Abstract: To establish the extraction technology of Glutathione from beer waste brewing yeast by high- pressure steam, the optimization was applied to discuss some factors including the pressure, extracting time, the ratio of solid to liquid and extracting times with Box-Behnken center-united design and the method of response surface analysis ( RSM) . Through establishing the second regression model, the optimal parameters were conformed as follows: the pressure 1.1MPa, extracting time 15 min, the ratio of solid to liquid 10∶1 and extracting times 1.Under this conditions, the yield of GSH was 4.63 mg /g. These results suggested that the high- pressure steam extraction technology was one of the effective ways to improve the yield of reduced glutathione.