The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Purification and Characterization of Fibrinolytic Enzyme from Lepista nuda

민자주방망이버섯으로부터 혈전용해효소의 정제 및 특성 연구

  • 김준호 (상지대학교 이공과대학 화학과)
  • Published : 2005.12.30
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Abstract

Fibrinolytic enzyme has been isolated and purified from the edible mushroom, Lepista nuda. The apparent molecular mass of purified enzyme was estimated to be 34 KDa by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the enzyme was Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X. It has a pH optimum at $7.0.{\sim}9.5$, suggesting that the purified enzyme is an alkaline protease. It shows the maximum fibrinolytic activity at $55^{\circ}C$. The fibrinolytic activity was inhibited by phenylmethylsulfonyl fluoride, indicating that the purified enzyme is a serine protease. The activity of the purified enzyme was totally inhibited by $Hg^{2+}$.

민자주방망이버섯으로부터 분리한 혈전용해효소의 비활성은 22.78 U/mg 이었으며, fibrin를 직접 용해하는 fibrinolytic enzyme 이었다. 15번째까지 N-terminal amino acid 서열 분석 결과, Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X로 지금까지 발표되지 않은 새로운 효소였다. 분자량은 34 KDa 이고 pH 7.0 부터 pH 9.5의 넓은 영역에서 높은 활성을 나타내는 alkaline protease 였으며, $55^{\circ}C$에서 가장 큰 활성을 보이는 이 효소는 serine protease 저해제인 phenylmethylsulfonyl fluoride를 첨가한 경우 효소의 활성이 전혀 나타나지 않는 것으로 미루어 높은 혈전용해 활성을 갖는 새로운 serine protease로 생각된다. 또한 $Hg^{2+}$의 금속이온을 첨가한 경우에도 효소의 활성은 완전히 사라졌다.

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