Abstract: Skatole is a substance that affects the acceptability of pig meat, and it is released from the cecum and colon of intact male pigs. In this study, skatole level was reduced by fermentation broth of Lactobacillus brevis 1.12 (L. brevis 1.12) in vitro, and response surface methodology (RSM) was used for optimization of the fermentation process and evaluation of interaction effects of different parameters. Optimal conditions for fermentation of L. brevis 1.12 were found to be 77mL/L (bacterial suspension:fermentation broth, v/v) , 107mL, 4d, 36.6℃ , and 5.61 for inoculum volume, De Man, Rogosa, and Sharpe medium (MRS) volume, fermentation time, temperature, and pH, respectively. The skatole removal rate under the optimal conditions was found to be 71.83%±0.07%. Furthermore, the removal ability of supernatant fluid of fermention broth from L. brevis 1.12 was the strongest and the removal rate was 17.25%±0.79% for 24h. The mode of skatole removal was not physical binding by cells of L. brevis 1.12.