Purification and characterization of fibrinolytic enzyme from Pseudoalteromonas sp., IND11 and its in vitro activity on blood clot

Abstract

Fibrinolytic enzymes are agents that dissolve fibrin clots. These fibrinolytic agents have potential use to treat cardiovascular diseases, such as heart attack and stroke. The aim of the study was to purify fibrinolytic enzyme from the marine isolate, Pseudoalteromonas sp., IND11. Enzyme was purified to electrophoretic homogeneity using ammonium sulphate precipitation, ion exchange and affinity chromatography. The SDS-PAGE showed that it was a monomeric protein with an apparent molecular weight of 64 kDa. The purified enzyme was active at pH 6.0-9.0 with an optimum pH of 8.0. It was stable upto 50°C, exhibiting maximum activity between 30 and 60°C. Among the ions, Na + and Ca 2+ activated enzyme activity. The Fe 2+ did not obviously activate or inhibit the enzyme activity. The ions such as Cu 2+, Hg 2+ and Zn 2+ strongly affected enzyme activity. This enzyme activated plasminogen and also had direct clot lytic activity. It digested the fibrin net of blood clot, suggests its potential as an effective thrombolytic agent. This study explores new sources of fibrinolytic enzymes to treat and prevent CVDs.