Purification, characterization, and mode of action of a novel bacteriocin BM173 from Lactobacillus crustorum MN047 and its effect on biofilm formation of Escherichia coli and Staphylococcus aureus.

There is an increasing demand for dairy products, but the presence of food-spoilage bacteria seriously affects the development of the dairy industry. Bacteriocins are considered to be a potential antibacterial or antibiofilm agent that can be applied as a preservative. In this study, bacteriocin BM173 was successfully expressed in the Escherichia coli expression system and purified by a 2-step method. Furthermore, it exhibited a broad-spectrum antibacterial activity, high thermal stability (121°C, 20 min), and broad pH stability (pH 3-11). Moreover, the minimum inhibitory concentration values of BM173 against E. coli ATCC 25922 and Staphylococcus aureus ATCC 25923 were 14.8 µg/mL and 29.6 µg/mL, respectively. Growth and time-kill curves showed that BM173 exhibited antibacterial and bactericidal activity. The results of scanning electron microscopy and transmission electron microscopy demonstrated that BM173 increased membrane permeability, facilitated pore formation, and even promoted cell lysis. The disruption of cell membrane integrity was further verified by propidium iodide uptake and lactic dehydrogenase release. In addition, BM173 exhibited high efficiency in inhibiting biofilm formation. Therefore, BM173 has promising potential as a preservative used in the dairy industry.

Characterization and antibacterial action mode of bacteriocin BMP32r and its application as antimicrobial agent for the therapy of multidrug-resistant bacterial infection.

Multidrug-resistant (MDR) bacterial infection still poses a serious threat to public health, therefore, effective and safe antimicrobial agents are urgently needed. In this study, recombinant bacteriocin BMP32 (BMP32r) prepared by the Escherichia coli expression system had a broad-spectrum antibacterial activity even against some MDR bacteria and its minimum inhibitory concentration ranged from 9.2 to 36.8 mg/L. Furthermore, BMP32r showed good stable performance in heat, pH and storage. Moreover, the scanning electron microscope and transmission electron microscope revealed that BMP32r killed indicator strains through cell wall destruction, pore formation, and the membrane permeability increasing which was proved by propidium iodide uptake investigation. The wound healing of an animal MDR S. aureus infected model was promoted by BMP32r, and the safety was verified by the cytotoxicity assay that the viability of HFF cells remained 87.3% in even when the concentration of BMP32r was as high as 147.2 mg/L. In addition, no abnormalities or damages to major organs was found in vivo assessments after treatment with BMP32r. In conclusion, BMP32r has great potential to be developed as a safe antimicrobial agent to treat MDR bacterial infections.