Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Characterization of a Fibrinolytic Enzyme from Pickled Anchovy

멸치젓갈유래의 혈전용해호소에 대한 특성

  • Yang Woong-Suk (Department of Life Science and Biotechnology, Dongeui University) ;
  • Lim Hak-Seob (Institute of BIO, MILLENNIUM PROMISE CO., LTD.,) ;
  • Chung Kyung Tae (Department of Life Science and Biotechnology, Dongeui University) ;
  • Kim Young-Hee (Department of Life Science and Biotechnology, Dongeui University) ;
  • Huh Man Kyu (Molecular Biology Dongeui University) ;
  • Choi Byung Tae (College of Oriental Medicine, Dongui University) ;
  • Choi Yung Hyun (College of Oriental Medicine, Dongui University) ;
  • Jeong Yong Kee (Department of Life Science and Biotechnology, Dongeui University)
  • 양웅석 (동의대학교 자연과학대 생명응용과학과) ;
  • 임학섭 (㈜천년약속 바이오연구소) ;
  • 정경태 (동의대학교 자연과학대 생명응용과학과) ;
  • 김영희 (동의대학교 자연과학대 생명응용과학과) ;
  • 허만규 (동의대학교 자연과학대 분자생물학과) ;
  • 최병태 (동의대학교 자연과학대 분자생물학과) ;
  • 최영현 (동의대학교 한의학대과) ;
  • 정영기 (동의대학교 자연과학대 생명응용과학과)
  • Published : 2005.06.01
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Abstract

In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

멸치 액젖을 이용하여 ammonium sulfate침전, ion exchange, gel filtration, ethanol침전등의 과정을 거쳐 혈전분해 효소(myulchikinase, MK)를 분리 및 정제하였다. 이 정제된 효소를 SDS-PACE gel 전기영동한 결과 분자량은 약 28 kDa 이었다. MK의 활성에 대한 특성을 조사한 결과 NaCl $30\%$까지 활성이 $80\%$이상 유지되는 것으로 보아 내염성 효소로 판단된다. 온도에 대한 효소활성을 조사한 결과, MK의 온도에 대한 안정성은 $40^{\circ}C$까지는 안정하였으나 $50^{\circ}C$이상의 온도에서는 급격하게 활성 떨어졌고, 최적온도는 $40^{\circ}C$였다 MK의 활성은 $pH6\~9$범위에서 매우 안정하였고, 최적 pH는 8이였다. 또한 2가 금속 양이온에 대한 효과는 $Hg^{2+} (1mM)$에 의하여 완전히 활성저해를 보였고, $Zn^{2+}(1mM)$에 의하여 약 $50^{\circ}C$의 저해되는 것을 알았다. Fibrinogen-rich plate와 Fibrinogen-free plate에서 활성을 측정 한 결과, Fibrinogen-rich plate에서는 fibrin 분해능이 있었지만 Fiberinogen-free Plate에서는 분해활성이 없는 것으로 보아 본 효소는 plasminogen activator type의 혈전용해효소로 사료된다.

Keywords

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