Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Purification and Biochemical Characteristics of a 45 kDa Fibrinolytic Enzyme from a Halophile

호염성균 유래 45 kDa 혈전용해효소의 순수분리와 생화학적 특성

  • 김도형 (식품의약품안정청) ;
  • 박정욱 (동아대학교 의약생명공학과) ;
  • 서민정 (동아대학교 의약생명공학과) ;
  • 김민정 (동아대학교 생물공학과) ;
  • 이혜현 (동아대학교 생물공학과) ;
  • 최영현 (동의대학교 한의과대학 생화학교실) ;
  • 주우홍 (창원대학교 생물학과) ;
  • 정영기 (동아대학교 의약생명공학과)
  • Received : 2010.01.10
  • Accepted : 2010.02.18
  • Published : 2010.02.28
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Abstract

A fibrinolytic enzyme producing Bacillus sp. J-19 was isolated from the popular Korean seasoning, pickled anchovy. The fibrinolytic enzyme was purified to homogeneity by chromatographic methods including ethanol precipitation and gel-filtration using Sephadex G-50. Compared to the crude enzyme extract, the specific activity of the enzyme increased 1021-fold with a recovery of 23%. The purified enzyme was estimated to be approximately 45 kDa by SDS-PAGE. Especially, the amidolytic activity in the presence of the synthetic substrate for serine protease (H-D-Ile-Pro-Arg-pNA, S-2288) represented approximately 17 U/mg. In addition, more than the 60% activity of the 45 kDa fibrinolytic activity was maintained in the presence of up to 30% (w/v) sodium chloride. These findings could provide a unique fibrinolytic enzyme, leading to a potential thrombolytic agent.

혈전용해효소를 생산하는 Bacillus sp. J-19가 보편적인 한국의 조미제인 멸치젓갈에서 분리되었다. 그 혈전용해 효소는 에탄올 침전, Sephadex G-50을 이용한 젤 여과법 등을 포함하는 일련의 크로마토그래피 방법으로 순수분리 되었다. 조효소 추출액과 비교해서, 그 효소의 비활성은 1021배 증가하였고, 수율은 23%이었다. 순수분리한 효소의 분자량은 SDS-PAGE 상 약 45 kDa이었다. 특히, 합성기질인 serine protease (H-D-Ile-Pro-Arg-pNA,S-2288)에 대한 아미드활성은 약 17 U/mg이었다. 또한, 그 45 kDa 혈전용해효소의 60% 이상의 활성이 30%(w/v) sodium chloride 의 존재 하에서도 유지되었다. 이러한 발견들은 특이한 혈전용해효소를 제공해서, 실용적인 혈전용해제 개발을 유도할 수 있다.

Keywords

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