Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Fibrinolytic Enzyme from Bacillus amyloliquefaciens D4-7 Isolated from Chungkook-Jang; It′s Characterization and Influence of Additives on Thermostability.

청국장으로부터 분리한 Bacillus amyloliquefaciens D4-7이 분비하는 혈전용해효소의 특성 및 열안정성에 미치는 첨가물의 효과

  • 김상숙 (이지바이오시스템 생물자원연구소) ;
  • 이주훈 (이지바이오시스템 생물자원연구소) ;
  • 안용선 (이지바이오시스템 생물자원연구소) ;
  • 김정환 (서울보건대학 조리예술과) ;
  • 강대경 (서울보건대학 조리예술과)
  • Published : 2003.09.01
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Abstract

Bacillus amyloliquefaciens D4, which produces a strongly fibrinolytic enzyme, was isolated from Chungkook-Jang, a traditional Korean soybean-fermented food. B. amyloliquefaciens D4 was mutated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to yield a series of mutants with increasing levels of fibrinolytic enzyme production. After mutation, a mutant D4-7 was obtained with fibrinolytic activity about eight times stronger than the parent strain. The fibrinolytic activity of B. amyloliquefaciens D4-7, reached a maximum, when the producer was cultivated in 2% Isolated Soy Protein (ISP) broth for 48 h at $37^{\circ}C$. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of B. amyloliquefaciens D4-7 showed stronger activity than plasmin and streptokinase. The optimum temperature and pH were $50^{\circ}C$ and 10.0 and thermostability was increased by the addition of glycerol, glucose, and NaCl.

한국의 전통 대두발효식품인 청국장에서 혈전용해능이 우수한 미생물을 분리하였으며, 이를 동정한 결과 B.amyloliquefaciens D4로 명명하였다. 혈전용해효소의 대량분비를 위해 N-methyl-N-nitro-N-nitrosoguanidine을 사용한 돌연변이를 유도하여 B. amyloliquefaciens D4-7 변이주를 얻었으며, plasmin 1 U/ml 6.5배 정도의 높은 혈전분해능을 나타내었다. B. amyloliquefaciens D4-7는 분리대두단백배지에서 혈전용해효소 분비능이 가장 우수하였다. B. amyloliquelaciens D4-7가 생산하는 혈전용해효소의 최적 활성조건은 pH 10, 5$0^{\circ}C$이었고, pH 7.0 에서 pH 11 사이에서 상대적으로 안정하였으며, $50^{\circ}C$에서 30분간 방치하였을 경우 50%의 활성이 유지되었다. 또한, NaCl, glycerol 또는glucose 첨가에 의해 혈전용해효소의 저장안정성이 높아지는 것을 확인할 수 있었다.

Keywords

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