Abstract: This study aimed to investigate the effects of complex enzymatic hydrolysis on the volatile flavor compounds and sensory characteristics of thermal reaction products derived from beef hydrolysate, providing a scientific basis for optimizing the production process and elucidating the flavor formation mechanism of beef flavorings. Using beef shank as the raw material, beef hydrolysates were prepared via either single-enzyme or complex enzymatic hydrolysis, followed by the Maillard reaction to generate beef flavorings. The differences in flavor composition between the flavorings produced by the two hydrolysis methods were compared. Results showed that, compared to single-enzyme hydrolysis, complex enzymatic hydrolysis significantly enhanced the degree of protein hydrolysis (DH increased from 8.07% to 12.76%), promoted the release of free amino acids (content from 7.009 mg/mL to 11.939 mg/mL), and led to an intensified browning reaction (absorbance: 0.790 to 0.904). Electronic nose analysis showed enhanced responses for the complex enzyme group on W2S, W1W, and W2W sensors, indicating a richer content of thiols, olefins, and aromatic compounds. Headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) analysis identified 57 volatile compounds in the complex enzyme group and 33 in the single-enzyme group. Furthermore, the number of key aroma compounds with odor activity values (OAV)>1 was significantly higher in the complex enzyme group (12 types) than in the single-enzyme group (2 types). Compounds such as dimethyl disulfide (OAV=816.75), 3-methylbutanal (OAV=130), and furfural (OAV=6.37) were identified as key aroma contributors in the complex enzyme group. Sensory evaluation results confirmed that the complex enzyme group received higher scores than the single-enzyme group for meaty, roasted, beef-specific, umami, and sweet attributes. In conclusion, complex enzymatic hydrolysis can effectively enhance the complexity and authenticity of beef aroma, providing valuable insights and practical guidance for the industrial production and optimization of natural beef flavorings.