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

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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.

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Acknowledgments

This work was supported by the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), National Natural Science Foundation of China (No. 31000005, 31170605), the Project of Outstanding Scientific and Technological Innovation Team for Higher Education Institutions in Jiangsu Province (Pre-development of medical microbiology) and University-Industry Cooperation Program of Jiangsu Province (No. BY2009116).

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  1. The Key Laboratory of Biotechnology for Medicinal Plant of Jiangsu Province, Jiangsu Normal University, Xuzhou, 221116, China

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

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  1. Xiuyun Ju
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  2. Xiaoying Cao
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  3. Yong Sun
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  4. Zhe Wang
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  6. Jinjuan Liu
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Correspondence to Jihong Jiang.

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Ju, X., Cao, X., Sun, Y. et al. Purification and characterization of a fibrinolytic enzyme from Streptomyces sp. XZNUM 00004. World J Microbiol Biotechnol 28, 2479–2486 (2012). https://doi.org/10.1007/s11274-012-1055-9

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  • DOI: https://doi.org/10.1007/s11274-012-1055-9

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