Abstract: By studying the fermentation characteristics of Saccharomyces cerevisiae WBG3, we aim at obtaining a good 2,3-Butanediol (2,3-BD) producing strain. Shake flask fermentation method was used in this study. The substrate concentrations, the dissolved oxygen and the concentration of acid-base in the fermentation process were changed. The key intermediate products (pyruvate, α - acetolactate, acetoin) contents in the 2,3- BD synthesis pathway and the concentrations of fermentation products were detected with the method of UV spectrophotometry and HPLC. The results showed that the S. cerevisiae WBG3 grew well. Under the condition of 80 g/L glucose, 30℃ and 200 r/min, the content of pyruvate,α -acetolactate and acetoin was 0.10±0.03 g/L, 3.38±0.06 g/L and 0.17±0.02 g/L, respectively, and the minimum ethanol conversion reached 0.44±0.02 g/L. After adding acetic acid, the yield of glycerol and the conversion rate of ethanol were reduced by 9.5% and 10%, compared with the control group, and the final yield of 2,3-BD was 0.36±0.02 g/L. Thus,S. cerevisiae WBG3 has the potential for producing 2,3-BD.