Abstract: Using wheat B starch as raw material, optimize the one-step enzymatic hydrolysis process for preparing fermentable sugars and subsequent fermentation of alcohol by brewing yeast to improve its utilization value. Initially, the elemental composition of Wheat B starch was quantified. A composite enzymatic catalyst containing α-amylase and saccharase were then employed to generate fermentable sugars from wheat B starch through enzymatic hydrolysis efficiently. Various parameters, including composite enzyme concentration, solid-liquid ratio, enzyme temperature, enzymatic duration, and pH, were thoroughly investigated to assess their effects on reducing sugar yield. An orthogonal optimization approach was conducted to determine the optimal conditions for enzymatic hydrolysis. Results showed that wheat B starch contained 85.61% starch, with approximately 2% protein and mineral content, and pentosan composition nearing 5%. Following the orthogonal optimization, the most effective parameters for enzymatic saccharification were identified: A composite enzyme dosage of 4 mL, a solid-liquid ratio of 1:25 g/mL, an enzymatic digestion temperature of 60 ℃, an enzymatic digestion duration of 10 h, and a pH of 5.0. Under these specific conditions, the yield of reducing sugar reached 94.6%. Subsequently, wheat B starch was converted into alcohol through yeast-mediated fermentation. This process led to a final alcohol concentration 12.76 g/L, accompanied by an alcohol yield of 31.90%. In conclusion, wheat B starch holded remarkable promise for large-scale bioethanol production, providing valuable insights for maximizing the utilization of this resource on a broad scale.