Saccharomyces cerevisiae Y3401 with high-yield for ethyl caproate was selected， and the fermentation conditions of ethyl caproate from it were optimized. Firstly， the medium conditions (including initial pH value and sugar content) and induction conditions (including temperature， rotation speed， inoculation amount， ethanol addition amoun， caproic acid addition amount， pre-induced period， and post-induced period) were optimized by single factor design. Then， five significant factors for ethyl caproate production were screened by Plackett-Burman design， and the maximum response area was determined by the steepest ascent design. Finally， the optimal fermentation conditions of Y3401 for producing ethyl caproate were obtained by response surface methodology. The optimal conditions were as follows: sugar content 14 Brix， initial pH 7， inoculum size of 3.5%， ethanol content 8%， caproic acid content 0.059%， pre-induced period 30 h， and post-induced period 31 h incubated at 25 ℃ with 180 r/min. Under the condition， the yield of ethyl caproate reached 10.1 mg/L. These results are helpful for the application of S. cerevisiae Y3401 in Baijiu brewing.