Abstract:[Objective] Higher alcohols is an important part of liquor trace aroma components, but higher alcohols will have negative effects on wine flavor as well as the quality when its content is too high. Higher alcohols in liquor are mainly produced by yeast during alcoholic fermentation and alcoholic fermentation is mainly processed by Saccharomyces cerevisiae, it means that looking for higher alcohol synthesis functional proteins is of great importance for liquor production. [Methods] Two-dimensional electrophoresis was used to compare the differences of intracellular proteins between the Saccharomyces cerevisiae mutant ARTP5, whose higher alcohols production is much less than the origin strain, and the origin strain CF4. Comparative analysis of the two strains’ intracellular proteomic differences helps to find higher alcohols synthesis functional proteins. [Results] Compared with the origin strain CF4, the higher alcohols production of mutant strain ARTP5 was reduced by 20% and it indicates that 45 spots (expression ratio≥2) were expressed differently. Among them, 29 proteins were identified by MALDITOF MS-mass spectrometry, including proteins in carbohydrate and energy metabolism, stress response, protein translation and folding, amino acids metabolism, higher alcohol metabolism progress, etc. It is amazing to find out that the protein ILV5 involved in branched chain amino acid metabolism was up-regulated and the protein ADH1 involved in higher alcohol synthesis was down-regulated in strain ARTP5 compared with those in strain CF4, this could possibly be the reason that the higher alcohols production decreased in strain ARTP5. [Conclusion] The finding of the higher alcohols synthesis related protein ILV5 and ADH1 has important significance for the revealing of higher alcohol synthesis mechanism in Saccharomyces cerevisiae alcohol fermentation process.