Characterization of Antibacterial Substance - Producing Bacillus subtilis Isolated from Traditional Doenjang

전통 된장으로부터 분리한 향균물질 생산 Bacillus subtilis의 특성

  • Ryu, Hyun-Soon (Korea Fermented Food Research Institute, Department of Microbiological Engineering, Jinju National University) ;
  • Shon, Mi-Yae (Korea Fermented Food Research Institute) ;
  • Cho, Soo-Jeong (Department of Microbiological Engineering, Jinju National University) ;
  • Park, Seok-Kyu (Korea Fermented Food Research Institute, Department of Food and Nutrition, Sunchon National University) ;
  • Lee, Sang-Won (Korea Fermented Food Research Institute, Department of Microbiological Engineering, Jinju National University)
  • 유현순 (한국전통발효식품연구소, 진주산업대학교 미생물공학과) ;
  • 손미예 (한국전통발효식품연구소) ;
  • 조수정 (진주산업대학교 미생물공학과) ;
  • 박석규 (한국전통발효식품연구소, 순천대학교 식품영양학과) ;
  • 이상원 (한국전통발효식품연구소, 진주산업대학교 미생물공학과)
  • Published : 2007.06.30

Abstract

A bacterium which has high enzymatic activities such as amylase, cellulase and protease was isolated from Korean traditional soybean food, doenjang. The isolated bacterium was identified to Bacillus subtilis HS25 by the test of morphological and biochemical properties according to Bergey's Manual of Systematic Bacteriology and API 50 CHL kit, and by the 16S rDNA sequence. The isolated B. subtilis HS25 had a potent antibacterial activity against food born causative or pathogenic bacteria. B. subtilis HS25 is endospore forming cell and contained flagella and abundant viscous material at the out layer of cell wall. It was rod type bacterium $(0.5{\sim}0.8{\times}3{\sim}5{\mu}m)$ having biochemical characteristics such as gram staining(+), catalase(+), oxidase(-) and hydrolysis of esculin(+). The optimal medium compositions for production of antibacterial substance in the B. subtilis HS25 were 1% of soluble starch, 0.5% of yeast extract, 0.5% of peptone and 0.05% of MgCl$_2{\cdot}6H_{2}O$. The optimum temperature and pH of the growth of the B. subtilis HS25 was 35$^{\circ}C$ and pH 7.5, respectively. The antibacterial activity was more high in neutral to a little alkaline pH (6.5-10.5) than in acidic pH. The optimal shaking speed to grow and to produce antibacterial substance of the B. subtilis HS25 was 160${\sim}$200 rpm. The optimal culture time for antibacterial activities of the bacterium were shown to be in the range of 12-36 hr.

전통 장류 발효식품의 발효도와 저장성을 증진시키기 위하여 전통 콩 발효식품으로부터 cellulase, amylase, protease 활성 및 항균활성 물질 생성이 우수한 바실러스 HS25 균주를 분리하여 그 배양학적 특성을 검토하였다. HS25 균주는 편모와 내생포자를 가지며, 다량의 점질물을 형성하는 그람양성 간균으로 catalase 양성, oxidase 음성으로 esculin을 가수분해하였고, 16S rDNA분석 등을 통하여 Bacillus subtilis로 밝혀져 B. subtilis HS25로 명명하였다. B. subtilis HS25 균주의 생육 및 항균물질 생산을 위하여 최적 배지조성은 soluble starch 1%, yeast extract 0.5%, peptone 0.5% 및 $MgCl_2{\cdot}6H_{2}O$ 0.05%이었으며, 최적 배양온도는 $25{\sim}45^{\circ}C$, 초기 pH는 $6.5{\sim}9.5$, 최적 교반속도는 $160{\sim}200$ rpm이었고, 항균활성이 가장 높게 나타나는 배양 시간 범위는 $12{\sim}36$시간째였다.

Keywords

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