Purification, characterization, and antimicrobial mechanism of a novel broad-spectrum bacteriocin produced by Lactiplantibacillus plantarum SCA12 from Traditional Chinese Fermented Mustard greens

Abstract

In this study, Lactiplantibacillus plantarum SCA12, a producer of bacteriocins, derived from traditional Chinese fermented mustard greens was isolated. A novel L. plantarum SCA12-derived bacteriocin, LPsca12, was purified through a multi-step protocol involving macroporous adsorption resin, cation exchange, Sephadex G-25 column chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). The molecular mass of LPsca12 was determined to be 1197.0 Da through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The amino acid sequence of LPsca12 was characterized as GLFPSLVRGPR via liquid chromatography-tandem mass spectrometry (LC-MS/MS). Antimicrobial assessment was then carried out, identifying LPsca12 as a broad-spectrum bacteriocin active against both Gram-negative and Gram-positive bacteria, particularly Salmonella paratyphi A. It demonstrates excellent thermal stability (121 °C, 15 min) and maintains its antibacterial activity under acidic conditions (pH 2.0–6.5). Additionally, LPsca12 is sensitive to digestion by pepsin and trypsin. The antimicrobial action of LPsca12 involves compromising the cell membrane integrity, forming pores, and promoting leakage of intracellular contents, which dissipates both the membrane potential (ΔΨ) and cytoplasmic transmembrane pH gradient (ΔpH) of the target cells. These findings suggest that LPsca12 holds promise as a natural biopreservative in food processing.