This study aimed to evaluate the effect of a partial substitution of chicken breast meat with oyster mushroom stalk powder on the quality attributes of mushroom-chicken burgers. Chicken breast eat was substituted with oyster mushroom stalk powder at different levels ranging from 2.5 to 10%, with an interval increase of 2.5%. The substitution level had a significant effect on the different characteristics studied. A decrease in both crude protein and crude lipid was observed as the substitution level increased from 2.5 to 10%. However, a contradictory trend was observed for crude fiber, ash, carbohydrates, antioxidant properties, water-binding capacity, and browning index. The cooking loss and shrinkage were reduced by about 81.6% and 72.6%, respectively, for the formulation prepared with 10% compared to the control sample. The formulation prepared with 2.5% showed the lowest hardness, chewiness, and shear force, which increased as the substitution level increased to 10%. The oyster mushroom stalk powder can be used at a level ranging from 2.5 to 5% to substitute chicken breast meat in the preparation of healthier and more functional chicken burgers