The article explores the potential of lignocellulosic waste as a renewable feedstock for biohydrogen production through dark fermentation processes. Hydrogen is considered a promising alternative energy source that could aid in reducing dependence on fossil fuels and facilitating the shift towards decarbonization in the energy sector. Agricultural biomass waste, such as willow, hay, wheat, or barley, rich in carbon content, can be effectively pretreated to serve as an excellent substrate for microorganisms. In this study, strains like Shewanella oneidensis MR-1, Cellvibrio japonicus Ueda107, and Sorangium cellulosum So ce27 were evaluated for their ability to produce biohydrogen through dark fermentation of lignocellulosic waste. Results indicated significant biohydrogen production rates, with S. oneidensis generating 787.6 ± 69.3 mL H2/L from willow in 12 h and C. japonicus producing 851.6 ± 20.8 mL H2/L from hay in 36 h. The main metabolites produced varied among the strains, with S. oneidensis primarily yielding acetic acid and C. japonicus and S. cellulosum producing isobutyric acid in different media. These findings suggest the potential of utilizing diverse feedstocks and microbial strains to enhance biohydrogen and metabolite production, showcasing the feasibility of sustainably revalorizing agricultural waste for energy generation. The study signifies a step towards leveraging biohydrogen as a renewable energy source and highlights the importance of exploring innovative approaches for sustainable energy production.