Fibrinolytic characteristics of Bacillus subtilis G8 isolated from Natto

Abstract

Due to the high prevalence of vascular obstructive diseases, discovering potent, safe, and affordable fibrinolytic agents is of importance. There is particular interest concerning the use of functional foods that have a fibrinolytic activity, such as natto, a Japanese fermented soy-based product made with Bacillus subtilis (natto) strain BEST195. We recently isolated another bacterial strain from natto commercialized in Indonesia, Bacillus subtilis G8 , which has proven to exert fibrinolytic activity. Herein, a further characterization of Bacillus subtilis G8 was assessed through a comparison with commercialized nattokinase, the major fibrinolytic enzyme of Bacillus subtilis, by utilizing various in vitro fibrinolytic assays, namely whole blood clot lysis, euglobulin clot lysis, the fibrin plate method, and zymography. Both nattokinase and Bacillus subtilis G8 were able to dissolve both whole blood and euglobulin clots. Furthermore, both nattokinase and Bacillus subtilis G8 were able to lyse blood clots, presumably due to their ability to directly lyse fibrin. Finally, a crude extract of Bacillus subtilis G8 displayed six zymogram bands of approximately 42.0, 35.5, 30.8, 26.7, 20.0, and 13.7 kDa, with the strongest activity observed at 20.0 kDa. This indicates that Bacillus subtilis G8 contained several fibrinolytic enzymes, which might have comprised nattokinase and other fibrinolytic enzymes. In summary, we demonstrated that a crude extract of Bacillus subtilis G8 has potent fibrinolytic activity and that the activity was mediated by various fibrinolytic enzymes.