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, 27 (3), 695-703
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Purification, Characterization and Action Mechanism of Plantaricin JY22, a Novel Bacteriocin Against Bacillus cereus Produced by Lactobacillus plantarum JY22 From Golden Carp Intestine

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Purification, Characterization and Action Mechanism of Plantaricin JY22, a Novel Bacteriocin Against Bacillus cereus Produced by Lactobacillus plantarum JY22 From Golden Carp Intestine

Xinran Lv et al. Food Sci Biotechnol.

Abstract

A novel bacteriocin-producing strain, Lactobacillus plantarum JY22 isolated from golden carp intestine, was screened and identified by its physiobiochemical characteristics and 16S rRNA gene sequence analysis. This bacteriocin, named plantaricin JY22, was purified using ethyl acetate extraction and gel filtration. Its molecular weight was approximately 4.1 kDa by SDS-PAGE analysis. The partial amino acid sequence of plantaricin JY22 was DFGFDIPDEV. It was highly heat-stable and remained active at pH range from 2.5 to 5.5, but was sensitive to protease. Plantaricin JY22 had a bactericidal mode. Scanning electron microscope analysis indicated that plantaricin JY22 damaged the morphology of cells and spores for Bacillus cereus. Moreover, the plantaricin JY22 destroyed cell membrane integrity as confirmed by the leakage of electrolytes, the losses of Na+K+-ATP, AKP, nucleic acids (OD260nm) and proteins. SDS-PAGE of B. cereus proteins further demonstrated that plantaricin JY22 had a remarkable effect on bacterial proteins.

Keywords: Action mechanism; Bacillus cereus; Bacteriocin; Lactobacillus plantarum; Purification and characterization.

Figures

Fig. 1
Fig. 1
(A) The phylogenetic tree of Lb. plantarum JY22 based on 16S rRNA; (B) purification of bacteriocin by Sephadex G-50 chromatogram; (C) determination of molecular weight of the purified bacteriocin by SDS-PAGE, lane 1: gel overlaid with LB soft agar containing B. cereus; lane 2: plantaricin JY22; lane 3: protein Marker
Fig. 2
Fig. 2
Effect of plantaricin JY22 on cell growth of B. cereus (A). OD600 was observed with 1 MIC (filled circle), 3 MIC (filled triangle) and without (filled square) plantaricin JY22, viable cell counts (log CFU/ml) were observed with 1 MIC (circle), 3 MIC (triangle) and without (square) plantaricin JY22. Effect of plantaricin JY22 on permeability of cell membrane of B. cereus (B). Electrical conductivity was observed with 1 MIC (filled circle), 3 MIC (filled triangle) and without (filled square) plantaricin JY22
Fig. 3
Fig. 3
Scanning electron micrograph of B. cereus cells without plantaricin JY22 (A) and treated with 1 MIC (B) and 3 MIC (C), spores of B. cereus without plantaricin JY22 (D) and treated with 1 MIC (E) and 3 MIC (F)
Fig. 4
Fig. 4
SDS-PAGE profiles of B. cereus total soluble proteins treated with plantaricin JY22. Lane 1: Marker bands; lane 2: untreated B. cereus; 3: B. cereus treated with 1 MIC plantaricin JY22; 4: B. cereus treated with 3 MIC plantaricin JY22

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