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Purification and Characterization of a Novel Bacteriocin Pediocin Z-1 Produced by Pediococcus Pentosaceus Z-1 Screened From Jinhua Ham

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Abstract Background: Lactic acid bacteria (LAB) can produce bacteriostatic substances, among which bacteriocins attracted wide attention in food preservation for decades. Up to date, nisin (class I bacteriocins) has been considered to be the only bacteriocin produced by Lactococcus lactis strains for commercial use. Moreover, there are many other reports concerning the isolated bacteriocins for potential application in food industry while some exhibited a narrow bactericidal spectrum, thermal stability and low acid-base stability. Jinhua ham is a representative of traditional dry-cured meat product in China. The microbial community structure and diversity was speculated to be responsible for flavor and quality formation of Jinhua ham, protecting from the spoilage microorganisms contamination. However, no studies was performed to investigate the bacteriocin from LAB in Jinhua ham. Thus, the objective of this study was to screen out high-efficiency, safe and non-toxic bacteriocin-producing lactic acid bacteria (LAB) from Jinhua Ham and subsequently perform the purification, identification and characterization of its bacteriocin. Results: The bacteriocin-producing LAB was screened from Jinhua ham and then designated as Pediococcus pentosaceus Z-1 by colony morphology and 16S rDNA sequencing. The bacteriocin was then crudely extracted from the bacterial cell-free supernatant by pH adsorption, and further purified by cellulose DEAE-52 ion exchange and Sephadex G-50 chromatography columns. The tricine-SDS-PAGE electrophoresis showed a highly purified protein band with 8227.35 Da with 60 amino acids identified by MALDI-TOF-MS analysis. The bacteriocin was named as pediocin Z-1 and its antibacterial activity exhibited an acid-base stability between pH 2-10 and a thermal stability at a range of 50-110°C. The pediocin Z-1 was sensitive to proteases and showed an inhibitory effect against Gram-positive bacteria and Gram-negative bacteria, including Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, Salmonella potsdam and Escherichia coli. Conclusions: Our data suggest that pediocin Z-1 has a comparable effect of antibacterial activity with nisin and it is promising that pediocin Z-1 has great potential for application prospect in the food industry.
Title: Purification and Characterization of a Novel Bacteriocin Pediocin Z-1 Produced by Pediococcus Pentosaceus Z-1 Screened From Jinhua Ham
Description:
Abstract Background: Lactic acid bacteria (LAB) can produce bacteriostatic substances, among which bacteriocins attracted wide attention in food preservation for decades.
Up to date, nisin (class I bacteriocins) has been considered to be the only bacteriocin produced by Lactococcus lactis strains for commercial use.
Moreover, there are many other reports concerning the isolated bacteriocins for potential application in food industry while some exhibited a narrow bactericidal spectrum, thermal stability and low acid-base stability.
Jinhua ham is a representative of traditional dry-cured meat product in China.
The microbial community structure and diversity was speculated to be responsible for flavor and quality formation of Jinhua ham, protecting from the spoilage microorganisms contamination.
However, no studies was performed to investigate the bacteriocin from LAB in Jinhua ham.
Thus, the objective of this study was to screen out high-efficiency, safe and non-toxic bacteriocin-producing lactic acid bacteria (LAB) from Jinhua Ham and subsequently perform the purification, identification and characterization of its bacteriocin.
Results: The bacteriocin-producing LAB was screened from Jinhua ham and then designated as Pediococcus pentosaceus Z-1 by colony morphology and 16S rDNA sequencing.
The bacteriocin was then crudely extracted from the bacterial cell-free supernatant by pH adsorption, and further purified by cellulose DEAE-52 ion exchange and Sephadex G-50 chromatography columns.
The tricine-SDS-PAGE electrophoresis showed a highly purified protein band with 8227.
35 Da with 60 amino acids identified by MALDI-TOF-MS analysis.
The bacteriocin was named as pediocin Z-1 and its antibacterial activity exhibited an acid-base stability between pH 2-10 and a thermal stability at a range of 50-110°C.
The pediocin Z-1 was sensitive to proteases and showed an inhibitory effect against Gram-positive bacteria and Gram-negative bacteria, including Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, Salmonella potsdam and Escherichia coli.
Conclusions: Our data suggest that pediocin Z-1 has a comparable effect of antibacterial activity with nisin and it is promising that pediocin Z-1 has great potential for application prospect in the food industry.

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