Membraneless microbial electrolysis cells (MECs) are potentially considered to produce biohydrogen (bioH2) in a green manner and simultaneously minimize agricultural and wastewater facility wastes. However, effective, sustainable, and cost-effective system configuration and improvement of operating variables, working at ambient conditions, are needed to make the MEC a sustainable process. Therefore, this study investigates the bioH2 production from poplar leaves and anaerobic sludge mixture by incorporating nanomaterials comprising Al2O3, MgO, and Fe2O3 metal oxides at various dosages. Moreover, the effects of applied cell voltage (0.5–1.5 V) and inoculum amount (20–40 mL) on bioH2 production and organic matter removal performance are evaluated. The maximum bioH2 production value is 417 mL at an applied voltage of 1.5 V with a chemical oxygen demand (COD) removal efficiency of 37.6 % under operating times of 5 min using 40 ml of inoculum. The bioH2 production of the MEC system is reduced with the decrease in inoculum amount. The highest bioH2 production of 828 mL is obtained at improved conditions in the presence of 1 g of Fe2O3 metal oxide. Overall, this study provides the potentiality of simultaneous waste minimization and bioH2 production under ambient conditions that highlight the waste-to-energy pathway for membraneless and green bioelectrochemical process.