백만송이 버섯 균사체 배양액을 이용한 혈전 용해 효소의 분리 및 특성

Abstract

The effect of fibrinolytic activity of Lyophyllum shemeji mushroom mycelial was observed in culture medium containing extracted potato and sugar in the controlling at 25˚C in pH 4.0. In addition, the maximum production of mycelial dry weight was 7.5 mg/ml after 10 days under the optimal conditions. A novel fibrinolytic enzyme was isolated from Lyophyllum shemeji mycelial culture media. The enzyme was purified using combination of anion exchange chromatography on a Mono Q 5/5 column and size exclusion gel filtration chromatography on Superdex 200 100/300 column. The molecular weight of the purified enzyme was estimated to be 21 kDa by SDS-PAGE the N-terminal amino acid sequence were confirmed by protein sequencing analysis, which is dissimilar from that of known fibrinolytic enzymes. The purified enzyme was a protease with an optimal reaction pH 4-6 and temperature of 25°C, respectively. The relative fibrinolytic activity of 1 μg purified enzyme have 1.5 fold more activity than 1 unit/ml of plasmin on fibrin plate. Furthermore, purified enzyme preferentially hydrolyzed the Aα-chain followed by the Bβ- and γ-chain of fibrinogen, which is precursor of fibrin. Therefore, these data suggests that the fibrinolytic enzyme derived from edible mushroom, lyophyllum shemeji mushroom mycelial might be useful for thrombolytic therapy and preventing thrombotic related variable diseases.