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Purification and characterization of a fibrinolytic enzyme from Streptomyces sp. XZNUM 00004
文章作者: 发布时间: 2013-07-04 访问次数: 325

Volume 28, Number 7 (2012), 2479-2486, DOI: 10.1007/s11274-012-1055-9 

Purification and characterization of a fibrinolytic enzyme from Streptomyces sp. XZNUM 00004

Xiuyun Ju, Xiaoying Cao, Yong Sun, Zhe Wang, Chengliang Cao, Jinjuan Liu and Jihong Jiang

 

Abstract

A fibrinolytic enzyme (SFE1) from Streptomyces sp. XZNUM 00004 was purified to electrophoretic homogeneity with the methods including ammonium sulfate precipitation, polyacrylamide gel, DEAE-Sepharose Fast Flow anion exchange and gel-filtration chromatography. The molecular weight of SFE1 was estimated to be 20 kDa by SDS-PAGE, fibrin zymography, and gel filtration chromatography. The isoelectric point was 4.9. K m and V max values were 0.96 mg/ml and 181.8 unit/ml, respectively. It was very stable at pH 5.0–8.0 and below 65 °C. The optimum pH for enzyme activity was 7.8. The optimum temperature was 35 °C. The fibrinolytic activity of SFE1 was enhanced by Na+, K+, Mn2+, Mg2+, Zn2+ and Co2+. Conversely, Cu2+ showed strong inhibition. Furthermore, the fibrinolytic activity was strongly inhibited by PMSF, and partly inhibited by EDTA and EGTA. SFE1 rapidly hydrolyzed the Aα-chain of fibrinogen, followed by the Bβ-chain and finally the γ-chain. The first 15 amino acids of the N-terminal sequence were APITLSQGHVDVVDI. Additionally, SFE1 directly digested fibrin and not by plasminogen activators in vitro. SFE1 can be further developed as a potential candidate for thrombolytic therapy.

Purification and characterization of a fibrinolytic enzyme.pdf

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