The article explores a novel approach to biohydrogen production by utilizing vegetable residue (VR) and wine lees as substrates in anaerobic fermentation processes, particularly dark fermentation. It addresses the challenges posed by increasing waste production and the demand for alternative energy sources. The study, conducted in Campania, Italy, focused on evaluating the potential of utilizing wine lees as an effective inoculum for biohydrogen production from VR. By assessing various VRs such as tomatoes, bell peppers, beans, zucchini, peas, and eggplants, the research aimed to model the biohydrogen production curve and identify the microbial community involved. The results demonstrated that wine lees showed promising biohydrogen production efficiency compared to other substrates. The physical-chemical characteristics of the inoculum and substrates played a significant role in the success of the fermentation processes. The study highlighted the importance of sustainable waste management practices and the potential of wine lees as a valuable source of microorganisms for eco-friendly biohydrogen production. Overall, the research presents a positive outlook on the use of innovative strategies for renewable energy production and waste valorization.