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KMID : 0644119980110020178
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Journal of Oriental Gynecology
1998 Volume.11 No. 2 p.178 ~ p.192
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A Study on characteristics of the purified thrombolytic enzyme from Holotrachia extracts
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姜東輝/Kang, Dong Hwi
成娟受/洪永旭/李泰均/Seong, Yeon Su/Hong, Young Yook/Lee, Tae Kyun
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Abstract
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A thrombus is a mass formed from the constituents of the blood within the vessels or the heart during life. The process of its formation is known as thrombosis. In the recent years, many efforts to seek natural fibrinolytic enzymes have been carried out through herbs medicines. It has been generally accepted that Holotrichia is an useful medicine for thrombosis.
In previous studies, Holotrichia extracts had a fibrinolytic activity. After isolation of the fibrinolytic enzymes by column chromatography and electroelution, each enzyme was applied on SDS-PAGE and 3 protein bands having the fibrinolytic activity were detected with molecular weights of 30,000, 45,000 and 75,000 dalton, respectively.
Thus, in this study, a thrombolytic enzyme having a molecular weight of 75,000 Dalton on SDS-PAGE and gel filtration has been purified.
The rate of fibrinolysis of the purified enzyme continued high level for 10 days. The maximum fibrinolytic activity of the purified enzyme was detected after 10 days of incubation at 37℃. Heat and pH stabilities of the purified enzyme on fibrinolysis is highly dependent on temperature. The optimal activity was observed between pH 5 and pH 9 at 37℃. Optimal pH of the purified enzyme on fibrinolysis was between pH 7.0 and pH 8.5 and it showed relatively broad pH range for enzyme
activity.
In experiments of various inhibitors of the purified enzyme from Holotrichia extracts on fibrinolytic activity, the enzyme was strongly inhibited by SBTI and Aprotinin, and weakly by D and t-AMCHA, without any effect by PSMB and TLCK. These results suggested that the fibrinolytic enzymes of Holotrichia extract are belonged to serine protease group.
When the purified enzyme was mixed with fibrinogen and action pattern was observed by Electron microscopy, it, shows a partially erosive-shaped fibrinolytic pattern.
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