The medium with high sporulation rate and quantity was obtained by selecting medium component, single factor experiment and orthogonal experiment, and the pH and temperature was optimized for the optimal culture condition. The medium and condition were verified in 20 L fermenter and the controlling of pH and fed-batch fermentation was explored. The optimal culture medium was: wheat bran 5 g/L, corn flour 3 g/L, NaCl 5 g/L, KH2PO4 1 g/L, MnSO4·H2O 0.2 g/L and K2HPO4 1.5 g/L. Cultured in shake flask with pH 6, 200 r/min, at 37 ℃ for 21 h, the sporulation rate reached to 91% and sporulation quantity 5.2×10 10 CFU/mL. In larger experiment, cultured in 20 L fermenter with constant pH and fed-batch fermentation, the sporulation rate reached to 95% and sporulation quantity 1.38×10 11 CFU/mL. The production process of the ZEN-degrading strain ASAGW-10 was obtained, which provided the guarantee for the subsequent study on the degrading bacteria and application.