biotechnology, Trichoderma, gourd flour, thrombolytic therapy

 

 

The present study describes the production, purification and characterization of a fibrinolytic protease obtained from the Trichoderma sp. UFPIT04 strain, focusing on the biotechnological application of the enzyme, mainly in the area of thrombosis treatment. The protease was produced extracellularly by submerged fermentation, using gurgueia flour as substrate. The maximum protease activity achieved was 52.92 U/mL, which indicates good enzyme production. The protease purification was performed by FPLC using a Sepharose CL4B column. This method allowed the isolation of the protease, with a peak of protease activity, confirming the purification with 160 mAU. The study demonstrated that the ideal pH for the purified enzymatic activity was 7.0 and the optimum temperature was 40°C. Furthermore, the demonstrated enzyme was classified as a metalloprotease, being inhibited by metal ions such as Zn²⁺, Mg²⁺ and Ca²⁺. The molecular mass of the protease was determined as 33 kDa. The process used to purify the fibrinolytic protease from Trichoderma sp. UFPIT04 results in an effective study, with low cost and significant speed, which makes the production of the enzyme viable and accessible to the industry, contributing to the reduction of operational costs and the decrease in the use of chemical processing, evidencing its possible application and use in the treatment of thrombosis and encouraging further research for its in-vivo application.