Hydrogen production from water hyacinth through dark- and photo- fermentation

This article discusses the method of producing hydrogen from water hyacinth. Water hyacinth was pretreated with microwave heating and alkali to enhance the enzymatic hydrolysis and hydrogen production in a two-step process of dark- and photo- fermentation. Water hyacinth with various concentrations of 10–40 g/l was pretreated with four methods: (1) steam heating; (2) steam heating and microwave heating/alkali pretreatment; (3) steam heating and enzymatic hydrolysis; (4) steam heating, microwave heating/alkali pretreatment and enzymatic hydrolysis. Water hyacinth (20 g/l) pretreated with method 4 gave the maximum reducing sugar yield of 30.57 g/100 g TVS, which was 45.6% of the theoretical reducing sugar yield (67.0 g/100 g TVS). The pretreated water hyacinth was used to produce hydrogen by mixed H₂-producing bacteria in dark fermentation. The maximum hydrogen yield of 76.7 ml H₂/g TVS was obtained at 20 g/l of water hyacinth. The residual solutions from dark fermentation (mainly acetate and butyrate) were used to further produce hydrogen by immobilized Rhodopseudomonas palustris in photo fermentation. The maximum hydrogen yield of 522.6 ml H₂/g TVS was obtained at 10 g/l of water hyacinth. Through a combined process of dark- and photo- fermentation, the maximum hydrogen yield from water hyacinth was dramatically enhanced from 76.7 to 596.1 ml H₂/g TVS, which was 59.6% of the theoretical hydrogen yield.     

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.     

Combination of dark- and photo-fermentation to enhance hydrogen production and energy conversion efficiency

In this study, we investigated a two-phase process of combining the dark- and photo-fermentation methods to reutilize the residual solution derived from dark fermentation and increase the hydrogen yield (HY) from glucose. In dark fermentation, an orthogonal experimental design was used to optimize the culture medium for Clostridium butyricum (C. butyricum). The optimal culture medium composition was determined as glucose 20 g/l, NaCl 3 g/l, MgCl₂ 0.1 g/l, FeCl₂ 0.1 g/l, K₂HPO₄ 2.5 g/l, l-cysteine 0.5 g/l, vitamin solution 10 ml/l, and trace element solution 10 ml/l. In this method, the maximum HY increased from 1.59 to 1.72 mol H₂/mol glucose and hydrogen production rate (HPR) from 86.8 to 100 ml H₂/l/h. The metabolite byproducts from dark fermentation, mostly containing acetate and butyrate, were inoculated with Rhodopseudomonas palustris (R. palustris) and reutilized to produce hydrogen in photo-fermentation. In photo-fermentation, the maximum HY was 4.16 mol H₂/mol glucose, and the maximum removal ratios of acetate and butyrate were 92.3% and 99.8%, respectively. Combining dark fermentation and photo-fermentation caused a dramatic increase of HY from 1.59 to 5.48 mol H₂/mol glucose. The conversion efficiency of heat value in dark fermentation surged from 13.3% to 46.0% in the two-phase system.     

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.     

Batch and continuous biohydrogen production from starch hydrolysate by Clostridium species

In this study, hydrogen gas was produced from starch feedstock via combination of enzymatic hydrolysis of starch and dark hydrogen fermentation. Starch hydrolysis was conducted using batch culture of Caldimonas taiwanensis On1 able to hydrolyze starch completely under the optimal condition of 55 °C and pH 7.5, giving a yield of 0.46–0.53 g reducing sugar/g starch. Five H₂-producing pure strains and a mixed culture were used for hydrogen production from raw and hydrolyzed starch. All the cultures could produce H₂ from hydrolyzed starch, whereas only two pure strains (i.e., Clostridium butyricum CGS2 and CGS5) and the mixed culture were able to ferment raw starch. Nevertheless, all the cultures displayed higher hydrogen production efficiencies while using the starch hydrolysate, leading to a maximum specific H₂ production rate of 116 and 118 ml/g VSS/h, for Cl. butyricumCGS2 and Cl. pasteurianum CH5, respectively. Meanwhile, the H₂ yield obtained from strain CGS2 and strain CH5 was 1.23 and 1.28 mol H₂/mol glucose, respectively. The best starch-fermenting strain Cl. butyricum CGS2 was further used for continuous H₂ production using hydrolyzed starch as the carbon source under different hydraulic retention time (HRT). When the HRT was gradually shortened from 12 to 2 h, the specific H₂ production rate increased from 250 to 534 ml/g  VSS/h, whereas the H₂ yield decreased from 2.03 to 1.50  mol H₂/mol glucose. While operating at 2 h HRT, the volumetric H₂ production rate reached a high level of 1.5 l/h/l.     

Recent developments of hydrogen production from sewage sludge by biological and thermochemical process

Hydrogen is a kind of clean effective resource. Sewage sludge is regarded as a promising material for hydrogen production because it owns a wide range of sources and the methods are consistent with the goal of sustainable development. This work reviews existing hydrogen production technologies from sewage sludge, including photo-fermentation, dark-fermentation, sequential dark- and photo-fermentation, pyrolysis, gasification, and supercritical water gasification (SCWG). Overall comparison for the involving approaches is conducted based on their inherent features and current development status along with the technical and environmental aspect. Results show that sequential dark- and photo-fermentation and pyrolysis have improved hydrogen yields, but the emissions of carbon dioxide are also remarkable. Biological processes have an advantage in cost, but the reaction rates are inferior to those of thermochemical method. Enhancing methods and improvements are proposed to guide future research on hydrogen production from sewage sludge and promote the effectiveness both technically and economically.     

Biological hydrogen production from steam-exploded straw by simultaneous saccharification and fermentation

Hydrogen was produced by simultaneous saccharification and fermentation from steam-exploded corn straw (SECS) using Clostridium butyricum AS1.209. Effect of various process parameters, such as solid to liquid ratio, enzyme loading and initial pH, etc., were examined with respect to maximum hydrogen productivity which was obtained by fitting the cumulative hydrogen production data to a modified Gompertz equation. Maximum specific hydrogen production rate and maximal hydrogen yield were 126 ml/g VSS d and 68 ml/g SECS, respectively. The yield of soluble metabolites was 197.7 mg/g SECS. Acetic acid accounted for 46% of the total was the most abundant product and this shows that hydrogen production from SECS was essentially acetate-type fermentation. Hydrogen production by simultaneous saccharification and fermentation of SECS has the predominance of short lag-stage and high maximum specific hydrogen production rate and it was a promising method for hydrogen production and straw biomass conversion.     

Bioconversion of corncob to hydrogen using anaerobic mixed microflora

Biohydrogen production from corncob using natural anaerobic microflora was reported for the first time. The optimum pretreatment condition for the corncob was determined to be 100 °C, 30 min, and 1% HCl (w/w). The maximum hydrogen yield of 107.9 ml/g-TVS and hydrogen production rate of 4.20 ml/g-TVS h⁻¹ was obtained under the condition of 10 g/l substrate concentration and initial pH 8.0. Butyrate and acetate were the dominant metabolic by-products of hydrogen fermentation. Chemical composition analysis, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to study the mechanism of degrading corncob for hydrogen production. The amorphous domains of cellulose and hemicellulose were hydrolyzed into fermentable saccharides through acid pretreatment and the microorganisms had a devastating effect on the crystallinity of the cellulose. The hydrogen yield from pretreated corncob was much higher than from raw corncob. Therefore, the acid pretreatment played a crucial role on hydrogen production from corncob.     

Fermentative bio-hydrogen production from cellulose by cow dung compost enriched cultures

The performance of hydrogen production from cellulose by the cow dung compost enriched continuously in defined medium containing cellulose was investigated. In the initial experiments, batch-fermentation was carried out to observe the effects of different substrate concentration conditions on the rate of cellulose-degrading, growth of bacteria and the capability of hydrogen-producing from cellulose. The result showed that the cellulose degradation decreased from 55% at 5 g/l to 22% at 30 g/l. The maximum cumulative hydrogen production and the rate of hydrogen production first increased from 828 ml/l at 5 g/l to 1251 ml/l at 10 g/l then remained constant beyond 10 g/l. The maximum hydrogen production potential, the rate of hydrogen production and the yield of hydrogen was 1525 ml/l, 33 ml/l.h, and 272 ml/g-cellulose (2.09 mol/mol-hexose) was obtained at substrate concentration 10 g/l, the hydrogen concentration in biogas was 47–50%(v/v) and there was no methane observed. During the conversion of cellulose into hydrogen, acetate and butyrate were main liquid end-products in the metabolism of hydrogen fermentation. These results proposed that cow dung compost enriched cultures were ideal microflora for hydrogen production from cellulose.     

Comparative study on bio-hydrogen production from corn stover: Photo-fermentation,dark-fermentation and dark-photo co-fermentation

In this study, three different fermentation methods, such as photo-fermentation (PF), dark-fermentation (DF) and dark-photo co-fermentation (DPCF) for bio-hydrogen production from corn stover were compared in terms of hydrogen production, substrate consumption, by-products formation and energy conversion efficiency. A modified Gompertz model was applied to perform the kinetic analysis of hydrogen production. The maximum cumulative hydrogen yield of 141.42 mL·(g TS)⁻¹ was achieved by PF, DF with the minimum cumulative hydrogen yield of 36.08 mL· (g TS)⁻¹ had the shortest lag time of 4.33 h, and DPCF had the maximum initial hydrogen production rate of 1.88 mL· (g TS)⁻¹·h⁻¹ and maximum initial hydrogen content of 44.40%. The results also indicated PF was an acid-consuming process with a low total VFAs concentration level of 2.90–4.19 g·L⁻¹, DF was a process of VFAs accumulation with a maximum total VFAs concentration of 12.66 g·L⁻¹, and DPCF was a synergistic process in which the total VFAs concentration was significantly reduced and the hydrogen production efficiency was effectively improved compared with DF. The energy conversion efficiency of PF, DF and DPCF were 10.12%, 2.58% and 6.45%, respectively.     

Comparison of biohydrogen production processes

For hydrogen to be a viable energy carrier, it is important to develop hydrogen generation routes that are renewable like biohydrogen. Hydrogen can be produced biologically by biophotolysis (direct and indirect), photo-fermentation and dark-fermentation or by combination of these processes (such as integration of dark- and photo-fermentation (two-stage process), or biocatalyzed electrolysis, etc.). However, production of hydrogen by these methods at commercial level is not reported in the literature and challenges regarding the process scale up remain. In this scenario net energy analysis (NEA) can provide a tool for establishing the viability of different methods before scaling up. The analysis can also be used to set targets for various process and design parameters for bio-hydrogen production.     

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.     

Control strategies for hydrogen production through co-culture of Ethanoligenens harbinense B49 and immobilized Rhodopseudomonas faecalis RLD-53

This study evaluated hydrogen production by co-culture of Ethanoligenens harbinense B49 and immobilized Rhodopseudomonas faecalis RLD-53 with different control strategies. To enhance cooperation of dark and photo-fermentation bacteria during hydrogen production process, the glucose concentration, phosphate buffer concentration and initial pH were controlled at 6 g/l, 50 mmol/l and 7.5, respectively. The maximum yield and rate of hydrogen production were 3.10 mol H₂/mol glucose and 17.2 mmol H₂/l/h, respectively. Ethanol from E. harbinense B49 in acetate medium can enhance hydrogen production by R. faecalis RLD-53 except the ratio of ethanol to acetate (R_E/A) among 0.8 to 1.0. Control of the proper phosphate buffer concentration (50 mmol/l) not only increased acetic acid production by E. harbinense B49, but also maintained stable pH of co-culture system. Therefore, the results showed that co-culture of E. harbinense B49 and immobilized R. faecalis RLD-53 was a promising way of converting glucose into hydrogen.     

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.     

Comparison between heterofermentation and autofermentation in hydrogen production from Arthrospira (Spirulina) platensis wet biomass

Hydrogen production from Arthrospira (Spirulina) platensis wet biomass through heterofermentation by the [FeFe] hydrogenase of hydrogenogens (hydrogen-producing bacteria) and autofermentation by the [NiFe] hydrogenase of Arthrospira platensis was discussed under dark anaerobic conditions. In heterofermentation, wet cyanobacterial biomass without pretreatment was hardly utilized by hydrogenogens for hydrogen production. But the carbohydrates in cyanobacterial cells released after cell wall disruption were effectively utilized by hydrogenogens for hydrogen production. Wet cyanobacterial biomass was pretreated with boiling and bead milling, ultrasonication, and ultrasonication and enzymatic hydrolysis. Wet cyanobacterial biomass pretreated with ultrasonication and enzymatic hydrolysis achieved the maximum reducing sugar yield of 0.407 g/g-DW (83.0% of the theoretical reducing sugar yield). Different concentrations (10 g/l to 40 g/l) of pretreated wet cyanobacterial biomass were used as substrate to produce fermentative hydrogen by hydrogenogens, which were domesticated with the pretreated wet cyanobacterial biomass as carbon source. The maximum hydrogen yield of 92.0 ml H₂/g-DW was obtained at 20 g/l of wet cyanobacterial biomass. The main soluble metabolite products (SMPs) in the residual solutions from heterofermentation were acetate and butyrate. In autofermentation, hydrogen yield decreased from 51.4 ml H₂/g-DW to 11.0 ml H₂/g-DW with increasing substrate concentration from 1 g/l to 20 g/l. The main SMPs in the residual solutions from autofermentation were acetate and ethanol. The hydrogen production peak rate and hydrogen yield at 20 g/l of wet cyanobacterial biomass in heterofermentation showed 110- and 8.4-fold increases, respectively, relative to those in autofementation.     

Cogeneration of hydrogen and methane from the pretreated biomass of algae bloom in Taihu Lake

In order to efficiently utilize the biomass waste of algae bloom in Taihu Lake, China and improve energy conversion efficiency, a three-stage process comprising dark hydrogen fermentation with acid-domesticated hydrogenogens, photohydrogen fermentation, and methanogenesis was undertaken. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses showed that algal cells pretreated by microwave heating with diluted acid were degraded into smaller fragments (<5 μm) than those pretreated by steam heating with diluted acid. The microwave pretreatment of algae resulted in higher saccharification efficiency. The domesticated hydrogenogens in presence of acids improved the dark hydrogen production from microwave-pretreated algae biomass and led to a total combined dark and photofermentation hydrogen yield of 283.4 mL/g-total volatile solid (TVS). The energy conversion efficiency of steam-pretreated algae biomass remarkably increased to 47.0% by cogenerating 256.7 mL/g-TVS hydrogen and 253.5 mL/g-TVS methane in the three-stage process: dark-fermentation, photofermentation, and methanogenesis.     

Optimization of biohydrogen production from beer lees using anaerobic mixed bacteria

Beer lees are the main by-product of the brewing industry. Biohydrogen production from beer lees using anaerobic mixed bacteria was investigated in this study, and the effects of acidic pretreatment, initial pH value and ferrous iron concentration on hydrogen production were studied at 35 °C in batch experiments. The hydrogen yield was significantly enhanced by optimizing environmental factors such as hydrochloric acid (HCl) pretreatment of substrate, initial pH value and ferrous iron concentration. The optimal environmental factors of substrate pretreated with 2% HCl, pH = 7.0 and 113.67 mg/l Fe²⁺ were observed. A maximum cumulative hydrogen yield of 53.03 ml/g-dry beer lees was achieved, which was approximately 17-fold greater than that in raw beer lees. In addition, the degradation efficiency of the total reducing sugar, and the contents of hemicellulose, cellulose, lignin and metabolites are presented, which showed a strong dependence on the environmental factors.     

Biohydrogen production from apple pomace by anaerobic fermentation with river sludge

The biological hydrogen (bio-H₂) production from apple pomace (AP) by fermentation using natural mixed microorganisms in batch process was studied under various experimental conditions. The river sludge was used as a seed after being boiled for 15 min. The results show that the optimal pretreatment for AP was to soak it in the ammonia liquor of 6% for 24 h at room temperature. An optimal fermentation condition for bio-H₂ production was proposed that the pretreated AP at 37 °C, the initial pH of 7.0 and the fermentation concentration of 15 g/l could produce a maximum cumulative H₂ yield (CHYm) of 101.08 ml/g total solid (TS) with an average H₂ production rate (AHPR) of 8.08 ml/g TS/h. During the conversion of AP into H₂, acetic acid, ethanol, propionic acid and butyric acid were main liquid end-products.