Inhibitory effect of ethanol,acetic acid,propionic acid and butyric acid on fermentative hydrogen production

The inhibitory effect of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production by mixed cultures was investigated in batch tests using glucose as substrate. The experimental results showed that, at 35 °C and initial pH 7.0, during the fermentative hydrogen production, the substrate degradation efficiency, hydrogen production potential, hydrogen yield and hydrogen production rate all trended to decrease with increasing added ethanol, acetic acid, propionic acid and butyric acid concentration from 0 to 300 mmol/L. The inhibitory effect of added ethanol on fermentative hydrogen production was smaller than those of added acetic acid, propionic acid and butyric acid. The modified Han–Levenspiel model could describe the inhibitory effects of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production rate in this study successfully. The modified Logistic model could describe the progress of cumulative hydrogen production.     

Effect of operation parameters on anaerobic fermentation using cow dung as a source of microorganisms

Anaerobic fermentation by microorganisms is a promising method of hydrogen production for it can be conducted at mild conditions. In this paper, a series of tests were carried out to investigate the effect of pH, hydraulic retention time (HRT), temperature (T) and substrate concentration on anaerobic dark fermentation. Glucose was utilized as model substrate. The Taguchi orthogonal array was applied in the experimental design and a verification experiment was tested. The results showed the optimal parameters for hydrogen production were pH 5.0, HRT 8.34 h, T 33.5 °C and substrate concentration of 14 g/L, with hydrogen yield of 2.15 mol H₂/mol glucose. Butyric-type fermentation occurred in most tests. According to the analysis of effluent contents, at pH 5.5, 5.0, 4.0, the effluent contained mostly butyric acid (43.1–56.6%), followed by acetic acid (24.6–29.8%); at HRT 4.17, 6.26, 8.34 h, the effluent contained mostly butyric acid (43.0–53.6%). Increasing temperature from 29 to 39.5 °C resulted in the decrease of butyric acid percentage but increase of ethanol percentage. Substrate concentration had little effect on product constitution.     

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.     

Revivability of fermentative hydrogen producing bioreactors

In this study we investigated the revivability of a continuous biological hydrogen producing reactor after a period of feed interruption. Before the feed interruption, the hydrogen production yield was 1.36 mol H₂/mol glucose with butyric acid and acetic acid as the main metabolic products. However, after feed interruption, butyric acid formation completely stopped and the hydrogen yield decreased to 0.29 mol H₂/mol glucose. Lactic acid, ethanol and acetic acid became the main metabolites after re-start up. Reduction of organic loading rate together with increasing the pH after the feed interruption resulted in an increase in the hydrogen yield to 0.7 mol H₂/mol glucose. The microbial community dynamics showed complete elimination of Clostridium affiliated strains and predominance of Lactobacillus affiliated strains after the re-start up of the reactor.     

生物制氢反应器产氢产乙酸菌群对挥发酸的转化

采用间歇培养的方式,利用取自生物制氢反应器的厌氧活性污泥考察了活性污泥中产氢产乙酸菌群对乙醇、乙酸、丙酸、丁酸、戊酸和乳酸的转化和产氢。结果表明,培养时间为44h时,厌氧活性污泥发酵葡萄糖的累计产气量为356mL,累计产氢量为209mL,氢气含量为58.7%。发酵产物的组成成分乙醇为427.1mg/L、乙酸为716.5mg/L、丙酸为172.5mg/L、丁酸为689.4mg/L、戊酸为123.6mg/L。发酵生物制氢反应器厌氧活性污泥中产氢产乙酸菌群能够对乙醇和乳酸进行产氢产乙酸转化,厌氧污泥转化乙醇形成的乙酸含量约为270mg/L,累计产氢量为15mL;转化乳酸形成的乙酸含量约为190mg/L,累计产氢量为7mL。厌氧污泥不能对乙酸、丙酸、丁酸和戊酸进行产氢产乙酸转化,培养过程中也没有气体生成,分析认为产氢产乙酸菌群对挥发酸的转化不是发酵生物制氢反应器产氢的主要途径。     

Hydrogen production potentials and fermentative characteristics of various substrates with different heat-pretreated natural microflora

Batch tests were carried out to investigate the effects of heat-pretreated inocula on the fermentative hydrogen production characteristics of various types of substrates. A total of 8 different inocula and 4 different substrates (starch, glycerol, oil and peptone) were used. Heat pretreatment of the inocula was conducted in order to harvest spore-forming clostridial bacteria. Significant hydrogen production potentials were observed from starch (20.5–174.4 ml H₂/g-COD_starch) and glycerol (11.5–38.1 ml H₂/g-COD_glycerol); however, almost no hydrogen was produced from oil and peptone. When starch was used as a substrate, two different fermentation patterns were observed, according to the inocula: butyric acid-type and ethanol-type fermentation. Polymerase chain reaction combined with denaturing gradient gel electrophoresis (PCR-DGGE) analysis was conducted to compare the bacterial structures cultivated on the starch medium. Different species of clostridial bacteria were observed between the butyric acid-type and ethanol-type fermentation cultures. When glycerol was used as a substrate, 1,3-propanediol was the main by-product with each inoculum. The results of the present study suggest that simultaneous production of ethanol or 1,3-propanediol in addition to hydrogen is a more promising strategy than conventional hydrogen production in acidogenesis.     

Influence of pH on fermentative hydrogen production from sweet sorghum extract

The present study focused on the influence of pH on the fermentative hydrogen production from the sugars of sweet sorghum extract, in a continuous stirred tank bioreactor. The reactor was operated at a Hydraulic Retention Time of 12 h and a pH range of 3.5–6.5. The maximum hydrogen production rate and yield were obtained at pH 5.3 and were 1752 ± 54 mL H₂/d or 3.50 ± 0.07 L H₂/L reactor/d and 0.93 ± 0.03 mol H₂/mol glucose consumed or 10.51 L H₂/kg sweet sorghum, respectively. The main metabolic product at this pH value was butyric acid. The hydrogen productivity and yield were still at high levels for the pH range of 5.3–4.7, suggesting a pH value of 4.7 as optimum for hydrogen production from an economical point of view, since the energy demand for chemicals is lower at this pH. At this pH range, the dominant fermentation product was butyric acid but when the pH culture sharply decreased to 3.5, hydrogen evolution ceased and the dominant metabolic products were lactic acid and ethanol.     

Dark fermentative biohydrogen production by mesophilic bacterial consortia isolated from riverbed sediments

Dark fermentative bacterial strains were isolated from riverbed sediments and investigated for hydrogen production. A series of batch experiments were conducted to study the effect of pH, substrate concentration and temperature on hydrogen production from a selected bacterial consortium, TERI BH05. Batch experiments for fermentative conversion of sucrose, starch, glucose, fructose, and xylose indicated that TERI BH05 effectively utilized all the five sugars to produce fermentative hydrogen. Glucose was the most preferred carbon source indicating highest hydrogen yields of 22.3 mmol/L. Acetic and butyric acid were the major soluble metabolites detected. Investigation on optimization of pH, temperature, and substrate concentration revealed that TERI BH05 produced maximum hydrogen at 37 °C, pH 6 with 8 g/L of glucose supplementation and maximum yield of hydrogen production observed was 2.0–2.3 mol H₂/mol glucose. Characterization of TERI BH05 revealed the presence of two different bacterial strains showing maximum homology to Clostridium butyricum and Clostridium bifermentans.     

Anaerobic fluidized bed reactor with expanded clay as support for hydrogen production through dark fermentation of glucose

This study evaluated hydrogen production in an anaerobic fluidized bed reactor (AFBR) fed with glucose-based synthetic wastewater. Particles of expanded clay (2.8–3.35 mm) were used as a support material for biomass immobilization. The reactor was operated with hydraulic retention times (HRT) ranging from 8 to 1 h. The hydrogen yield production increased from 1.41 to 2.49 mol H₂ mol⁻¹ glucose as HRT decreased from 8 to 2 h. However, when HRT was 1 h, there was a slight decrease to 2.41 mol H₂ mol⁻¹ glucose. The biogas produced was composed of H₂ and CO₂, and the H₂ content increased from 8% to 35% as HRT decreased. The major soluble metabolites during H₂ fermentation were acetic acid (HAc) and butyric acid (HBu), accounting for 36.1–53.3% and 37.7–44.9% of total soluble metabolites, respectively. Overall, the results demonstrate the potential of using expanded clay as support material for hydrogen production in AFBRs.     

Fermentative hydrogen production by diverse microflora

In this study, hydrogen production with activated sludge, a diverse bacterial source has been investigated and compared to microflora from anaerobic digester sludge, which is less diverse. Batch experiments were conducted at mesophilic (37 °C) and thermophilic (55 °C) temperatures. The hydrogen production yields with activated sludge at 37 °C and 55 °C were 0.56 and 1.32 mol H₂/mol glucose consumed, respectively. While with anaerobically digested sludge hydrogen yield was 2.18 mol H₂/mol glucose consumed at 37 °C and 1.25 mol H₂/mol glucose consumed at 55 °C. The results of repeated batch experiments for 615 h resulted in average yields of 1.21 ± 0.62 and 1.40 ± 0.16 mol H₂/mol glucose consumed for activated sludge and anaerobic sludge, respectively. The hydrogen production with activated sludge was not stable during the repeated batches and the fluctuation in hydrogen production was attributed to formation of lactic acid as the predominant metabolite in some batches. The presence of lactic acid bacteria in microflora was confirmed by PCR-DGGE.     

Pretreatment of sweet sorghum bagasse for hydrogen production by Caldicellulosiruptor saccharolyticus

Pretreatment of sweet sorghum bagasse, an energy crop residue, with NaOH for the production of fermentable substrates, was investigated. Optimal conditions for the alkaline pretreatment of sweet sorghum bagasse were realized at 10% NaOH (w/w dry matter). A delignification of 46% was then observed, and improved significantly the efficiency of enzymatic hydrolysis. Under hydrolysis conditions without pH control, up to 50% and 41% of the cellulose and hemicellulose contained in NaOH-pretreated sweet sorghum bagasse were converted by 24 h enzymatic hydrolysis to soluble monomeric sugars. The extreme thermophilic bacterium Caldicellulosiruptor saccharolyticus showed normal growth on hydrolysates of NaOH-pretreated biomass up to a sugar concentration of 20 g/L. Besides hydrogen, the main metabolic products detected in the fermentations were acetic and lactic acid. The maximal hydrogen yield observed in batch experiments under controlled conditions was 2.6 mol/mol C6 sugar. The maximal volumetric hydrogen production rate ranged from 10.2 to 10.6 mmol/(L h). At higher substrate concentrations the production of lactic acid increased at the expense of hydrogen production.     

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.     

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.     

Non-sterile bio-hydrogen fermentation from food waste in a continuous stirred tank reactor (CSTR): Performance and population analysis

Bio-hydrogen production from food waste by anaerobic mixed cultures was conducted in a continuous stirred tank reactor (CSTR). The hydraulic retention time (HRT) was optimized in order to maximize hydrogen yield (HY) and hydrogen production rate (HPR). The maximum hydrogen content (38.6%), HPR (379 mL H₂/L. d) and HY (261 mL H₂/g-VS_added) were achieved at the optimum HRT of 60 h. The major soluble metabolite products were butyric and acetic acids which indicated a butyrate-acetate type fermentation. Operation of CSTR at HRT 60 h could select hydrogen producing bacteria and eliminate lactic acid bacteria and acetogenic bacteria. The microbial community analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) revealed that the predominant hydrogen producer was Clostridium sp.     

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.     

Improving biohydrogen production in a carrier-induced granular sludge bed by altering physical configuration and agitation pattern of the bioreactor

Our newly developed carrier-induced granular sludge bed (CIGSB) bioreactor was shown to be very effective in hydrogen production. However, since mechanical agitation was not employed to enable sludge granulation, the CIGSB system might encounter problems with poor mass transfer efficiency during prolonged operations. This work was undertaken to improve mixing efficiency of CIGSB for better biomass-substrate contact by adjusting the height to diameter (H/D) ratios of the reactor and by implementing appropriate agitation device. Three H/D ratios (4, 8, and 12) resulting in liquid upflow velocities (v_up) of 0.057–1.32 m/h were examined as the CIGSB reactor was carried out at a descending hydraulic retention time (HRT) from 4 to 0.5 h. The results show that decreasing HRT resulted in increases in the H₂ production rate, regardless of the H/D ratios. Reactors with a H/D ratio of 8 gave better H₂ production performance with a H₂ production rate of 6.87 l/h/l and a H₂ yield of 3.88 mol H₂/mol sucrose, suggesting that the effectiveness of H₂ production in the CIGSB system can be enhanced by using a proper v_up and physical configuration of the reactor. Supply of additional mechanical agitation for CIGSB reactor (H/D=12) alleviated the phenomena of sludge piston floatation, leading to further increases in the H₂ production rate and H₂ yield to 9.31 l/h/l and 4.02 mol H₂/mol sucrose, respectively. The major soluble metabolite was butyric acid, followed by acetic acid, propionic acid, and ethanol. The former two accounted for nearly 67–76% of total soluble microbial products, indicating the presence of favorable pathways in the CIGSB culture from the aspect of H₂-producing metabolism.     

The role of surface reactions on the active and selective catalyst design for bioethanol steam reforming

In order to study the role of surface reactions involved in bioethanol steam reforming mechanism, a very active and selective catalyst for hydrogen production was analysed. The highest activity was obtained at 700 °C, temperature at which the catalyst achieved an ethanol conversion of 100% and a selectivity to hydrogen close to 70%. It also exhibited a very high hydrogen production efficiency, higher than 4.5 mol H₂ per mol of EtOH fed. The catalyst was operated at a steam to carbon ratio (S/C) of 4.8, at 700 °C and atmospheric pressure. No by-products, such as ethylene or acetaldehyde were observed. In order to consider a further application in an ethanol processor, a long-term stability test was performed under the conditions previously reported. After 750 h, the catalyst still exhibited a high stability and selectivity to hydrogen production. Based on the intermediate products detected by temperature programmed desorption and reaction (TPD and TPR) experiments, a reaction pathway was proposed. Firstly, the adsorbed ethanol is dehydrogenated to acetaldehyde producing hydrogen. Secondly, the adsorbed acetaldehyde is transformed into acetone via acetic acid formation. Finally, acetone is reformed to produce hydrogen and carbon dioxide, which were the final reaction products. The promotion of such reaction sequence is the key to develop an active, selective and stable catalyst, which is the technical barrier for hydrogen production by ethanol reforming.     

Continuous biohydrogen production from liquid swine manure supplemented with glucose using an anaerobic sequencing batch reactor

Liquid swine manure supplemented with glucose (10 g/L) was used as substrate for hydrogen production using an anaerobic sequencing batch reactor at 37 ± 1 °C and pH 5.0 under different hydraulic retention times (HRTs). Decreasing HRT from 24 to 8 h caused an increasing hydrogen production rate from 0.05 to 0.15 L/h/L. Production rates of both total biogas and hydrogen were linearly correlated to HRT with R² being 0.993 and 0.997, respectively. The hydrogen yield ranged between 1.18 and 1.63 mol-H₂/mol glucose and the 12 h HRT was preferred for high production rate and efficient yield. For all the five HRTs examined, the glucose utilization efficiency was over 98%. The biogas mainly consisted of carbon dioxide and hydrogen (up to 43%) with no methane detected throughout the experiment. Ethanol and organic acids were the major aqueous metabolites produced during fermentation, with acetic acid accounting for 56–58%. The hydrogen yield was found to be related to the acetate/butyrate ratio.     

光合产氢混合菌群的碳源代谢实验研究

以红螺菌科光合产氢混合菌群为研究对象,通过血清瓶培养实验,研究不同碳源对光合细菌生长和产氢过程的影响。结果表明:光合细菌能有效利用乙酸和丁酸快速增殖和产氢,其中以乙酸最佳,促使光合产氢混合菌群增殖的最佳乙酸浓度为80mmol/L,最佳产氢浓度为40mmol/L。光合产氢混合菌群利用乳酸增殖产氢的能力较低,而乙醇则对其表现为抑制效应。