Purification and Characterization of Plantaricin JLA-9: A Novel Bacteriocin against Bacillus spp. Produced by Lactobacillus plantarum JLA-9 from Suan-Tsai, a Traditional Chinese Fermented Cabbage

J Agric Food Chem. 2016 Apr 6;64(13):2754-64. doi: 10.1021/acs.jafc.5b05717. Epub 2016 Mar 24.

Abstract

Bacteriocins are ribosomally synthesized peptides with antimicrobial activity produced by numerous bacteria. A novel bacteriocin-producing strain, Lactobacillus plantarum JLA-9, isolated from Suan-Tsai, a traditional Chinese fermented cabbage, was screened and identified by its physiobiochemical characteristics and 16S rDNA sequence analysis. A new bacteriocin, designated plantaricin JLA-9, was purified using butanol extraction, gel filtration, and reverse-phase high-performance liquid chromatography. The molecular mass of plantaricin JLA-9 was shown to be 1044 Da by MALDI-TOF-MS analyses. The amino acid sequence of plantaricin JLA-9 was predicted to be FWQKMSFA by MALDI-TOF-MS/MS, which was confirmed by Edman degradation. This bacteriocin exhibited broad-spectrum antibacterial activity against Gram-positive and Gram-negative bacteria, especially Bacillus spp., high thermal stability (20 min, 121 °C), and narrow pH stability (pH 2.0-7.0). It was sensitive to α-chymotrypsin, pepsin, alkaline protease, and papain. The mode of action of this bacteriocin responsible for outgrowth inhibition of Bacillus cereus spores was studied. Plantaricin JLA-9 had no detectable effects on germination initiation over 1 h on monitoring the hydration, heat resistance, and 2,6-pyridinedicarboxylic acid (DPA) release of spores. Rather, germination initiation is a prerequisite for the action of plantaricin JLA-9. Plantaricin JLA-9 inhibited growth by preventing the establishment of oxidative metabolism and disrupting membrane integrity in germinating spores within 2 h. The results suggest that plantaricin JLA-9 has potential applications in the control of Bacillus spp. in the food industry.

Keywords: Bacillus spp.; Lactobacillus plantarum; bacteriocin; characterization; purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anti-Bacterial Agents / isolation & purification
  • Anti-Bacterial Agents / pharmacology*
  • Bacillus / drug effects*
  • Bacteriocins / pharmacology*
  • Brassica / chemistry*
  • Brassica / metabolism
  • Chromatography, Reverse-Phase
  • Chymotrypsin / metabolism
  • Fermentation
  • Gram-Negative Bacteria / drug effects
  • Hydrogen-Ion Concentration
  • Lactobacillus / metabolism
  • Lactobacillus plantarum / chemistry*
  • Lactobacillus plantarum / metabolism
  • Molecular Weight
  • Picolinic Acids
  • Pyridines / analysis
  • Pyridines / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Anti-Bacterial Agents
  • Bacteriocins
  • Picolinic Acids
  • Pyridines
  • Chymotrypsin
  • alpha-chymotrypsin
  • dipicolinic acid