Saccharomyces cerevisiae and Hansenula anomalies as yellow rice wine fermentation strains， the fermentation characteristics of the two yeasts on the 18 °Bx saccharification solution were investigated respectively. It was found that the alcohol-producing capacity of Saccharomyces cerevisiae (9.9% vol) was stronger than that of Hansenula anomala (6.8% vol)， and the volatile ester content (2.41 g/L) of Hansenula anomalies was significantly higher than that of Saccharomyces cerevisiae (0.53 g/L). Further comparing the mixed fermentation pathways， the alcohol content of sequential mixed fermentation (8.1%vol) was slightly lower than that of simultaneous mixed fermentation alcohol (8.7% vol)， and the volatile ester content (1.59 g/L) was significantly higher than that of simultaneous mixed fermentation (0.56 g/L) (P<0.05). Combined with the physical and chemical indicators of the fermentation process and sensory evaluation， it is more appropriate to choose a sequential mixed fermentation route. Through the combined experimental design of the single-factor and Box-Benhnken Center， the response surface of the volatile ester content of the sequentially mixed fermented rice wine products was optimized by selecting three factors: the main fermentation temperature， pH value and yeast inoculation amount. The results showed that the optimal parameters for sequential mixing of standing fermented yellow rice wine were: fermentation temperature of 31.5 ℃， inoculation amount of 1.0×107 cells/mL， and pH of 4.2. The verification experiments showed that the volatile ester content of sequentially mixed yellow rice wine under this condition was (1.66±0.028) g/L， which was not much different from the predicted value of 1.70 g/L， which was 213% higher than the fermentation of Saccharomyces cerevisiae alone. In this study， the fermentation pathway and application effect were elucidated for the application of non-Saccharomyces cerevisiae in yellow rice wine fermentation.