Abstract: A method for preparing IPP- and VPP-rich hydrolysate from wheat gluten protein by enzymatic hydrolysis is disclosed, comprising the following steps: (1) dissolving wheat gluten protein in an alkaline solution, and performing ultrasonic treatment; (2) adding the alkaline protease to the wheat gluten protein solution to hydrolyze; (3) adjusting the pH value of the alkaline hydrolysate to neutral or weakly acidic, and adding protease to hydrolyze, and inactivating the enzyme at a high temperature; (4) centrifuging the hydrolysate to obtain the supernatant, and ultrafiltrating with an ultrafiltration membrane; (5) decolorizing and debitterizing the permeate using a macroporous resin; (6) after the debittered and decolorized liquid is adsorbed by an ion exchange resin, eluting the ion exchange resin and drying, to obtain a powder of IPP- and VPP-rich hydrolysate. The preparation method has a high protein recovery rate, with rich IPP and VPP in the hydrolysate.