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Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce

간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제

  • Jung, Sung-Sub (Interdisciplinary Program in Biotechnology, Graduate School, Changwon National University) ;
  • Choi, Jung-I (Interdisciplinary Program in Biotechnology, Graduate School, Changwon National University) ;
  • Joo, Woo-Hong (Department of Biology, Changwon National University) ;
  • Suh, Hyun-Hyo (Department of Environmental Engineering, Jinju National University) ;
  • Na, Ae-Sil (Department of Biochemistry and Health Science, Changwon National University) ;
  • Cho, Yong-Kweon (Department of Biochemistry and Health Science, Changwon National University) ;
  • Moon, Ja-Young (Department of Biochemistry and Health Science, Changwon National University) ;
  • Ha, Kwon-Chul (Department of Biochemistry and Health Science, Changwon National University) ;
  • Paik, Do-Hyeon (Department of Biochemistry and Health Science, Changwon National University) ;
  • Kang, Dae-Ook (Department of Biochemistry and Health Science, Changwon National University)
  • 정성엽 (창원대학교 대학원 생명공합협동과정) ;
  • 최정이 (창원대학교 대학원 생명공합협동과정) ;
  • 주우홍 (창원대학교 생물학과) ;
  • 서현효 (진주산업대학교 환경공학과) ;
  • 나애실 (창원대학교 보건의학과) ;
  • 조용권 (창원대학교 보건의학과) ;
  • 문자영 (창원대학교 보건의학과) ;
  • 하권철 (창원대학교 보건의학과) ;
  • 백도현 (창원대학교 보건의학과) ;
  • 강대욱 (창원대학교 보건의학과)
  • Published : 2009.07.30

Abstract

A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

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