Purification, characterization, and mode of action of Paracin 54, a novel bacteriocin against Staphylococci

Abstract

Staphylococci belong to conditionally pathogenic bacteria, and the pathogenicity of Staphylococcus aureus is the strongest among them. Enterotoxin produced by it can contaminate food and cause food poisoning. Bacteriocin is a kind of polypeptide with antibacterial activity synthesized by some bacteria during metabolism. In this study, we report on purification, characterization, and mode of action of the bacteriocin named Paracin 54, produced by Lactobacillus paracasei ZFM54. Paracin 54 was purified by precipitation with 80% ammonium sulfate, strong cation-exchange chromatography, G-25 gel column, and reversed-phase high-performance liquid chromatography (HPLC). The molecular weight of Paracin 54 (5718.1843 Da) was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Paracin 54 showed broad-spectrum inhibitory activity. It had a strong inhibitory effect on Staphylococci with minimum inhibitory concentration values of 3.00-4.50 μg/mL. Paracin 54 was heat-stable and active only in acidic pH range (2-6). After treatment with proteases, the activity was lost. The results of mode of action showed Paracin 54 damaged the cell membrane and cell wall of Staphylococcus aureus, and then the cytoplasm leaked out, leading to death of the bacteria. These properties make Paracin 54 a promising candidate to prevent the growth of spoilage bacteria and control food poisoning caused by Staphylococci. KEY POINTS: • Paracin 54 was purified from Lactobacillus paracasei ZFM54 with good biochemical characteristics. • Paracin 54 had a strong effect against Staphylococci, making it a promising preservative to prevent the growth of Staphylococci in food. • The mode of action of Paracin 54 on Staphylococcus aureus was revealed.

Keywords: Bacteriocin; Mode of action; Paracin 54; Purification; Staphylococci.