Abstract: This study aimed to optimize the sterilization process for soft-pack canned stuffed shiitake mushrooms, maximizing the preservation of their edible and nutritional qualities while achieving commercial sterility. This study investigated the effects of sterilization conditions—specifically F-values of 9, 12, and 15 minutes and sterilization temperatures of 110, 115, 121, and 126 ℃—on the quality of soft canned shiitake mushroom stuffed meat products. First, by measuring indicators such as pH, color, texture, sensory evaluation scores, and commercial sterility, we investigated the relationship between product quality and F-value to determine the optimal F-value. Building upon this foundation, we further investigated the effects of varying sterilization temperatures on the edible and nutritional qualities of stuffed shiitake mushroom products under identical sterilization intensity (F=9 min). Key indicators included electronic nose analysis, TBARS values, total carbonyl content, MRI imaging, moisture distribution, protein secondary structure, and essential nutritional components. This study aimed to determine optimal sterilization conditions for stuffed shiitake mushroom products. Results indicated that as sterilization intensity (F-value) increased, the product's pH first decreased then increased, while textural properties (hardness, chewiness) first improved then deteriorated. Sensory scores significantly declined (P<0.05). A sterilization duration of 9 minutes represented the optimal process parameter for achieving both commercial sterility requirements and synergistic optimization of superior overall product quality. As sterilization temperature increased, oxidation of fats and proteins in ground meat intensified; ordered protein secondary structures (α-helices) transformed into disordered coils; and volatile flavor compound response values rose. The sterilization condition of 121 ℃ for 23 minutes resulted in a larger peak area for the non-flowing water A₂₂ group, indicating greater water retention, superior water-holding capacity, and higher retention of essential nutrients. After comprehensive evaluation, the sterilization conditions of 121 ℃ for 23 minutes yielded the optimal hardness and highest sensory scores for the stuffed shiitake mushroom products, making them the ideal sterilization parameters for this product.