In this study,the response surface methodology(RSM) was used to optimize the fermentation conditions for the novel antimicrobial peptide by Bacillus sp.fmbJ224.First,Plackett-Burman design was undertaken to evaluate the effects of the six factors.By the statistical regression analysis,the significant factors affecting the antimicrobial peptide in fermentation by Bacillus sp.fmbJ224 were determined as follows: temperature,broth content,fermentation time.In the second phase of the optimization process,a response surface methodology(RSM) was employed to optimize the process variables of the above critical internal factors.Result showed that the optimal concentration of the variables were determined as: temperature 33.08℃,broth content 71.93 mL,fermentation time 43.45h.The content of the antimicrobial peptide was increased by 199.19μg/mL.The experimental data under various conditions have validated the theoretical values.