Bacillus lentimorbus WJ5의 분비 전용 Srb Homologue 발현과 항진균 활성

Expression of Secretion-dedicated Srb Homologue and Antifungal Activity of Bacillus lentimorbus WJ5

  • 장유신 (한국원자력연구소 방사선이용연구부) ;
  • 이영근 (한국원자력연구소 방사선이용연구부) ;
  • 김재성 (한국원자력연구소 방사선이용연구부) ;
  • 조규성 (한국원자력연구소 방사선이용연구부) ;
  • 장병일 (한국원자력연구소 방사선이용연구부)
  • 발행 : 2003.09.01

초록

Bacillus sp.는 다량의 효소와 기능성 펩타이드들을 세포외로 분비하는 것으로 알려져 있다. Signal recognition particle (SRP)과 SRP receptor는 세포외 분비 단백질의 이동에 있어서 중심적 역할을 담당한다. B. lentimorbus WJ5의 DNA microarray 결과, 감마선 조사로 유도된 항진균 활성 결핍 돌연변이체인 WJ5m12에서 B. subtilis srb homologue (srbL)의 발현이 감소되는 것을 관찰할 수 있었다. SRP receptor와 항진균 활성 사이의 연관성을 고찰하기 위하여 B. lentimorbus WJ5의 srbL을 PCR로 증폭하여 pQE30 vector에 클로닝 하였으며, B. lentimorbus WJ5m12에 형질전환 시켰다. 형질전환된 B. lentimorbus WJ5m12::srbL은 항진균 활성이 복원되었다. 이차원 전기영동 결과, 수 종의 세포외 분비 단백질의 전구체들이 항진균 활성 결핍 돌연변이 균주에서 축적되었고 B.lentimorbus WJ5m12::srbL에서는 야생형 균주의 단백질 수준까지 감소되는 현상을 관찰할 수 있었다. 이는 B.lentimorbus WJ5의 항진균 활성 관련 물질의 이동에 있어서 srbL이 중요한 역할을 한다는 것을 암시한다.

Bacillus sp. secretes high levels of extracellular enzymes into the culture medium. The signal recognition particle (SRP) and the SRP receptor play a central role in targeting pre secretory proteins to the translocase. By the analysis of the DNA microarray of B. lentimorbus WJ5, it was detected that WJ5m12, antifungal activity deficient mutant induced by gamma radiation, had a down-regulated expression of the SRP receptor gene (B. subtitis srb homologue, srbL). To determine the relationship of SRP receptor to antifungal activity, srbL of B. lentimorbus WJ5 was amplified by PCR and ligated into pQE30 vector, and then transferred into WJ5m12. The transformant, WJ5m12::srbL, recovered the antifungal activity. From the 2-DE analysis, the several presecretory proteins accumulated in the mutant cell and decreased to a level of the wild type in WJ5m12::srbL. It seems that the srbL could play an important role in the secretion of the antifungal activity related proteins of B. lentimorbus WJ5.

키워드

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