Production and purification of a fibrinolytic enzyme from Bionectria sp.

JOSÉ I. ROVATI; VICTOR G. ARNAU; LUCÍA I. C. DE FIGUEROA; JULIA I. FARIÑA

S.M. Tucumán, Argentina

Congreso; XLV Reunión Anual SAIB; 2009

SAIB

Several thrombolytic agents such as urokinase, alteplase (t-PA), and streptokinase (SK) are currently available for clinical usage, although they still suffer significant shortcomings, including requeriment of large therapeutic dose, short plasma half-life, limited fibrin specificity, reocclusion and bleeding complications. Among microbial sources, strains of Bacillus, Aspergillus, Fusarium and Streptomyces have been reported as fibrinolytic enzyme producers. In the present study we isolated a novel Bionectria sp. LY 4.1 fibrinolytic enzyme producer from Las Yungas forest. After cultivation at shake-flask scale, the supernatant was precipitated with cold acetone, and purified by gel filtration chromatography with Sephacryl S-300 HR. Fibrinolytic activity was monitored by the fibrin plate method, and quantified by the amount of tyrosine released from fibrin clots per min. The effect of pH and incubation temperature was studied. Finally, in vitro activity over human blood clots was also analyzed to verify thrombolytic effects. Results confirmed that the purified fibrinolytic enzyme obtained from cultures of Bionectria sp. LY 4.1 showed thrombolytic effect over human blood clots and constitude a promising candidate for treating thrombolytic diseases with the additional benefits of its more specific mode of action (plasminogen independent) and probably associated to less side effects.