Anaerobic co-digestion is a promising alternative to manage agri-food waste rather than landfilling, composting or incineration. But improvement of methane yield and biodegradability is often required to optimize its economic viability. Biomethanization of agri-food solid waste presents the disadvantage of a slow hydrolytic phase, which might be enhanced by adding a readily digestible substrate such as glycerol. In this study, strawberry extrudate, fish waste and crude glycerol derived from biodiesel manufacturing are mixed at a proportion of 54:5:41, in VS (VS, total volatile solids), respectively. The mesophilic anaerobic co-digestion at lab-scale of the mixture was stable at loads lower than 1.85 g VS/L, reaching a methane yield coefficient of 308 L CH4/kg VS (0 °C, 1 atm) and a biodegradability of 96.7%, in VS. Moreover, the treatment capacity of strawberry and fish waste was increased 16% at adding the crude glycerol. An economic assessment was also carried out in order to evaluate the applicability of the proposed process. Even in a pessimistic scenario, the net balance was found to be positive. The glycerol adding implied a net saving in a range from 25.5 to 42.1 €/t if compared to landfill disposal.