Production of antimicrobial activity by Bacillus sp. P11 was tested on different byproducts of food industry, like fish meal, grape waste, an industrial fibrous soybean residue, soybean meal, and cheese whey. Bacillus sp. P11 produced the higher amount of antimicrobial activity on soybean meal, followed by fish meal and fibrous soybean residue. Soybean meal was the selected medium to determine the effect of three variables (temperature, initial pH, and substrate concentration) on bacteriocin activity by response surface methodology, using a 23 factorial design. Statistical analysis showed good adequacy to the model (R 2 of 0.8268). In the range studied, temperature and initial pH of the medium have a significant effect on bacteriocin production, and substrate concentration has no significant effect. Response surface data showed maximum bacteriocin production at initial pH between 7.0 and 8.5 and temperature between 39 and 42 °C. In the optimum conditions (initial pH 7.0 and 42 °C), production of bacteriocin activity by Bacillus sp. P11 was compared using a commercial medium (BHI broth) and soybean meal. Maximum activity achieved with the soybean meal-based medium was similar to that obtained with BHI, indicating that soybean meal may be a cost-effective substrate for production of antimicrobial activity by Bacillus sp. P11.