Purification, molecular characterization of Lactocin 63 produced by Lactobacillus coryniformis FZU63 and its antimicrobial mode of action against Shewanella putrefaciens

Abstract

Bacteriocins derived from lactic acid bacteria (LAB) are well recognized as promising food preservative due to high safety and potent antibacterial activity against foodborne pathogens and spoilage bacteria. In this study, an antimicrobial agent-producing strain FZU63 from Chinese sauerkraut was identified as Lactobacillus coryniformis based on physio-biochemical characterization and 16S rDNA sequence analysis. In addition, a bacteriocin was purified from the culture supernatant of L. coryniformis FZU63, and its molecular mass was determined as 1493.709 Da. Moreover, the amino acid sequence of the bacteriocin was predicted to be RQQPMTLDYRW-NH₂ using nanoliter/microliter liquid chromatography combined with triple quadrupole-linear ion trap tandem mass spectrometry and was named as Lactocin 63. Furthermore, Lactocin 63 displays potent antimicrobial activity against the tested Gram-positive and negative bacteria based on the results of determining MICs. Subsequently, the action mode of Lactocin 63 against Shewanella putrefaciens was investigated. The results demonstrated that Lactocin 63 targets and is adsorbed onto the bacterial cell wall and membrane and then disrupts cytoplasmic membrane, which is leading to leakage of cytoplasm according to the results of flow cytometry analysis and the observation of cellular ultra-structure using confocal laser microscopy and atomic force microscopy. Collectively, these results are helpful and providing the theoretical base for developing and applying LAB-derived bacteriocins as promising bio-preservatives to combat foodborne pathogens and spoilage bacteria in seafood industries.Key points• A bacteriocin-producing strain Lactobacillus coryniformis was isolated.• A novel bacteriocin produced by Lactobacillus coryniformis FZU63 was characterized.• Action mechanism of the bacteriocin against S. putrefaciens was elucidated in vitro.

Keywords: Bacteriocin; Lactobacillus coryniformis; Lactocin 63; Molecular characterization; Shewanella putrefaciens.