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There are two steps (alcoholic fermentation and acetic acid fermentation) in the production of fruit vinegar by liquid fermentation. The yeast alcoholic fermentation step plays an important role in the quality of apple vinegar. In this work, Candida tropicalisand aromatizing yeast were used in mixed alcoholic fermentation to improve the flavor of the apple vinegar. The total organic acid contents of apple cider and vinegar in mixed cultures were all higher than those in pure culture (Candida tropicalis). Umami and sweet free amino acid levels in mixed-culture apple vinegar (MCAV; 1236.71 and 858.25 mg/L, respectively) were significantly higher than those in pure-culture apple vinegar (PCAV; 1214.69 and 820.37 mg/L, respectively). The total esters, total alcohols, and total phenolics were also significantly increased in MCAV (282.36 g/L, 254.22 g/L and 47.49 g/L, respectively), fruit flavor and floral aromas in MCAV were higher than that in PCAV. In the principal component analysis (PCA), the integrative score for MCAV was higher than that for PCAV. Therefore, mixed cultures of Candida tropicalisand aromatizing yeast in alcoholic fermentation can effectively improve the flavor and quality of apple cider vinegar; more details about the mixed culture need to be investigated in the future.