The objective of this research is to recover yeast protein from pot ale, a whisky by-product. Different yeast disruption methods were tested: enzymatic disruption, glass beads, the commercial reagent Y-PER plus, autolysis, sonication, homogenization and combination of mechanical and non-mechanical methods. This work was focused on the enzymatic disruption and combined method; the effect of different enzymes, enzyme concentration and time on protein recovery was studied. The optimal extraction method was the pot ale incubation with 1000 ppm Rohalase® BX and 0.015M EDTA followed by homogenization at 500 bars, which increases the protein recovery yield up to 50% compared with homogenization. The hydrolysis of yeast in pot ale permits the recovery of high quality proteins, which can be used as food or feed grade.