Integrated biological hydrogen production

Biological systems offer a variety of ways by which to generate renewable energy. Among them, unicellular green algae have the ability to capture the visible portion of sunlight and store the energy as hydrogen (H₂). They hold promise in generating a renewable fuel from nature's most plentiful resources, sunlight and water. Anoxygenic photosynthetic bacteria have the ability of capturing the near infrared emission of sunlight to produce hydrogen while consuming small organic acids. Dark anaerobic fermentative bacteria consume carbohydrates, thus generating H₂ and small organic acids. Whereas efforts are under way to develop each of these individual systems, little effort has been undertaken to combine and integrate these various processes for increased efficiency and greater yields. This work addresses the development of an integrated biological hydrogen production process based on unicellular green algae, which are driven by the visible portion of the solar spectrum, coupled with purple photosynthetic bacteria, which are driven by the near infrared portion of the spectrum. Specific methods have been tested for the cocultivation and production of H₂ by the two different biological systems. Thus, a two-dimensional integration of photobiological H₂ production has been achieved, resulting in better solar irradiance utilization (visible and infrared) and integration of nutrient utilization for the cost-effective production of substantial amounts of hydrogen gas. Approaches are discussed for the cocultivation and coproduction of hydrogen in green algae and purple photosynthetic bacteria entailing broad utilization of the solar spectrum. The possibility to improve efficiency even further is discussed, with dark anaerobic fermentations of the photosynthetic biomass, enhancing the H₂ production process and providing a recursive link in the system to regenerate some of the original nutrients.     

Truncated chlorophyll antenna size of the photosystems—a practical method to improve microalgal productivity and hydrogen production in mass culture

Unicellular microalgae hold the promise of commercial exploitation in mass culture for hydrogen and biomass production. In any microalgal production system, the achievable photosynthetic productivity and light utilization efficiency of the algae are the single most important factors in the determination of cost. Microalgal mass cultures growing under full sunlight have a low per chlorophyll (Chl) productivity since, at high photon flux densities, the rate of photon absorption by the Chl antenna far exceeds the rate at which photons can be utilized for photosynthesis. Excess photons are dissipated as fluorescence or heat. Up to 80% of absorbed photons could thus be wasted, reducing light conversion efficiencies and cellular productivity to fairly low levels. This shortcoming could possibly be alleviated by the development of microalgal strains with a limited number of Chl molecules in the light-harvesting antenna of their photosystems, i.e., strains that have a truncated Chl antenna size. It is expected that individually, such microalgae will not be able to saturate rates of photosynthesis and, thus, will not be subject to wasteful dissipation of excitation energy. In turn, the productivity of the mass culture will be improved. The method of choice to reach the objective of a “truncated light-harvesting Chl antenna” size (tla) employed DNA insertional and chemical mutagenesis of the unicellular green algae Chlamydomonas reinhardtii and Dunaliella salina, followed by a rigorous screening protocol to identify mutants with a smaller light-harvesting Chl antenna size. Molecular and genetic analyses of isolated tla strains were performed. Biochemical and physiological analyses in terms of photosynthetic productivity and light conversion efficiencies are presented. The results show that a truncated Chl antenna size of PSII is more important than that of PSI in terms of the photosynthetic productivity of a mass culture. A list of genes that confer a “truncated light-harvesting Chl antenna” size to green algae is being compiled for future application in algal hydrogen and biomass production.     

Photobiological production of hydrogen

This literature survey of photobiological hydrogen production covers the period from its discovery in relatively pure cultures during the early 1930s through 1978. The focus is hydrogen production by phototrophic organisms (and their components) which occurs at the expense of light energy and electron-donating substrates. The survey covers the major contributions in the area; however, in many cases, space has limited the degree of detail provided. Among the topics included is a brief historical overview of hydrogen metabolism in photosynthetic bacteria, eucaryotic algae, and cyanobacteria (blue-green algae). The primary enzyme systems, including hydrogenase and nitrogenase, are discussed along with the manner in which they are coupled to electron transport and the primary photochemistry of photosynthesis. A number of in vivo and in vitro photobiological hydrogen evolving schemes including photosynthetic bacterial, green algal, cyanobacterial, two-stage, and cell-free systems are examined in some detail. The remainder of the review discusses specific technical problem areas that currently limit the yield and duration of many of the systems and research that might lead to progress in these specific areas. The final section outlines, in broadest terms, future research directions necessary to develop practical photobiological hydrogen-producing systems. Both whole cell (near- to mid-term) and cell-free (long-term) systems should be emphasized. Photosynthetic bacteria currently show the most promise for near-term applied systems.     

Hydrogen production from biological systems under different illumination conditions

Hydrogen is a natural by-product of several microbial driven biochemical reactions, mainly in anaerobic fermentation processes. In addition, certain microorganisms produce enzymes by which H₂ from water may be obtained if an outside energy source, like sunlight, is provided. Biophotolysis is a biological process which involves solar energy and algae clusters to convert water into hydrogen. Algae pigments absorb solar energy and enzymes in the cell act as catalysts to split water into hydrogen and oxygen. There are many research activities studying hydrogen production from biological systems cyanobacteria and green algae and some studies present a complete outline of the main available pathways to improve the photosynthetic H₂ production [1] and [2].Efficiency (energy produced from hydrogen divided by solar energy) of such processes can be estimated up to 10%. This value has to be increased for a large-scale hydrogen production. The effect of different artificial illumination conditions on H₂ production was studied for green algae cultures (Chlamydomonas reinhardtii). Results will be used to design a high-efficiency photobioreactor for a large-scale hydrogen production.     

Production of H2 by steam reforming in schizochytrium algae oil of cell disruption and extraction via ultrasound method

Algae species like Schizochytrium sp. are resilient photosynthetic factories that produce large quantities of fatty acids that can be converted to biodiesel. These lipids must be extracted from the raw biomass before they can be used as biofuel feedstocks or other bioproducts such as hydrogen. In this paper, hydrogen production by steam reforming of schizochytrium algae oil are studied. The effects of continuous microwave and intermittent microwave, freezing and thawing times of different mass fractions, time of alkali heat and other factors on the ratio of cell disruption and ultrasonic-assisted ethanol extraction are considered. The results show that the highest lipid yield can reach by ultrasonic-assisted ethanol extraction from the schizochytrium algae which are pretreated by alkali heat method. The hydrogen production from steam reforming of the lipids, which are extracted by ethanol with ultrasonic-assisted from algae after alkali thermal pretreated is proposed.     

Growth characteristics,biohydrogen production and photochemical activity of photosystems in green microalgae Parachlorella kessleri exposed to nitrogen deprivation

The biohydrogen (H₂) generation in green microalgae is associated with electron transfer during photosynthesis. In this study, the ability of Parachlorella kessleri RA-002 isolated in Armenia to generate biohydrogen (H₂) in TAP and Tamiya media with and without nitrogen deprivation have been determined. Nitrogen deprivation led to a suppression of the growth rate and a decrease in the content of photosynthetic pigments in algae, as well as to increase of H₂ yield. The highest H₂ yield was found during the growth of algae in TAP and Tamiya media under nitrogen-deprived conditions, which was 4–5 times more compared to control cells grown without nitrogen deprivation. Diuron inhibited this process, indicating that H₂ generation was due to PS II activities. The analysis of the effect of nitrogen-deprived conditions on the activity of photosystem I and II has been carried out. Nitrogen deprivation suppressed of photosynthetic activity of PS II (up to 20% compared to control) that facilitated the formation of anaerobic conditions increasing the total yield of H₂.     

Introducing pyruvate oxidase into the chloroplast of Chlamydomonas reinhardtii increases oxygen consumption and promotes hydrogen production

In an anaerobic environment, the unicellular green algae Chlamydomonas reinhardtii can produce hydrogen (H₂) using hydrogenase. The activity of hydrogenase is inhibited at the presence of molecular oxygen, forming a major barrier for large scale production of hydrogen in autotrophic organisms. In this study, we engineered a novel pathway to consume oxygen and correspondingly promote hydrogen production in Chlamydomonas reinhardtii. The pyruvate oxidase from Escherichia coli and catalase from Synechococcus elongatus PCC 7942 were cloned and integrated into the chloroplast of Chlamydomonas reinhardtii. These two foreign genes are driven by a HSP70A/RBCS2 promoter, a heat shock inducing promoter. After continuous heat shock treatments, the foreign genes showed high expression levels, while the growth rate of transgenic algal cells was slightly inhibited compared to the wild type. Under low light, transgenic algal cells consumed more oxygen than wild type. This resulted in lower oxygen content in sealed culture conditions, especially under low light condition, and dramatically increased hydrogen production. These results demonstrate that pyruvate oxidase expressed in Chlamydomonas reinhardtii increases oxygen consumption and has potential for improving photosynthetic hydrogen production in Chlamydomonas reinhardtii.     

Enhancement of biological hydrogen production using green alga Chlorococcum minutum

It is estimated that the fossil fuel reserves are going to deplete continuously due to extensive usage. In order to cope with this crisis, it is necessary to increase the efforts towards production of biofuels such as biological hydrogen (H₂). It is well-known fact that the biological hydrogen is a clean and ideal energy and liberates high amount of energy per unit mass. Several groups are working for the large scale production of H₂ chemically and also using photosynthetic organisms, but output is not satisfactory. The best way to achieve enhancement of H₂ is through altering the photosynthetic process by applying various stress conditions or by natural selection. In the process of selection, Chlorococcum minutum was found with improved H₂ output when compared to model green alga Chlamydomonas reinhardtii in a massively parallel and competitive high-throughput screen of different green algae. Both the species belongs to class chlorophyceae of green algae and live in fresh water conditions. In extent various light, pH and temperature conditions were applied and achieved the enhancement of H₂ production in this species under in vitro settings. Augmented hydrogenase activity was found in Chlorococcum minutum when compared to model alga and this may be one of the reason behind improved H₂ output. Hence this species may be considered as one of the best species with respect to H₂ production and also this work may be useful for future renewable energy research.     

Mathematical modelling and simulation of the thermo-catalytic decomposition of methane for economically improved hydrogen production

The demand for low-emission hydrogen is set to grow as the world transitions to a future hydrogen economy. Unlike current methods of hydrogen production, which largely derive from fossil fuels with unabated emissions, the thermo-catalytic methane decomposition (TCMD) process is a promising intermediate solution that generates no direct carbon dioxide emissions and can bridge the transition to green hydrogen whilst utilising existing gas infrastructure. This process is yet to see widespread adoption, however, due to the high catalyst turnover costs resulting from the inevitable deactivation of the catalyst, which plays a decisive role in the feasibility of the process. In this study, a feasible TCMD process was identified and a simplified mathematical model was developed, which provides a dynamic estimation for the hydrogen production rate and catalyst turnover costs over various process conditions. The work consisted of a parametric study as well as an investigation into the different process modes. Based on the numerous simulation results it was possible to find the optimal process parameters that maximise the hydrogen production rate and minimise the catalyst turnover costs, therefore increasing the economic potential of the process and hence its commercial viability.     

Improvement of hydrogen yield of lba-transgenic Chlamydomonas reinhardtii caused by increasing respiration and impairing photosynthesis

The transgenic alga lba of Chlamydomonas reinhardtii yielded H₂ with 50%–180% higher than the control strain. Further experiments showed that photosynthetic rates and photosynthetic reaction center II's photochemical capacities of the transgenic algae obviously decreased 33.4%–85.9% and 30.0%–51.7%, respectively, compared with those of the control. On the contrary, respiration rates of the transgenic algae significantly increased, with 40.0%–200.0% higher than those of the control. Furthermore, starch contents of the transgenic algae were also improved significantly by 79.1%–592.8% compared with the control. Therefore, the reason of H₂ yield improvement of the transgenic alga lba is not only due to its decrease of photosynthetic capacity and increase of the respiration rate, but also due to the metabolic changes related to starch metabolism, photosynthesis and respiration which is possibly caused by hetero-expression of lba gene in chloroplasts of C. reinhardtii, indicating the potential of utilization of lba gene to improve hydrogen yield of micro-green algae.     

Process simulation and optimization for enhanced biophotolytic hydrogen production from green algae using the sulfur deprivation method

This article explores the modeling, simulation and optimization of a biophotolytic cyclic process for enhanced hydrogen production from microalgae, employing the sulfur deprivation method. To achieve sulfur deprivation, each process cycle contained two temporally separated steps of sulfur-controlled algae growth and sulfur-deprived anaerobic hydrogen production.Reaction kinetics were modeled via an empirical logistic model. Reaction times, sulfate concentrations, and medium pH levels of each cycle were controlled to optimize the rate and yield of hydrogen production. Consequently, 65% and 23% improved values were obtained, respectively, with a smaller total process time (?11%), higher ratio of algae growth-to-hydrogen production time (29% vs. 21%), buffered pH (7.8), controlled sulfate injection and intermediary algae concentrations. Two- and 15-times higher hydrogen yields were obtained for 2- and 12-times lower initial algae concentrations. The proposed method is a significant tool for the design and optimization of a process for enhanced hydrogen production from microalgae.     

Temporal phenomena of hydrogen photobioproduction

The photobioproduction of hydrogen through in water alga systems has been studied as a suitable way for clean hydrogen generation from renewable solar energy and renewable bio-sources. There is evidence of such hydrogen path metabolism involving some algae types in stress conditions and it has been reported by several authors. In this paper some results of hydrogen production are shown for different stress conditions carried out in not full aseptic environment stabilized by antibiotics, Chlamydomonas reinhardtii has got resistance to. Oscillations and temporal phenomena of hydrogen production have been observed and studied by means of Fourier analysis. Their nature can be related to the variation of hydrogen production rate usually reflected on the cumulative hydrogen curves by the presence of shoulders or accentuated changes of slope.     

Environmental sustainability assessment of large-scale hydrogen production using prospective life cycle analysis

The need for a rapid transformation to low-carbon economies has rekindled hydrogen as a promising energy carrier. Yet, the full range of environmental consequences of large-scale hydrogen production remains unclear. Here, prospective life cycle analysis is used to compare different options to produce 500 Mt/yr of hydrogen, including scenarios that consider likely changes to future supply chains. The resulting environmental and human health impacts of such production levels are further put into context with the Planetary Boundaries framework, known human health burdens, the impacts of the world economy, and the externality-priced production costs that embody the environmental impact. The results indicate that climate change impacts of projected production levels are 3.3–5.4 times higher than the allocated planetary boundary, with only green hydrogen from wind energy staying below the boundary. Human health impacts and other environmental impacts are less severe in comparison but metal depletion and ecotoxicity impacts of green hydrogen deserve further attention. Priced-in environmental damages increase the cost most strongly for blue hydrogen (from ～2 to ～5 USD/kg hydrogen), while such true costs drop most strongly for green hydrogen from solar photovoltaic (from ～7 to ～3 USD/kg hydrogen) when applying prospective life cycle analysis. This perspective helps to evaluate potentially unintended consequences and contributes to the debate about blue and green hydrogen.     

Nature inspired artificial photosynthesis technologies for hydrogen production: Barriers and challenges

Hydrogen drives the big wheel of nature. Hydrogen nuclear fusion in the sun produces light and heat. Solar flux reaching the earth's surface in an hour is far more than global annual energy demand. Photosynthesis traps 100-TW solar energy annually into biomass on land at 0.1% efficiency that is about six times more than global yearly energy demand. All photosynthetic organisms (photoautotroph) annually convert 100-billion tons of carbon in the atmosphere into biomass. The rampant rise in energy demand requires to replicate natural photosynthesis process artificially to convert solar energy and Carbon dioxide (CO₂) in liquid and burnable gaseous fuels. Chemists, physicists and biologists are collaborating to develop suitable catalysts for artificial photosynthesis. There is a consensus the sun can fuel transport sector by hydrogen and power grid by photo-electricity. It is well in time to develop a full spectrum of solar technologies instead of keeping ourselves plugged to hydrocarbon honey. Photocathodes and catalysts can mediate water splitting using nature-inspired artificial photosynthesis. Economic hydrogen production can accomplish the grand energy transition from fossil fuels to sustainable and renewable energy sources. This paper reviews the recent advances in artificial photosynthesis technologies and presents our work on the microbial fuel cell for hydrogen production and points out technical barriers and operational challenges.     

Low-emission hydrogen production via the thermo-catalytic decomposition of methane for the decarbonisation of iron ore mines in Western Australia

The Pilbara, located in Western Australia is one of the largest iron ore-mining regions in the world and will need to achieve significant emission reductions in the short term to conserve the limited carbon budget and abide by the Paris Agreement targets. Green hydrogen has been communicated as the desired solution, however, the high production cost limits the deployment of these systems. The thermo-catalytic methane decomposition (TCMD) process is an alternative solution, which could be implemented as a bridge technology to produce low-emission hydrogen at a potentially lower cost. This is especially attractive for iron ore mines due to the utilisation of iron ore as a process catalyst, which reduces the catalyst turnover costs and can increase the grade of spent iron ore catalyst. In this study, a preliminary techno-economic assessment was carried out in comparison with green hydrogen to determine the feasibility of the TCMD process for the decarbonisation of iron ore mine sites in the Pilbara. The results show that the TCMD process had a CO₂ abatement cost between 25 and 40% less than green hydrogen, however, the magnitude of these costs was lowest for mining operations >60 Mt/yr at approximately $150 and $200 USD/t CO₂ respectively. Since green hydrogen is expected to have significant cost reductions in the future, integrating renewables already into the mine could reduce emissions in the short term, which could then be extended for green hydrogen production once it becomes viable. The TCMD process, therefore, only has a narrow window of opportunity, although considering the uncertainty of the process and that green hydrogen is a proven technology with greater emission-reduction potential, green hydrogen may be the most suitable solution despite the model results presented in this work.     

Experimental verification of various methods for biological hydrogen production

The aim of the work was to compare two different biological methods for hydrogen production: fermentative and photosynthetic based upon the modality of batch cultures. For testing of fermentative bio-hydrogen production four mixed cultures representing anaerobic microorganisms (dominant strain Clostridium) were selected. The kinetic parameters on the intensity of bio-hydrogen production were established. The efficiency coefficient of transformation ranged from 1.65 mol H₂/mol glucose in the pectin culture up to 2.45 in the mixed culture. The bio-hydrogen concentration never exceeded 30%. The carbon dioxide was produced in a ratio of CO₂ to H₂ (0.5–0.67)/1. The testing of green algae proved that the most effective was the algae species Scenedesmus. High bio-hydrogen purity was analytically verified. The fermentative method of H₂ production is more efficient; it does not need light, has a longer efficiency of one charge and enables effective use of different biological wastes.     

A newly isolated green alga, Tetraspora sp. CU2551, from Thailand with efficient hydrogen production

A novel unicellular hydrogen-producing green alga was isolated from fresh water pond in Pathumthani province, Thailand. Under light microscope, this alga was identified as belonging to the genus Tetraspora. Phylogenetic analysis of 18S rRNA sequence revealed that the green alga, identified as Tetraspora sp. CU2551, is closely related to other unicellular green algal species. Tetraspora sp. CU2551 had the shortest doubling time when grown in Tris-acetate-phosphate (TAP) medium under a light intensity of 48–92 μE/m²/s and a temperature of 36 °C. Hydrogen production increased with increasing pH from 5.75 to 9.30; however, almost no production was observed at a pH of 5.25. Addition of 0.5 mM β-mercaptoethanol to the TAP medium stimulated hydrogen production about two-fold. During the hydrogen production phase, the use of TAP medium lacking both nitrogen and sulfur resulted in about 50% increase in the hydrogen production. This was in contrast to only a small increase in the production when either nitrogen or sulfur was omitted in TAP medium. The stimulation of hydrogen production by 0.5 mM β-mercaptoethanol under nitrogen- and sulfur-deprived conditions occurred only when the cells were grown at a light intensity lower than 5 μE/m²/s with no effects at higher intensities. Maximal calculated hydrogen production, 17.3–61.7 μmol/mg Chl a/h, is a very high production rate compared to other green algae and makes Tetraspora sp. CU2551 an interesting model strain for photobiological hydrogen production.     

Artificial chloroplast: Au/chloroplast-morph-TiO2 with fast electron transfer and enhanced photocatalytic activity

The unique structures and functional features of chloroplasts in green plants provide a promising blueprint for greatly improving solar energy utilization efficiency. In this paper, a prototype of artificial chloroplast, Au/chloroplast-morph-TiO₂ with natural chloroplasts' nanostructure and analogous functional features, is provided. The nano-layered structures of chloroplast template inherited in the chloroplast-morph-TiO₂ lead to a large reaction area and fast photo-induced electron transfer; cocatalyst Au nanoparticles which work as reaction centers promote photo-induced charge separation and improve the overall photocatalytic activity in hydrogen production; Nitrogen and phosphorus self-doped from the bio-template increase visible light absorption, similar to the antenna pigment. With this new inorganic artificial photosynthetic system, we achieve effective light utilization, fast photo-induced charge separation, high electron transfer, enhanced photocatalytic activity for dye degradation rate and improved H₂ evolution efficiency. This concept provides the inspiration for constructing efficient photocatalysts by imitating the photosynthesis process from both structures and functions.     

Potassium deficiency,a “smart” cellular switch for sustained high yield hydrogen production by the green alga Scenedesmus obliquus

Hydrogen is considered to be the future optimal energy carrier, and is expected to contribute to the growth of the world's economy by facilitating a stable supply of energy. The ability of green algae to produce hydrogen was discovered 74 years ago. Since then, several attempts were made, to increase hydrogen production yields, sulfur starvation being the best known. The main concern during these attempts was that the achievable increase in yield was not sustainable. In this contribution, potassium deficiency is presented as a biochemical/bioenergetic switch for a sustained high yield of hydrogen production via the photosynthetic apparatus. Potassium can partially be replaced by sodium in the majority of biochemical processes and as a result the system remains functional. However, sodium cannot replace potassium in the conversion of glucose to starch. This fact significantly increased the yield of hydrogen production through the Photosystem II independent pathway, since electrons originating from the metabolism of glucose are used in the continuous donation to the plastoquinone-pool of the photosynthetic electron chain. Additionally, PSII inactivation (and therefore the inhibition of O₂-production), the further synthesis and over activation of Photosystem I and plastidic hydrogenase, generated a sustained increase in hydrogen production, mainly through the PSII-independent pathway. The self regulation of these multistage processes in hermitically closed static systems of Scenedesmus obliquus cultivation, permitted the establishment of anoxic conditions and the continuous electron supply to highly activated hydrogenase, resulting in the sustained high yield hydrogen production and paving the way for future usage in an industrial scale application.