Photo-hydrogen production rate of a PVA-boric acid gel granule containing immobilized photosynthetic bacteria cells

The polyvinyl alcohol (PVA)-boric acid gel granule facilitates the light penetration and mass transport as it has the features of the transparency and adequate porous structure. In this work, a hydrogen production bioreactor with the indigenous photosynthetic bacteria (PSB) Rhodopseudomonas palustris CQK 01 immobilized in a PVA-boric acid gel granule is developed to enhance the rate of photo-hydrogen production. Particular attention is paid to exploring the effects of illumination wavelength and intensity, as well as the effects of concentration, flow rate, pH, and temperature of influent substrate solution on the hydrogen production rate. The immobilized PSB gel granule exhibited the maximum hydrogen production rate of 3.6 mmol/g cell dry weight/h in all tests. The experimental results show that the hydrogen production rate of an immobilized PSB granule illuminated at 590 nm is distinctly higher than that at 470 and 630 nm. Photo-inhibition of the gel granule occurs as the long-wavelength illumination intensity exceeds 7000 lux. In addition, there exists an optimal pH of 7.0 and temperature of 30 °C for PSB immobilized in the granule to produce hydrogen. More importantly, the feasibility of PSB immobilized in the PVA-boric acid gel granule for the enhancement of the photo-hydrogen production is demonstrated.     

Bubble behavior and photo-hydrogen production performance of photosynthetic bacteria in microchannel photobioreactor

A transparent microchannel photobioreactor was manufactured to visualize the colony formation of photosynthetic bacteria (PSB), Rhodopseudomonas palustris CQK 01, as well as the biogas bubble behavior within the microstructure. The results showed that the formation of PSB colony in the interior of microchannels can be divided into four stages: bacteria absorption, bacteria reproduction, morphological transformation and colony formation. It was founded that the microchannel vents immobilized by PSB colony was the favorable sites for the emergence of biogas bubbles. In this work, the effects of substrate concentration and flow rate of the influent solution as well as illumination wavelength and intensity on the photo-hydrogen production performance of the bioreactor were also investigated. The microchannel photobioreactor exhibited a maximal hydrogen production rate of 1.48 mmol/g cell dry weight/h, maximal hydrogen yield of 0.91 mol H₂/mol glucose in all tests at an optimal inlet medium flow rate of 2.8 ml/h and substrate concentration of 50 mmol/l. In addition, photobioreactor showed a highest performance of hydrogen production and substrate consumption at 590 nm illumination wavelength and 5000 lx illumination intensity.     

Mathematical modeling of two-phase flow and transport in an immobilized-cell photobioreactor

A one-dimensional two-phase flow and transport model is presented for a packed bed photobioreactor with transparent gel granules containing immobilized photosynthetic bacterial cells. The inherently coupled two-phase flow and mass transport, along with the biochemical reactions occurring in the photobioreactor are taken into account. The source term in the species conservation equation of the substrate is derived from a local transport model for a single gel granule. Model predictions of the glucose consumption efficiency and hydrogen production rate are in good agreement with experimental data. The results show that the photoinhibition of immobilized cells appears at incident light intensities higher than 6000 lux. It is the most suitable for photo-hydrogen production under neutral conditions and 30 °C of the influent substrate solution. Moreover, a high influent substrate solution flow rate results in a large hydrogen production rate due to the improved substrate transport from the bulk solution to gel granules.     

Hydrogen production by Rhodopseudomonas palustris CQK 01 in a continuous photobioreactor with ultrasonic treatment

An ultrasonic treatment technique was applied to a continuous photobioreactor with Rhodopseudomonas palustris CQK 01 suspension to enhance photo-hydrogen production performance. After the start-up period, R. palustris CQK 01 suspension in the photobioreactor was intermittently agitated by ultrasonic wave with a frequency of 20 kHz and then the hydrogen production performance was evaluated. The ultrasonic agitation significantly dropped the hydrogen concentration in the suspension from 300 μmol/L to 50 μmol/L and thus increased the hydrogen production rate and hydrogen yield by nearly 2 times as compared with conventional photobioreactor. Furthermore, the effects of the ultrasonic power and time, influent substrate flow rate and concentration were investigated and discussed, respectively. The maximum hydrogen production performance of the continuous photobioreactor with ultrasonic treatment was obtained under the conditions of ultrasonic power 40 W, agitation/interval time 3/7 s, substrate concentration 75 mmol/L and substrate flow rate 40 ml/h, leading to the hydrogen production rate of 1.12 mmol/L/h and hydrogen yield of 0.23 mol-H₂/mol-glucose.     

Performance of a groove-type photobioreactor for hydrogen production by immobilized photosynthetic bacteria

The biofilm technique has been proved to be an effective cell immobilization method for wastewater biodegradation but it has had restricted use in the field of photobiological H₂ production. In the present study, a groove-type photobioreactor was developed and it was shown that a groove structure with large specific surface area was beneficial to cell immobilization and biofilm formation of the photosynthetic bacteria on photobioreactor surface as well as light penetration. A series of experiments was carried out on continuous hydrogen production in the groove-type photobioreactor illuminated by monochromatic LED lights and the performance was investigated. The effects of light wavelength, light intensity, inlet glucose concentration, flow rate and initial substrate pH were studied and the results were compared with those obtained in a flat panel photobioreactor. The experimental results show that the optimum operational conditions for hydrogen production in the groove-type photobioreactor were: inlet glucose concentration 10 g/L, flow rate 60 mL/h, light intensity 6.75 W/m², light wavelength 590 nm and initial substrate pH 7.0. The maximum hydrogen production rate, H₂ yield and light conversion efficiency in the groove-type photobioreactor were 3.816 mmol/m²/h, 0.75 molH₂/molglucose and 3.8%, respectively, which were about 75% higher than those in the flat panel photobioreactor.     

Photo fermentative hydrogen production using dominant components (acetate, lactate, and butyrate) in dark fermentation effluents

Engineering strategies were applied to promote the phototrophic H₂ production of an indigenous purple nonsulfur bacterium Rhodopseudomonas palustris WP3-5 using major components (i.e., acetate, butyrate, and lactate) of dark fermentation effluents as carbon sources. First, performance of cell growth and photo-H₂ production on each carbon source was examined individually. It appeared that acetate was the most effective carbon source for photo-H₂ production, giving an overall H₂ production rate and H₂ yield of 12.68 ml/h/l and 67.1%, respectively. Next, the effect of substrate concentration of each carbon source on photo-hydrogen production was investigated. Kinetic models were developed to describe the correlation between maximum specific growth rate/specific H₂ production rate and the substrate concentration. The results show that using acetate and lactate as the carbon source, the kinetics for the cell growth and photo-hydrogen production can be described by Monod-type and Michaelis–Menten models, respectively, whereas substrate inhibition occurred when using butyrate as the carbon source. The continuous cultures were also conducted at a hydraulic retention time of 96 h using synthetic dark fermentation soluble metabolites (with a 5 and 10 fold dilution) as the influent. The phototrophic H₂ production efficiency was stably maintained for over 30 days with an overall H₂ yield 10.30 and 11.97 mol H₂/mol sucrose, when using 5-fold and 10-fold diluted dark fermentation effluent, respectively, as the substrate for dark fermentation. This demonstrates the feasibility of using the sequential dark and photo fermentation for high-yield biohydrogen production.     

Microbial diversity analysis of long term operated biofilm configured anaerobic reactor producing biohydrogen from wastewater under diverse conditions

This communication provides an insight into the composition of the microbial community survived in the biofilm configured anaerobic reactor operated for biohydrogen (H₂) production using wastewater as substrate under diverse conditions for past four years. PCR amplified 16S rDNA product (at variable V3 region using universal primers 341F and 517R) was separated by using denaturing gradient gel electrophoresis (DGGE) to identify the diversity in microbial population survived. The phyologenetic profile of the bioreactor showed significant diversity in the microbial community where major nucleotide sequences were affiliated to Class Clostridia followed by Bacteroidetes, Deltaproteobacteria and Flavobacteria. Clostridium were found to be dominant in the microbial community observed. The controlled growth conditions, application of pre-treatment to biocatalyst, operation with specific pH and variation in substrate composition are reasoned for the robust acidogenic culture identified in the bioreactor. Most of the operational taxonomic units (OTUs) observed in the bioreactor are capable to undergo acetate producing pathway, feasible for effective H₂ production.     

Improvement of hydrogen production with Rhodopseudomonas palustris CQK-01 by Ar gas sparging

For improving photo-biohydrogen production, a novel gas bubble column photobioreactor with Ar gas sparging was developed for biohydrogen production by purple non-sulfur phototrophic bacteria, Rhodoseudomonas palustris CQK-01. The dissolved hydrogen concentration was in-situ measured by a hydrogen microsensor. Experimental results demonstrated that Ar gas sparging dramatically decreased the dissolved hydrogen concentration, resulting in an improvement in the photo-biohydrogen production performance. Furthermore, effects of the gas flow rate and the time interval of gas sparging were investigated. The results showed that with an increase in the gas flow rate, the hydrogen production performance increased initially due to the reduced dissolved hydrogen concentration and enhanced mass transport, and then it decreased as a result of an increased shear stress. Meanwhile, the short sparging time interval resulted in a low accumulation of dissolved hydrogen in the bioreactor, hence high hydrogen production performance. The optimal hydrogen production rate (5.86 mmol/l/h) and hydrogen yield (3.38 mol H₂/mol glucose) were obtained at the gas flow rate of 10 ml/min, respectively.     

Enhancement of phototrophic hydrogen production by Rhodobacter sphaeroides ZX-5 using fed-batch operation based on ORP level

In this study, a new outer-cycle flat-panel photobioreactor was designed for an anaerobic, photo-fermentation process by Rhodobacter sphaeroides ZX-5. In order to obtain the high hydrogen yield, photo-hydrogen production by fed-batch culture with on-line oxidation-reduction potential (ORP) feedback control was investigated. Meanwhile, the effects of feeding malic acid concentration and pH adjustment on the growth and hydrogen production of R. sphaeroides ZX-5 were studied. In the entire fed-batch culture, biomass (i.e., OD₆₆₀) rapidly increased up to 1.79 within 18 h, and then OD₆₆₀ value stayed constant within a range of 1.85-2.18 until the end of the photo-fermentation. The cumulative hydrogen volumes in each phase of fed-batch process were 2339, 1439, 1328, and 510 ml H₂/l-culture, respectively. Throughout the entire repeated fed-batch photo-fermentation, the maximum substrate conversion efficiency of 73.03% was observed in the first fed-batch process, obviously higher than that obtained from batch culture process (59.81%). In addition, compared to the batch culture, a much higher maximum hydrogen production rate (102.33 ml H₂/l h) was achieved during fed-batch culture. The results demonstrated that photo-hydrogen production using fed-batch operation based on ORP feedback control is a favorable choice of sustainable and feasible strategy to improve phototrophic hydrogen production efficiency.     

Enhancing continuous hydrogen production by photosynthetic bacterial biofilm formation within an alveolar panel photobioreactor

Photofermentative hydrogen production at higher rate is desirable to make the technology of biological hydrogen production in practical application. An easy fabricating alveolar panel photobioreactor with high surface-to-volume ratio was proposed in this study to realize biofilm formation and used for developing a continuous bioprocess of hydrogen production. Effects of key operating parameters, i.e. variation in intensity of incident light, initial concentration of carbon substrate and flow rate on the rate of nitrogenase-based H₂ production were investigated using response surface methodology (RSM) with Box-Behnken design. Surface and contour plots of the fitted regression model revealed that optimum H₂ production rate of 57.6 mL/h/L was obtained at 125.9 μE/m²/s incident light intensity at 590 nm light wavelength, 52.4 mM initial concentration of carbon substrate and 209 mL/h flow rate. Regular groove surfaces within this photobioreactor were considered to have mutual effects on enhancement of continuous hydrogen production by enriching bacterial cell density, enhancing mass transfer of carbon substrate to facilitate release of protons and electrons, enhancing removal of molecular H₂, and uniformly distribution of incident light within the photobioreactor for sufficient conversion into ATPs.     

光合细菌生物膜制氢反应器的产氢特性

对光合细菌生物膜制氢反应器的产氢性能进行了实验研究,探讨了光照度、光谱和底物浓度对反应器产氢性能的影响.实验结果表明:光合细菌生物膜反应器内最适底物浓度为0.12mol/L,最佳光照度为5000 lx,过高或过低的底物浓度和光照度均对光合细菌产氢具有抑制作用;而光的波长对光合细菌产氢的最适底物浓度和最佳光照度均无影响.当底物浓度为0.12mol/L,光照度为5000 lx,波长为590 nm时,反应器的产氢率达到最高,为3.54mg/h.     

The kinetic characterization of photofermentative bacterium Rhodopseudomonas faecalis RLD-53 and its application for enhancing continuous hydrogen production

In this work, the kinetic characterization of hydrogen production by the photofermentative bacteria Rhodopseudomonas faecalis RLD-53 was investigated at different growth phase. During entire fermentation, 89.30% of total biomass was accumulated in exponential growth phase, while hydrogen yield was only 1.82 mol H₂/mol acetate at the expense of 51.25% substrate. In the stationary phase, biomass synthesis was minimal (7.51%), and 38.17% of the substrate was directly converted into hydrogen. As a result, hydrogen (59.19%) was mainly produced in stationary phase with highest hydrogen yield of 3.67 mol H₂/mol acetate. Consequently, bacteria in stationary phase were most effective for hydrogen production. Based on these findings, a novel membrane photobioreactor was developed to retain bacteria during stationary phase in reactor through membrane separation. Maximum rate (32.82 ml/l/h) and yield (3.27 mol H₂/mol acetate) of hydrogen production were achieved using membrane photobioreactor under the continuous operation. Therefore, using bacteria in stationary phase as hydrogen producer can offer considerable benefits for enhancing photo-hydrogen production.     

Effects of vitamins (nicotinic acid, vitamin B1 and biotin) on phototrophic hydrogen production by Rhodobacter sphaeroides ZX-5

The effects of vitamins (nicotinic acid, vitamin B₁ and biotin) on the growth and hydrogen production of Rhodobacter sphaeroides ZX-5 were investigated by batch culture in this study. The results showed that nicotinic acid, as a precursor of NAD⁺/NADH, plays a crucial role in effectively enhancing the phototrophic hydrogen synthesis during photo-fermentation process. Lack of nicotinic acid in hydrogen production medium resulted in the failure of photo-hydrogen production. In addition, though vitamin B₁ and biotin do not have direct impact on photo-hydrogen production, they are still essential and must exist in either growth medium or hydrogen production medium. Without either of them, photo-hydrogen production decreased seriously, regardless of the existence of nicotinic acid.     

Enhancement of hydrogen production by adsorption of Rhodoseudomonas palustris CQK 01 on a new support material

To enhance bacterial adhesion and biofilm activity, a new support material named as SiO₂-Chitosan-Medium Sol is proposed for hydrogen production by photosynthetic bacteria. The physicochemical properties of the support material are comparatively investigated with other four different materials in terms of topography, surface energy and composition. And the biocompatibilities to Rhodoseudomonas palustris CQK 01 for these five support materials are experimentally studied by quantifying the initially adhered cell numbers, biofilm dry-weights, protein and polysaccharide in EPS of the biofilms and the average H₂ production rates. Experimental results show that the proposed support material shows favorable properties like rough surface, amine group and nutrients. The new support material enhances the cell adhesion capacity, reduces the biofilm formation time and improves the biofilm activity. The average H₂ production rate of the bioreactor with proposed support material was improved by 80% compared with the rate of the bioreactor with the unmodified material.     

Enhanced photo-fermentative hydrogen production of Rhodopseudomonas sp. nov. strain A7 by biofilm reactor

To achieve stable and efficient photo-fermentative hydrogen production, this work investigated photo-fermentative hydrogen production by forming biofilm on the surface of carrier in the biofilm reactor (BR). Results showed the hydrogen production performance was greatly improved by formed biofilm. The time of hydrogen production and efficiency of substrate utilization were enhanced obviously compared to the control reactor (CR). When the CR was used, hydrogen production stopped at 7th day and maximum cumulative hydrogen volume and hydrogen yield were 1730 ± 87 mL/L and 1.44 ± 0.07 mol H₂/mol acetate, respectively. However, in the BR hydrogen production volume of 3028 ± 150 mL/L and hydrogen yield of 2.52 ± 0.13 mol H₂/mol acetate were obtained, which were enhanced about 75% compared to that of the CR. The time of hydrogen production extended from 7 days of CR to 12 days of BR and the substrate conversion efficiency increased from 36% of CR to 63% of BR. It was worth noting at 8th day that substrate was almost utilized completely but hydrogen production still lasted for 4 days. This suggested that the formation of biofilm in BR was favorable to continuous hydrogen production and substrate utilization with high efficiency. Results demonstrated the BR can get a more stable and consistent operating process and it was a proper and potential way to produce hydrogen by photo-fermentative bacteria (PFB).     

Critical assessment of anaerobic processes for continuous biohydrogen production from organic wastewater

Production of biohydrogen using dark fermentation has received much attention owing to the fact that hydrogen can be generated from renewable organics including waste materials. The key to successful application of anaerobic fermentation is to uncouple the liquid retention time and the biomass retention time in the reactor system. Various reactor designs based on biomass retention within the reactor system have been developed. This paper presents our research work on bioreactor designs and operation for biohydrogen production. Comparisons between immobilized-cell systems and suspended-cell systems based on biomass growth in the forms of granule, biofilm and flocs were made. Reactor configurations including column- and tank-based reactors were also assessed. Experimental results indicated that formation of granules or biofilms substantially enhanced biomass retention which was found to be proportional to the hydrogen production rate. Rapid hydrogen-producing culture growth and high organic loading rate might limit the application of biofilm biohydrogen production, since excessive growth of fermentative biomass would result in washout of support carrier. It follows that column-based granular sludge process is a preferred choice of process for continuous biohydrogen production from organic wastewater, indicating maximum hydrogen yield of 1.7 mol-H₂/mol-glucose and hydrogen production rate of 6.8 L-H₂/L-reactor h.     

紫花苜蓿酶解光合生物制氢工艺正交优化实验

为了研究紫花苜蓿酶解液作为光合制氢原料时的最佳产氢工艺条件,对产氢影响较为显著的温度、光照强度、初始pH值三种因素设计了三因素三水平L9(33)正交实验,并对实验结果进行直观分析与方差分析,以获取最佳产氢工艺条件。实验结果表明:在所选水平范围内,各因素对能源草产氢的影响主次顺序为温度→光照强度→初始pH值;由方差分析可知,温度和光照强度对能源草光合产氢性能影响为显著;初始pH值为不显著;由各因素水平值的均值可见,能源草光合生物制氢的最佳工艺水平为30℃、pH=7、光照度为3 000 lx。     

Light shade bands for the improvement of solar hydrogen production efficiency by Rhodobacter sphaeroides RV

At mid-day, the sunlight provides excessive energy for photo-hydrogen production by photosynthetic bacteria. Conversion efficiency from light energy to hydrogen decreased under high illumination. To overcome this problem, we examined a method to spatial dispersion of the high illumination. The new photobioreactor using the light shade bands set on the surface of the reactor was developed for efficient hydrogen production. Spatial dispersion gave remarkable effect on conversion efficiency under the excessive light condition. Indoors, the 1.0 cm width of light shade bands gave the best conversion efficiency (2.1%). Actual use of the sunlight, the 1.5 cm width of light shade bands provided the best conversion efficiency (1.4%). Light inhibition was successfully suppressed by the light shade bands in both experiments. The dispersed light energy could be used for other energy conversion device as solar cells.     

Optimal operational conditions for biohydrogen production from sugar refinery wastewater in an ASBR

The performance of biohydrogen production in an anaerobic sequencing batch reactor (ASBR) was evaluated with respect to variations in the key operational parameters – pH, hydraulic retention time HRT, and organic loading rate OLR using sugar refinery wastewater as substrate. Analysis of variance (ANOVA) indicated HRT had less significant influence on hydrogen content and yield in comparison to pH and OLR, whereas OLR has much impact on hydrogen production rate. Taxonomic analysis results showed that diverse bacterial species contributed to hydrogen production and the dominant species in the bioreactor were governed by all operational parameters. Even without pretreatment of the seed sludge, a high proportion of Clostridium spp. over the other bacterial species was observed at pH 5.5, and this is compatible with the high hydrogen productivity. Consequently, pH 5.5, HRT 10 h, and OLR 15 kg/m³ d were delineated as the optimal operational conditions for an ASBR fed with sugar refinery wastewater.     

Hydrogen in education—a biological approach

Hydrogen gas is regarded as a potential candidate for a future energy economy. The change towards new energy systems with hydrogen gas as an energy carrier will have an immense impact on society. Thus, an integrate part of current research and development must be the inclusion of the new technology into public education. By means of a model bioreactor for light-dependent (photobiological) production of hydrogen gas with green algae, we try to serve this goal in biological education.Various simple photo-bioreactor types (closed batch, open batch) were analyzed for their capability to produce hydrogen under different conditions. The focus laid on functionality and simplicity rather than on high efficiency. Easy-to-handle systems that can be used in the classroom are presented. In a more sophisticated version a proton exchange membrane (PEM-) fuel cell was connected to a continuous gas flow tube bioreactor. We developed a software interface, designed to read light intensity, temperature and power generation by the bioreactor and the connected fuel cell, respectively. Thus, this bioreactor is specially aimed at integrative teaching in natural science and computer technology at middle and high school level.