Abstract: To develop a novel tobacco flavoring, different amino acids (proline, glutamic acid, and glycine) were added to bayberry extracts to prepare flavorings via the Maillard reaction. Volatile compounds were qualitatively and quantitatively analyzed using gas chromatography-mass spectrometry (GC-MS). Multivariate statistical analysis was conducted to identify and analyze key differential aroma compounds. Molecular docking was used to assess the binding affinity between differential aroma compounds and olfactory receptors. Finally, the flavorings were added to cigarettes and evaluated through olfactory testing. The results showed that 74 volatile compounds were identified by GC-MS in the three flavorings, with 56 compounds common to all. The flavoring produced from proline and bayberry extract showed the highest volatile content (2283.82 μg/mL). Cluster analysis based on key differential aroma compounds grouped the proline-bayberry Maillard flavoring separately. Seventeen key differential volatile compounds were identified. In the proline-bayberry Maillard flavoring, compounds with high odor activity values (OAV) included ethyl cinnamate (OAV=583.43), 5-methyl-2(5H)-furanone (OAV=516.25), ethyl maltol (OAV=501.62), methyl cinnamate (OAV=259.00), and leaf alcohol (OAV=165.50), contributing fruity, roasted, and sweet aromas. In the glycine-bayberry Maillard flavoring, methyl cinnamate (OAV=840.00) had the highest contribution, while in the glutamic acid flavoring, ethyl tiglate (OAV=523.73) was dominant. Molecular docking results showed that 12 key differential aroma compounds had average binding energies with 11 olfactory receptor proteins ranging from −6.19 to −4.20 kcal·mol⁻¹, indicating spontaneous and perceptible binding. Cigarette flavoring tests demonstrated that the proline-bayberry Maillard flavoring provided the best effects, significantly improving aroma, smoke density, off-flavors, and richness. Olfactory evaluation indicated dominant fruity, sweet, and floral notes, with secondary smoky and roasted aromas. Compared to glutamic acid and glycine, proline was more suitable for preparing bayberry Maillard tobacco flavorings. This study provides theoretical support for developing fruit-type tobacco flavorings.