Abstract: Furfural, one of the main inhibitory compounds in lignocellulosic hydrolytes, inhibits the growth and ethanol production rate of yeast. To get a global view of the dynamic expression pattern of proteins in S. cerevisiae during the fermentation with introduction of 8 g/L furfural, the protein samples were taken before the addition of furfural, during the initial phase of furfural conversion, and immediately after the conversion of furfural for comparative proteomic analysis with iTRAQ on a LC-ESI-MS/MS instrument. A comparison of the temporal expression pattern of 107 proteins related to protein synthesis between the reference cultures and the furfural-treated cultures showed that a temporal downregulation of these proteins was retarded after the addition of furfural. The expression levels of 20 enzymes in glucose fermentation and 5 enzymes in the TCA cycle were reduced by furfural, with notably delayed temporal downregulations of Glk1p, Tdh1p, Eno1p and Aco1p, which is correlated to the reduced ethanol formation rate and glucose consumption rate by 66.7% and 60.4% respectively. In the presence of furfural, proteins catalyzing the upper part of the super pathway of sulfur amino acid biosynthesis were repressed at all time points, which is related to the inhibited growth of furfural-treated yeast. The expressions of 18 proteins related to stress response showed increased trends, including several highly induced heat shock proteins and proteins related to cellular signaling pathways.