Influence of initial anolyte pH and temperature on hydrogen production through simultaneous saccharification and fermentation of lignocellulose in microbial electrolysis cell

An investigation on the performance of hydrogen production by simultaneous saccharification and fermentation (SSF) in a dual-chamber microbial electrolysis cell (MEC) was carried out to consider different anolyte pH levels and culture temperatures, and the influences of anolyte pH value and culture temperature on changes of current, organic acid and pH value were also evaluated. The maximal hydrogen production rate (HPR) of 2.46 mmol/L/D (hydrogen energy recovery 219.02%) was obtained at the initial anolyte pH of 6.5. Within the range of the tested operation temperatures (30–50 °C), the optimal temperature for hydrogen production by SSF in the MEC systems was 35 °C. Moreover, the contents of organic acids and reducing sugar significantly changed with varying in initial anolyte pH and temperature levels. The result indicates that a low initial anolyte pH value and high culture temperature was beneficial to hydrolysis of cellulose, and a high initial anolyte pH value and a moderate culture temperature to hydrogen production.     

Biohydrogen production from sugar industry effluents using nickel based electrode materials in microbial electrolysis cell

Biohydrogen production from sugar industry effluents in a dual chamber microbial electrolysis cell (MEC) was investigated in this study. The MEC reactor was operated with different effluents as a substrate from cane sugar and raw sugar reprocessing units of sugar industry. The biohydrogen production was investigated using different cathode materials of Nickel plate, Nickel foam, Stainless Steel mesh. The performance of MEC was tested based on the production of hydrogen, coloumbic efficiency, hydrogen recovery and COD removal efficiency respectively. The MEC hydrogen productions revealed that cane sugar effluent was more effective as compared to raw sugar effluent. The experimental results showed that at an applied voltage of 1.0 V, Ni-foam exhibited maximum hydrogen production of 1.59 and 1.43 mmol/L/D in cane sugar and raw sugar effluents respectively, which was about twice than SS-mesh and 1.2 times Ni-plate. This study shows that Ni-foam is one of the potential candidate as low cost electrode for improving hydrogen production in MEC technology with the treatment of industrial effluents.     

Surfactant-assisted green liquor dregs pretreatment to enhance the digestibility of paper mill sludge

This study optimizes a novel surfactant-assisted green liquor dregs (GLD) pretreatment of paper mill sludge (PMS), both of which are wastes from the kraft pulping industry, using a combined Response Surface Methodology (RSM) design. Optimized conditions give a maximal reducing sugar release of 16.38 g/L. A substantial reduction in heavy metals aluminum, chromium, cobalt, arsenic, lead, and copper after pretreatment illustrates the enhancement of substrate digestibility by reducing toxic elements. Separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) for hydrogen production are assessed. SSF produced a hydrogen yield of 3.72 mL/g, displaying a 36.26% increase from pretreated PMS compared to SHF. These findings provide insights into possible methods of reducing process duration, energy input, and costs incurred with waste disposal within the paper industry. Furthermore, improved hydrogen yield using an SSF process demonstrates the potential beneficiation of pulp and paper GLD and PMS wastes.     

Enhancing biohydrogen production from sugar industry wastewater using metal oxide/graphene nanocomposite catalysts in microbial electrolysis cell

Biohydrogen production through Microbial Electrolysis Cell (MEC) has drifted towards the development of suitable cost-effective cathode catalysts. In this study, two graphene hybrid metal oxide nanocomposites were used as catalysts to investigate hydrogen production in the MEC operated with sugar industry wastewater as substrate against phosphate buffer catholyte. Electrochemical characterizations exposed the better performance of NiO.rGO coated cathode which showed lesser overpotential at 600 mV and overall lowest resistance in the Nyquist plots than Ni-foam and Co₃O₄.rGO cathodes. The experimental results showed that at an applied voltage 1.0 V, NiO.rGO nanocomposite had exhibited maximum hydrogen production rate of 4.38 ± 0.11 mmol/L/D, Coloumbic efficiency of 65.6% and Cathodic hydrogen recovery of 20.8% respectively. The MEC performance in terms of biohydrogen production was 1.19 and 2.68 times higher than Co₃O₄.rGO and uncoated Ni-Foam. Hence, economical hybrid nanocomposite catalysts were demonstrated in MEC using industrial effluent for energy and environment sustainability.     

Converting waste molasses liquor into biohydrogen via dark fermentation using a continuous bioreactor

This study investigated the effects of substrate concentration, HRT (hydraulic retention time), and pre-treatment of the substrate molasses on biohydrogen production from waste molasses (condensed molasses fermentation solubles, CMS) with a CSTR (continuously-stirred tank reactor). First, the hydrogen production was performed with various CMS concentrations (40–90 g COD/L, total sugar 8.7–22.6 g/L) with 6 h HRT. The results show that the maximal hydrogen production rate (HPR) occurred at 80 g COD/L substrate (19.8 g ToSu/L, ToSu: Total Sugar), obtaining an HPR of 0.417 mol/L/d. However, maximum hydrogen yield (HY) of 1.44 mol H₂/mol hexose and overall hydrogen production efficiency (HPE) of 25.6% were achieved with a CMS concentration of 70 g COD/L (17.3 g ToSu/L). The substrate inhibition occurred when CMS concentration was increased to 90 g COD/L (22.6 g ToSu/L). Furthermore, it was observed that the optimal HPR, HY, and HPE all occurred at HRT 6 h. Operating at a lower HRT of 4 h decreased the hydrogen production performance because of lower substrate utilization efficiency. The employment of pre-heating treatment (60 °C for 1 h) of the substrate could markedly enhance the fermentation performance. With 6 h HRT and substrate pre-heating treatment, the HPE raised to 29.9%, which is 18% higher than that obtained without thermal pretreatment.     

Hydrogen gas production with Ni,Ni–Co and Ni–Co–P electrodeposits as potential cathode catalyst by microbial electrolysis cells

The development of efficient and economical cathode, operating at ambient temperature and neutral pH is a crucial challenge for microbial electrolysis cell (MEC) to become commercialize hydrogen production technology. In the present work, eight different electrodes are prepared by the electroplating of Ni, Ni–Co and Ni–Co–P on two base metals i.e., Stainless Steel 316 and Copper separately to use as cathode in MEC. Electrodeposited cathode materials have been characterized by XRD, XPS, FESEM, EDX and linear voltammetry. The fabricated cathodes show higher corrosion stability with improved electro-catalytic performance for the hydrogen production in the MECs as compared to the bare cathodes (SS316 and Cu). Data obtained from linear voltammetry and MEC experiments show that developed cathode possess four times higher intrinsic catalytic activity in comparison to bare cathode. Electrodeposited cathodes are intensively examined in membrane-less MEC, operating under applied voltage of 0.6 V in batch mode at 30 ± 2 °C temperature, in neutral pH with acetate as substrate and activated sludge as inoculum. Ni–Co–P electrodeposit on Stainless Steel 316 cathode gives maximum hydrogen production rate of 4.2 ± 0.5 m³(H₂)m⁻³d⁻¹, columbic efficiencies 96.9 ± 2%, overall hydrogen recovery 90.3 ± 4%, overall energy efficiency 241.2 ± 5%, volumetric current density 310 ± 5 Am⁻³. The net energy recovery and COD removal are 4.25 kJ/gCOD and 61%, respectively. Prepared cathodes show stable performance for continuous 5 batch cycle operations in MEC.     

Co-fermentation of a mixture of glucose and xylose to hydrogen by Thermoanaerobacter thermosaccharolyticum W16: Characteristics and kinetics

Glucose and xylose co-fermentation is crucial to maximize hydrogen yield from waste lignocellulose. In this study, cell growth, sugar consumption, and hydrogen production profiles of Thermoanaerobacter thermosaccharolyticum W16 feeding with a range of glucose and xylose were experimental investigated coupled with kinetic analysis. Results showed although T. thermosaccharolyticum W16 could use both glucose and xylose for hydrogen production, a maximum cell growth rate of 0.27 g/L/h and hydrogen production rate of 14.53 mmol/L/h was found with glucose as sole substrate, the value was 92.8% and 49.8% higher than using xylose as the only carbon source. Further interpolation analysis and experimental demonstration suggested when glucose content in the mixed substrate higher than 58.2%, the inhibitory effect on xylose utilization was increased, but when glucose concentration fell below 21.7%, its utilization will be subject to a certain degree of feedback inhibition. Coupling experimental results with kinetic analysis in this study provides a powerful evidence to further develop the potential of T. thermosaccharolyticum W16 as a biocatalyst for hydrogen production from lignocellulosic biomass.     

Impact of applied cell voltage on the performance of a microbial electrolysis cell fully catalysed by microorganisms

The effect of the operating voltage on the performance of a microbial electrolysis cell (MEC) equipped with both a bioanode and a biocathode for hydrogen production is reported. Chronoamperometry tests ranged between 0.3 and 2.0 V were carried out after both bioelectrodes were developed. A maximum current density up to 1.6 A m⁻² was recorded at 1.0 V with hydrogen production rate of nearly 6.0 ± 1.5 L m⁻² cathode day⁻¹. Trace amounts of methane, acetone and formate were detected in cathode's headspace and catholyte which followed the same trend as hydrogen production rate. Meanwhile substrate consumption in anolyte also followed the trend of hydrogen production and current density changes. The bioanode could utilise up to 95% of acetate in the tested voltage ranges, however, at a cell voltage of 2.0 V the bioanode's activity stopped due to oxygen evolution from water hydrolysis. Cyclic voltammograms revealed that the bioanode activity was vital to maintain the functionality of the whole system. The biocathode relied on the bioanode to maintain its potential during the hydrogen evolution. The overall energy efficiency recovered from both bioanode and external power in terms of hydrogen production at the cathode was determined as 29.4 ± 9.0%, within which substrate oxidation contributed up to nearly 1/3 of the total energy marking the importance of bioanode recovering energy from wastewater to reduce the external power supply.     

Enhancement of batch biohydrogen production from prehydrolysate of acid treated oil palm empty fruit bunch

Carbohydrates from hydrolyzed biomass has been a potential feedstock for fermentative hydrogen production. In this study, oil palm empty fruit bunch (OPEFB) was treated by sulfuric acid in different concentrations at 120 °C for 15 min in the autoclave. The optimal condition for pretreatment was obtained when OPEFB was hydrolyzing at 6% (w/v) sulfuric acid concentration, which gave the highest total sugar of 26.89 g/L and 78.51% of sugar production yield. However, the best conversion efficiency of OPEFB pretreatment was 39.47 at sulfuric acid concentration of 4%. A series of batch fermentation were performed to determine the effect of pH in fermentation media and the potential of this prehydrolysate was used as a substrate for fermentative hydrogen production under optimum pretreatment conditions. The prehydrolysate of OPEFB was efficiently converted to hydrogen via fermentation by acclimatized mixed consortia. The maximum hydrogen production was 690 mL H₂ L⁻¹ medium, which corresponded to the yield of 1.98 molH₂/mol_xylose achieved at pH 5.5 with initial total sugar concentration of 5 g/L. Therefore, the results implied that OPEFB prehydrolysate is prospective substrate for efficient fermentative hydrogen conducted at low controlled pH. No methane gas was detected throughout the fermentation.     

Integrated process to produce biohydrogen from wheat straw by enzymatic saccharification and dark fermentation

In this study, different pretreatment methods, including lyophilization, hydrothermal pretreatment, and ultrasound combined with dilute alkali post-cooking, were investigated to enhance the efficiency of enzymatic saccharification and biohydrogen production of the wheat straw. All pretreatment methods could effectively remove lignin and hemicellulose while retaining cellulose, further enhancing the biomass accessibility for subsequently enzymatic saccharification and biohydrogen production. A reducing sugar concentration of 13.18 g/L was acquired when wheat straw was treated with ultrasound and dilute alkali cooking (R_U). The sequential fermentative hydrogen yield of the substrate R_U was 133.6 mL/g total solids (TS), which was 5.6-fold larger than that of the raw material (23.9 mL/g TS). The study confirmed that ultrasound combined with dilute alkali cooking was an effective method, which not only provided significant guideline for improving biohydrogen production but also presented helpful direction for the efficient pretreatment of other lignocellulosic biomass.     

Enhanced hydrogen production in a UASB reactor by retaining microbial consortium onto carbon nanotubes (CNTs)

Biohydrogen and subsequent biomethane generation from biomass is a promising strategy for renewable energy supply, because this combination can lead to higher energy recovery efficiency and faster fermentation than single methane fermentation. Microbial consortium control by retaining hydrogen-producers through the addition of microbial carriers is an alternative to constructing hydrogen-producing reactors. Here we report the use of carbon nanotubes (CNTs) as microbial carriers to enhance microbial retention and the production of biohydrogen. Laboratory-scale upflow anaerobic sludge blanket (UASB) reactors with CNTs at 100 mg/L achieved a maximal hydrogen production rate of 5.55 L/L/d and a maximal hydrogen yield of 2.45 mol/mol glucose. Compared to frequently used activated carbon (AC) particles, CNTs resulted in quicker startup and better performance of hydrogen fermentation in UASB reactors. Scanning electron microscopy (SEM) and pyrosequencing results revealed that the reactor with CNTs led to a high proportion of hydrogen-producing bacteria among the microbial consortium, which endowed the microbes with strong flocculation capacity and hydrogen productivity.     

Maximizing biohydrogen production from water hyacinth by coupling dark fermentation and electrohydrogenesis

Biohydrogen production via dark fermentation has shown immense potential for simultaneous energy generation and waste remediation. However, the low substrate conversion rates limit its practical feasibility. Therefore, the present work attempts to develop a single chamber microbial electrolysis cell (MEC) as an additional means for biohydrogen production. Different organic substrates including simple sugars and volatile fatty acids were demonstrated as potential substrates for H₂ production in MEC. The use of water hyacinth as sole substrate for H₂ production was examined. Furthermore, the feasibility of using MEC for second stage energy recovery after dark fermentation was explored. The two-stage process exhibited improved performance as compared to single stage MEC process with overall hydrogen yield of 67.69 L H₂/kg COD_consumed, COD removal of 70.33% and energy recovery of 46%. These results suggest that coupled dark fermentation-MEC process can be a promising means for obtaining high yield biohydrogen from water hyacinth.     

Production of biohydrogen from waste wheat in continuously operated UPBR: The effect of influent substrate concentration

Utilization of waste materials is one of the most economical approaches to biohydrogen production. Continuous generation of biohydrogen in a bioreactor makes the process more economical with respect to the conventional physical and chemical method. The two main parameters that affect the biohydrogen production in a continuously operated bioreactor are hydraulic retention time (HRT) and influent substrate concentrations. The effect of influent substrate concentration on biohydrogen generation in an up-flow packed bed reactor (UPBR) at HRT = 3 h was investigated in this study. The substrate was waste wheat which was acid hydrolyzed in H₂SO₄ by adjusting the pH value to pH = 2, under high temperature as T = 90 °C in an autoclave to obtain fermentable sugar solution. A natural and porous support particle namely, aquarium biological sponge (ABS) was the microbial immobilization surface in the reactor. Total and hydrogen gas volumes, hydrogen percentage, influent and effluent substrate concentrations, VFA concentrations were monitored. The influent substrate concentration (TS_o) was varied between TS_o = 10 g/L and TS_o = 35 g/L. The process performance was evaluated as biohydrogen volume, percentages, rate and yield under varying operating conditions. The production volume (4275 ml/day) and the rate (3.05 L H₂/L day) were maximum at influent sugar concentration of TS_o = 25 g/L, but the yield reached to its maximum value as Y = 1.22 mol H₂/mol glucose at TS_o = 19 g/L. Substrate limitation and inhibitions were observed at influent concentrations of TS_o = 10 g/L and TS_o = 35 g/L, respectively. The results indicated that ABS could be suggested as a microbial support particle for hydrogen generation in immobilized systems.     

Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bio-ethanol production

A major constraint in the enzymatic saccharification of biomass for ethanol production is the cost of cellulase enzymes. Production cost of cellulases may be brought down by multifaceted approaches which include the use of cheap lignocellulosic substrates for fermentation production of the enzyme, and the use of cost efficient fermentation strategies like solid state fermentation (SSF). In the present study, cellulolytic enzymes for biomass hydrolysis were produced using solid state fermentation on wheat bran as substrate. Crude cellulase and a relatively glucose tolerant BGL were produced using fungi Trichoderma reesei RUT C30 and Aspergillus niger MTCC 7956, respectively. Saccharification of three different feed stock, i.e. sugar cane bagasse, rice straw and water hyacinth biomass was studied using the enzymes. Saccharification was performed with 50 FPU of cellulase and 10 U of β-glucosidase per gram of pretreated biomass. Highest yield of reducing sugars (26.3 g/L) was obtained from rice straw followed by sugar cane bagasse (17.79 g/L). The enzymatic hydrolysate of rice straw was used as substrate for ethanol production by Saccharomyces cerevisiae. The yield of ethanol was 0.093 g per gram of pretreated rice straw.     

Multi-electrode continuous flow microbial electrolysis cell for biogas production from acetate

Most microbial electrolysis cells (MECs) contain only a single set of electrodes. In order to examine the scalability of a multiple-electrode design, we constructed a 2.5 L MEC containing 8 separate electrode pairs made of graphite fiber brush anodes pre-acclimated for current generation using acetate, and 304 stainless steel mesh cathodes (64 m²/m³). Under continuous flow conditions and a one day hydraulic retention time, the maximum current was 181 mA (1.18 A/m², cathode surface area; 74 A/m³) within three days of operation. The maximum hydrogen production (day 3) was 0.53 L/L-d, reaching an energy efficiency relative to electrical energy input of η_E = 144%. Current production remained relatively steady (days 3–18), but the gas composition dramatically shifted over time. By day 16, there was little H₂ gas recovered and methane production increased from 0.049 L/L-d (day 3) to 0.118 L/L-d. When considering the energy value of both hydrogen and methane, efficiency relative to electrical input remained above 100% until near the end of the experiment (day 17) when only methane gas was being produced. Our results show that MECs can be scaled up primarily based on cathode surface area, but that hydrogen can be completely consumed in a continuous flow system unless methanogens can be completely eliminated from the system.     

Improved hydrogen recovery in microbial electrolysis cells using intermittent energy input

Microbial electrolysis cell (MEC) is a bioelectrochemical technology that can produce hydrogen gas from various organic waste/wastewater. Extra voltage supply (>0.2 V) is required to overcome cathode overpotential for hydrogen evolution. In order to make MEC system more sustainable and practicable, it is necessary to minimize the external energy input or to develop other alternative energy sources. In this study, we aimed to improve the energy efficiency by intermittent energy supply to MECs (setting anode potential = −0.2 V). The overall gas production was increased up to ∼40% with intermittent energy input (on/off = 60/15sec) compared to control reactor. Cathodic hydrogen recovery was also increased from 62% for control MEC to 69–80% for intermittent voltage application. Energy efficiency was increased by 14–20% with intermittent energy input. These results show that intermittent voltage application is very effective not only for energy efficiency/recovery but also for hydrogen production as compared with continuous voltage application.     

Pretreatment of sweet sorghum bagasse for hydrogen production by Caldicellulosiruptor saccharolyticus

Pretreatment of sweet sorghum bagasse, an energy crop residue, with NaOH for the production of fermentable substrates, was investigated. Optimal conditions for the alkaline pretreatment of sweet sorghum bagasse were realized at 10% NaOH (w/w dry matter). A delignification of 46% was then observed, and improved significantly the efficiency of enzymatic hydrolysis. Under hydrolysis conditions without pH control, up to 50% and 41% of the cellulose and hemicellulose contained in NaOH-pretreated sweet sorghum bagasse were converted by 24 h enzymatic hydrolysis to soluble monomeric sugars. The extreme thermophilic bacterium Caldicellulosiruptor saccharolyticus showed normal growth on hydrolysates of NaOH-pretreated biomass up to a sugar concentration of 20 g/L. Besides hydrogen, the main metabolic products detected in the fermentations were acetic and lactic acid. The maximal hydrogen yield observed in batch experiments under controlled conditions was 2.6 mol/mol C6 sugar. The maximal volumetric hydrogen production rate ranged from 10.2 to 10.6 mmol/(L h). At higher substrate concentrations the production of lactic acid increased at the expense of hydrogen production.     

Kinetics of cotton cellulose hydrolysis using concentrated acid and fermentative hydrogen production from hydrolysate

The kinetics of cotton cellulose hydrolysis using concentrated sulfuric acid and the performance of fermentative hydrogen production from the hydrolysate in the batch system was carried out in this study. Effects of sulfuric acid concentrations, cotton cellulose concentrations and operating temperatures on the cotton cellulose hydrolysis were investigated. It was found that cotton cellulose can dissolve completely in sulfuric acid concentration above 55% (by volume) at room temperature. The reduced sugar yields were varied from 64.3 to 73.9% (g R-sugar/g cotton cellulose) with the initial cotton cellulose concentrations of 30-70 g/L at a temperature of 40 °C.The reduced sugar concentrations and the initial pH of biohydrogen production were investigated at 37 °C. It was found that the optimal values of the hydrogen yield and substrate utilization were 0.95 mol H₂/mol R-sugar and 98% with an initial pH of 8.2, when substrate concentration was fixed at 20 g R-sugar/L. The maximum hydrogen yield was 0.99 mol H₂/mol R-sugar at a substrate concentration of 15 g R-sugar/L. Using the Gompertz Equation Model simulation, the maximum hydrogen production rate was 253 mL H₂/h/L at a substrate of 30 g/L and initial pH of 8.4.     

Biohydrogen production from pure and natural lignocellulosic feedstock with chemical pretreatment and bacterial hydrolysis

Pretreatment and saccharification of lignocellulosic materials is the key technology affecting the efficiency of cellulosic biohydrogen production. In this work, two pure cellulosic materials (i.e., carboxymethyl-cellulose (CMC) and xylan) were directly hydrolyzed (without pretreatment) by a cellulolytic isolate Cellulomonas uda E3-01 able to release extracellular cellulolytic enzymes. Natural cellulosic feedstock (i.e., sugarcane bagasse) was chemically pretreated prior to the bacterial hydrolysis.A temperature-shift strategy (35 °C for cellulolytic enzymes production and 45 °C for hydrolysis reaction) was used to increase the production of reducing sugars during the bacterial hydrolysis. The hydrolysates of CMC, xylan, and bagasse were efficiently converted to H₂ via dark fermentation with Clostridium butyricum CGS5. The maximum hydrogen yield was 8.80 mmol H₂/g reducing sugar (i.e., 1.58 mol H₂/mol hexose) for CMC, 6.03 mmol H₂/g reducing sugar (i.e., 0.91 mol H₂/mol pentose) for xylan, and 6.01 mmol H₂/g reducing sugar for bagasse.     

Electrodeposition of nickel particles on a gas diffusion cathode for hydrogen production in a microbial electrolysis cell

Gas diffusion cathodes with electrodeposited nickel (Ni) particles have been developed and tested for hydrogen production in a continuous flow microbial electrolysis cell (MEC). A high catalytic activity of electrodeposited Ni particles in such a MEC was obtained without a proton exchange membrane, i.e. under direct cathode exposure to anodic liquid. Co-electrodeposition of Pt and Ni particles did not improve any further hydrogen production. The maximum hydrogen production rate was 5.4 L/L_R/day, corresponding to Ni loads between 0.2 and 0.4 mg cm⁻². Continuous MEC operation demonstrated stable hydrogen production for over one month. Owing to the fast hydrogen transport through the cathodic gas diffusion layer, the loss of hydrogen production to methanogenic activity was minimal, generally with less than 5% methane in the off-gas. Overall, gas diffusion cathodes with electrodeposited Ni particles demonstrated excellent stability for hydrogen production compared to expensive Pt cathodes.