Characterization of the Bacteriocin (J4) Produced by Bacillus amyloliquefaciens J4 Isolated from Korean Traditional Fermented Soybean Paste

Title
Characterization of the Bacteriocin (J4) Produced by Bacillus amyloliquefaciens J4 Isolated from Korean Traditional Fermented Soybean Paste
Author(s)
김상달임종희정희영[정희영]
Keywords
LISTERIA-MONOCYTOGENES; PURIFICATION; LACTOCOCCUS; FOOD; LACTOBACILLUS; DOENJANG; ENZYMES; CEREUS; STRAIN; KIMCHI
Issue Date
201106
Publisher
KOREAN SOC APPLIED BIOLOGICAL CHEMISTRY
Citation
JOURNAL OF THE KOREAN SOCIETY FOR APPLIED BIOLOGICAL CHEMISTRY, v.54, no.3, pp.468 - 474
Abstract
In order to isolate the antibacterial bacteriocin-producing bacteria against Listeria monocytogenes, one of noxious and prevalent food-poisoning bacterium, 117 strains of indigenous ferment bacteria were isolated from Korean traditional fermented soybean paste (Doenjang) in Korea. Among the isolated strains, 4 isolates inhibited the growth of L. monocytogenes. Strain J4, showing the strongest inhibition activity against L monocytogenes, was identified as a Bacillus amyloliquefaciens by 16S rDNA sequencing. The strain had a broad spectrum of antibacterial activity against various food-borne pathogens such as Micrococcus luteus, Vibrio parahaemolyticus, and Staphylococcus aureus besides L. monocytogenes. The bacteriocin (J4) was purified by 60% ammonium sulfate precipitation, carboxymethyl (CM)-Sephadex column chromatography, and Sephadex G-100 gel filtration. The purified bacteriocin (J4) was estimated to be 39 kDa molecular weight by tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Bacterocin (J4) had antimicrobial activity in a wide pH range (2.0-12.0) and was heat-stable at 80 degrees C for 20 min, but the activity of bacterocin was inactivated by heating at 100 degrees C for 15 min. The antimicrobial activity of the bacteriocin (J4) was completely abolished by treatment with proteolytic enzymes such as protease (type I and IX), papain, trypsin, proteinase K, and pepsin, but not when treated with alpha-amylase, alpha-glucosidase, and beta-glucosidase. The encoding gene of the bacteriocin was included in chromosomal DNA, not plasmid. The bacteriocin had a bacteriolytic mechanism, resulting in cell wall degradation of L. monocytogenes.
URI
http://hdl.handle.net/YU.REPOSITORY/25124http://dx.doi.org/10.3839/jksabc.2011.072
ISSN
1738-2203
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생명공학부 > 생명공학부 > Articles
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