In this study, to improve the quality and shelf life of hamburgers, sesame meal protein hydrolysates (SPH) were produced using two enzymes of alcalase and flavourzyme and then four hamburger treatments: T1: control (10% soybean), T2: 1% SPH + soybean 9%, T3: 2% SPH + soybean 8%, and T4: 3% SPH + soybean 7% were prepared. Physicochemical properties were analyzed at the beginning of the storage period; microbial and chemical quality was evaluated at intervals of 0, 4, 8, 12, and 16 days. The results of SPH showed that alcalase enzyme can produce a SPH with a higher antioxidant properties (DPPH, FRAP, and beta-carotene-linoleic acid) (P < 0.05); therefore, this SPH was used for hamburger properties. According to the results, with the addition of SPH, moisture, fat, texture firmness decreased, protein, and brightness increased (P < 0.05), and all treatments had the allowable range.SPH replacement with soybean slowed down the increasing trend of oxidation and microbial spoilage (P < 0.05). In general, better results were observed in T3 and T4, which had a permissible range chemical and microbial index until the end of the storage period, as well as these treatments inhibited the growth of Staphylococcus aureus and Escherichia coli. Only T3 was approved by the evaluators.