Abstract: In this study,culture medium for thermostable β- glucanase was optimized using response surface methodology.The results indicated that the highest production of β- glucanase could be obtained in the medium containing( g / L) : barley powder 43.48,soybean powder 34.40,sodium chloride 2.40,potassium dihydrogen phosphate 2.40 and dipotassium hydrogen phosphate 12.50. Compared with the initial fermentation medium,the product of β-glucanase cultivated by optimized fermentation medium increased by 11%,which was( 110 ± 2.67) U / m L.Barley powder and soybean powder were chosen as carbon and nitrogen sources in optimization fermentation medium,which are cheap materials. All of this can significantly reduce the costs of β- glucanase,and has good practical value.