The optimum conditions for the preparation of sorghum gliadin by ultrasonic assisted enzymatic method were studied by single factor and response surface test. The effects of standing time and temperature on the oil retention and water holding capacity of gliadin were studied. The results showed that the optimum conditions were as followed: enzyme dosage 1%, enzyme action time 3 h, ultrasonic action time 19 min, ethanol concentration 60%, the maximum extraction rate under the conditions of (7.19±0.36)%. At the temperature of 35 ℃and standing time of 40 min, the maximum oil holding capacityof sorghum gliadin was (2.84±0.15) g/g and the maximum water holding capacity was (2.78±0.09) g/g.