Abstract: In order to promote the separation of high amylose wheat starch（HAWS） and gluten protein, liquid and solid viscosity-reducing enzymes consisting of xylanase, cellulase, and β-glucanase were selected to study the effect of viscosity-reducing enzymes on the separation of high-amylose wheat starch（HAWS） in the Martin and batter process. The properties of the isolated HAWS were analyzed. The results showed that viscosity-reducing enzymes significantly improved the separation of starch from gluten in Martin process, increasing concentration of viscosity-reducing enzymes significantly decreased the protein content of HAWS from 2.70% to 1.72%, improved the gluten yield from 14.02% to 15.25%, and elevated the protein content of gluten from 80.83% to 82.00%. Addition of viscosity-reducing enzymes to the high-amylose wheat flour（HAWF） dough significantly improved the separation efficiency of HAWS from gluten, elevated the water absorption of gluten, and increased the gluten index silghtly. In batter process, adding viscosity-reducing enzymes to the batter significantly increased the yield of A-starch from 48.00% to 52.93%, reduced the protein content of A-starch from 1.09% to 0.93%, decreased the yield and protein content of B-starch, and reduced the viscosity of batter from 130.00 mPa·s to 45.80 mPa·s. Gluten yield and protein content increased with the concentrations increasing of viscosity-reducing enzymes. There was no significant difference between liquid and solid viscosity-reducing enzymes on the isolation of starch and gluten protein in high-amylose wheat flour. In the Martin and batter process, HAWS isolated using viscosity-reducing enzymes showed similar amylose content, pasting properties, thermal properties, in vitro starch digestibility properties, and granule morphology. These results suggest that the viscosity-reducing enzymes could be applied for industrial isolation of HAWS. This study provides valuable insights into enzyme-assisted HAWS isolation.