FIBRINOLYTIC PROTEASES OF ENTOMOPATHOGENIC MICROMYCETES: PRODUCTION AND CHARACTERISTICS

Abstract

The search for new fibrinolytic enzymes is of great interest, as they are widely used in the treatment of thromboembolic conditions. However, existing treatments are limited and may cause side effects. The aim of the study is to find out whether entomopathogenic micromycetes can produce fibrinolytic enzymes and to study their properties. Primary screening of proteolytic enzyme production was performed among
15 strains of entomopathogenic micromycetes. After cultivation of the producer strain and precipitation with ammonium sulfate, followed by purification by dialysis, enzyme extracts were obtained. Fractionation was then performed using isoelectric focusing to isolate individual samples for the analysis of the proteolytic activity of fungal strains.

Three most promising strains from the Cordycipitaceae and Ophiocordycipitaceae families were selected. The most suitable cultivation conditions were determined, and the producing strain was selected. Micromycetes have demonstrated a significant ability to break down protein molecules. Consequently, a comprehensive analysis of proteolytic activity was conducted in 15 entomopathogenic strains.
The dynamics of protease accumulation in the nutrient medium was investigated in three strains after submerged cultivation. A preparation of a proteolytic enzyme from the micromycete Akanthomyces cf. aculeatus, exhibiting both overall proteolytic and fibrinolytic activities, was obtained. The results indicate that the proteolytic agent under consideration is a protein with a molecular weight estimated at approximately 35 kDa and an isoelectric point of 2.15. Moreover, it has been confirmed that this enzyme can cleave both fibrin and fibrinogen according to SDS-PAGE analysis.