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 mainly of α-helices. Selectivity for N-succinyl-Ala–Ala–Pro–Phe-p-nitroanilide as substrate suggests that this enzyme is a chymotrypsin-like serine protease, whose activity was enhanced by the addition of Cu2+, Mg2+, and Fe2+. The enzyme showed a fibrinolytic activity of 22.53 U/mL at 40 °C and its contact with polyethylene glycol did not lead to any significant alteration of its secondary structure. This protein represents an important example of a novel fibrinolytic enzyme with potential use in the treatment of thromboembolic disorders such as strokes, pulmonary emboli, and deep vein thrombosis.
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Acknowledgements
The authors would like to thank the Projects RENNORFUN/CNPq process No. 563382/2010-4 and SISBIOTA/FACEPE process No. APQ-0086-2.12/11 for financial support of the present work. The authors also appreciate FACEPE and CNPq for the scholarship of the first author and the Laboratory of Bioactive Technology (LABTECBIO/UFRPE) that provided the necessary structure for the research.
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Nascimento, T.P., Sales, A.E., Porto, T.S. et al. Purification, biochemical, and structural characterization of a novel fibrinolytic enzyme from Mucor subtilissimus UCP 1262. Bioprocess Biosyst Eng 40, 1209–1219 (2017). https://doi.org/10.1007/s00449-017-1781-3
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DOI: https://doi.org/10.1007/s00449-017-1781-3