Saccharomyces cerevisiae Et 20 could tolerant ethanol volume fraction of 20% was acquired by ethanol acclimatization. The effects of different concentration of fatty acids, amino acids, inorganic ions, trehalose and inositol on ethanol tolerance of Et 20 were studied. Results showed that all the additives could improve ethanol tolerance and fermentation performance of Et 20. The order of different additives in enhancing ethanol tolerance of Et 20 was: fatty acids > amino acids > inorganic ions > trehalose > inositol. While the order of different additives in promoting fermentative performance of Et 20 was: trehalose > fatty acids > amino acids > inorganic ions > inositol. Under the stress of 10% ethanol, adding trehalose improved 39.04% of total fermentation performance. However, effect of trehalose addition on ethanol tolerance exhibited no outstanding superiority, which only improved 15.81% of strain biomass. The impact of stearic acid on fermentation performance was next only to trehalose, and adding stearic acid could improve 26.13% of fermentation performance. What’s more, adding stearic acid improved 40% of microorganism biomass under ethanol tolerance of 10%. The study can lay the groundwork for further improving ethanol tolerance and fermentation performance of Saccharomyces cerevisiae.