Wickerhamomyces anomalus and Saccharomyces cerevisiae were mixed by co-inoculation or sequential inoculation, and the physicochemical properties, electronic sensory characteristics, and aromatic characteristics of longan (Dimocarpus longan Lour.) wine were evaluated to analyze the effects of mixed fermentation on wine quality. The results demonstrate that mixed fermentation obtained by co-inoculation or sequential inoculation decreases the alcohol content of longan wine. Furthermore, mixed fermentation also leads to the reduction of the electronic sensory acidity and richness of longan wine. Moreover, the two mixed inoculation methods resulted in different effects on the aromatic characteristics of longan wine. The varieties of aldehyde and ketone aromatic compounds increase in longan wine fermented by co-inoculation, with increasing amounts of acids, aldehydes, ketones, and other compounds, and a decrease in the amounts of ester compounds. However, the variety of ester aromatic compounds and the amounts of acids, aldehydes, and ketones increase when using sequential inoculation. Therefore, the application of mixed fermentation can regulate the physicochemical properties, as well as the electronic sensory characteristics and aromatic characteristics of longan wine, and this contributes to the enrichment of the different types of longan wine. Practical Application Two methods were used to ferment longan wine, consisting of sequential inoculation or co-inoculation with -Wickerhamomyces anomalus and Saccharomyces cerevisiae. The basic physicochemical indexes, electronic sensory characteristics, and aromatic substances were then analyzed to determine the influence of the two inoculation methods on the qualities of the wine. Our study provides a theoretical basis for the application of both inoculation methods in the production of longan wine and a reference for the production of other fruit wines.