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. 2018 Oct 1:9:2165.
doi: 10.3389/fmicb.2018.02165. eCollection 2018.

Purification and Partial Characterization of Bacteriocin Lac-B23, a Novel Bacteriocin Production by Lactobacillus plantarum J23, Isolated From Chinese Traditional Fermented Milk

Jianming Zhang  1 , Yanyan Yang  1 , Hui Yang  2 , Yushan Bu  2 , Huaxi Yi  2 , Lanwei Zhang  2 , Xue Han  1 , Lianzhong Ai  3
Affiliations

Purification and Partial Characterization of Bacteriocin Lac-B23, a Novel Bacteriocin Production by Lactobacillus plantarum J23, Isolated From Chinese Traditional Fermented Milk

Jianming Zhang et al. Front Microbiol. .

Abstract

The exploration and evaluation of bacteriocin-producing lactic acid bacteria (LAB) have been one of the powerful means to food preservation. A total of 300 strains were isolated from Chinese traditional fermented milk products. A bacteriocin-producing LAB, named Lactobacillus plantarum J23, was screened and identified. Bacteriocin Lac-B23 from L. plantarum J23 was purified by 80% ammonium sulfate precipitation, cation-exchange chromatography, and reverse-phase high-performance liquid chromatography. Molecular weight of bacteriocin Lac-B23 was determined to be approximately 6.73 kDa by tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, and it was confirmed as a novel bacteriocin by liquid chromatography-mass spectrometry. Moreover, bacteriocin Lac-B23 showed thermal stability when heated at below 100°C for 30 min, pH stability between pH 2.0 and 12.0, and sensitivity to trypsin, proteinase K, and proteinase E. The antimicrobial activity of bacteriocin could be enhanced by addition of Fe2+, Mn2+, and ethyl alcohol, and inhibited by Cu2+, K+, Ca2+, Zn2+, Mg2+, and sodium chloride. The results suggested bacteriocin Lac-B23 to have potential application prospects in the food industry.

Keywords: Lactobacillus plantarum; antimicrobial activity; bacteriocin Lac-B23; characterization; food industry; purification.

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Figures

FIGURE 1
FIGURE 1
Phylogenetic tree based on the 16S rDNA sequences of strain J23.
FIGURE 2
FIGURE 2
The anti-Listeria monocytogenes activity of bacteriocin produced with different ammonium sulfate precipitations: treated with (a) 40% ammonium sulfate, (b) 50% ammonium sulfate, (c) 60% ammonium sulfate, (d) 70% ammonium sulfate, and (e) 80% ammonium sulfate.
FIGURE 3
FIGURE 3
Purification of bacteriocin Lac-B23 using SP Sepharose Fast Flow column.
FIGURE 4
FIGURE 4
Analytical HPLC chromatography of bacteriocin Lac-B23.
FIGURE 5
FIGURE 5
(A) Tricine SDS–PAGE analysis of purified bacteriocin Lac-B23; (B) gel overlaid.
FIGURE 6
FIGURE 6
Partial characterization of bacteriocin Lac-B23. (A) Effect of heat treatment; (B) Effect of pH; (C) Effect of metal ion; (D) Effect of ethyl alcohol; and (E) Effect of sodium chloride, on the antimicrobial activity.
FIGURE 7
FIGURE 7
Effect of different proteases on Bacteriocin Lac-B23: (a) cell-free supernatants with no treatment; treated with (b) proteinase E; (c) papain; (d) pepsin; (e) trypsin; (f) proteinase K; and (g) blank control.
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