To address the limitations in existing cell disruption technologies, which make it difficult to balance extraction efficiency and flavor in the industrial production of yeast extract, the cell disruption process for preparing high-umami yeast extract was optimized. Three cell disruption methods, namely colloid milling, ultrasonic disruption, and ethyl acetate addition, were systematically compared. Multimodal analysis of the yeast extract after cell disruption and enzymatic hydrolysis was performed by integrating sensory evaluation, electronic nose, electronic tongue, and electroencephalography (EEG) sensing technologies. The results showed that the cell disruption rates for colloid milling (15 min, 50 ℃ ), ultrasonic disruption (80 min, 70 W), and ethyl acetate assistance (mass fraction 7.5%, 9 h) were 90.33%, 82.13%, and 65.12%, respectively. After enzymatic hydrolysis, the overall sensory flavor evaluation scores of the samples were 8.33, 8.30, and 7.73, respectively, with significant umami characteristics. The results of the electronic nose and electronic tongue further verified the sensory evaluation results, with umami values of 18.03, 17.32, and 17.12, respectively. The sample treated with colloid milling had the best taste and coherent flavor, while the sample treated with ethyl acetate had a more intense aroma. EEG analysis (focused on δ rhythm) showed that the whole-brain δ-wave energy density of the sample treated with colloid milling was significantly higher than that of other groups, and this was associated with the highest umami intensity. Colloid milling for cell disruption achieves both high extraction efficiency and optimal flavor quality, and the resulting enzymatic hydrolysate exhibited a strong umami flavor and good consistency. This study provides a technical foundation and theoretical support for the development of high-flavor yeast extract based on multimodal sensory technology.