The current concerns on global energy security, climate change, and environmental pollution represent some of the major elements of the growing interest on renewable energy. In this framework agro-food energy systems are at the center of a twofold debate: on the one hand they represent a key option for energy production while on the other their sustainability is threatened by the expansion of the bioenergy market that could lead to negative social and environmental consequences. The aim of this work is to evaluate—through a case study—the technical and economic feasibility of the replacement of energy crops (ECs) with used cooking oil (UCO) in an anaerobic digestion (AD) full-scale plant. At this purpose, a full-scale plant performing AD was monitored for two years. Three scenarios were developed and compared to evaluate the impacts and the potential benefits in terms of land saving in case of a substitution of ECs with UCO. Results highlighted a reduction of land use of over 50% if UCO is introduced in co-digestion with ECs. The lack of an appropriate legislative framework limits the utilization of used cooking oils (UCOs) in AD with a consequently missed opportunity for biogas owners that could find an important alternative in UCO.