Objective： This study aims to optimize the adsorption of enzymatic hydrolysis of Pueraria protein by macroporous resin and maximize the antioxidant properties in vitro. Methods: The Box-Behnken response surface method was used to determine the optimization of adsorption of enzymatic hydrolysis of Pueraria protein. Using V_C as a control, the antioxidant activity of Kase was determined before and after purification under the optimal purification process. Results： The optimal adsorption-desorption process of macroporous resin is: the mass concentration of the sample solution was 10.0 mg/mL, the flow rate of the eluent was 2.6 mL/min, and the volume fraction of ethanol in the eluent was 74%. After adsorption, the content of enzymatic hydrolysis of Pueraria protein increased to 37.19%. The scavenging rate of DPPH, ABTS⁺, and hydroxyl radicals of Pueraria Mirifica protease after adsorption was stronger than before adsorption. Conclusion： This study shows that enzymatic hydrolysis of Pueraria protein after adsorption of macroporous resin has good antioxidant effect, which can be used as a potential protein polypeptide in food, providing data reference for further research of enzymatic hydrolysis of Pueraria protein.