Objective To solve the problems of single strain, limited fermentation capability, and weak environmental tolerance of yeast Fuqu.Methods The yeast community of Jiangxiang Daqu is utilized as the strain source, and bran is used as the raw material to prepare a multi-yeast mixed Fuqu with fortified function. The preparation conditions of yeast Fuqu are optimized by single factor tests combined with orthogonal tests. Then, the acid, ethanol, and high-temperature resistance are systematically evaluated, while the microbial community structure is analyzed via internal transcribed spacer (ITS) rDNA high-throughput sequencing.Results The optimal process parameters are a moisture content of 45%, an incubation time of 48 h, and an inoculum size of 5%. Under these conditions, the yeast count in the fortified Fuqu reaches 7.98×10⁹ CFU/g, which is two orders of magnitude higher than that of the traditional Jiangxiang Daqu (P<0.05). Maintaining robust growth activity in high-acid, high-ethanol, and high-temperature environments, the Fuqu, with high microbial community purity, successfully enriches Saccharomycopsis and Saccharomyces.Conclusion The multi-yeast mixed Fuqu with high yeast biomass, strong fermentation function, and potent environmental adaptability can be prepared using the yeast community of Jiangxiang Daqu.