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Grain stillage from the liquor industry was pretreated by using microwave-assisted hydrothermal pretreatment, fungal pretreatments, and their combination to enable efficient enzymatic hydrolysis for sugar production. The microwave-assisted hydrothermal (MH) pretreatment was optimized by using a response surface methodology, and the respective maximum reducing sugar yield and saccharification efficiency of 17.59 g/100 g and 33.85%, respectively, were achieved under the pretreatment conditions of microwave power = 120 W, solid-to-liquid ra