Organic waste can provide energy in the form of biogas, but processes should be improved. There is little knowledge on sequential pretreatment of extrusion and enzymatic hydrolysis for the production of biogas from biomass. Here, sequential extrusion and enzymatic hydrolysis were tested as pretreatment for methane production from vine trimming shoots. Results show that methane production by anaerobic digestion from extruded and hydrolyzed material, of 144.3 L CH4/kg volatile solids after 13 days, was faster than from untreated material, of 139.1 L CH4/kg volatile solids after 32 days. The final volume produced from extruded and hydrolyzed material exceeded in a 40% the production from untreated material. Thus, the sequence of extrusion and hydrolysis as pretreatment was positive for methane production from vine trimming shoots.