Isolation, purification and functional characterization of fibrinolytic Protease from an Earthworm Eisenia foetida

Abstract

Isolation, purification and functional characterization of fibrinolytic Protease from an Earthworm Eisenia foetida.Abstract Thrombosis (development of clots in blood vessels) may cause vascular blockade culminating into serious consequences broadly known as cardiovascular diseases (CVDs). CVDs are causing more than 31% of total deaths globally. 'Thrombolysis' is the term used for dissolution of such blood clots. Numerous protein molecules called thrombolytics mediate the process of thrombolysis. Such thrombolytics are not only expensive but also cause non-specific fibrinolysis as the main side-effect, instigating excessive internal bleeding (haemorrhage) and substantial blood loss ultimately leading to death. On the other hand, lumbrokinase (LK), an earthworm-derived fibrinolytic agent being very specific, dissolves fibrin itself or convert plasminogen to plasmin by inducing endogenous tissue plasminogen activator (t-PA) to degrade fibrin clots without any ill-effects. In present study we describe the isolation of a fibrinolytic protease namely Lumbrokinase from indigenous earthworm Eisenia foetida. Protein precipitation was carried out by using various concentrations of (NH4)2SO4, desalted and concentrated by dialysis and purified by gel filtration chromatography. Furthermore, its kinetic characterization and in vitro estimation of caseinolytic activity was carried out. Characterization of earthworm-derived fibrinolytic enzyme reveals that it can serve as an ideal therapeutic molecule for oral delivery in CVD patients.

Bioevaluation of Pheretima vulgaris Antithrombotic Extract, PvQ, and Isolation, Identification of Six Novel PvQ-Derived Fibrinolytic Proteases

“…It has been used as a traditional medicine in China, Japan, and southeast Asian countries for centuries, and possesses outstanding bioactivities in terms of fibrinolysis and anti-thrombosis [ 6 ]. Although various fibrinolytic extracts have been discovered from different earthworm species, such as Eisenia fetida [ 7 , 8 , 9 ], Lumbricus rubellus [ 10 , 11 ], among others [ 12 , 13 , 14 ], since the first report of lumbrokinase by Mihara’s team in 1991 [ 15 ], few fibrinolytic activity studies related to P. vulgaris are reported. Moreover, the complexity of medicinal animal compositions has led to the previous reports being either evaluations of the activity of crude extracts [ 11 , 16 ] or vague descriptions of mixtures [ 17 ], with few reporting specifically on functional monomers.…”