Abstract: To develop high-quality cigarette flavor and enhance the quality of cigarette, the aroma-producing strain Yarrowia lipolytica was utilized for the fermentation of tobacco extract, and the aroma components were analyzed using solid-phase microextraction-gas chromatography-mass spectrometry. The single-factor and orthogonal experiments were performed to optimize the fermentation conditions with the key aroma component as an indicator, and the aroma-improving effect of cigarette flavor obtained under the optimal conditions was evaluated. The results showed that the fermentation of tobacco extract by Y. lipolytica could promote the degradation of macromolecules such as sugars, amino acids and proteins, as well as generate the high contents of limonene and other characteristic aroma components including phenylethanol, citronellol, α-terpineol, dihydroactinidiolide, and ethyl palmitate. The optimal condition for limonene (key flavor component) production through tobacco extract fermentation by Y. lipolytica was inoculum dosage of 1%, fermentation time of 72 h, fermentation temperature of 28 ℃, and rotational speed of 150 r/min. Under these conditions, the content of limonene reached up to 204.40 μg/L, which was 103.55 μg/L higher than before optimization and 185.15 μg/L higher than that before fermentation. The application of tobacco flavor obtained under the optimal conditions to cigarettes could effectively improve the smoking quality of cigarettes, endow cigarettes with citrus-like aroma, enhance the freshness and sweetness of the smoke, and reduce the irritation.