Spent mushroom compost is one of the main potentials for biogas production. In recent years, several studies employed adding nanoparticles and alkaline pretreatment for improving biogas production. The present study is one of the pioneer studies that employ hybrid alkaline pretreatment (0, 5, and 15 mg of NaOH) and zero-valent iron nanoparticles (0, 10, 20, 30, and 40 mg) for improving the co-digestion of spent mushroom compost and rice straw. According to the results, retention time (RT) and nanoparticle (NP) concentrations have the most significant impact on biomethane production (significant at 1% probability level), while the NaOH concentration has the lowest impact on biomethane production (significant at 5% probability level) in comparison with RT and NP concentration. Also, the maximum biomethane production is related to NP40Na15 (about 200% higher than the control). The minimum cumulative biomethane production is related to NP0Na15 (about 30% lower than the control). The lowest relative environmental midpoint impact is related to NP40Na15, which was on average about 60% lower than the control. Adding NPs at high concentrations of NaOH reduces the midpoint impacts. The results of the study could lead to new, ecologically friendly biomethane production methods that make better use of agricultural and organic wastes.