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

  • Xiuyun Ju
  • Xiaoying Cao
  • Yong Sun
  • Zhe Wang
  • Chengliang Cao
  • Jinjuan Liu
  • Jihong Jiang
Original Paper

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. Km and Vmax 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.

Keywords

Fibrinolytic enzyme Purification Streptomyces Thrombolytic therapy 

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Xiuyun Ju
    • 1
  • Xiaoying Cao
    • 1
  • Yong Sun
    • 1
  • Zhe Wang
    • 1
  • Chengliang Cao
    • 1
  • Jinjuan Liu
    • 1
  • Jihong Jiang
    • 1
  1. 1.The Key Laboratory of Biotechnology for Medicinal Plant of Jiangsu ProvinceJiangsu Normal UniversityXuzhouChina

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