In this study, alternative uses for lignocellulosic waste by considering them a source of eco-friendly and renewable energy generation with the application of the anaerobic digestion of treated and untreated waste for biogas and biohydrogen generation were investigated. The diluted sulfuric acid method was used for both the substrates and inoculum. Hydrogen production was absent when untreated spent coffee grounds (SCG) and alcohol waste (AW) were both used with the inoculum at pH 5.5 and pH 7.5. Meanwhile, the highest biogas yield of 320 dm³ kg V.S⁻¹ was obtained when using AW at pH 7.5, with a 190 dm³ kg V.S⁻¹ yield of methane. Instead, hydrogen production was observed when initially 4% (w/v) and 6% (w/v) SCG-containing hydrolysates were used as the substrates at pH 5.5, yielding 2.9 ± 0.09 dm³ kg V.S⁻¹ and 3.85 ± 0.12 dm³ kg V.S⁻¹, respectively. The further optimization of pretreatment technologies and pH control could lead to increased and prolonged hydrogen production.

After brewing roasted coffee spent coffee grounds (SCG) are generated being one of the daily wastes emerging in dominant countries with high rate and big quantity. E. coli BW25113 wild type strain, mutants with defects in hydrogen (H₂) producing/oxidizing four hydrogenases (Hyd) (ΔhyaB ΔhybC, ΔhycE, ΔhyfG) and septuple mutant (ΔhyaB ΔhybC ΔhycA ΔfdoG ΔldhA ΔfrdC ΔaceE) were investigated by measuring change of external pH, bacterial growth and H₂ production during the utilization of SCGs hydrolysate. In wild type, H₂ was produced with rate of 1.28 mL H₂ (g sugar)⁻¹ h⁻¹ yielding 30.7 mL H₂ (g sugar)⁻¹ or 2.75 L (kg SCG)⁻¹ during 24 h. In septuple mutant H₂ production yield was 72 mL H₂ (g sugar)⁻¹ with rate of 3 mL H₂ (g sugar)⁻¹ h⁻¹. H₂ generation was absent in hycE single mutant showing the main role of Hyd-3 in H₂ production. During utilization of SCG wild type specific growth rate was 0.72 ± 0.01 h⁻¹ with biomass yield of 0.3 g L⁻¹. Genetic modifications, control of external parameters during growth could lead to prolonged and enhanced microbiological H₂ production by organic wastes, which will aid more efficiently global sustainable energy needs resulting in diversification of mobile and fixed energy sources.

This study investigates the potential of sugar beet pulp (SBP), a lignocellulosic by-product of sugar production, as a low-cost substrate for biohydrogen and biomass generation using Escherichia coli under dark fermentation conditions. Two strains—BW25113 wild-type and a genetically engineered septuple mutant—were employed. SBP was pretreated via thermochemical hydrolysis, and the effects of substrate concentration, dilution, and glycerol supplementation were evaluated. Hydrogen production was highly dependent on substrate dilution and nutrient balance. The septuple mutant achieved the highest H₂ yield in 30 g L⁻¹ SBP hydrolysate (0.75% sulfuric acid) at 5× dilution with glycerol, reaching 12.06 mmol H₂ (g sugar)⁻¹ and 0.28 mmol H₂ (g waste)⁻¹, while the wild type under the same conditions yielded 3.78 mmol H₂ (g sugar)⁻¹ and 0.25 mmol H₂ (g waste)⁻¹. In contrast, undiluted hydrolysates favored biomass accumulation over H₂ production, with the highest biomass yield (0.3 g CDW L⁻¹) obtained using the septuple mutant in 30 g L⁻¹ SBP hydrolysate without glycerol. These findings highlight the potential of genetically optimized E. coli and optimized hydrolysate conditions to enhance the valorization of agro-industrial waste, supporting future advances in sustainable hydrogen bioeconomy and integrated waste biorefineries.

Various pretreatment methods for the valorization of sunflower husks (SHs) for H₂ gas generation through fermentation by Escherichia coli were investigated. We analyzed thermal treatment (TT), acid hydrolysis (AH), and alkaline hydrolysis (AlkH) at different substrate concentrations (50 g L⁻¹, 75 g L⁻¹, 100 g L⁻¹, and 150 g L⁻¹) and dilution levels (undiluted, 2× diluted, and 5× diluted). A concentration of 75 g L⁻¹ SH that was acid-hydrolyzed and dissolved twice in the medium yielded optimal microbial growth, reaching 0.3 ± 0.1 g cell dry weight (CDW) L⁻¹ biomass. The highest substrate level enabling effective fermentation was 100 g L⁻¹, producing 0.37 ± 0.13 (g CDW) × L⁻¹ biomass after complete fermentation, while 150 g L⁻¹ exhibited pronounced inhibitory effects. It is worth mentioning that the sole alkaline treatment was not optimal for growth and H₂ production. Co-fermentation with glycerol significantly enhanced both biomass formation (up to 0.42 ± 0.15 (g CDW) × L⁻¹)) and H₂ production. The highest H₂ yield was observed during batch growth at 50 g L⁻¹ SH hydrolysate with 5× dilution, reaching up to 5.7 mmol H₂ (g sugar)⁻¹ with glycerol supplementation. This study introduces a dual-waste valorization strategy that combines agricultural and biodiesel industry residues to enhance clean energy generation. The novelty lies in optimizing pretreatment and co-substrate fermentation conditions to maximize both biohydrogen yield and microbial biomass using E. coli, a widely studied and scalable host.