Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus

  • Sung Wook Hong (Technology Innovation Research Division, World Institute of Kimchi) ;
  • Jong-Hui Kim (National Institute of Animal Science, Rural Development Administration) ;
  • Hyun A Cha (National Institute of Animal Science, Rural Development Administration) ;
  • Kun Sub Chung (Division of Biological Science and Technology, Yonsei University) ;
  • Hyo Ju Bae (National Institute of Animal Science, Rural Development Administration) ;
  • Won Seo Park (National Institute of Animal Science, Rural Development Administration) ;
  • Jun-Sang Ham (National Institute of Animal Science, Rural Development Administration) ;
  • Beom-Young Park (National Institute of Animal Science, Rural Development Administration) ;
  • Mi-Hwa Oh (National Institute of Animal Science, Rural Development Administration)
  • Received : 2022.08.05
  • Accepted : 2022.10.11
  • Published : 2022.11.28
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Abstract

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0-70℃ and pH 2-10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

Keywords

Acknowledgement

This work was carried out with the support of "Cooperative Research Program for Agricultural Science & Technology Development (Project No. PJ009221012014)" Rural Development Administration and was financially supported by the Ministry of Trade, Industry and Energy (MOTIE) and Korea Institute for Advancement of Technology (KIAT) through the National Innovation Cluster R&D program(P0015309), Republic of Korea.

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