摘要：[Objective] The paper was to ascertain the fermentation conditions and medium formula of bacillus subtilis B-332. [Method] Using single factor and orthogo- nal experimental designs, the fermentation medium and culture conditions for B. sub- i tills B-332 were optimized through shaking flask culture. [Result] The optimal culture and inoculation time was 18 h, and the optimized medium formula was as follows： soybean powder 0.60 g/L, sucrose 0.25 g/L, ammonium sulfate 0.07 g/L, trisodium citrate 0.03 g/L, dipotassium hydrogen phosphate 0.003 g/L, magnesium sulfate 0.005 g/L, ferrous sulfate 0.000 5%; the fermentation condition was as follows： tem- perature 30 ℃, shaking bed speed 180 r/min, shaking bottle volume 80 ml/500 ml. In the optimal fermentation conditions, the fermented spore quantity was 1.43×10^11 cfu/ml, which was 34.48 times higher than the spore quantity of 4.03×10^9 cfu/ml in initial condition. [Result] The study laid the foundation for factory production of the strain.