Evaluation of the effect of aerobic–anaerobic conditions on photohydrogen and chlorophyll a production by environmental Egyptian cyanobacterial and green algal species

In this study, we tested four algal species (Synechocystis sp. PCC 6803 and three novel algal species isolated from Egyptian paddy rice soil, having high 16S rRNA gene sequence identity to Nostoc spongiaeforme, Parachlorella kessleri SAG 211-11 and Nostoc sp. PCC 7524) under aerobic, anaerobic and 3% CO₂-supplemented anaerobic condition. Significant changes in photohydrogen production, morphology, chlorophyll a and protein content/pattern were observed in all species, when grown in these different conditions. H₂ production was higher in anaerobic condition in all species with the highest H₂ production rate of 4 mmol H₂ mg Chla^?1 h^?1 at 24 h in Synechocystis sp. In contrast, Chla content and protein content decreased (%) in N. spongiaeforme, (29%, 58%), P. kessleri (47%, 7%) and Nostoc sp. (59%, 65%).The anaerobic condition with 3% CO₂ stimulated early production of H₂ in all species except Synechocystis sp. Our results compared all selected algal species under different growth conditions for the screening of a superior H₂-producing algal species that can help to address engineering challenges in the field of large-scale H₂ photoproduction by microalgae.     

Performance of batch solid state fermentation for hydrogen production using ground wheat residue

Ground wheat (21 g) was subjected to batch solid state dark fermentation for bio-hydrogen production. Clostridium acetobutylicum (B-527) was used as the culture of dark fermentation bacteria at mesophilic conditions. Effects of moisture content on the rate and yield of bio-hydrogen formation were investigated. The highest CHF (1222 ml), hydrogen yield (63 ml H₂ g^?1 starch), formation rate (10.64 ml H₂ g^?1 starch h^?1) and specific hydrogen formation rate (0.28 ml H₂ g^?1 biomass h^?1) were obtained with a moisture content of 80%. Nearly complete starch hydrolysis and glucose fermentation were achieved with more than 80% moisture content and the highest substrate conversion rate (21.9 mg L^?1 h^?1) was obtained with 90% moisture content at batch solid state fermentation producing volatile fatty acids (VFA) and H₂.     

Prolongation of H2 production during mixed carbon sources fermentation in E. coli batch cultures: New findings and role of different hydrogenases

Hydrogen (H₂) gas production in batch cultures was studied upon utilization of the mixture of glucose, glycerol and formic acid by Escherichia coli BW25113 wild type (wt) at pH of 5.5–7.5. At pH 7.5H₂ was continuously produced during 240 h but at pH 6.5 and 5.5 it was detected till 168 h and 120 h, respectively. Specific growth rate (μ) of wt was the highest (1.05 h^?1) at pH 6.5. Moreover, at pH 5.5 in hycE μ decreased by ～4.14 fold compared to wt, suggesting major role of Hyd-3 in cell growth. H₂ yield (8.8 mmol H₂ L^?1) was the highest at pH 7.5. In hybC H₂ yield was increased ～1.62 fold than in wt. These data might be applied for biomass and biohydrogen production from various organic wastes where mixtures of carbon sources are present.     

Scale-up studies for stable,long-term indoor and outdoor production of hydrogen by immobilized Rhodobacter capsulatus

Photofermentative hydrogen production by immobilized Rhodobacter capsulatus YO3 was carried out in a novel photobioreactor in sequential batch mode under indoor and outdoor conditions. Long-term H₂ production was realized in a 1.4 L photobioreactor for 64 days using Rhodobacter capsulatus YO3 immobilized with 4% (w/v) agar on 5 mM sucrose and 4 mM glutamate. The highest hydrogen yield (19 mol H₂/mol sucrose) and hydrogen productivity (0.73 mmol H₂ L^?1 h^?1) were achieved indoors on 5 mM sucrose. The effect of initial sucrose concentration (5 mM, 10 mM, and 20 mM) on hydrogen production was also investigated. Sustained hydrogen production was carried out under natural, outdoor conditions as well. For the outdoor experiments, the highest hydrogen productivity and yield were obtained as 0.87 ± 0.06 mmol H₂ L^?1 h^?1 and 6.1 ± 0.2 mol H₂/mol sucrose, respectively on 10 mM sucrose. Furthermore, this system prevented sudden pH drops and fluctuations caused by the utilization of sucrose throughout the process. These results demonstrate that a proper immobilization setup can lead to long-term efficient and robust hydrogen production even under naturally varying conditions.     

Biological hydrogen production from sugar beet molasses by agar immobilized R. capsulatus in a panel photobioreactor

Biohydrogen production from sugar beet molasses was investigated by using agar immobilized R. capsulatus YO3. A panel photobioreactor (1.4 L) was employed for a long-term hydrogen production in both indoor and outdoor conditions. The impact of several initial molasses concentrations on hydrogen production, yield and productivity were assessed. Indoor studies revealed that initial sucrose concentration in molasses should be kept below 20 mM to prevent inhibition of hydrogen production. The highest hydrogen productivity of 0.64 ± 0.06 mmol H₂ L^?1 h^?1 and yield of 12.2 ± 1.5 mol H₂/mol sucrose were obtained in indoors throughout 20 days of operation. For outdoors, hydrogen production continued for 40 days including consecutive 10 rounds under natural outdoor conditions. In outdoor conditions, the maximum hydrogen productivity and yield were 0.79 ± 0.04 mmol H₂ L^?1 h^?1 and 5.2 ± 0.4 mol H₂/mol sucrose respectively. These results indicate that the proposed system is promising for biohydrogen production from molasses at large-scale natural conditions.     

Isochrysis sp. IOAC724S,a newly isolated,lipid-enriched,marine microalga for lipid production,and optimized cultivation conditions

An oleaginous, unicellular, marine microalga termed IOAC724S was isolated from the South China Sea. Morphology and genetic analyses indicated it belongs to the genus Isochrysis. Gas chromatography (GC) results showed that more than 10 types of fatty acids existed in Isochrysis sp. IOAC724S and that 90% of them were suitable for lipid production. The culture conditions suitable for cell growth were progressively optimized through photosynthetic and respiratory analyses. The optimal culture conditions were: photon flux 200–500 μmol m⁻² s⁻¹, temperature 35 °C during daytime and 24 °C at night, pH value between 7 and 8, NaNO₃ 160 g m⁻³ and NaH₂PO₄·2H₂O 80 g m⁻³ for starting culture. When microalgal cultures were exposed to these optimal conditions, the specific growth rate reached to 0.26 d⁻¹ on average and 1.0 d⁻¹ in MAX. Lipid production was optimized through nutrient starvation processes, including nitrate or phosphate deprivation and simultaneous nitrate and phosphate deprivation. The highest lipid mass fraction of dry cell weight (about 55.6%) was obtained after the stationary phase algal culture was transferred into phosphate-free medium for 3 days. GC data demonstrated that the enhancement of lipid accumulation in algal cells maintained under nutrient starvation came mainly from an increase of C16:0 and C18:1 fatty acids; however, the lipids with a chain length appropriate for fuel use (C14 to C18) were unchanged at 90% mass fraction of the dry cell weight. Based on these good characteristics, Isochrysis sp. IOAC724S appeared to be a strong candidate for lipid production.     

Hydrogen-rich gas production from pyrolysis of wet sludge in situ steam agent

A series of wet sludge samples with different moisture contents were pyrolyzed in situ steam in a bench-scale fixed bed reactor in order to examine the influence of moisture and temperature on product distribution and gas composition. The results demonstrated that inherent moisture in wet sludge had a great effect on the product yield. The pyrolysis of wet sludge (43.38% moisture content) at 800 °C exhibited maximum H₂ yield (7.76 mol kg^?1 dry basis wet sludge) and dry gas yield (0.61 Nm³ kg^?1) and H₂ content of 42.13 vol%. When the moisture exceeded 43.38%, H₂ yield and gas yield both tended to decline. It was also shown that the elevated temperature exhibited a significant influence on gas content increase and tar reduction; at the same time, H₂ yield and H₂ content were increased from 1.83 mol kg^?1 dry basis wet sludge and 16.67 vol% to 9.15 mol kg^?1 dry basis wet sludge and 45.67 vol%, respectively, as temperature increased from 600 °C to 850 °C. LHV of fuel gas varies from 15.49 MJ Nm^?3 to 11.65 MJ Nm^?3 because of decrease in CH₄ and C₂H₄ content as temperature increasing. In conclusion, hydrogen rich gas production by pyrolysis of wet sludge which avoided pre-drying process and utilized in situ steam agent from wet sludge is an economic method.     

Continuous photosynthetic biohydrogen production from acetate-rich wastewater: Influence of light intensity

Photo-biohydrogen by microalgae is attractive sustainable energy caused by the utilization of solar energy and water. However, due to oxygen (O₂) sensitive hydrogenase (HydA) activity, effective control of O₂ and light intensity is critical for achieving sustainable photosynthetic hydrogen (H₂) production. Here we demonstrate continuous algal H₂ production using acetate-enriched fermenter effluent, achieving the complete O₂ cessation without sulfur depletion. Average H₂ production of 108 ± 4 μmol L^?1 for Chlamydomonas reinhardtii and 88 ± 7 μmol L^?1 for Chlorella sorokiniana at 100 μmol m^?2 s^?1 were observed for 15 days, respectively. The highest light energy to H₂ energy conversion efficiency (LHCE) of 1.61% for C. reinhardtii and 1.06% for C. Sorokiniana was obtained under low light intensity (50 μmol m^?2 s^?1) but the LHCE decreased with the increase of light intensity followed by photoinhibition, which led to a decrease of HydA activity and H₂ production. Low H₂ production was observed at 50 μmol m^?2 s^?1 under the highest LHCE, in which microalgae exhibited photoinhibition biomass growth kinetics to produce chlorophyll a (Chl a) for electron generation. These results demonstrate that light is a feasible strategy for producing electron for H₂ production under anoxygenic photosynthesis.     

Photoautotrophic hydrogen production by nitrogen-deprived Chlamydomonas reinhardtii cultures

In this study we have demonstrated the possibility of phototrophic hydrogen production in C. reinhardtii under N-deprived conditions. When tested under air + CO₂, and Ar + CO₂ N-deprived C. reinhardtii demonstrated decrease in PSII activity mainly due to over reduction of PQ, in addition no ascorbate accumulation was observed in cells. Under air + CO₂ atmosphere cells accumulated excessive amounts of starch. When incubated under Ar + CO₂ atmosphere cells accumulated starch as nitrogen replete cultures and no hydrogen production was observed. Hydrogen production (86 ml H₂ per one l of culture) occurred under Ar + CO₂ atmosphere when particular two-step illumination protocol was implicated. In oxygen producing and early oxygen consuming stage cells were illuminated under light intensity 169 μE m^?2 s^?1. When light was switched to 30 μE m^?2 s^?1, cultures quickly respired all oxygen and transient to anaerobic conditions with subsequent hydrogen production 2 h later. Actual quantum yield of C. reinhardtii cultures was measured in photobioreactor and maximal quantum efficiency of PSII of dark adapted cells together with JIP test were studied.     

Reinforced photocatalytic reduction of CO2 to fuel by efficient S-TiO2: Significance of sulfur doping

The photocatalytic reduction of CO₂ to valuable chemicals and fuels is an efficient approach to control the ever-rising CO₂ level in the atmosphere. The present paper describes a significant improvement in photoreduction of carbon dioxide (CO₂) using sulfur (S) doped titania (S-TiO₂) nanoparticles as a photocatalyst under UV-A and visible light irradiation. The sulfur doping was done by following a simple sonothermal method, and a series of photocatalysts were synthesized with the varied amount of S doping. Various characterization techniques were employed for the photocatalysts such as XRD, surface area, UV–Visible, SEM, TEM, and XPS. The XPS reveals that S is predominantly present as S⁴⁺ in S-TiO₂. The electronic structure for S-TiO₂ anatase was calculated with the Vienna ab initio simulation package (VASP) code in the framework of spin-polarized density functional theory. Additional states closer to the valence band are produced inside the band gap as a result of doping. In situ reductive reaction conditions can partially reduce the catalyst, and results in the shift of Fermi level into the conduction band. It is suggested that S-doping increases catalyst surface conductivity, improves the charge transfer rate and the rate of photocatalytic reactions. The prepared series of catalysts have shown excellent activity under UV-A and visible light for photocatalytic reduction of CO₂. The effect of the different base including K₂CO₃, Na₂CO₃, NaOH and KOH; catalyst amount; sulfur doping amount; and light wavelength were monitored. Methane, ethylene, propylene, and propane were observed as reaction products. In 24 h, S-TiO₂ exhibited the highest photoactivity in KOH aqueous solution with a maximum yield of 6.25 μmol g^?1 methane, 2.74 μmol g^?1 of ethylene, 0.074 μmol g^?1 of propylene and 0.030 μmol g^?1 of propane under UV-A irradiation. The catalysts were active in visible light and able to generate methane and methanol in acetonitrile-H₂O mixture with/without TEOA as sacrificial donor producing 846.5 μmol g^?1 of methane and 4030 μmol g^?1 of methanol for the former and 167.6 μmol g^?1 of methane and 12828.4 μmol g^?1 of methanol for the latter case. An estimate demonstrates that mass transfer does not limit the CO₂ reaction.     

Costless and huge hydrogen yield by manipulation of iron concentrations in the new bacterial strain Brevibacillus invocatus SAR grown on algal biomass

There is a growing global demand on bio-hydrogen production (BHP) using costless and wastes material. Herein we demonstrate the possibility to produce high yield of hydrogen using a new bacterial strain grown on acidic hydrolyzed cyanobacterial biomass as a costless carbon feedstock under various iron concentrations. We used E. coli DH701 mutant and new strain Brevibacillus invocatus SAR isolated from Assiut city soil samples. The mentioned new strain was identified morphologically, biochemically and by molecular analysis using 16S rDNA sequence. Limitation of iron induced BHP in tested cyanobacteria. Iron concentration (0.045 mM) enhanced hydrogenase activity and cumulative hydrogen evolution in the investigated cultures. B. invocatus yielded 3.3 mol H₂/mole glucose and 3.8 mol H₂/mole reducing sugar (algal biomass), while the mutant strain yielded 1.78 mol H₂/mole glucose and 3.4 mol H₂/mole reducing sugar (algal biomass). Thus, the use of algal biomass induced higher potency of BHP especially at 0.045 mM iron.     

Photohydrogen production from lactose and lactate by recombinant strains of Rhodobacter capsulatus: Modeling and optimization

Photofermentative hydrogen production from synthetic mixtures of lactose and lactate mimicking cheese whey was modeled and optimized using Design of Experiments and Response Surface Methodology. Five continuous parameters (light intensity, pH, lactose, lactate and glutamate concentrations) were studied as a function of buffer type (KPi or Borax) using two recombinant bacterial strains. For Rhodobacter capsulatus B10(lacZ), buffer type influenced the optimal parameter values but the optimal responses were similar in both buffers. In contrast, for R. capsulatus IR3(lacZ), responses were higher in Borax buffer than in KPi and were significantly higher than in strain B10(lacZ). Thus, the experimental optimized responses for specific volumetric H₂ production, volumetric H₂ production rate and substrate (lactose plus lactate) to H₂ conversion rate in Borax buffer, were 12,150 ml L^?1, 48.5 ml L^?1 h^?1 and 41.2%, respectively, for IR3(lacZ) compared to 6150 ml L^?1, 33.5 ml L^?1 h^?1 and 32.5%, respectively, for B10(lacZ).     

Inoculum pretreatment differentially affects the active microbial community performing mesophilic and thermophilic dark fermentation of xylose

The influence of different inoculum pretreatments (pH and temperature shocks) on mesophilic (37 °C) and thermophilic (55 °C) dark fermentative H₂ production from xylose (50 mM) and, for the first time, on the composition of the active microbial community was evaluated. At 37 °C, an acidic shock (pH 3, 24 h) resulted in the highest yield of 0.8 mol H₂ mol^?1 xylose. The H₂ and butyrate yield correlated with the relative abundance of Clostridiaceae in the mesophilic active microbial community, whereas Lactobacillaceae were the most abundant non-hydrogenic competitors according to RNA-based analysis. At 55 °C, Clostridium and Thermoanaerobacterium were linked to H₂ production, but only an alkaline shock (pH 10, 24 h) repressed lactate production, resulting in the highest yield of 1.2 mol H₂ mol^?1 xylose. This study showed that pretreatments differentially affect the structure and productivity of the active mesophilic and thermophilic microbial community developed from an inoculum.     

Assessment of the adequacy of different Mediterranean waste biomass types for fermentative hydrogen production and the particular advantage of carob (Ceratonia siliqua L.) pulp

The conversion of agro-industrial byproducts, residues and microalgae, which are representative or adapted to the Mediterranean climate, to hydrogen (H₂) by C. butyricum was compared. Five biomass types were selected: brewery’s spent grain (BSG), corn cobs (CC), carob pulp (CP), Spirogyra sp. (SP) and wheat straw (WS). The biomasses were delignified and/or saccharified, except for CP which was simply submitted to aqueous extraction, to obtain fermentable solutions with 56.2–168.4 g total sugars L^?1. In small-scale comparative assays, the H₂ production from SP, WS, CC, BSG and CP reached 37.3, 82.6, 126.5, 175.7 and 215.8 mL (g biomass)^?1, respectively. The best fermentable substrate (CP) was tested in a pH-controlled batch fermentation. The H₂ production rate was 204 mL (L h)^?1 and a cumulative value of 3.9 L H₂ L^?1 was achieved, corresponding to a H₂ production yield of 70.0 mL (g biomass)^?1 or 1.6 mol (mol of glucose equivalents)^?1_. The experimental data were used to foresight a potential energy generation of 2.4 GWh per year in Portugal, from the use of CP as substrate for H₂ production.     

Enhanced hydrogen production through co-cultivation of Chlamydomonas reinhardtii CC-503 and a facultative autotrophic sulfide-oxidizing bacterium under sulfurated conditions

Photoproduction of H₂ using microalgae has been considered as a promising approach for developing sustainable hydrogen energy. The algae C. reinhardtii CC-503 was co-cultured with a facultative autotroph sulfur-oxidizing bacterium Thuomonas intermedia BCRC 17547 to improve H₂ production. The maximum H₂ production of co-culture at sulfur deficiency conditions was 122 μmol/mg Chl with algae/bacteria ratio as 60:1, which was 2.8-fold higher than that of the pure algal culture. Na₂S₂O₃ treatment can result in a maximum H₂ photoproduction rate of 255 μmol/mg Chl, which was 5.9 and 2.1 times higher than those of pure algae culture and co-culture without Na₂S₂O₃. Co-cultivation under sulphate condition can also significantly increase the biomass, respiratory rate, starch content and hydrogenase activity of C. reinhardtii. By supplement of Na₂S₂O₃, persistent (52 days) H₂ production of bacteria/algae co-culture can be achieved. Our results demonstrated that co-culture of C. reinhardtii CC-503 and bacteria BCRC17547 is a cost-effective strategy for improving photobiological H₂ production.     

Improved photocatalytic H2 production assisted by aqueous glucose biomass by oxidized g-C3N4

Chemically modified g-C₃N₄ for the photocatalytic H₂ evolution from water was explored. Bulk g-C₃N₄ was treated in hot HNO₃ aqueous solution to obtain the oxidized material (o-g-C₃N₄), tested in water containing glucose as model water-soluble sacrificial biomass, using Pt as co-catalyst, under simulated solar light. The behaviour of o-g-C₃N₄ was studied in relation with catalyst amount, Pt loading, glucose concentration. Results showed that H₂ production is favoured by increasing glucose concentration up to 0.1 M and Pt loading up to 3 wt%, and it resulted strongly enhanced using small amount of o-g-C₃N₄ (0.25 g L^?1). o-g-C₃N₄ possesses superior photocatalytic activity (～26-fold higher) compared to pristine g-C₃N₄, with H₂ evolution further improved by ultrasound-assisted exfoliation and evolution rates up to ca. 1370 μmol h^?1 per gram of catalyst, with excellent reproducibility (RSD < 6%, n = 3). Significant production was observed also in river water and seawater, with results far better (up to ca. 2500 μmol g^?1 h^?1) compared to commercial AEROXIDE^® P25 TiO₂ under natural solar light.     

New cyanobacterial strains for biohydrogen production

Under certain conditions, cyanobacteria can switch from photosynthesis to hydrogen production, which is a good energy carrier. However, the biological diversity of hydrogen-releasing cyanobacteria has a great unexplored potential. This study is aimed to investigate the ability of new strains of cyanobacteria Cyanobacterium sp. IPPAS B-1200, Dolichospermum sp. IPPAS B-1213, and Sodalinema gerasimenkoae IPPAS B-353 to release H₂ and to evaluate the effects of photosystem II inhibitor 3-(3,4-dichlorphenyl)-1,1-dimethylurea (DCMU) on H₂ production under light and dark conditions. The results showed that cultures treated with DCMU produced several times more H₂ than untreated cells. The highest rate of H₂ photoproduction of 4.24 μmol H₂ (mg Chl a h)^?1 was found in a Dolichospermum sp. IPPAS B-1213 culture treated with 20 μM DCMU.     

Continuous production of biohydrogen from oil palm empty fruit bunch hydrolysate in tubular photobioreactors

Production of hydrogen by the photosynthetic bacterium Rhodobacter sphaeroides was compared in continuously operated tubular photobioreactors illuminated by natural outdoor sunlight (0.15–66 klux; diurnal cycle) and constant indoor artificial light (10 klux; tungsten lamps). In both cases the operating temperature was 35 °C and the organic carbon source was an acid hydrolysate of oil palm empty fruit bunch (EFB), an agroindustrial waste. In the outdoor photobioreactor, under the best production conditions, the daytime feeding rate of the mixed carbon substrate was 48 mL h^?1 and the average pseudo-steady state hydrogen production rate was 36 mL H₂ L^?1 medium h^?1. The cumulative hydrogen production was 430 mL H₂ L^?1 medium. For the indoor photobioreactor fed at the same rate as the outdoor system, the steady state average hydrogen production rate was 43 mL H₂ L^?1 h^?1 and the cumulative hydrogen production was 517 mL H₂ L^?1 medium. Reducing the feed rate to less than 48 mL h^?1, enhanced the biomass concentration, but reduced hydrogen production in both bioreactors. The sunlight-based cumulative hydrogen production was only about 17% less compared to the artificially lit system, but required only 22% of the electrical energy.     

Enhancement of fermentative hydrogen production by Thermotoga maritima through hyperthermophilic anaerobic co-digestion of fruit-vegetable and fish wastes

In this work, different proportions of model fruit and vegetable wastes (MFVW) and acid hydrolyzed fish wastes (AHFW) were used for hydrogen production in a minimum culture medium based on seawater. Experiments were performed in pH-controlled Stirred Tank Reactor (STR) with or without the addition of nitrogen and sulfur sources. The total H₂ production and the maximum hydrogen productivity of T. maritima in the culture medium, containing MFVW and AHFW (45 mmol L^?1 carbohydrates) at a C/N ratio of 12, were 132 mmol L^?1 and 15 mmol h^?1 L^?1, respectively. However, tripling the concentration of carbohydrates to reach a C/N ratio of 22, has increased two times the maximum H₂ productivity (28 mmol h^?1 L^?1) due to the improvement in nutrient balance. The cumulative H₂ production was 285 mmol L^?1, yielding a potential energy generation of 0.1210³ MJ ton^?1 wastes, which could be an interesting alternative for energy recovery.     

Single-stage photo-fermentative hydrogen production from hydrolyzed straw biomass using Rhodobacter sphaeroides

The hydrogen production utilizing photosynthetic and anaerobic bacteria in two-stage approach has many drawbacks, such as shortage of raw materials and complexity of operations. Accordingly, we aimed to develop a simple one-stage H₂ production protocol using the depolymerization of maize straw cellulose as a cheap carbon source. R. sphaeroides HY01 and its mutant (Hup^?) were studied regarding their H₂ production under different culture conditions. Further study using two model sugars, their combination, and straw hydrolysate as carbon sources was conducted to determine the effects of substrate on H₂ production. When using the straw hydrolysate as carbon source, the pH remained in a range of 7.1–7.6, whereas it dropped to 5.4–7.4 when using the model sugars, and the former biomass value was greater. The H₂ production performance of the mutant was significantly better than that of HY01. One-step photo-fermentative H₂ production was superior when using straw hydrolysate as opposed to the simple model sugars, and its yield was up to 4.62 mol H₂·mol^?1 reducing sugar.