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This research focused on the effect of temperature (25-75 °C), extraction time (40-120 min) and solvent/solid ratio (15-45 mL/g) and the enzyme assisted extraction on protein extraction efficiency from by-product of sugar beet. 3 different methods were applied in protein isolation and the highest protein yield (34.55%) was obtained by the isoelectric-ammonium sulfate precipitation method. At the optimized condition of temperature 54.25 °C, 81.35 min, and solvent/solid ratio of 27.65 mL/g, the protein yield was found 55.15%. The protein yie