A high-value biohythane production: Feedstocks, reactor configurations, pathways, challenges, technoeconomics and applications.

In recent years, the demand for high-quality biofuels from renewable sources has become an aspirational goal to offer a clean environment by alternating the depleting fossil fuels to meet future energy needs. In this aspect, biohythane production from wastes has received extensive research interest since it contains superior fuel characteristics than the promising conventional biofuel i.e. biogas. The main aim is to promote research and potentials of biohythane production by a systematic review of scientific literature on the biohythane production pathways, substrate/microbial consortium suitability, reactor design, and influential process/operational factors. Reactor configuration also decides the product yield in addition to other key factors like waste composition, temperature, pH, retention time and loading rates. Hence, a detailed emphasis on different reactor configurations with respect to the type of feedstock has also been given. The technical challenges are highlighted towards process optimization and system scale up. Meanwhile, solutions to improve product yield, technoeconomics, applications and key policy and governance factors to build a hydrogen based society have also been discussed.

Application of Doehlert design in optimizing the solid-state hydrogenogenic stage augmented with biomass fly ash in a two-stage biohythane production process.

This study aimed to optimize the solid-state hydrogenogenic stage supplemented with biomass fly ash in a two-stage anaerobic digestion (AD) process for biohythane production from the organic fraction of municipal solid waste (OFMSW). Doehlert's experimental design was used to obtain the optimal set of two investigated variables, namely total solids (TS) content and biomass fly ash dosage in the defined ranges of 0-20 g/L and 20-40%, respectively. Applying the optimal conditions of TS content (29.1%) and fly ash dosage (19.2 g/L) in the first stage led not only to a total H2 yield of 95 mL/gVSadded, which was very close to the maximum H2 yield predicted by the developed model (97 mL/gVSadded), but also to a high CH4 yield of 400 mL/gVSadded (76% of the theoretical CH4 yield). Moreover, the biohythane obtained from the optimized two-stage process met the standards of a biohythane fuel with an H2 content of 19% v/v.

Biohythane Production in Hydrogen-Oriented Dark Fermentation of Aerobic Granular Sludge (AGS) Pretreated with Solidified Carbon Dioxide (SCO2).

Though deemed a prospective method, the bioconversion of organic waste to biohydrogen via dark fermentation (DF) has multiple drawbacks and limitations. Technological difficulties of hydrogen fermentation may, in part, be eliminated by making DF a viable method for biohythane production. Aerobic granular sludge (AGS) is a little-known organic waste spurring a growing interest in the municipal sector; its characteristics indicate the feasibility of its use as a substrate for biohydrogen production. The major goal of the present study was to determine the effect of AGS pretreatment with solidified carbon dioxide (SCO2) on the yield of H2 (biohythane) production during anaerobic digestion (AD). It was found that an increasing dose of SCO2 caused an increase in concentrations of COD, N-NH4+, and P-PO43- in the supernatant at the SCO2/AGS volume ratios from 0 to 0.3. The AGS pretreatment at SCO2/AGS ratios within the range of 0.1-0.3 was shown to enable the production of biogas with over 8% H2 (biohythane) content. The highest yield of biohythane production, reaching 481 ± 23 cm3/gVS, was obtained at the SCO2/AGS ratio of 0.3. This variant produced 79.0 ± 6% CH4 and 8.9 ± 2% H2. The higher SCO2 doses applied caused a significant decrease in the pH value of AGS, modifying the anaerobic bacterial community to the extent that diminished anaerobic digestion performance.

Can digestate recirculation promote biohythane production from two-stage co-digestion of rice straw and pig manure?

Digestate recirculation is often considered an important way to improve system stability (system acidification, ammonia inhibition, hydrolysis limitations, etc.) and gas production performance. However, it is not clear how the promotion of biohythane production works in anaerobic co-digestion with digestate recirculation of rice straw (RS) and pig manure (PM). Two sets of laboratory-scale two-stage continuous stirred tank reactors were operated continuously for 95 d to investigate the performance of biohythane production in the first/second phase under mesophilic (M)/thermophilic (T) and digestate recirculation conditions. Firstly, biohythane was not produced by PM with RS under digestate recirculation. The main reasons were: 1) Digestive recirculation promoted the growth of hydrogenotrophic methanogenic bacteria; and 2) limitations in hydrolysis. Secondly, digestate recirculation has positive effects on the removal rates (removal rates of TS, VS, polysaccharide, protein and TCOD increased by 30.4%, 22.3%, 9.9%, 31.4%, and 11.9%, respectively) and energy yield (up to 68.7%). Finally, there was a higher abundance of hydrogen-producing bacteria (Fervidobacterium [44.9%] and Coprothermobacter [18.8%]) in T2, accounting for >80% of the total, and of which the huge hydrogen production potential cannot be ignored. The results provide new ideas for alleviating the energy crisis and developing green energy in the future.

Two-Stage anaerobic digestion in agroindustrial waste treatment: A review.

The anaerobic digestion is a process widely recognized as an interesting alternative for the treatment and stabilization of residual organic substrates. However, several technical limitations were observed based on the characteristics of the organic matter submitted to the process, such as the presence of high concentrations of soluble sugars or fats. The technology of anaerobic digestion in multiple stages is described as a viable option in the control of variables, optimizing the environmental conditions of the main microorganisms involved in the process, assuring high solid removal and methane production, besides allowing a higher energy yield through the generation of molecular fuel hydrogen. Several studies reviewed the process of anaerobic digestion in multiple stages in the treatment of food waste, although few report its use applied directly to agroindustrial residues. Thus, the present work aims to review the literature evaluating the scenario and viability of the multi-stage anaerobic digestion process applied to agroindustrial effluents. Effluents such as manipueira, vinasse, and dairy wastewater are substrates that present high yields when treated by AD processes with stage separation. The high concentration of easily fermentable sugars results in a high production of molecular hydrogen (co-product of the production of volatile acids in the acid phase) and methane (methanogenic phase). The great challenges related to the development of the sector are focused on the stability of the composition and yield of hydrogen in the acid phase, besides the problems resulting from the treatment of complex residues. Thus, the present study suggests that future works should focus on the technologies of new microorganisms and optimization of process parameters, providing maturation and scale-up of the two-stage anaerobic digestion technique.

Effects of sudden shock load on simultaneous biohythane production in two-stage anerobic digestion of high-strength organic wastewater.

The two-stage anaerobic digestion (AD) for biohythane production is a sustainable solution, but it is sensitive to organic shock load that disrupts reactors and inhibits biohythane production. This study investigated biohythane production, reactor performance, and the possibility of post-failure restoration in a two-stage AD system designed for treating high-strength organic wastewater. Sudden shock load was applied by increasing the OLR threefold higher after reaching steady state phase. During shock load phase, hydrogen content, hydrogen yield and methane production rate (MPR) reached its peak values of 62.61 %, 1.641 mol H2/mol glucose, and 1.003 L CH4/L⋅d respectively before declining significantly. Interestingly, during the restorative phase, hydrogen production sharply declined to nearly zero, while methane production exhibited a resilience and reached its peak methane content of 52.2 %. The study successfully demonstrated the system's resilience to sudden shock load, ensuring stable methane production, while hydrogen production did not exhibit the same capability.

Potential of biofuel production from leather solid wastes: Indian scenario.

India is one among the major leather-producing countries in the world which shares close to one-fourth of the world's leather solid wastes and most of these wastes are not effectively utilized. These wastes are rich in protein and lipids that could be a potential feedstock for biofuel production, i.e., biogas, biodiesel, etc. Among the 150,000 tons of daily leather solid wastes in India, approximately 87,150 tons are shared by pre-tanning operations (i.e., raw trimmings, fleshing, and hair wastes) while the rest of the 62,850 tons are shared by tanning, post-tanning, and finishing operations (i.e., wet blue trimmings, chrome splits, shavings, buffing dust, crust trimming wastes). This review article shows that there is considerable bioenergy potential for the use of leather solid wastes as a green fuel. The biogas potential of leather solid wastes is estimated to be 40,532.9 m3/day whereas the biodiesel potential is estimated as 15,452.6 L/day. The bio-oil and bio-char potential of leather solid wastes is estimated to be 80,513.0 L/day and 45.8 tons/day, respectively. Several factors influence the biofuel process efficacy, which needs to be taken into consideration while setting up a biofuel recovery plant. The overall biofuel potential of leather solid wastes shows that this feedstock is an untapped resource for energy recovery to add commercial benefits to India's energy supply. Furthermore, in addition to the economic benefits for investors, the use of leather solid wastes for biofuel production will yield a positive environmental impact.

Two-stage biohydrogen and methane production from sugarcane-based sugar and ethanol industrial wastes: A comprehensive review.

The transition to renewable energy sources is crucial to ensure a sustainable future. Although the sugar and ethanol industries benefit from this transition, there are untapped opportunities to utilize the waste generated from the sugar and ethanol process chains through two-stage anaerobic digestion (TSAD). This review comprehensively discusses the utilization of various sugarcane-based industrial wastes by TSAD for sequential biohydrogen and methane production. Factors influencing TSAD process performance, including pH, temperature, hydraulic retention time, volatile fatty acids and alkalinity, nutrient imbalance, microbial population, and inhibitors, were discussed in detail. The potential of TSAD to reduce emissions of greenhouse gases is demonstrated. Recent findings, implications, and promising future research related to TSAD, including the integration of meta-omics approaches, gene manipulation and bioaugmentation, and application of artificial intelligence, are highlighted. The review can serve as important literature for the implementation, improvement, and advancements in TSAD research.

Coproduction of amino acids and biohythane from microalgae via cascaded hydrothermal and anaerobic process.

In search of the candidate for animal feed and clean energy, a new vision of algal biorefinery was firstly proposed to coproduce amino acids and biohythane via hydrothermal treatment and two-stage anaerobic fermentation. This study focused on the comprehensive analysis of amino acids recovered from Chlorella sp. and the subsequent biohythane production from microalgal residues. The content and recovery rate of amino acids were in the range of 2.07-27.62 g/100 g and 3.65 %-48.66 % with increasing temperature due to more cell wall disruptions. Furthermore, it was rich in essential amino acids for livestock, including leucine, arginine, isoleucine, valine and phenylalanine. A comparable hydrogen production (9 mL/g volatile solids (VS)) was reached at 70 °C and 90 °C, while it reduced to 5.84 mL/gVS at 150 °C. The group at 70 °C got the maximum methane generation of 311.9 mL/gVS, which was 16.67 %, 24.94 %, 38.38 % and 46.49 % higher than that of other groups. Microalgal residues at lower temperature contained more organics, which was the reason for the better biohythane production. The coproduction of amino acids and biohythane at 130 °C was favorable, which led to 43.71 % amino acids recovery and 93.82 mL biohythane production from per gVS of Chlorella sp. The improved microalgal biorefinery could provide an alternative way to mitigate the crisis of food and energy, but animal experimentations and techno-economic assessments should be considered for further study.

Improved Biohythane Production from Rice Straw in an Integrated Anaerobic Bioreactor under Thermophilic Conditions.

This study evaluated the feasibility of continuous biohythane production from rice straw (RS) using an integrated anaerobic bioreactor (IABR) at thermophilic conditions. NaOH/Urea solution was employed as a pretreatment method to enhance and improve biohythane production. Results showed that the maximum specific biohythane yield was 612.5 mL/g VS, including 104.1 mL/g VS for H2 and 508.4 mL/g VS for CH4, which was 31.3% higher than the control RS operation stage. The maximum total chemical oxygen demand (COD) removal stabilized at about 86.8%. COD distribution results indicated that 2% of the total COD (in the feed) was converted into H2, 85.4% was converted to CH4, and 12.6% was retained in the effluent. Furthermore, carbon distribution analysis demonstrated that H2 production only diverted a small part of carbon, and most of the carbon flowed to the CH4 fermentation process. Upon further energy conversion analysis, the maximum value was 166.7%, 31.7 times and 12.8% higher than a single H2 and CH4 production process. This study provides a new perspective on lignocellulose-to-biofuel recovery.

Biohythane: a Potential Biofuel of the Future.

Today, the world is becoming more dependent on fossil fuels. The major drawbacks of these non-renewable energy resources include an extreme environmental pollution and an extinction threat. Several technologies including microalgal biodiesel production, biomass gasification, and bioethanol production have been explored for the generation of renewable energy especially, biofuels. One such promising research has been carried out in the generation of biohythane which has the potential to become an alternative fuel to the existing non-renewable ones. It has been reported that biohydrogen can be produced from organic wastes or agricultural feedstocks with the help of acidogens. Dark fermentation can be carried out by acidogens to produce biohydrogen under anaerobic conditions by utilizing lignocellulosic biomass or sugarcane feedstocks in the absence of light. The spent medium contains volatile short-chain fatty acids like acetate, butyrate, and propionate that can serve as substrates for acetogenesis followed by methane biosynthesis by methanogens. Therefore, the sequential two-stage anaerobic digestion (AD) involves a production of biohydrogen followed by the biosynthesis of methane. This combined process is termed as a single eponym "Biohythane" (hydrogen + methane). Several studies have demonstrated about the effectiveness of biofuel, and it is believed to have a greater energy recovery, environmental friendliness, and shorter fermentation time. Biohythane can serve as an alternative future green biofuel and solve the present energy crisis in India as well as the entire world.

Gradient electro-processing strategy for efficient conversion of harmful algal blooms to biohythane with mechanisms insight.

Globally eruptive harmful algal blooms (HABs) have caused numerous negative effects on aquatic ecosystem and human health. Conversion of HABs into biohythane via dark fermentation (DF) is a promising approach to simultaneously cope with environmental and energy issues, but low HABs harvesting efficiency and biohythane productivity severely hinder its application. Here we designed a gradient electro-processing strategy for efficient HABs harvesting and disruption, which had intrinsic advantages of no secondary pollution and high economic feasibility. Firstly, low current density (0.888-4.444 mA/cm2) was supplied to HABs suspension to harvest biomass via electro-flocculation, which achieved 98.59% harvesting efficiency. A mathematic model considering coupling effects of multi-influencing factors on HABs harvesting was constructed to guide large-scale application. Then, the harvested HABs biomass was disrupted via electro-oxidation under higher current density (44.44 mA/cm2) to improve bioavailability for DF. As results, hydrogen and methane yields of 64.46 mL/ (g VS) and 171.82 mL/(g VS) were obtained under 6 min electro-oxidation, along with the highest energy yield (50.1 kJ/L) and energy conversion efficiency (44.87%). Mechanisms of HABs harvesting and disruption under gradient electro-processing were revealed, along with the conversion pathways from HABs to biohythane. Together, this work provides a promising strategy for efficient disposal of HABs with extra benefit of biohythane production.

Integrative biohydrogen- and biomethane-producing bioprocesses for comprehensive production of biohythane.

The production of biohythane, a combination of energy-dense hydrogen and methane, from the anaerobic digestion of low-cost organic wastes has attracted attention as a potential candidate for the transition to a sustainable circular economy. Substantial research has been initiated to upscale the process engineering to establish a hythane-based economy by addressing major challenges associated with the process and product upgrading. This review provides an overview of the feasibility of biohythane production in various anaerobic digestion systems (single-stage, dual-stage) and possible technologies to upgrade biohythane to hydrogen-enriched renewable natural gas. The main goal of this review is to promote research in biohythane production technology by outlining critical needs, including meta-omics and metabolic engineering approaches for the advancements in biohythane production technology.

Fundamentals and recent progress in bioelectrochemical system-assisted biohythane production.

Biohythane, a balanced mixture of 10%-30% v/v of hydrogen and 70%-90% v/v of methane, could be the backbone of an all-purpose future energy supply. Recently, bioelectrochemical systems (BES) became a new sensation among environmental biotechnology processes with the potential to sustainably generate biohythane. Therefore, to unleash its full potential for scaling up, researchers are consistently improving microbial metabolic pathways, novel reactors, and electrode designs. This review presents a detailed analysis of recently discovered fundamental mechanisms and science and engineering intervention of different strategies to improve the biohythane composition and production rate from BES. However, several milestones are to be achieved, for instance, improving electrode kinetics using efficient catalysts, engineered microbial communities, and improved reactor configurations, for commercializing this sustainable technology. Thus, a future perspective section is included to recommend novel research lines, mainly focusing on the microbial communities and the efficient electrocatalysts, to enhance reactor performance.

Biohythane production from food waste in a two-stage process: assessing the energy recovery potential.

ABSTRACTBiohythane (hydrogen + methane) production in a two stage dark fermentation (DF) and anaerobic digestion (AD) process from food waste (FW) has been studied. This paper investigated the effect of operation temperature, i.e. mesophilic (34 °C) and thermophilic (55 °C) , on biohythane yield and total energy recovery carried out at the initial culture pH 5.5 and pH 7, respectively for DF and AD batch tests. The mesophilic DF tests gave a higher hydrogen yield of 53.5 (±4) mL H2/g VS added compared to thermophilic DF tests, i.e. 37.6 (±1) mL H2/g VS added. However, higher methane yields, i.e. 307.5 (± 10) mL CH4/g VS, were obtained at thermophilic AD tests compared to mesophilic AD, i.e. 276.5 (±4.3) mL CH4/g VS. The total energy recovery from thermophilic DF + AD was higher (11.4 MJ/kg VS) than the mesophilic (10.4 MJ/kg VS) combined process. Interestingly, the analysis of kinetic parameters of mesophilic tests, determined from the Modified Gompertz equation, showed that mesophilic DF had faster H2 production kinetics, which can be attributed to a faster adaptation of the heat-shocked inoculum used in the tests to the incubation temperature. However, thermophilic AD tests exhibited faster kinetics for methane production.

Combination of ultrasonic and acidic pretreatments for enhancing biohythane production from tofu processing residue via one-stage anaerobic digestion.

Tofu processing residues (TPR) have received more attention as a source of bioenergy. However, their low solubility has hindered biohythane generation. Consequently, the ultrasonic and H2SO4 pretreatments were combined and compared for the first time to improve the hydrolysis of organic matter and carbohydrate and increase free amino nitrogen generation from TPR. Besides, the impact of pretreatments on biohythane generation was investigated. Under the optimal conditions of 7.54% substrate level, 8% H2SO4 concentration, 80 °C and 50 min, the coincident ultrasonic-H2SO4 pretreatment enriched the contents of soluble chemical oxygen demand, reducing sugar, and free amino nitrogen to 49675 mg/L, 26 g/L, and 1721 mg/L, respectively, greater than individual pretreatments. Also, Biohythane yield increased by 4.24-13.61% over control (389.42 ± 23.7 ml/g-VSfed). Furthermore, hydrogen yield at 42.5 ± 2.08 and 28.1 ± 1.07 ml/g-VSfed and sulfate removal efficiency at 93 and 92% were significantly improved with ultrasonic-H2SO4 and H2SO4 pretreatments, respectively, indicating acidogenic and sulfidogenic activity enhancement.

Progress in biohythane production from microalgae-wastewater sludge co-digestion: An integrated biorefinery approach.

Recent advances in microalgae to biohythane (bio-H2 and bio-CH4) conversion have achieved growing attention due to their eco-friendly and energy-efficient nature. Although microalgae are considered a potential 3rd - 4th generation biomass, their low C/N ratio and cell-wall biopolymers are challenging for biohythane production. This study emphasizes the solutions to mitigate the adverse effects of microalgae-based biohythane production using co-digestion with wastewater sludge. Wastewater sludge, an emerging environmental concern, is reviewed to be an effective co-substrate with microalgae to establish a biorefinery approach. The future trends and prospects of this biorefinery approach is critically reviewed to attain a profitable process. This study also reviewed the advantages of microalgae-wastewater co-cultivation and the application of activated sludge for bio-flocculation as a cost-effective solution for microalgae cultivation and harvesting. Microalgae-wastewater co-cultivation is also recommended to be effective for biohythane purification. The liquid digestate is suggested to be used as a culture media to enhance microalgal growth; whereas, the solid digestate could be transformed into resources through hydrothermal processes as a solution of digestate management. A practical biorefinery approach combining the synergistic benefits of microalgae-wastewater sludge and its biological conversion to biohythane would be an adjoining link to the beginning of a sustainable future.

Effect of mixing ratio and total solids content on temperature-phased anaerobic codigestion of rice straw and pig manure: Biohythane production and microbial structure.

Long-term semi-continuous experiments were carried out under three feedstock conditions to study the effects of mixing ratio and total solids (TS) content on temperature-phased anaerobic codigestion of rice straw (RS) and pig manure (PM). The results showed that biohythane only produced from the mixture with 6% TS content and its average content were 12.83 ± 1.19% (hydrogen) and 23.68 ± 1.12% (methane). Increasing mixture TS content and decreasing its RS ratio increased biohythane production and organic matter removal by creating a suitable process pH and increasing the anaerobic reaction rates. The highest biohythane production of the mixture reached 73.09 ± 3.03 ml/g VS (hydrogen) and 235.81 ± 9.30 ml/g VS (methane) at a mixing ratio of 5:1 and TS content of 6%. A variety of hydrogen-producing bacteria were found in the thermophilic reactor and Clostridium_sensu_stricto_1 played an important role. Butyric acid fermentation is the main hydrogen-producing pathway. Methanobacterium and Methanosaeta were dominant archaea in the mesophilic reactor.

Long-term in situ bioelectrochemical monitoring of biohythane process: Metabolic interactions and microbial evolution.

Microbial stability and evolution are a critical aspect for biosensors, especially in detecting dynamic and emerging anaerobic biohythane production. In this study, two upflow air-cathode chamber microbial fuel cells (UMFCs) were developed for in situ monitoring of the biohydrogen and biomethane reactors under a COD range of 1000-6000 mg/L and 150-1000 mg/L, respectively. Illumina MiSeq sequencing evidenced the dramatic shift of dominant microbial communities in UMFCs from hydrolytic and acidification bacteria (Clostridiaceae_1, Ruminococcaceae, Peptostreptococcaceae) to acetate-oxidizing bacteria (Synergistaceae, Dysgonomonadaceae, Spirochaetaceae). In addition, exoelectroactive bacteria evaluated from Enterobacteriaceae and Burkholderiaceae to Desulfovibrionaceae and Propionibacteriaceae. Especially, Hydrogenotrophic methanogens (Methanobacteriaceae) were abundant at 93.41% in UMFC (for monitoring hydrogen reactor), which was speculated to be a major metabolic pathway for methane production. Principal component analysis revealed a similarity in microbial structure between UMFCs and methane bioreactors. Microbial network analysis suggested a more stable community structure of UMFCs with 205 days' operation.

Onset Investigation on Dynamic Change of Biohythane Generation and Microbial Structure in Dual-chamber versus Single-chamber Microbial Electrolysis Cells.

Biohythane is alternative fuel to replace fossil fuel for car combustion, and biohythane generation could be potential pathway for energy recovery from wastewater treatment. Microbial electrolysis cell (MEC) is electrochemical technique to convert waste to methane and hydrogen gas for biohythane generation, but the feasibility and stability of MEC needs further investigation to assure sustainable energy recovery. System configuration is paramount factor for electrochemical reaction and mass transfer, and this study was to investigate the configuration impact (single vs dual chamber) of MEC for biohythane generation rate and stability. This study showed that dual-chamber MEC could separate methane and hydrogen gas production in the anode and cathode, and combined both together to produce biohythane. To reduce ohmic resistance for higher current, cation exchange membrane (CEM) was removed from dual-chamber to single-chamber MEC. However, free hydrogen diffusion was allowed in the single chamber since CEM was removed. The diffused hydrogen and substrate towards the cathode would favor the methanogen growth, and thus the hydrogen was consumed to reduce the biohythane generation and energy recovery efficiency (i.e., 7.5 × 10-3 reduced to 5.7 × 10-3 kWh kg-1 degraded COD day-1 after converting dual-chamber to single-chamber MEC). Absolute abundance of methanogen in single-chamber MEC was greatly boosted, as Methanosarcina and Methanobacteriale on the anode surface, increased by 132% and 243%, respectively, while the original dual-chamber MEC could maintain Geobacter growth for high current generation. This is the keystone study to demonstrate the importance of dual-chamber MEC for the feasibility and stability for the biohythane generation, building up the foundation to use electrochemical device to convert the organic waste to the alternative biohythane.