Optimization of Culture Conditions and Batch Process Control for the Augmented Production of Bacteriocin by Bacillus Species

Abstract

The emergence of antibiotic-resistant microorganisms poses a significant threat to human health worldwide. Recent advances have led to the discovery of molecules with potent antimicrobial activity from environmental sources. In this study, fifteen bacterial isolates were obtained from agricultural and polluted soil samples collected from different areas of the cities of Jizan and Jeddah. These isolates were screened for antagonistic activity against a set of human pathogenic bacterial strains. The results showed that two Bacillus strains, identified as Bacillus atrophaeus and Bacillus amyloliquefaciens based on 16S rDNA, synthesized bacteriocin with strong antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 33591, Pseudomonas aeruginosa ATCC 9027, Salmonella typhimum ATCC 14028, carbapenem-resistant E. coli, and MRSA 2. To optimize bacteriocin production, the effects of medium composition, incubation period, temperature, and pH were investigated. Nutrient broth and Mueller–Hinton broth were chosen as the optimal original media for bacteriocin production. The optimal incubation period, temperature, and pH were found to be 48 h at 37 °C and 7 pH in Bacillus atrophaeus and 72 h at 37 °C and 8 pH in Bacillus amyloliquefaciens. Batch cultures of Bacillus atrophaeus and Bacillus amyloliquefaciens were grown in a 10 L benchtop bioreactor, and pH control was found to significantly increase the production of bacteriocin by two-fold compared to uncontrolled conditions. The time course of growth, substrate consumption, pH, and enzyme production were investigated. This study demonstrates the potential of optimizing culture conditions and batch process control to enhance bacteriocin production by Bacillus spp.