Enhanced Biohydrogen Production Research Articles

Biohydrogen production enhancement from organic solid waste using consecutive intermittent feeding strategies in a sequencing batch reactor

Abstract The organic fraction of municipal solid waste (OFMSW) can be valorized for bioenergy production by dark fermentation (DF) using sequencing batch reactors (SBR). Alternative feeding strategies such as fed-batch have shown increased biogas production. Since fed-batch operation with OFMSW is difficult because of the viscosity and density of its substrate, this work proposes the use of a feeding strategy that operates intermittently to improve the biohydrogen (H2) production from OFMSW in an SBR. The consecutive intermittent feeding strategy consisted in supplying the influent volume with a given number of equal feeding pulses in the first 16 h. Two, four, eight, and frequent pulses were tested in three consecutive cycles. Single pulse feeding (i.e., conventional batch) was performed before and after each feeding strategy for comparison. The four feeding strategies had a change in H2 and metabolite composition, cumulative volume, productivity, and yield. Intermittent feeding also diminished the lag phase for H2 production (λ ≤ 0.62 h). The frequent pulse strategy showed the best performance (365.5 ± 10.8 NmL, 340.3 ± 0.7 NmL L−1∙d−1, and 26.1 ± 0.7 NmL gVSadded for accumulated hydrogen production, hydrogen productivity, and yield, respectively), and it also increased caproate production (up to 1.12 gCOD L−1). Significant correlations between the production of organic acids and specific microbial genera were observed, highlighting the complex microbial community interactions present during biological hydrogen production. Graphical Abstract

Self-regulation of system buffering through flow rate optimization in continuous acidogenic fermentation for enhanced biohydrogen production

Self-regulation of system buffering through flow rate optimization in continuous acidogenic fermentation for enhanced biohydrogen production

Deep learning based modelling and control of a microbial electrolysis cell for enhanced bio hydrogen production

Deep learning based modelling and control of a microbial electrolysis cell for enhanced bio hydrogen production

Microwave-Assisted Acid and Alkali Pretreatment of Napier Grass for Enhanced Biohydrogen Production and Integrated Biorefinery Potential

Napier grass, a promising lignocellulosic energy crop, presents a complex composition that limits its bioconversion into fermentable products. To address this challenge, we applied microwave (MW) pretreatment assisted by acid and alkali, using varying chemical concentrations (0.5–1% w/v) and pretreatment times (3–10 min). Acid-catalyzed MW pretreatment achieved a maximal hemicellulose removal of 69.8%, while alkali-catalyzed MW pretreatment resulted in significant lignin removal of 65.5%. Without chemical catalysis, the pretreated hydrolysate significantly increased hydrogen yield to 38.0 ± 2.9 mL H2/g volatile solid (VS), five times greater than that obtained from untreated biomass. Hydrogen yield was further enhanced when the MW-pretreated solid underwent simultaneous saccharification and fermentation. The highest hydrogen yield of 89.2 ± 7.2 mL H2/g VS was achieved from alkali-catalyzed MW pretreated solid (0.5% w/v NaOH, 5 min), with a chemical oxygen demand (COD) solubilization of 62.6%. Increasing the NaOH concentration to 1% (w/v) slightly decreased hydrogen yield but significantly increased COD solubilization to 85.8%. The high carbohydrate content facilitated rapid cellulase hydrolysis, producing and accumulating a high concentration of fermentable sugars. However, this accumulation subsequently led to a shift towards lactic acid formation. The improved hydrogen yield and increased COD solubilization, along with the shift towards lactic acid production, suggest the possibility of optimizing this process for simultaneous production of multiple valuable products in an integrated biorefinery approach, potentially enhancing the economic viability of biomass conversion.

Self-adaptable HAc/NaAc buffer system enhanced biohydrogen production from dark fermentation of cellulose

ThepHdecrease caused by potential accumulation and dissociation of organic acidsis considereda major challenge hindering stable and constant operation in hydrogen production. In this study, a self-adaptableHAc/NaAc buffer system was investigated based on batch dark fermentation hydrogen production (DFHP) metabolic typesto controlthe pH of fermentation process. Resultsshowedthat increasing substrate concentration resulted in lower H2 production yield, especially when the substrate concentration exceeded 10 g/L. A maximum H2yield of2326.25 mL/L was achieved at the HAc/NaAc-buffered group; productions were 2.84 times and 57.7 % higher than the control and NaOH control groups. Our buffersystem retardedthe decrease of pH, enhanced the selectivemetabolic flux of acetic acid production, promoted the growth of microorganisms, enhanced microbial secretion of cellulase, andregulatedthe ratio of NADH/NAD+. The research provided a preliminary understanding and reference for the buffer regulatory strategy on organic waste for DFHP.

Exploring Optimal Pretreatment Approaches for Enhancing Biohydrogen and Biochar Production from Azolla filiculoides Biomass

Exploring Optimal Pretreatment Approaches for Enhancing Biohydrogen and Biochar Production from Azolla filiculoides Biomass

Incorporating concrete waste as additive in acidic fermentation–A novel approach for enhanced biohydrogen production and concrete mass reduction

Acidic fermentation (AF) of organic waste/wastewater generates valuable byproducts, including hydrogen, volatile fatty acids, and ethanol; however, its sensitivity to pH drops limits the stability and efficiency of the process. A reversal process, the acidic chemical treatment of concrete waste (CW) to recycle its aggregates, generates calcium hydroxide, which can serve as buffering agent. Meanwhile, integrating both processes can introduce a sustainable win-win approach, which is the aim of this study. To assess this approach, a series of batch AF experiments were conducted at 50 °C and pH 5.5, using glucose as the substrate. Cementitious specimen (1.5 × 1.5 × 0.8 cm) was supplemented to each 200 mL-fermenter. Unamended fermenters, with and without additional NaHCO3-buffering, were used as controls to compare with the amended ones. Introducing cementitious specimens to AF increased H2-production by 2-fold and 1.7-fold compared to controls with and without NaHCO3-addition, respectively, after three consecutive feed-cycles. The surface analysis of incorporated specimen confirmed the Ca, Al, Mg, and Si leaching. The AF efficiency and resulting cementitious mass reduction were further assessed at different organic loads and specimens’ volume, surface area, and porosity (by changing water-to-cement [W/C] ratio). Increasing the organic load from 10 to 20 g-glucose/L resulted in lower H2-production, higher specimen mass reduction (up to ∼32%), and higher Ca2+ release (up to 2 g/L); however, no significant effect was observed when using specimens with higher W/C ratio or surface area. Moreover, the presence of cementitious specimens significantly influenced the microbial composition, leading to notable developments in the abundant genera Thermoanaerobacterium and Bacillus. This study presents a novel approach to sustainably enhancing AF process using CW as both an additive and a treatable substance, with reusable aggregates as a byproduct. It provides valuable insights for optimizing the process and guiding future practical applications. This includes considering various concrete compositions, adjusting organic load conditions, and evaluating long-term stability in larger-scale systems.

Synergising hydrothermal pre-treatment and biological processes for enhancing biohydrogen production from palm oil mill effluent

The high quantity of nutrient-rich palm oil mill effluent (POME) waste in the industry will have a severe negative environmental impact and a high cost of treatment. However, POME waste could be converted into bioenergy via environmentally sustainable processes. Several studies have explored using hydrothermal carbonisation for solid biomass products in biofuel production, but the potential of the liquid phase produced during these processes has received less attention. Therefore, this study aims to assess the potential of biohydrogen production from treated POME as a substrate by hydrothermal process. This study is presented in two phases: the first phase involves substrate pre-treatment using a hydrothermal process to improve biomass properties at different temperatures, and the second phase explores the potential for biohydrogen production from treated POME through dark fermentation. Substrate pre-treatment was conducted at 180, 210, and 240 °C using 100 % raw POME. Next, the treated POME was incubated for biohydrogen production at 50 °C for 24 h. A microbial analysis was conducted to determine the most dominant species present in the sample. Our findings show that at 180 °C, the total chemical oxygen demand (COD) removal efficiency was 80 %, and acetic acid concentration was 28 %. Compared to raw POME, treated POME generated a maximum hydrogen yield and rate (HPR) of 52.19 mL H2 g−1 CODrem and 0.59 mL H2 mL POME−1 day−1 with a 2.32-fold and 1.59-fold increase, respectively. Meanwhile, Clostridium was a dominant bacterial species identified in the treated POME. These findings demonstrated the feasibility of implementing a hydrothermal process to treat POME and improve its biohydrogen production efficiency. The treated POME from the hydrothermal process is more homogenous and readily consumable by microorganisms used in dark fermentation. Hydrothermal pre-treatment could potentially increase the rate and efficiency of microbial digestion, leading to enhanced hydrogen production. The high COD removal efficiency during the process significantly reduces the environmental impact of POME discharge, and converting POME into a valuable resource through the hydrothermal and dark fermentation process aligns with sustainable waste management practices.

Application of artificial intelligence and red-tailed hawk optimization for boosting biohydrogen production from microalgae

Enhancing biohydrogen production from microalgae is crucial in addressing environmental and energy challenges. It provides a sustainable, clean energy source while reducing greenhouse gas emissions. Moreover, it advances microalgae-based biotechnology, enabling innovative biofuel production and ecological revitalization. The main target of this study is to develop a robust ANFIS model to simulate the biohydrogen production process from microalgae within photobioreactors. The study focuses on enhancing hydrogen yield by optimizing three critical process parameters: sulfur concentration (%), run time (hours), and wet biomass concentration (g/L). Initially, an adaptive neuro-fuzzy inference system (ANFIS) model for biohydrogen production process is constructed based on empirical data. Subsequently, the red-tailed hawk algorithm (RTH) is used to determine the optimal values for the process parameters, corresponding to maximum hydrogen yield. The performance of ANFIS model in predicting hydrogen yield is assessed using root mean square error (RMSE) and coefficient-of-determination (R2) values. The obtained RMSE values for training and testing data are 2.8477 × 10−05 and 1.2807, respectively, while the corresponding R2 values are 1.0 and 0.9911 for training and testing. The introduction of fuzzy logic into the model significantly improves its predictive accuracy, as evidenced by the drop in RMSE from 10.79 with ANOVA to 0.7159 with ANFIS, representing a substantial 93.4 % decrease. The remarkable precision of the ANFIS model, indicated by its low RMSE and high R2 values, underscores the success of the modeling stage. The combination between ANFIS with the RTH technique yields impressive results, leading to a hydrogen yield enhancement of 6.87 % and 26.65 % when compared to both measured data and ANOVA.

Enhancing biohydrogen production: A comparative analysis of utilization of Jerusalem artichoke and bakery waste by dark fermentation

This study investigates the potential of using Jerusalem artichoke (JA) and bakery waste (BW) for biogas and biohydrogen production through dark fermentation. The experiment included four series with different ratios of JA to BW (100:0, 0:100, 50:50, 75:25) under mesophilic conditions at 39 °C. The highest biogas (1.46 L/L.d) and hydrogen production (0.508 L/L.d) was achieved with BW alone. A synergistic effect was observed with the 50:50 mixture, resulting in the highest production of volatile fatty acids (VFA), which reached up to 22252 mg/L. This shows the advantages of combining these substrates for optimized energy production. Spearman rank correlation analysis identified ammonium-nitrogen (N-NH4) as the most influential factor and showed a strong positive correlation with butyric acid/acetic acid (B/A) ratio (ρ = 0.85, p < 0.001). This indicates that maintaining optimal ammonium-nitrogen levels is critical for maximizing yields. In addition, total solids (TS) and volatile solids (VS) showed moderate positive correlations with specific VFAs, indicating their significant role in VFA dynamics. These results highlight the importance of substrate optimization and maintaining stable fermentation conditions for sustainable and efficient energy production.

Enhanced biohydrogen production from thermally hydrolysed pulp and paper sludge via Al2O3 and Fe3O4 nanoparticles

The growing demand for sustainable and green energy sources has led to increasing interest in biohydrogen production from renewable biomass feedstocks. In this study, pulp and paper sludge (PPS), a widely available waste residue, was thermally treated at different temperatures (90°C, 130°C, and 165°C) for varying durations (15, 30, and 60 min). Thermal hydrolysis of PPS increased the chemical oxygen demand (COD) solubilization from 11 % to 24.7 %, and volatile suspended solids (VSS) solubilization up to 15 % with increasing both hydrolysis temperature and reaction time. The resulting thermally treated samples were then evaluated for biohydrogen production through a batch assay. Among the different thermal treatment conditions, the sample treated at 165°C for 60 min exhibited the highest biohydrogen production potential and yield (1287 mL-H2 and 201 mL-H2/g volatile solids (VS)), which is 72 % higher the control untreated PPS (747 mL-H2 and 117 mL-H2/gVS). To further enhance the biohydrogen yield, this optimal sample was mixed with two types of chemically synthesized nanoparticles, namely aluminium oxide (Al2O3) and magnetite (Fe3O4), at various concentrations (50, 100, and 200 mg/g VS). The addition of nanoparticles significantly influenced the biohydrogen production from the thermal-treated PPS. Remarkably, the batch assay mixed with 200 mg of Fe3O4 nanoparticles per gram of VS demonstrated the highest biohydrogen production potential, compared to the thermally treated PPS (1577 vs. 1226 mL-H2). This finding suggests that the presence of Fe3O4 nanoparticles enhances the biohydrogen production process, possibly through improved microbial activity and substrate accessibility. The results of this study highlight the potential of utilizing PPS, an abundant waste product, as a valuable feedstock for biohydrogen production. Overall, this study contributes to the advancement of green energy technologies and underscores the potential of biohydrogen as a renewable and sustainable energy source.

The intracellular accumulation of iron coincides with enhanced biohydrogen production by Thermoanaerobacterium thermosaccharolyticum

The aim of this study was to investigate the effects of different doses (10–200 mg/L) of magnetite (FeO·Fe2O3), Fe0, Ni0, and FeCl3 on the efficiency of dark fermentative H2 production from cheese whey using Thermoanaerobacterium thermosaccharolyticum SP-H2. Energy conversion efficiency and hydrogen yield increased significantly by 22.7 % and 34.8 %, respectively, when 10 mg/L of magnetite was added, and hydrogenase activity increased by 23.6 % when 100 mg/L of Fe0 was added compared to the control (p < 0.001). Using scanning electron microscopy and energy dispersive X-ray spectroscopy, it was found that during the early exponential phase of hydrogen evolution, T. thermosaccharolyticum cells accumulated up to 11.2 times more Fe (up to 2.8 wt%) in the presence of optimal concentrations of Fe additives compared to the control. Intracellular iron accumulation and hydrogen yield were positively correlated with acetate concentration (p < 0.001), suggesting an efficient acetate pathway for dark fermentation. During the stationary growth phase, with almost no H2 production, the intracellular iron content decreased by up to 67.9 %. The phenomenon of changes in the amount of metal in cells, depending on the phase of hydrogen production, was demonstrated for the first time. The findings suggest that intracellular metals can be used in cell metabolism to produce H2, and the mechanisms behind this process need further investigation. Overall, this study provides valuable insights into the optimal forms and doses of metals for enhancing biohydrogen production, as well as possible research directions for the potential use of metals by microorganisms during additive-assisted dark fermentation of carbohydrate-rich wastes.

Regulating the metabolites through SiC, ZnO, and SnO2 nano-photocatalysts for elevating efficiency of photo fermentative bioH2 from Arundo donax L.

Photo-fermentation offers a promising roadmap for hydrogen production through their ability to harness light energy, however, its lower conversion efficiency is the major bottleneck for its realization. This study investigates the catalytic role of wideband gap nano-photocatalysts (SiC, ZnO, and SnO2) in enhancing hydrogen generation from Arundo donax L. Physio-optical characteristics of SiC, ZnO, and SnO2 nano-photocatalysts were assessed through SEM, XRD, and FT-IR which confirmed that incorporated nano-photocatalysts are spherical with uniform size distribution and there is no secondary phase. Hydrogen production experiments with varying nano-photocatalyst concentrations revealed that the maximum hydrogen yield of 104 and 98.83 mL/g was achieved with SiC and ZnO, respectively, at loading concentration of 200 mg/L, whereas 69.30 mL/g were observed with 100 mg/L of SnO2. The incorporation of SIC, ZnO, and SnO2 nano-photocatalyst permitted to increase the hydrogen content in overall gas production by 41 %, 37.34 %, and 18.26 % as compared to the control group. The increase in hydrogen production is related to the enhanced e-h pair separation lifetime of SiC, confirmed from PL spectroscopy. Addition of SiC caused a longer availability of electrons that helped to increase the metabolic rate by 38 % and 40.29 % in energy conversion efficiency (ECE). ANOVA results revealed significant differences between the control and maximum values of CHP, HC%, byproducts, and ECE. Tukey pairwise comparisons showed that SiC and ZnO had non-significant differences at 200 mg/L but both significantly outperformed SnO2 at 100 mg/L. Thus, the selection of suitable catalysts photocatalysts with enhanced e-h pair separation lifetime may offer promising prospects for sustainable energy technologies by enhancing biohydrogen production.

Enhancing biohydrogen production from xylose at low temperature (20 °C) using natural FeS2 Ore: Thermodynamic analysis and mechanistic insights

This study investigates the efficacy of pyrite in enhancing biohydrogen production from xylose at low temperature (20 °C). Higher hydrogen yield rates (Rm) and reduced lag time (λ) were achieved across initial xylose concentrations ranging from 2-10 g/L. At an optimal xylose concentration of 5 g/L, pyrite reduced λ by 2.5 h and increased Rm from 1.3 to 2.7 mL h−1. These improvements are attributed to pyrite’s ability to enhance the secretion of extracellular polymeric substance and flavins, facilitate NADH and NAD+ generation and transition, and favor biohydrogen production. Thermodynamic analyses and Gibbs free energy calculations further elucidated pyrite’s role in the full reaction process and rate-limiting steps at low temperature. This study offers valuable insights into improving the efficiency of biohydrogen production at low temperature, with significant implications for energy conservation.

Effects of carbon source variability on enhanced Bio-hydrogen production

This study investigated how glucose, starch, and rapeseed oil, three common food waste components with diverse molecular and physicochemical characteristics, influenced hydrogen production and microbial communities in dark fermentation under varying carbon/nitrogen (C/N) ratios. The results indicated that glucose and starch groups, significantly increased hydrogen yields to 235 mL H2/gVS (C/N = 40) and 234 mL H2/gVS (C/N = 40), respectively, while rapeseed oil, with a lower yield of 30 mL H2/gVS (C/N = 20), demonstrated a negative impact. Additionally, an accumulation of propionate was observed with increasing carbon source complexity, suggesting that simpler carbon sources favored hydrogen production and bacterial growth. Conversely, lipid-based materials required rigorous pre-treatment to mitigate their inhibitory effects on hydrogen generation. Overall, this study underscores the importance of carbon source selection, especially glucose and starch, for enhancing hydrogen production and microbial growth in dark fermentation, while highlighting the challenges posed by lipid-rich substrates that require intensive pre-treatment to optimize yields.

Enhancing Biohydrogen Production: The Role of Iron-Based Nanoparticles in Continuous Lactate-Driven Dark Fermentation of Powdered Cheese Whey

Here, a comprehensive investigation was conducted under various operational strategies aimed at enhancing biohydrogen production via dark fermentation, with a specific focus on the lactate metabolic pathway, using powdered cheese whey as a substrate. Initially, a batch configuration was tested to determine both the maximum hydrogen yield (100.2 ± 4.2 NmL H2/g CODfed) and the substrate (total carbohydrates) consumption efficiency (94.4 ± 0.8%). Subsequently, a transition to continuous operation was made by testing five different operational phases: control (I), incorporation of an inert support medium for biomass fixation (II), addition of carbon-coated, zero-valent iron nanoparticles (CC-nZVI NPs) at 100 mg/L (III), and supplementation of Fe2O3 nanoparticles at concentrations of 100 mg/L (IV) and 300 mg/L (V). The results emphasized the critical role of the support medium in stabilizing the continuous system. On the other hand, a remarkable increase of 10% in hydrogen productivity was observed with the addition of Fe2O3 NPs (300 mg/L). The analysis of the organic acids’ composition unveiled a positive correlation between high butyrate concentrations and improved volumetric hydrogen production rates (25 L H2/L-d). Moreover, the presence of iron-based NPs effectively regulated the lactate concentration, maintaining it at low levels. Further exploration of the bacterial community dynamics revealed a mutually beneficial interaction between lactic acid bacteria (LAB) and hydrogen-producing bacteria (HPB) throughout the experimental process, with Prevotella, Clostridium, and Lactobacillus emerging as the predominant genera. In conclusion, this study highlighted the promising potential of nanoparticle addition as a tool for boosting biohydrogen productivity via lactate-driven dark fermentation.

Comparative Analysis of Various Seed Sludges for Biohydrogen Production from Alkaline Pretreated Rice Straw

The present work studied the effects of alkali pretreatment on the cellulosic biomass of rice straw. The improvement in the cellulose content and reduction in the lignin and hemicellulose percentage was observed with alkali pretreatment. Fourier transformation infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM) analysis confirm the modification in the surface structure of alkali rice straw. Further, the study investigated the potential of different types of seed sludge as inoculum sources for dark fermentative biohydrogen production. In comparison to other sludge samples (beverage industry, food industry, and sewage treatment plant sludge), the mixed culture of sewage treatment plant sludge had the highest cumulative volume of biohydrogen (90.52 mL), as well as the highest hydrogen production yield (0.75 moleH2/mole) with the substrate utilization of 86.72%. The results provide information on the best sludge source for enhancing biohydrogen production in the dark fermentation method.

Improved biohydrogen production by co-fermentation of corn straw and excess sludge: Insights into biochemical process, microbial community and metabolic genes

The global climate change mainly caused by fossil fuels combustion promotes that zero-carbon hydrogen production through eco-friendly methods has attracted attention in recent years. This investigation explored the biohydrogen production by co-fermentation of corn straw (CS) and excess sludge (ES), as well as comprehensively analyzed the internal mechanism. The results showed that the optimal ratio of CS to ES was 9:1 (TS) with the biohydrogen yield of 101.8 mL/g VS, which was higher than that from the mono-fermentation of CS by 1.0-fold. The pattern of volatile fatty acids (VFAs) indicated that the acetate was the most preponderant by-product in all fermentation systems during the biohydrogen production process, and its yield was improved by adding appropriate dosage of ES. In addition, the content of soluble COD (SCOD) was reduced as increasing ES, while concentration of NH4+-N showed an opposite tendency. Microbial community analysis revealed that the microbial composition in different samples showed a significant divergence. Trichococcus was the most dominant bacterial genus in the optimal ratio of 9:1 (CS/ES) fermentation system and its abundance was as high as 41.8%. The functional genes prediction found that the dominant metabolic genes and hydrogen-producing related genes had not been significantly increased in co-fermentation system (CS/ES = 9:1) compared to that in the mono-fermentation of CS, implying that enhancement of biohydrogen production by adding ES mainly relied on balancing nutrients and adjusting microbial community in this study. Further redundancy analysis (RDA) confirmed that biohydrogen yield was closely correlated with the enrichment of Trichococcus.

Enhanced bio-hydrogen production by photo-fermentation of corn stalk using Fe-doped CaTiO3 photocatalyst

The aim of this study was to analyze the effects of CaTiO3 photocatalysts doped with different concentrations of Fe on the biohydrogen production process by photo-fermentation using corn stover as the substrate. The experimental results showed that the maximum cumulative hydrogen production and the maximum rate of hydrogen production were obtained when the doping concentration was 6 % (g/g) (380.8 mL, 26.4 mL/h), which were 63.4 % and 203.4 % higher, respectively, than those of the blank control group (233 mL, 8.7 mL/h). The substrate conversion rate was 32.82 %. Iron-doped CaTiO3 lowered the oxygen reduction potential in the fermentation system to provide a suitable reduction environment for the growth and fermentation of photosynthetic bacteria. The lowest redox potential observed was −393.5 mV for 6 % iron-doped CaTiO3. Thermogram analysis revealed a strong correlation between cumulative hydrogen production and redox potential. Additionally, the analysis of soluble small molecules showed that iron-doped CaTiO3 could inhibit the production of ethanol from glycolysis during fermentation and promote the utilization of small molecule acids.

Screening of enhanced biohydrogen production from anaerobic fermentation amended with nano-sized barium ferrite supported by aluminum oxide

Anaerobic fermentation has attracted wide attention to effectively reduce carbon footprint and alleviate potential energy shortages, which is being regarded as a sustainable and promising technology to obtain high biohydrogen production from widespread organic wastes. However, biohydrogen commercial uptake is now dominantly inhibited by the low hydrogen yield and process instability. In this work, Al2O3/BaFe2O4 nanoparticles (NPs) are successfully applied to anaerobic fermentation for investigating their effects on hydrogen yield, activity of bacteria and microbial community structure. The benefits of Al2O3/BaFe2O4 NPs with respect to microbial metabolism, reactivity of enzymes and process optimization are systematically researched through batch experiments. In addition, various characterizations are used to verify the physical and chemical properties of Al2O3/BaFe2O4 NPs. The results show that the activity of hydrogenases and microbe metabolism can be significantly inhibited by excess addition of Al2O3/BaFe2O4 NPs (150 mg/L and 200 mg/L) while the optimum addition 100 mg/L of Al2O3/BaFe2O4 NPs can boost the biohydrogen yield by 49% via facilitating microorganism growth and enhancing key enzymatic activities, which indicate that the Al2O3/BaFe2O4 NPs display a beneficial influence on electron transfer efficiency and process stability. The existence of Fe3+, Al3+and Ba2+ derived from the corrosion of Al2O3/BaFe2O4 NPs by organic acids synergistically improve the microbial transmembrane transportation and substrate utilization, which contribute to higher conversion rate of glucose into acetate, butyrate and H2. Meanwhile, the complementary functions of Fe3+ and Al3+ can powerfully shorten the lag phase and promote the glycolysis, which finally accelerate the dark fermentation process. Compared with the control group, Clostridium_sensu_stricto_1 is selectively enriched by the addition of Al2O3/BaFe2O4 NPs, which keep heavily involved in the synthesis of H2. On balance, this study provides an effective strategy to enhance hydrogen productivity and maintain process stability, which offer the valuable information for the sustainable utilization of sewage sludge.