Abstract: In order to explore the fermentation characteristics of Saccharomyces cerevisiae in strong-flavoured liquor fermentation, Saccharomyces cerevisiae was inoculated in the fermentative grains and fermented for 50 days, then main ingredients and flavor substances were detected, and microbial flora was analyzed by high-throughput sequencing.It found that owing to the enhancement of Saccharomyces cerevisiae, the main flavor substances of strong-flavoured liquor in fermented grains decreased, especially the production of esters reduced, and ethyl acetate was significantly decreased by 71.0%. Furthermore, Saccharomyces cerevisiae obviously reduced the diversity of bacterial and fungal in fermentative grains, Lactobacillus and Pseudomonas were still the dominant genus in bacteria flora, while the frequency of Aspergillus was increased from 1.8% to60.4% in the fungal flora, and Aspergillus became new dominant fungi instead of Zygosaccharomyces.The results indicated that Saccharomyces cerevisiae had more negative effects on the fermentation of strong-flavoured liquor than expected benefits.