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

• Published in: World journal of microbiology & biotechnology Vol. 28; no. 7; pp. 2479 - 2486
• Main Authors: Ju, Xiuyun, Cao, Xiaoying, Sun, Yong, Wang, Zhe, Cao, Chengliang, Liu, Jinjuan, Jiang, Jihong
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.07.2012; Springer Nature B.V
• Subjects: Amino acids; Ammonium; Anion exchange; Applied Microbiology; Biochemistry; Biomedical and Life Sciences; Biomedical research; Biotechnology; Chromatography; Environmental Engineering/Biotechnology; Enzymatic activity; Enzyme Stability; Enzymes; Fibrinogen - metabolism; Fibrinolytic Agents - chemistry; Fibrinolytic Agents - metabolism; Filtration; Fungal Proteins - chemistry; Fungal Proteins - metabolism; Herbal medicine; Humans; Hydrogen-Ion Concentration; Isoelectric Point; Life Sciences; Microbiology; Molecular weight; Original Paper; Proteins; Streptomyces; Streptomyces - enzymology; Studies; Temperature; Thrombosis; China; Fibrinolytic enzyme; Purification; Thrombolytic therapy
• ISSN: 0959-3993; 1573-0972
• DOI: 10.1007/s11274-012-1055-9

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 + , Mn 2+ , Mg 2+ , Zn 2+ and Co 2+ . Conversely, Cu 2+ showed strong inhibition. Furthermore, the fibrinolytic activity was strongly inhibited by PMSF, and partly inhibited by EDTA and EGTA. SFE1 rapidly hydrolyzed the A α-chain of fibrinogen, followed by the B β-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.