In this study， the fermentation condition of glucanase production by Burkholderia pyrrocinia B1213 has been optimized. Firstly， cultural conditions (variety of carbon sources， particle size and mass concentration of carbon source， variety of nitrogen sources， mass concentration of nitrogen sources， shaking speed， temperature， liquid medium volume， inoculum volume， initial pH， variety of surfactants， mass concentration of surfactant and fermentation time) of B1213 were optimized by single-factor experiments. Then， four factors， signally affect production of glucanase by B1213 fermentation， were obtained by Plackett-Burman tests. The maximum response area of glucanase activity was determined by steepest ascent. Finally， Box-Behnken Design were used to ascertain the optimal condition of glucanase production. The optimal condition were: 40-60 mesh of wheat bran 50 g/L， yeast extract 8 g/L， initial pH 6.6， liquid medium volume 30 mL/250 mL， inoculum volume 0.15%， Triton X-100 25 g/L， incubating temperature 30 ℃， shaking speed 160 r/min and fermentation time 83 h. Under the optimal condition， the activity of glucanase produced by B1213 were achieved 1 230 U/mL. This study provides academic value for application of bacteria B1213 in Baijiu brewing from the aspect of enzymes related to degradation of raw material for Baijiu brewing.