Purification, Characterization, and Mode of Action of Plantaricin GZ1-27, a Novel Bacteriocin against Bacillus cereus

J Agric Food Chem. 2018 May 9;66(18):4716-4724. doi: 10.1021/acs.jafc.8b01124. Epub 2018 May 1.

Abstract

Bacillus cereus is an opportunistic pathogen that causes foodborne diseases. We isolated a novel bacteriocin, designated plantaricin GZ1-27, and elucidated its mode of action against B. cereus. Plantaricin GZ1-27 was purified using ammonium sulfate precipitation, gel-filtration chromatography, and RP-HPLC. MALDI-TOF/MS revealed that its molecular mass was 975 Da, and Q-TOF-MS/MS analysis predicted the amino acid sequence as VSGPAGPPGTH. Plantaricin GZ1-27 showed thermostability and pH stability. The antibacterial mechanism was investigated using flow cytometry, confocal laser-scanning microscopy, scanning and transmission electron microscopy, and RT-PCR, which revealed that GZ1-27 increased cell membrane permeability, triggered K+ leakage and pore formation, damaged cell membrane integrity, altered cell morphology and intracellular organization, and reduced the expression of genes related to cytotoxin production, peptidoglycan synthesis, and cell division. These results suggest that plantaricin GZ1-27 effectively inhibits B. cereus at both the cellular and the molecular levels and is a potential natural food preservative targeting B. cereus.

Keywords: Bacillus cereus; antibacterial mechanism; natural food preservative; plantaricin.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification*
  • Anti-Bacterial Agents / metabolism
  • Anti-Bacterial Agents / pharmacology*
  • Bacillus cereus / drug effects*
  • Bacillus cereus / growth & development
  • Bacteriocins / chemistry
  • Bacteriocins / isolation & purification*
  • Bacteriocins / metabolism
  • Bacteriocins / pharmacology*
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Lactobacillus plantarum / chemistry
  • Lactobacillus plantarum / classification
  • Lactobacillus plantarum / genetics
  • Lactobacillus plantarum / metabolism
  • Microbial Viability / drug effects
  • Molecular Weight
  • Phylogeny

Substances

  • Anti-Bacterial Agents
  • Bacteriocins