This study investigated the physicochemical properties and antioxidant activity of vinegar added with different levels (0%, 1%, 3%, 5%, and 7%) of Akebia quinata fruit during two-step fermentation. The physicochemical properties of vinegar evaluated were pH, total acidity, alcohol, and total sugar and amino acid contents. The antioxidant activities were based on ABTS radical scavenging activity, SOD-like activity, and reducing power. During alcohol fermentation, total acidity and alcohol contents of vinegar increased, but total sugar contents decreased. During acid fermentation, total acidities of vinegar increased. Vinegar added with 7% A. quinata fruit showed the highest total sensory score. Total polyphenol contents of vinegar added with 0% and 1% A. quinata fruit were not significantly different. However, vinegar added with 3, 5, and 7% A. quinata fruit showed significantly higher total polyphenol contents of 136.6, 381.59, and 415.35 mg/100 g, respectively, after 13 days of fermentation. Further, total flavonoid contents of vinegar added with 0～7% A. quinata fruit significantly increased to 21.73, 15.79, 15.15, 26.19, and 26.87 mg/100 g, respectively, after 13 days of fermentation. In addition, tannin contents of vinegar added with 0～7% A. quinata fruit significantly increased to 0.2042, 0.2004, 0.1255, 0.1384, and 0.1255 mg/100 g, respectively, after 13 days of fermentation. Moreover, ABTS radical scavenging activities of vinegar added with 0～7% A. quinata fruit significantly increased to 5. 87, 12.59, 25.63, 34.02, and 35.25, respectively, after 13 days of fermentation at a concentration of 5 mg/mL. Additionally, SOD-like activities of vinegar added with 0～7% A. quinata fruit significantly increased to 8.22, 17.49, 16.86, 16.89, and 15.68%, respectively, after 13 days of fermentation. Reducing power of 7% A. quinata fruit was 0.527 after 1 day and 1.539 at the end of fermentation. Our results demonstrate that antioxidant activity significantly increased during fermentation according to the content of A. quinata. Further, the total polyphenol, flavonoid, and tannin contents were shown to be closely related with antioxidant activities. Thus, A. quinata could be effectively used as a vinegar and functional food material based on its antioxidant activity.