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.     

Effect of changing temperature on anaerobic hydrogen production and microbial community composition in an open-mixed culture bioreactor

The temperature effect (37–65 °C) on H₂ production from glucose in an open-mixed culture bioreactor using an enrichment culture from a hot spring was studied. The dynamics of microbial communities was investigated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). At 45 and 60 °C the H₂ production was the highest i.e. 1.71 and 0.85 mol H₂/mol glucose, respectively. No H₂ was produced at temperatures 50 and 55 °C. At 37–45 °C, H₂ production was produced by butyrate type fermentation while fermentation mechanism changed to ethanol type at 60 °C. Clostridium species were dominant at 37–45 °C while at 50–55 °C and 60 °C the culture was dominated by Bacillus coagulans and Thermoanaerobacterium, respectively. In the presence of B. Coagulans the metabolism was directed to lactate production. The results show that the mixed culture had two optima for H₂ production and that the microbial communities and metabolic patterns promptly changed according to changing temperatures.     

Performance and population analysis of hydrogen production from sugarcane juice by non-sterile continuous stirred tank reactor augmented with Clostridium butyricum

Non-sterile operation of continuous stirred tank reactor (CSTR) augmented with Clostridium butyricum and fed with sugarcane juice was studied at various hydraulic retention time (HRT). The maximum hydrogen production rate and yield of 3.38 mmol H₂/L/h and 1.0 mol H₂/mol hexose consumed, respectively, were achieved at HRT 4 h. The relationship of the augmented microorganism and normal flora in the fermentation system under non-sterile condition were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Initially, at HRT 36 h, other species related to Lactobacillus harbinensis and Klebseilla pneumoniae were present as a major group in the reactor. When HRT was decreased to 12, 6 and 4 h, C. butyricum was present with a competition between L. harbinensis and K. pneumoniae. Results indicated that augmented C. butyricum could compete with contaminated microorganisms during non-sterile operation at low HRT (12-4 h) with the support of normal flora (K. pneumoniae).     

Hydrogen and ethanol fermentation of various carbon sources by immobilized Escherichia coli (XL1-Blue)

In this study, hydrogen and ethanol production by a facultative anaerobic bacterium Escherichia coli XL1-Blue immobilized in calcium-alginate beads have been investigated. Batch fermentations were carried out at mesophilic temperature (35 °C) and an initial cultivation pH of 6.5. Firstly, the influence of biomass concentration in terms of dry cell weight (expressed in g DCW/L, range 0.2–1.0) was investigated using fructose (5 g/L) as a carbon source. The peak hydrogen yield (HY) of 1.17 mol-H₂/mol-fructose_utilized was obtained at an initial cell concentration of 0.4 g DCW/L. The hydrogen production potential of other simple carbon sources (glucose and xylose) was evaluated at this optimized cell concentration and peak HY values were attained as 0.96 mol-H₂/mol-glucose_utilized and 0.69 mol-H₂/mol-xylose_utilized, respectively. In addition, utilization of the beverage wastewater (BWW) showed the peak cumulative hydrogen production and ethanol concentration of 120 mL and 5.65 g/L, attained at the substrate concentration of 20 g_{(glucose equivalent)}/L. However, peak HY (1.65 mol-H₂/mol-_{glucose eqivalent utilized}) was observed at low substrate concentration of 5 g_{(glucose equivalent)}/L. The percentage of sugar utilization of BWW was ranged between 80 and 96.     

Rapid detection and quantification methodology for genus Megasphaera as a hydrogen producer in a hydrogen fermentation system

Important issues in the microbiology of biological reactors include bacterial composition and cell counts of bacterial communities. Because oligonucleotide probes targeting a particular phyletic group are available for studying hydrogen fermentation, fluorescence in situ hybridization (FISH) could be an attractive research technique. Although we reported the importance of the Megasphaera spp. for a simple hydrogen fermentation system, fundamental information like population and composition remains unclear. Thus, here, we established the use of Megasphaera-specific oligonucleotide Mega-X for FISH and the optimum conditions for rapid single-cell level identification. The culture isolated from a simple hydrogen fermentation process was identified as Megasphaera elsdenii and was shown to be significantly dominant (30%–58% of the microflora) by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and FISH. The combination of Megasphaera-specific PCR-RFLP and FISH is likely to become an important methodology for understanding the global distribution and behaviour of Megasphaera spp. in the environment.     

Sugarcane vinasse as substrate for fermentative hydrogen production: The effects of temperature and substrate concentration

The present study aimed to evaluate the hydrogen production of a microbial consortium using different concentrations of sugarcane vinasse (2–12 g COD L⁻¹) at 37 °C and 55 °C. In mesophilic tests, the increase in vinasse concentration did not significantly impact the hydrogen yield (HY) (from 1.72 to 2.23 mmol H₂ g⁻¹ COD_influent) but had a positive effect on the hydrogen production potential (P) and hydrogen production rate (R_m). On the other hand, the increase in the substrate concentration caused a drop in HY from 2.31 to 0.44 mmol H₂ g⁻¹ COD_influent in the tests performed at 55 °C with vinasse concentrations from 2 to 12 g COD L⁻¹. The mesophilic community was composed of different species within the Clostridium genus, and the thermophilic community was dominated by organisms affiliated with the Thermoanaerobacter genus. Not all isolates affiliated with the Clostridium genus contributed to a high HY, as the homoacetogenic pathway can occur.     

Glycerol fermentation to hydrogen by Thermotoga maritima: Proposed pathway and bioenergetic considerations

The production of biohydrogen from glycerol, by the hyperthermophilic bacterium Thermotoga maritima DSM 3109, was investigated in batch and chemostat systems. T. maritima converted glycerol to mainly acetate, CO₂ and H₂. Maximal hydrogen yields of 2.84 and 2.41 hydrogen per glycerol were observed for batch and chemostat cultivations, respectively. For batch cultivations: i) hydrogen production rates decreased with increasing initial glycerol concentration, ii) growth and hydrogen production was optimal in the pH range of 7–7.5, and iii) a yeast extract concentration of 2 g/l led to optimal hydrogen production. Stable growth could be maintained in a chemostat, however, when dilution rates exceeded 0.025 h⁻¹ glycerol conversion was incomplete. A detailed overview of the catabolic pathway involved in glycerol fermentation to hydrogen by T. maritima is given. Based on comparative genomics the ability to grow on glycerol can be considered as a general trait of Thermotoga species. The exceptional bioenergetics of hydrogen formation from glycerol is discussed.     

Biohydrogen production from purified terephthalic acid (PTA) processing wastewater by anaerobic fermentation using mixed microbial communities

Purified terephthalic acid (PTA) processing wastewater was evaluated as a fermentable substrate for hydrogen (H₂) production with simultaneous wastewater treatment by dark-fermentation process in a continuous stirred-tank reactor (CSTR) with selectively enriched acidogenic mixed consortia under continuous flow condition in this paper. The inoculated sludge used in the reactor was excess sludge taken from a second settling tank in a local wastewater treatment plant. Under the conditions of the inoculants not less than 6.3 gVSS/L, the organic loading rate (OLR) of 16 kgCOD/m³ d, hydraulic retention time (HRT) of 6 h and temperature of (35 ± 1) °C, when the pH value, alkalinity and oxidation–reduction potential (ORP) of the effluent ranged from 4.2 to 4.4, 280 to 350 mg CaCO₃/L, and −220 to −250 mV respectively, soluble metabolites were predominated by acetate and ethanol, with smaller quantities of propionate, butyrate and valerate. Stable ethanol-type fermentation was formed with the sum of ethanol and acetate concentration ratio of 70.31% to the total liquid products after 25 days operation. The H₂ volume content was estimated to be 48–53% of the total biogas and the biogas was free of methane throughout the study. The average biomass concentration was estimated to be 10.82 gVSS/L, which favored H₂ production efficiently. The rate of chemical oxygen demand (COD) removal reached at about 45% and a specific H₂ production rate achieved 0.073 L/gMLVSS d in the study. This CSTR system showed a promising high-efficient bioprocess for H₂ production from high-strength chemical wastewater.     

Enhancement in hydrogen production by co-cultures of Bacillus and Enterobacter

Defined co-cultures of hydrogen (H₂) producers belonging to Citrobacter, Enterobacter, Klebsiella and Bacillus were used for enhancing the efficiency of biological H₂ production. Out of 11 co-cultures consisting of 2–4 strains, two co-cultures composed of Bacillus cereus EGU43, Enterobacter cloacae HPC123, and Klebsiella sp. HPC793 resulted in H₂ yield up to 3.0 mol mol⁻¹ of glucose. Up-scaling of the reactor by 16-fold resulted in a corresponding increase in H₂ production with an actual evolution of 7.44 L of H₂. It constituted 58.2% of the total biogas. Continuous culture evolution of H₂ by co-cultures (B. cereus EGU43 and E. cloacae HPC123) immobilized on ligno-cellulosic materials resulted in 6.4-fold improvement in H₂ yield compared to free floating bacteria. This synergistic influence of B. cereus and E. cloacae can offer a better strategy for H₂ production than undefined or mixed cultures.     

High temperature water gas shift reaction over promoted iron based catalysts prepared by pyrolysis method

In this work the effects of different promoters (Cr, Al, Mn, Ce, Ni, Co and Cu) on the structural and catalytic properties of Nanocrystalline iron based catalysts for high temperature water gas shift reaction were investigated. The catalysts were prepared in active phase (Fe₃O₄) via a facile direct synthesis routs without any additive and characterized using X-ray diffraction (XRD), N₂ adsorption (BET), temperature-programmed reduction (TPR), transmission and scanning electron microscopies (TEM,SEM) techniques. The obtained results indicated that synergic effect of Mn and Ni promoters can lead to obtain a Cr-free catalyst with high activity. In addition, the effect of Ni content on the structural and catalytic properties of the Fe–Mn–Ni catalysts was investigated. It was found that Fe–Mn–Ni catalyst with Fe/Mn = 10 and Fe/Ni = 5 weight ratios showed the highest catalytic activity among the prepared catalysts and possessed a stable catalytic performance without any decrease during 10 h time on stream. Moreover, the effect of GHSV and steam/gas ratio on the catalytic performance of this catalyst was investigated.     

Parametric study of membrane reactors for hydrogen production via high-temperature water gas shift reaction

This study presents numerical studies of hydrogen production performance via water gas shift reaction in membrane reactor. The pre-exponential factor in describing the hydrogen permeation flux is used as the main parameter to account for the membrane permeance variation. The operating pressure, temperature and H₂O/CO molar ratio are chosen in the 1–20 atm, 400–600 °C and 1–3 ranges, respectively. Based on the numerical simulation results three distinct CO conversion regimes exist based on the pre-exponential factor value. For low pre-exponential factors corresponding to low membrane permeance, the CO conversion approaches to that obtained from a conventional reactor without hydrogen removal. For high pre-exponential factor, high CO conversion and H₂ recovery with constant values can be obtained. For intermediate pre-exponential factor range both CO conversion and H₂ recovery vary linearly with the pre-exponential factor. In the high membrane permeation case CO conversion and H₂ recovery approach limiting values as the operating pressure increases. Increasing the H₂O/CO molar ratio results in an increase in CO conversion but decrease in H₂ recovery due to hydrogen permeation driving force reduction. As the feed rate increases in the reaction side both the CO conversion and hydrogen recovery decrease because of decreased reactant residence time. The sweep gas flow rate has a significant effect on hydrogen recovery. Low sweep gas flow rate results in low CO conversion H₂ recovery while limiting CO conversion and hydrogen recovery can be reached for the high membrane permeance and high sweep gas flow rate cases.     

Stoichiometric analysis of biological hydrogen production by fermentative bacteria

In this study, biohydrogen production from glucose by two fermentative bacteria (Clostridium butyricum, a typical strictly anaerobic bacterium, and Klebsiella pneumoniae, a well-studied facultative anaerobic and nitrogen-fixing bacterium) are stiochiometrically analyzed according to energy (ATP), reducing equivalent and mass balances. The theoretical analysis reveals that the maximum yield of hydrogen on glucose by Clostridium butyricum is 3.26 mol/mol when all acetyl-CoA entering into the acetate pathway (α=1$\alpha =1$), which is higher than that by Klebsiella pneumoniae under strictly anaerobic conditions. In the latter case, the maximum yield by Klebsiella pneumoniae is 2.86 mol hydrogen per mol glucose when five sevenths of acetyl-CoA is transformed to acetate. However, under microaerobic condition the maximum yield of hydrogen on glucose by Klebsiella pneumoniae could reach 6.68 mol/mol if all acetyl-CoA entered into tricarboxylic acid (TCA) cycle (γ=1$\gamma =1$) and a quantity of 53% of the reducing equivalents generated in the metabolism were completely oxidized by molecular oxygen. On the other hand, the relationship between hydrogen production and biomass formation is distinct by Clostridium butyricum from that by Klebsiella pneumoniae.   The former yield of hydrogen on glucose increases as biomass. In contrast, the latter one decreases as biomass in a certain range of molar fraction of acetate in total acetyl-CoA metabolism (5/7?β?0$5/7?\beta ?0$). Microaerobic condition is favorable for high hydrogen production with low biomass formation by Klebsiella pneumoniae   in a certain range of the molar fraction of all reducing equivalents oxidized completely by molecular oxygen (0.53?δ?0.83$0.53?\delta ?0.83$).     

Photofermentative hydrogen production from volatile fatty acids present in dark fermentation effluents

In the present study, the growth and hydrogen production of Rhodobacter sphaeroides O.U. 001, was investigated in media containing five different volatile fatty acids (VFA) individually (malate, acetate, propionate, butyrate and lactate) and in media containing mixtures of these acids that reflect the composition of dark fermentation effluents. The highest hydrogen production rate was obtained in malate (24 ml_hydrogen/l_reactor h) and the highest biomass concentration was obtained in acetate containing media (1.65 g/l). The substrate conversion efficiencies for different volatile fatty acids were found to vary between 14 and 50%. The malate and butyrate consumption rates were first order with consumption rate constants of 0.026 h⁻¹ and 0.015 h⁻¹, respectively. In the case of substrate mixtures, it was observed that the bacteria consumed acetate first, followed by propionate and then butyrate. It was also found that the consumption rate of the main substrate significantly increased when the minor substrates were depleted.     

Three-dimensional numerical modeling on high pressure membrane reactors for high temperature water-gas shift reaction

This study presents a three-dimensional numerical model that simulates the H₂ production from coal-derived syngas via a water-gas shift reaction in membrane reactors. The reactor was operated at a temperature of 900 °C, the typical syngas temperature at gasifier exit. The effects of membrane permeance, syngas composition, reactant residence time, sweep gas flow rate and steam-to-carbon (S/C) ratio on reactor performance were examined. Using CO conversion and H₂ recovery to characterize the reactor performance, it was found that the reactor performance can be enhanced using higher sweep gas flow rate, membrane permeance and S/C ratio. However, CO conversion and H₂ recovery limiting values were found when these parameters were further increased. The numerical results also indicated that the reactor performance degraded with increasing CO₂ content in the syngas composition.     

Photofermentative hydrogen production using dark fermentation effluent of sugar beet thick juice in outdoor conditions

In the present study, photofermentative hydrogen production on thermophilic dark fermentation effluent (DFE) of sugar beet thick juice was investigated in a solar fed-batch panel photobioreactor (PBR) using Rhodobacter capsulatus YO3 (hup⁻) during summer 2009 in Ankara, Turkey. The DFE was obtained by continuous dark fermentation of sugar beet thick juice by extreme thermophile Caldicellulosiruptor saccharolyticus and it contains acetate (125 mM) and NH₄⁺ (7.7 mM) as the main carbon and nitrogen sources, respectively. The photofermentation process was done in a 4 L plexiglas panel PBR which was daily fed at a rate of 10% of the PBR volume. The DFE was diluted 3 times to adjust the acetate concentration to approximately 40 mM and supplemented with potassium phosphate buffer, Fe and Mo. In order to control the temperature, cooling was provided by recirculating chilled water through a tubing inside the reactor. Hydrogen productivity of 1.12 mmol/L_c/h and molar yield of 77% of theoretical maximum over consumed substrate were attained over 15 days of operation. The results indicated that Rb. capsulatus YO3 could effectively utilize the DFE of sugar beet thick juice for growth and hydrogen production, therefore facilitating the integration of the dark and photo-fermentation processes for sustainable biohydrogen production.     

Development of a simple bio-hydrogen production system through dark fermentation by using unique microflora

In order to ensure efficient functioning of hydrogen fermentation systems that use Clostridium as the dominant hydrogen producer, energy-intensive process such as heat pretreatment of inoculum and/or substrate, continuous injection, and control of anaerobic conditions are required. Here, we describe a simple hydrogen fermentation system designed using microflora from leaf-litter cattle-waste compost. Hydrogen and volatile fatty acid production was measured at various hydraulic retention times, and bacterial genera were determined by PCR amplification and sequencing. Although hydrogen fermentation yield was approximately one-third of values reported in previous studies, this system requires no additional treatment and thus may be advantageous in terms of cost and operational control. Interestingly, Clostridium was absent from this system. Instead, Megasphaera elsdenii was the dominant hydrogen-producing bacterium, and lactic acid-producing bacteria (LAB) were prevalent. This study is the first to characterize M. elsdenii as a useful hydrogen producer in hydrogen fermentation systems. These results demonstrate that pretreatment is not necessary for stable hydrogen fermentation using food waste.     

Effect of temperature on continuous hydrogen production of cellulose

The effect of temperature on the hydrogen fermentation of cellulose was evaluated by a continuous experiment using a mixed culture without pretreatment. The experiments were conducted at three different temperatures, which were mesophilic [37 ± 2 °C], thermophilic [55 ± 2 °C] and hyper-thermophilic [80 ± 2 °C], with an influent concentration of cellulose of 5 g/l and a hydraulic retention time [HRT] of 10 days. A stable hydrogen production was observed at each condition. At 37 ± 2 °C, the maximum hydrogen yield was 0.6 mmol H₂/g cellulose. However, at 55 ± 2 °C and 80 ± 2 °C, the maximum hydrogen yields were 15.2 and 19.02 mmol H₂/g cellulose, respectively. While 26% of the biogas was methane under the mesophilic temperature, no methane gas was detected under both the thermophilic and hyper-thermophilic temperatures. The results show that operational temperature is a key to sustainable bio-hydrogen production and that the thermophilic and hyper-thermophilic conditions produced better results than mesophilic condition.