Screening of proteases produced by Aspergillus micromycetes active against proteins of the hemostasis system

Background and Purpose: The primary cause of death is cardiovascular disease, hence accurate diagnosis and treatment are urgently required. For these intents, proteases are regarded as perspective agents. High substrate specificity is a need for an effective enzyme, which makes Aspergillus micromycetes—known for producing proteases with precise action—biotechnologically promising. This study’s major goal was to look at the possibilities of Aspergillus species, which had never been mentioned in terms of general proteolytics. Materials and Methods: Every species was cultivated in two-stages submerged conditions with two different nitrogen sources, whereupon, proteolytic activity in culture fluid was determined. Using chromogenic peptide substrates and fibrin plates, these species’ thrombin, plasmin, factor Xa, urokinase, protein C-like, and activating activities towards hemostasis proteins, as well as fibrinolytic and plasminogen-activating activities, were evaluated. Results: It was shown that A. aureolatus and A. tennesseensis are active proteolytics exhibiting plasmin-like activity (116.17 and 87.09 U×10-3, respectively), factor Xa-like activity (76.27 and 77.92 U×10-3, respectively) and urokinase activity (85.99 and 59.91 U×10-3, respectively). The thrombin-like activity was found for A. tabacinus (50.37 U×10-3), and protein C-like activity was noticeable for A. creber, A. jensenii, A. protuberus, and A. ruber (62.90, 65.51, 73.37, and 111.85 U×10-3, respectively). Additionally, more than half of species have the ability to directly activate plasminogen or operate as fibrinolytics.Conclusion: New proteolytic strains were discovered, offering hope for the therapy of cardiovascular disorders. Fungal enzymes’ high specificity and activity make them useful in a variety of fields, including medicine and diagnostics.