Characterization of novel broad spectrum bacteriocin produced by Lactobacillus plantarum from pickles

Abstract

Abstract: To screen Lactobacillus strains which can produce specific inhibition to food borne pathogens and spoilage bacteria, and to further study the characteristics of bacteriocin, so as to lay a foundation for its application in food antisepsis. One of 112 isolates (Hlh32) produced a bacteriocin, designated as BacH32, and was isolated from Pickle juice and identified as Lactobacillus plantarum. This bacteriocin reached the maximum production （1850Au/mL） at phase stationary phase after 18-24 h of growth in MRS broth at 35 ℃ against Staphylococcus aureus. It was purified sequentially by performing ammonium sulfate precipitation, DEAE Sepharose Fast Flow anion exchange chromatography, Sephadex G-15 gel filtration chromatography and reversed-phase HPLC separation. According to Trinice SDS-PAGE, the molecular mass of bacteriocin Hlh32 was 6.7 kDa, which was significantly different from previous reported bacteriocins produced by Lactobacillus plantarum strains. It was sensitive to proteolytic enzymes. Complete inactivation of antimicrobial activity was observed after treatment of crude extracts with proteinase K, papain, trypsin, and pepsin, and the bacteriocin was resistant to treatments with lipase, α-amylase and β-amylase, which confirmed its proteinaceous nature. BacH32 remained active after 4 h at pH 2-9 and temperature treatments after 30 min at 37, 60, 80, 100 ℃, and 15 min at 121 ℃. Bacteriocin was found to have extensive spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria, such as Escherichia coli、Salmonella typhi、Staphylococcus aureus、Micrococcus luteus、Enterococcus faecalis、Pseudomonas aeruginosa、Vibrio parahaemolyticus、Bacillus cereus、Bacillus subtilis、Saccharomyces cerevisiae、Listeria monocytogenes. In addition, Hlh32 exhibited antimicrobial activity against fungus such as Candida krusei、Saccharomyces cerevisiae and Candida albicans. Addition of BacH32 to a mid-log culture of Staphylococcus aureus inhibited growth. These results indicate the potential of the bacteriocin for use as a food bio-preservative.

Key words: Lactobacillus plantarum, Bacteriocin, Purification, Antimicrobial characteristic

CLC Number:

Q819

References

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