rate (12.95 mL h⁻¹) and the yield (200 mL H₂ g⁻¹ total sugar) were the highest with 4.2 g L⁻¹ total sugar concentration.     

Hydrogen production from cassava wastewater using an anaerobic sequencing batch reactor: Effects of operational parameters,COD:N ratio,and organic acid composition

In this work, hydrogen production from cassava wastewater using anaerobic sequencing batch reactors (ASBR) was investigated to determine the optimum number of cycles per day, chemical oxygen demand (COD) loading rate, and COD:N ratio. The system operated at a COD loading rate of 30 kg/m³d and 6 cycles per day provided maximum hydrogen production performance in terms of specific hydrogen production rate (SHPR) (388 ml H₂/g VSS d or 3800 ml H₂/l d) and hydrogen yield (186 ml H₂/g COD removed). The effect of nitrogen supplementation was also studied by adding NH₄HCO₃ into the system at the COD:N ratios of 100:2.2, 100:3.3, and 100:4.4 under the COD loading rate of 30 kg/m³d and 6 cycles per day. The maximum SHPR and hydrogen yield of 524 ml H₂/g VSS d (5680 ml H₂/l d) and 438 ml H₂/g COD removed, respectively, were obtained at the stoichiometric COD:N ratio of 100:2.2. An excess nitrogen was found to promote the productions of higher organic acids and ethanol, resulting in lowering hydrogen production efficiency.     

Hydrogen gas production from olive mill wastewater by electrohydrolysis with simultaneous COD removal

Diluted olive mill wastewater (OMW) was subjected to direct current (DC) voltages (0.5-4.0 V) for hydrogen gas production with simultaneous chemical oxygen demand (COD) removal by electrohydrolysis. The highest cumulative hydrogen production (3020 ml) and hydrogen yield (2500 ml H₂ g⁻¹ COD) were obtained with 3 V DC voltage while the highest current intensity (80 mA), percent hydrogen (95%) in the gas phase, hydrogen gas formation rate (614 ml d⁻¹), percent COD removal (44%) and energy conversion efficiency (95%) were realized with 2 V. Hydrogen gas production by electrolysis of water was negligible for all voltages. COD removal from OMW with no DC voltage application was usually lower than that obtained with DC power application. Hydrogen gas production by electrohydrolysis of OMW was proven to be a fast and effective method with simultaneous COD removal.