Outdoor biohydrogen production by thermotolerant Rhodopseudomonas pentothenatexigens KKU-SN1/1 in a cluster of ten bioreactors system.

In tropical regions, the viability of outdoor photo-fermentative biohydrogen production faces challenges arising from elevated temperatures and varying light intensity. This research aimed to explore how high temperatures and outdoor environments impact both biohydrogen production and the growth of purple non-sulfur bacteria. Our findings revealed the potential of Rhodopseudomonas spp. as a robust outdoor hydrogen-producing bacteria, demonstrating its capacity to thrive and generate biohydrogen even at 40 °C and under fluctuating outdoor conditions. Rhodopseudomonas harwoodiae NM3/1-2 produced the highest cumulative biohydrogen of 223 mL/L under anaerobic light conditions at 40 °C, while Rhodopseudomonas harwoodiae 2M had the highest dry cell weight of 2.93 g/L. However, R. harwoodiae NM3/1-2 demonstrated the highest dry cell weight of 3.99 g/L and Rhodopseudomonas pentothenatexigens KKU-SN1/1 exhibited the highest cumulative biohydrogen production of 400 mL/L when grown outdoors. In addition, the outdoor enhancement of biohydrogen production was achieved through the utilization of a cluster of ten bioreactors system. The outcomes demonstrated a notable improvement in biohydrogen production efficiency, marked by the highest daily biohydrogen production of 493 mL/L d by R. pentothenatexigens KKU-SN1/1. Significantly, the highest biohydrogen production rate was noted to be 17 times greater than that observed in conventional batch production methods. This study is the first to utilize R. pentothenatexigens and R. harwoodiae for sustained biohydrogen production at high temperatures and in outdoor conditions over an extended operational period. The successful utilization of a clustered system of ten bioreactors demonstrates potential to scale-up for industrial biohydrogen production.

Recent advancements in wastewater treatment via anaerobic fermentation process: A systematic review.

This manuscript delves into the realm of wastewater treatment, with a particular emphasis on anaerobic fermentation processes, especially dark, photo, and dark-photo fermentation processes, which have not been covered and overviewed previously in the literature regarding the treatment of wastewater. Moreover, the study conducts a bibliometric analysis for the first time to elucidate the research landscape of anaerobic fermentation utilization in wastewater purification. Furthermore, microorganisms, ranging from microalgae to bacteria and fungi, emphasizing the integration of these agents for enhanced efficiency, are all discussed and compared. Various bioreactors, such as dark and photo fermentation bioreactors, including tubular photo bioreactors, are scrutinized for their design and operational intricacies. The results illustrated that using clostridium pasteurianum CH4 and Rhodopseudomonas palustris WP3-5 in a combined dark-photo fermentation process can treat wastewater to a pH of nearly 7 with over 90% COD removal. Also, integrating Chlorella sp and Activated sludge can potentially treat synthetic wastewater to COD, P, and N percentage removal rates of 99%,86%, and 79%, respectively. Finally, the paper extends to discuss the limitations and future prospects of dark-photo fermentation processes, offering insights into the road ahead for researchers and scientists.

Nutritional influences on biomass behaviour and metabolic products by Chlamydomonas reinhardtii.

The recent works have shown the unicellular green alga Chlamydomonas reinhardtii is a relevant model for investigations of algal bioprocesses. In the current work, several media were evaluated in batch mode for a better understanding of C. reinhardtii metabolism. Nutrient-suppression using heterotrophic and mixotrophic conditions were performed. The findings showed C. reinhardtii metabolized lactose (from milk whey permeate) resulting in high biomass density (2.08 g/L) and total chlorophyll content (86.74 mg/m3). It was observed a specific growth rate of 0.023 h and 29 h for the doubling time. In sulfur-suppression, the algal growth (1.17 g/L) was reduced even though a carbon source (glucose) has been supplemented. Also, the specific growth rate (0.022 h) and the doubling time (31 h) was verified. The production of ethanol was slight and the acetic acid-suppression affected the C. reinhardtii performance providing slow cell growth (0.004 h) and high doubling time (154 h).

A comprehensive review of mathematical models of photo fermentation.

This work aims at analyzing and comparing the different modeling approaches used to date to simulate, design and control photo fermentation processes for hydrogen production and/or wastewater treatment. The study is directed to researchers who approach the problem of photo fermentation mathematical modeling. It is a useful tool to address future research in this specific field in order to overcome the difficulty of modeling a complex, not totally elucidate process. We report a preliminary identification of the environmental and biological parameters, included in the models, which affect photo fermentation. Based on model features, we distinguish three different approaches, i.e. kinetic, parametric and non-ideal reactors. We explore the characteristics of each approach, reporting and comparing the obtained results and underlining the differences between models, together with the advantages and the limitations of each of them. The analysis of the approaches indicates that Kinetic models are useful to describe the process from a biochemical point of view, without considering bio-reactor hydrodynamics and the spatial variations that Parametric Models can be utilized to study the influence and the interactions between the operational conditions. They do not take into account the biochemical process mechanism and the influence of reactor hydrodynamics. Quite the opposite, non-ideal reactors models focus on the reactor configuration. Otherwise, the biochemical description of purple non-sulfur bacteria activities is usually simplified. This review indicates that there still is a lack of models that fully describe photo fermentation processes.

Carbon catabolite repression occurrence in photo fermentation of ethanol-rich substrates.

The paper investigates the phenomenon of Carbon Catabolite Repression occurring during photo fermentation of ethanol-rich effluents, which usually contain ethanol as main carbon source, and glycerol as secondary one. The study was conducted using mixed phototrophic cultures, adopting, as substrate, the effluent produced by the alcoholic fermentation of sugar cane bagasse. In order to elucidate the phenomenon, experimental tests were carried out using two different ethanol to glycerol ratios. Results were compared with those resulting from pure ethanol and glycerol conversion. According to the obtained data, as a result of Carbon Catabolite Repression occurrence, the presence of glycerol negatively affects hydrogen production. Indeed, part of the ethanol source is converted to biomass and polyhydroxybutyrate rather than to hydrogen. In more details, the presence of glycerol determines a drop of the hydrogen production, which goes from 12 % to 32 %, according to the ethanol/glycerol ratio, compared to the production obtained from fermentation of ethanol alone. Therefore, to promote the hydrogen production, it is advisable to apply strategies to produce low glycerol concentrations in the ethanol production stage.

Biological hydrogen production: molecular and electrolytic perspectives.

Exploration of renewable energy sources is an imperative task in order to replace fossil fuels and to diminish atmospheric pollution. Hydrogen is considered one of the most promising fuels for the future and implores further investigation to find eco-friendly ways toward viable production. Expansive processes like electrolysis and fossil fuels are currently being used to produce hydrogen. Biological hydrogen production (BHP) displays recyclable and economical traits, and is thus imperative for hydrogen economy. Three basic modes of BHP were investigated, including bio photolysis, photo fermentation and dark fermentation. Photosynthetic microorganisms could readily serve as powerhouses to successively produce this type of energy. Cyanobacteria, blue green algae (bio photolysis) and some purple non-sulfur bacteria (Photo fermentation) utilize solar energy and produce hydrogen during their metabolic processes. Ionic species, including hydrogen (H+) and electrons (e-) are combined into hydrogen gas (H2), with the use of special enzymes called hydrogenases in the case of bio photolysis, and nitrogenases catalyze the formation of hydrogen in the case of photo fermentation. Nevertheless, oxygen sensitivity of these enzymes is a drawback for bio photolysis and photo fermentation, whereas, the amount of hydrogen per unit substrate produced appears insufficient for dark fermentation. This review focuses on innovative advances in the bioprocess research, genetic engineering and bioprocess technologies such as microbial fuel cell technology, in developing bio hydrogen production.

A comprehensive review on two-stage integrative schemes for the valorization of dark fermentative effluents.

This review provides the alternative routes towards the valorization of dark H2 fermentation effluents that are mainly rich in volatile fatty acids such as acetate and butyrate. Various enhancement and alternative routes such as photo fermentation, anaerobic digestion, utilization of microbial electrochemical systems, and algal system towards the generation of bioenergy and electricity and also for efficient organic matter utilization are highlighted. What is more, various integration schemes and two-stage fermentation for the possible scale up are reviewed. Moreover, recent progress for enhanced performance towards waste stabilization and overall utilization of useful and higher COD present in the organic source into value-added products are extensively discussed.

Fermentative hydrogen production from low-value substrates.

Hydrogen is a promising energy source that is believed to replace the conventional energy sources e.g. fossil fuels over years. Hydrogen production methods can be divided into conventional production methods which depend mainly on fossil fuels and alternative production methods including electrolysis of water, biophotolysis and fermentation hydrogen production from organic waste materials. Compared to the conventional methods, the alternative hydrogen production methods are less energy intensive and negative-value substrates i.e. waste materials can be used to produce hydrogen. Among the alternative methods, fermentation process including dark and photo-fermentation has gained more attention because these processes are simple, waste materials can be utilized, and high hydrogen yields can be achieved. The fermentation process is affected by several parameters such as type of inoculum, pH, temperature, substrate type and concentration, hydraulic retention time, etc. In order to achieve optimum hydrogen yields and maximum substrate degradation, the operating conditions of the fermentation process must be optimized. In this review, two routes for biohydrogen production as dark and photo-fermentation are discussed. Dark/photo-fermentation technology is a new approach that can be used to increase the hydrogen yield and improve the energy recovery from organic wastes.

Mechanisms for hydrogen production by different bacteria during mixed-acid and photo-fermentation and perspectives of hydrogen production biotechnology.

H2 has a great potential as an ecologically-clean, renewable and capable fuel. It can be mainly produced via hydrogenases (Hyd) by different bacteria, especially Escherichia coli and Rhodobacter sphaeroides. The operation direction and activity of multiple Hyd enzymes in E. coli during mixed-acid fermentation might determine H2 production; some metabolic cross-talk between Hyd enzymes is proposed. Manipulating the activity of different Hyd enzymes is an effective way to enhance H2 production by E. coli in biotechnology. Moreover, a novel approach would be the use of glycerol as feedstock in fermentation processes leading to H2 production. Mixed carbon (sugar and glycerol) utilization studies enlarge the kind of organic wastes used in biotechnology. During photo-fermentation under limited nitrogen conditions, H2 production by Rh. sphaeroides is observed when carbon and nitrogen sources are supplemented. The relationship of H2 production with H(+) transport across the membrane and membrane-associated ATPase activity is shown. On the other hand, combination of carbon sources (succinate, malate) with different nitrogen sources (yeast extract, glutamate, glycine) as well as different metal (Fe, Ni, Mg) ions might regulate H2 production. All these can enhance H2 production yield by Rh. sphaeroides in biotechnology Finally, two of these bacteria might be combined to develop and consequently to optimize two stages of H2 production biotechnology with high efficiency transformation of different organic sources.

Enhancement of biohydrogen production by photo-fermentation of corn stover via visible light catalyzed titanium dioxide/activated carbon fiber.

In this study, titanium dioxide/activated carbon fiber (TiO2/ACF) was synthesized by liquid-phase deposition method and the effect of TiO2/ACF on the performance of photo-fermentation biohydrogen production (PFHP) from corn stover under visible light catalysis was discussed. Results show the maximum cumulative hydrogen yield (CHY) obtained under the optimal conditions was 74.0 ± 1.3 mL/g TS with TiO2/ACF addition of 100 mg/L, which was twice that without TiO2/ACF addition (36.9 ± 1.0 mL/g TS). Initial pH value had the most significant effect on CHY. The addition of TiO2/ACF promoted the metabolic pathway of nitrogenase to reduce H+ produced by consuming acetic acid and butyric acid to hydrogen, and also shortened the photo-fermentation period. By scanning electron microscopy and X-ray diffraction analysis, the morphology and phase structure of TiO2/ACF after PFHP did not change significantly. This study laid the foundation for the reuse of TiO2 and its practical application in PFHP.

Impact of light spectra on photo-fermentative biohydrogen production by Rhodobacter sphaeroides KKU-PS1.

Purple non-sulphur bacteria can only capture up to 10 % light spectra and only 1-5 % of light is converted efficiently for biohydrogen production. To enhance light capture and conversion efficiencies, it is necessary to understand the impact of various light spectra on light harvesting pigments. During photo-fermentation, Rhodobacter sphaeroides KKU-PS1 cultivated at 30 °C and 150 rpm under different light spectra has been investigated. Results revealed that red light is more beneficial for biomass accumulation, whereas green light showed the greatest impact on photo-fermentative biohydrogen production. Light conversion efficiency by green light is 2-folds of that under control white light, hence photo-hydrogen productivity is ranked as green > red > orange > violet > blue > yellow. These experimental data demonstrated that green and red lights are essential for photo-hydrogen and biomass productions of R. sphaeroides and a clearer understanding that possibly pave the way for further photosynthetic enhancement research.

Light energy utilization and microbial catalysis for enhanced biohydrogen: Ternary coupling system of triethanolamine-mediated Fe@C-Rhodobacter sphaeroides.

This study investigated the mediating effect of Triethanolamine on Fe@C-Rhodobacter sphaeroides hybrid photosynthetic system to achieve efficient biohydrogen production. The biocompatible Fe@C generates excited electrons upon exposure to light, releasing ferrum for nitrogenase synthesis, and regulating the pH of the fermentation environment. Triethanolamine was introduced to optimize the electron transfer chain, thereby improving system stability, prolonging electron lifespan, and facilitating ferrum corrosion. This, in turn, stimulated the lactic acid synthetic metabolic pathway of Rhodobacter sphaeroides, resulting in increased reducing power in the biohybrid system. The ternary coupling system was analyzed through the regulation of concentration, initial pH, and light intensity. The system achieved the highest total H2 production of 5410.9 mL/L, 1.29 times higher than the control (2360.5 mL/L). This research provides a valuable strategy for constructing ferrum-carbon-based composite-cellular biohybrid systems for photo-fermentation H2 production.

Ammonia-dependent reducing power redistribution for purple phototrophic bacteria culture-based biohydrogen production.

The global energy crisis has intensified the search for sustainable and clean alternatives, with biohydrogen emerging as a promising solution to address environmental challenges. Leveraging photo fermentation (PF) process, purple phototrophic bacteria (PPB) can harness reducing power derived from organic substrates to facilitate hydrogen production. However, existing studies report much lower H2 yields than theoretical value when using acetate as carbon source and ammonia as nitrogen source, primarily attributed to the widely employed pulse-feeding mode which suffers from ammonia inhibition effect on nitrogenase. To address this issue, a continuous feeding mode was applied to avoid ammonia accumulation in this study. On the other hand, other pathways like carbon fixation and polyhydroxyalkanoate (PHA) formation could compete reducing power with H2 production. However, the reducing power allocation under continuous feeding mode is not yet clear. In this study, the reducing power allocation and hydrogen production performance were evaluated under various ammonia loading, using acetate as carbon source and infrared LED at around 50 W·m-2 as light source. The results show that (a) The absence of ammonia resulted in the best performance for hydrogen production, with 44 % of the reducing power distributed to H2 and the highest H2 volumetric productivity, while the allocation of reducing power to hydrogen production stopped when ammonia loading was above 7.6 mg NH4-N·L-1·d-1; (b) when PPB required to eliminate reducing power under ammonia limited conditions, PHA production was the preferred pathway followed by the hydrogen production pathway, but once PHA accumulation reached saturation, hydrogen generation pathway dominated; (c) under ammonia limited conditions, the TCA cycle was more activated rendering higher NADH (i.e. reducing power) production compared with that under ammonia sufficient conditions which was verified by metagenomics analysis, and all the hydrogen production, PHA accumulation and carbon fixation pathways were highly active to dissipate reducing power. This work provides the insight of reducing power distribution and PPB biohydrogen production variated by ammonia loading under continuous feeding mode.

Integrative biological hydrogen production: an overview.

Biological hydrogen (H2) production by dark and photo-fermentative organisms is a promising area of research for generating bioenergy. A large number of organisms have been widely studied for producing H2 from diverse feeds, both as pure and as mixed cultures. However, their H2 producing efficiencies have been found to vary (from 3 to 8 mol/mol hexose) with physiological conditions, type of organisms and composition of feed (starchy waste from sweet potato, wheat, cassava and algal biomass). The present review deals with the possibilities of enhancing H2 production by integrating metabolic pathways of different organisms-dark fermentative bacteria (from cattle dung, activated sludge, Caldicellulosiruptor, Clostridium, Enterobacter, Lactobacillus, and Vibrio) and photo-fermentative bacteria (such as Rhodobacter, Rhodobium and Rhodopseudomonas). The emphasis has been laid on systems which are driven by undefined dark-fermentative cultures in combination with pure photo-fermentative bacterial cultures using biowaste as feed. Such an integrative approach may prove suitable for commercial applications on a large scale.

Pretreatment of Arundo donax L. for photo-fermentative biohydrogen production by ultrasonication and ionic liquid.

Combined pretreatment methods were assumed to further enhance photo-fermentative biohydrogen production (PFHP) from lignocellulosic biomass. For this purpose, an ultrasonication assisted ionic liquid pretreatment was applied to Arundo donax L. biomass for PFHP. The optimal condition for the combined pretreatment was 16 g/L of 1-Butyl-3-methylimidazolium Hydrogen Sulfate ([Bmim]HSO4) combined with ultrasonication at a solid to liquid ratio (SLR) of 1:10 for 1.5 h under 60 °C. Under this condition, the maximum delignification of 22.9 % was obtained, in addition, the hydrogen yield (HY) and energy conversion efficiency (ECE) were enhanced by 1.5-fold and 46.4 % (p < 0.05) compared to untreated biomass, respectively. Moreover, heat map analysis was performed to evaluate the correlation between pretreatment conditions and corresponding results, suggesting pretreatment temperature had the strongest (absolute value of Pearson's r was 0.97) linear correlation with HY. Combined multiple energy production approaches might be useful for further improved ECE.

Effect of deep eutectic solvent pretreatment on biohydrogen production from corncob: pretreatment temperature and duration.

Deep eutectic solvent pretreatment with different temperatures and durations was applied to corncob to increase hydrogen yield via photo-fermentation. The correlation of composition, enzymatic hydrolysis, and hydrogen production in pretreated corncobs, as well as energy conversion was evaluated. Deep eutectic solvent pretreatment effectively dissolved lignin, retained cellulose, and enhanced both enzymatic hydrolysis and hydrogen production. The maximum cumulative hydrogen yield obtained under a pretreatment condition of 50°C and 12 h was 677.45 mL; this was 2.72 times higher than that of untreated corncob, and the corresponding lignin removal and enzymatic reduction of sugar concentration were 79.15% and 49.83 g/L, respectively; the highest energy conversion efficiency was 12.08%. The hydrogen production delay period was shortened, and the maximum shortening time was 18.9 h. Moreover, the cellulose content in pretreated corncob was positively correlated with both reducing sugar concentration and hydrogen yield and had the strongest influence on hydrogen production.

Enhancement of static magnetic field on biological hydrogen production via photo-fermentation of giant reed.

The effect of static magnetic field (SMF) on the system of photo-fermentation biological hydrogen production remains dimness. The goal of this study was to clarify the correlation between external SMF addition and hydrogen production via photo-fermentation from giant reed. SMF with 20 mT improved the cumulative H2 yield by 26.1% and reduced the lag time of hydrogen production by 56.7% compared with that of without external magnetic field. Moreover, 20 mT of SMF not only enhanced the activity of nitrogenase by 94.52%, but also obtained the maximum energy conversion efficiency of 27.27%. The distribution of volatile fatty acids proved that the concentration of acetic acid and butyric acid were 137% and 81% higher than that of without SMF, respectively. The results would help to trigger the positive interaction between SMF and microorganism and to avoid the possible negative interaction.

Modeling and optimization of photo-fermentation biohydrogen production from co-substrates basing on response surface methodology and artificial neural network integrated genetic algorithm.

The main aim of the present study was to establish a relationship model between bio-hydrogen yield and the key operating parameters affecting photo-fermentation hydrogen production (PFHP) from co-substrates. Central composite design-response surface methodology (CCD-RSM) and artificial neural network-genetic algorithm (ANN-GA) models were used to optimize the hydrogen production performance from co-substrates. Compared to CCD-RSM, the ANN-GA had higher determination coefficient (R2 = 0.9785) and lower mean square error (MSE = 9.87), average percentage deviation (APD = 2.72) and error (4.3%), indicating the ANN-GA was more suitable, reliable and accurate in predicting biohydrogen yield from co-substrates by PFHP. The highest biohydrogen yield (99.09 mL/g) predicted by the ANN-GA model at substrate concentration 35.62 g/L, temperature 30.94 °C, initial pH 7.49 and inoculation ratio 32.98 %(v/v), which was 4.20 % higher than the CCD-RSM model (95.10 mL/g).

[Enhancing fucoxanthin production in Phaeodactylum tricornutum by photo-fermentation].

The aim of this study was to develop a technical system for high-efficient production of fucoxanthin by photo-fermentation of Phaeodactylum tricornutum. In a 5 L photo-fermentation tank, the effects of initial light intensity, nitrogen source and concentration as well as light quality on biomass concentration and fucoxanthin accumulation in P. tricornutum were investigated systematically under mixotrophic condition. The results showed that the biomass concentration, fucoxanthin content and productivity reached the highest level of 3.80 g/L, 13.44 mg/g and 4.70 mg/(L·d) under the optimal conditions of initial light intensity of 100 µmol/(m2·s), 0.02 mol TN/L of tryptone: urea (1:1, N mol/N mol) as mixed nitrogen source, and a mixed red/blue (R: B=6:1) light, 1.41, 1.33 and 2.05-fold higher than that before optimization, respectively. This study developed a key technology for enhancing the production of fucoxanthin by photo-fermentation of P. tricornutum, facilitating the development of marine natural products.

A review on biological biohydrogen production: Outlook on genetic strain enhancements, reactor model and techno-economics analysis.

Modernisation of industrial and transportation sector would have not imaginable without the help of fossil fuels, but constant usage has led to many environmental concerns. As a step forward, for safer next generation living we are forced to look into green fuels like bio­hydrogen and higher alcohols. This review mainly focuses on bio­hydrogen production via biological pathways, genetic improvements, knowledge gap, economics, and future directions. Dark and photo fermentation process with the factor influence the process (pH regulation, temperature, hydraulic retention time, organic loading rate, Maintenance, Nutrient) is studied. Integration of dark fermentation and microbial electrolysis cell is the most trending progression for sustainable bio­hydrogen production. Genetic improvement of microbe for biohydrogen production via inactivation of hydrogenase (H2ase) and improve oxygen tolerant H2ase. In future, bioaugmentation, multidisciplinary integrated process and microbial electrolysis needs to be experimented in industrial level scale for successful commercialization. About 41.47 mmol H2/g DCW h at 40 g/L of optimum biohydrogen production was obtained through glycerol fermentation. From the studies, the cost of biohydrogen production was found to high with respect to the direct bio photolysis it cost around $7.24 kg-1; for indirect bio photolysis it cost around $7.54 kg-1 and for fermentation it cost around $7.61 kg-1.