In this study, a new pre-treatment method based on novel non-thermal plasma technology was developed to improve the enzymatic hydrolysis of brewer's spent grain (BSG) and subsequent bioethanol production. A submerged dielectric barrier discharge plasma reactor system was applied for this purpose. Pre-treatments were performed by taking into account variables including; voltages (22 kV, 25 kV and 28 kV), solvent (acid, alkali and water) and time (5, 10, 15 min). The resulting treated biomass was subjected to enzymatic hydrolysis. A 2.14-fold increase in yield of the reducing sugar was achieved post hydrolysis when the biomass was treated in water for 10 min at a voltage setting of 28 kV (162.90 mg/g of BSG) compared to control (75.94 mg/g of BSG). This research suggests that subjecting lignocellulose to plasma discharges can enhance the efficiency of enzymatic hydrolysis. A high ethanol titre was also obtained upon fermentation of the hydrolysate (25.062 g/l).