Characterization of Bacteriocin N15 Produced by Enterococcus faecium N15 and Cloning of the Related Genes
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概要
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Enterococcus faecium N15 was isolated from nuka(Japanese rice-bran paste), which is utilized as starter in the fermenting of vegetables, and was found to produce a bacteriocin that exhibited a broad spectrum of activity, including activity against Listeria monocytogenes and Bacillus circulans JCM2504. The bacteriocin was sensitive to proteases(α-chymotrypsin, proteinase K, trypsin, and pepsin)and α-amylase, but it was resistant to lipase. The bacteriocin was resistant to heat treatment at 100℃ for 2 h, but its activity was completely lost after autoclaving at 121℃ for 15 min. It was active over a wide pH range from 2.0 to 10.0. The bacteriocin showed bactericidal activity against Lactobacillus sake JCM1157 at a concentration of 40 AU/ml. Its molecular weight was estimated by SDS-PAGE to be about 3-5 kDa. PCR primers were designed based on the conserved amino acid sequences of class IIa bacteriocins. A 3-kb DNA fragment was amplified and three open reading frames(ORFs)were found. The first encodes a probable immunity protein of 103 amino acid residues and shows complete homelogy with the putative immunity protein of E.faecium DPC1146. The second and third ORFs respectively encode a probable transposase gene and an inducing factor. The upstream region of the immunity gene, in which the bacteriocin structural gene is located, was amplified. A homology search revealed that the bacteriocin produced by E.faecium N15 exhibits complete identity to enterocin A, a bacteriocin produced by E.faecium DPC1146. PCR using the primers designed in this study is a rapid and sufficient method of screening for bacteriocin-producing strains.
- 公益社団法人日本生物工学会の論文
- 2001-04-25
著者
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Suzuki Shingo
Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka
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Suzuki Shingo
Technology Osaka University
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Uchiyama K
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Uchiyama Keiji
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Nagahisa Keisuke
Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka
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Nagahisa Keisuke
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Shioya S
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Shioya Suteaki
Department Of Biotechnology Graduate School Of Engineering Osaka University:(present Office)departme
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Shioya Suteaki
Department Of Biotechnology Faculty Of Engineering Osaka University
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Shioya Suteaki
Department Of Fermentation Technology Osaka University
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Losteinkit C
Department Of Biotechnology Graduate School Of Engineering Osaka University
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LOSTEINKIT CHANVADEE
Department of Biotechnology, Graduate School of Engineering, Osaka University
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OCHI SHUICHIRO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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TAKAOKA TOMOYO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Takaoka Tomoyo
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Ochi Shuichiro
Department Of Biotechnology Graduate School Of Engineering Osaka University
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