2017
DOI: 10.1007/s00449-017-1781-3
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Purification, biochemical, and structural characterization of a novel fibrinolytic enzyme from Mucor subtilissimus UCP 1262

Abstract: Fibrinolytic proteases are enzymes that degrade fibrin. They provide a promising alternative to existing drugs for thrombolytic therapy. A protease isolated from the filamentous fungus Mucor subtilissimus UCP 1262 was purified in three steps by ammonium sulfate fractionation, ion exchange, and molecular exclusion chromatographies, and characterized biochemically and structurally. The purified protease exhibited a molecular mass of 20 kDa, an apparent isoelectric point of 4.94 and a secondary structure composed… Show more

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Cited by 30 publications
(25 citation statements)
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“…The fibrinolytic enzyme obtained from Cordyceps militaris was neither inhibited by EDTA nor by PMSF, but was completely inhibited by aprotinin and soybean trypsin inhibitor [41]. On the other hand, enzyme from Bacillus subtilis ICTF-1 [65] and Mucor subtillissimus UCP 1262 [68] were inhibited by PMSF (1 mM), while the enzyme from B. pumilus 2.g, was totally inhibited by EDTA (1 mM) [69]. However, complete loss in activity of fibrinolytic protease from Bacillus cereus RSA1 was observed with PMSF (0.99%) and DFP (0.39%), and there was ≈2-fold increase in the activity with β-mercaptoethanol (191.41%) and DTT (183.42%), which indicates it to be a thiol-dependent serine fibrinolytic protease ( Figure 8A).…”
Section: Effect Of Inhibitors/activators Metal Ions and Organic Solmentioning
confidence: 97%
“…The fibrinolytic enzyme obtained from Cordyceps militaris was neither inhibited by EDTA nor by PMSF, but was completely inhibited by aprotinin and soybean trypsin inhibitor [41]. On the other hand, enzyme from Bacillus subtilis ICTF-1 [65] and Mucor subtillissimus UCP 1262 [68] were inhibited by PMSF (1 mM), while the enzyme from B. pumilus 2.g, was totally inhibited by EDTA (1 mM) [69]. However, complete loss in activity of fibrinolytic protease from Bacillus cereus RSA1 was observed with PMSF (0.99%) and DFP (0.39%), and there was ≈2-fold increase in the activity with β-mercaptoethanol (191.41%) and DTT (183.42%), which indicates it to be a thiol-dependent serine fibrinolytic protease ( Figure 8A).…”
Section: Effect Of Inhibitors/activators Metal Ions and Organic Solmentioning
confidence: 97%
“…The preparation of the inoculum, the production of the fibrinolytic enzyme by solid state fermentation and the enzymatic extraction was performed as described by Nascimento et al, (2015). The fibrinolytic protease extracted as mentioned above was initially precipitated with 40-60% ammonium sulfate and then purified by DEAE-Sephadex A50 ion exchange chromatography as described by Nascimento et al (2017). The methodological support was based on the work of Nascimento et al (2015Nascimento et al ( , 2016Nascimento et al ( , 2017.…”
Section: Obtaining Fibrinolytic Proteasementioning
confidence: 99%
“…Fungi have proven to be a good source of these enzymes, with the possibility of large-scale production, facilitating the production and purification of this bioproduct. In recent years, special attention has been paid to the production, assessment of acute toxicity, genotoxicity and cytotoxicity in different cell lines of fibrinolytic enzymes that have a high potential to degrade the fibrin clot using the genus Mucor, especially the species Mucor subtilissimus, in addition to assess its fibrinogenolytic and hemolytic capacity (Nascimento et al, 2016;Nascimento et al, 2017;Silva et al, 2019).…”
Section: Introductionmentioning
confidence: 99%
“…Such methodologies are used either individually or in combinations, accompanied by chromatographic techniques for further purification. Chromatofocusing, fast protein liquid chromatography, high performance liquid chromatography, affinity column chromatography, gel filtration chromatography, ion exchange chromatography and hydrophobic interaction chromatography are commonly employed techniques for fibrinolytic enzyme purification [2,5,[100][101][102][103][104][105][106][107][108]110,[162][163][164][165][166][167][168]. Some recent purification studies of microbial fibrinolytic enzymes employed by researchers are discussed below.…”
Section: Recovery and Purification Of Fibrinolytic Enzymesmentioning
confidence: 99%