This study evaluated the effect of enzymatic hydrolysis on malt bagasse for fermentative sugar extraction in thermostatic and ultrasonic bath. The enzymatic reaction was performed using cellulase, α-amylase, and amyloglucosidase (in separate and combined) on malt bagasse to analyze the hydrolysis effect. For this, a central composite rotational design was applied to evaluate the effect of solid:liquid (malt bagasse w/v) versus enzymatic concentration, considering cellulase, α-amylase, and amyloglucosidase enzymes in mix solution, on fermentable sugars liberation. Then the kinetic fermentative sugars liberation was evaluated in a hydrolysis reaction using cellulase, α-amylase, and amyloglucosidase (in separate and combined) after exposure in thermostatic and ultrasonic bath. From enzymatic mix, the maximum total reducing sugar (TRS) yield was 408.8 and 219.9 mg⋅g −1 in thermostatic and in ultrasound bath, respectively. Kinetic studies showed a TRS variation of 6 to 7 g⋅L −1 for all enzymes. Amyloglucosidase was more promising in assimilation of malt bagasse, responsible for 43% of sugar releasing. Briefly, a low-cost procedure using thermostatic bath improved the hydrolysis reaction. The results from the experiments present a new application where malt bagasse can be employed showing the potential of this brewing industry residue in fomenting processes, as bioethanol production.