Korean Journal of Environmental Agriculture (한국환경농학회지)

The Korean Society of Environmental Agriculture (한국환경농학회)
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Two-Dimensional Electrophoresis Analysis of Proteins between Bacillus licheniformis DM3 and Its Antifungal Activity Deficient Mutant

이차원전기영동법을 이용한 길항세균 Bacillus licheniformis DM3와 이의 항진균 활성 결여 돌연변이균주간 단백질 비교 분석

  • Lee, Young-Keun (Radiation Application Research Division, Korea Atomic Energy Research Institute) ;
  • Kim, Jae-Sung (Radiation Application Research Division, Korea Atomic Energy Research Institute) ;
  • Chung, Hye-Young (Radiation Application Research Division, Korea Atomic Energy Research Institute) ;
  • Jang, Yu-Sin (Radiation Application Research Division, Korea Atomic Energy Research Institute) ;
  • Jang, Byung-Il (Radiation Application Research Division, Korea Atomic Energy Research Institute)
  • 이영근 (한국원자력연구소, 방사선이용연구부) ;
  • 김재성 (한국원자력연구소, 방사선이용연구부) ;
  • 정혜영 (한국원자력연구소, 방사선이용연구부) ;
  • 장유신 (한국원자력연구소, 방사선이용연구부) ;
  • 장병일 (한국원자력연구소, 방사선이용연구부)
  • Published : 2003.09.30
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Abstract

In the course of screening for antifungal agents, a bacterial strain, DM3 was isolated from a mud sample collected at Daechon in Chungnam province and identified as Bacillus licheniformis based on API 50CHB test. It has antifungal activity against 12 plant pathogenic fungi in paper disc assay. At the 95% lethal dose of gamma radiation ($^{60}Co$, 10 kGy, $D_{10}=2.32\;kGy$), the antifungal activity deficient mutant (mDM3) against Colletotrichum gloeosporioides was induced From 2-D electrophoresis analysis, serine hydroxymethyltransferase (45.0 kDa), hypothetical protein(40.7 kDa), NifU protein homolog(15.4 kDa), and resolvase(12.5 kDa) homologous proteins were detected only in B. licheniformis DM3. Lysozyme(18.1 kDa) and alkyl hydroperoxide reductase(15.6 kDa) homologous proteins were expressed uniquely in B. licheniformis mDM3. Further studies are needed to reveal that these proteins from B. licheniformis DM3 could be closely related to the antifungal activity against plant pathogenic fungi.

항진균 활성균주를 선발하는 과정에서 DM3 균주를 대천 바닷가에서 수집된 진흙 시료로부터 분리하였으며 API 50CHB kit를 이용하여 동정한 결과 Bacillus licheniformis로 동정되었다. 이 균주는 12종의 식물병원성 진균에 대해 항균활성을 나타내었다. 감마선($^{60}Co$)을 조사하여 항진균 활성 결핍 돌연변이체(mDMB)를 유도한 후, 이차원전기영동으로 단백질을 분석한 결과 DM3와 mDM3에만 존재하는 각각 4종과 3종의 단 단백질을 확인할 수 있었다. 2-DE 결과 B. licheniformis DM3에서 spot 1은 serine hydroxymethyltransferase(45.0kDa), spot 2는 hypothetical protein(40.7 kDa), spot 3는 NifU protein homolog(15.4 kDa), 그리고 spot 4는 resolvase(12.5 kDa)와 상동성을 지닌 단백질로 동정되었고 B. licheniformis mDM3에서만 발현된 spot 5는 lysozyme(18.1 kDa)과 spot 6, 7은 alkyl hydroperoxide reductase(15.6 kDa)으로 동정되었다. B. licheniformis DM3에서 이들 단백질들의 항진균 활성 관련 여부를 규명하기 위해서 더 연구가 진행되어야 할 것으로 사료된다.

Keywords

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