Fibrinogenolytic activity of serine proteases(s) from Cucumis dipsaceus

Abstract

Cucurbitaceae species plants have been widely used in traditional medicine for wound care management and hemostasis. This study aims to explore the possible involvement of Cucumis dipsaceus fruit extract on the blood coagulation pathway. The aqueous Cucumis dipsaceus dialysate fraction (AqCDF) hydrolyzes casein and exhibiting a protease activity in a dose-dependent manner. Casein zymography further reveals the presence of high molecular weight protease(s). The inhibition of proteolytic activity by phenylmethanesulfonyl fluoride (PMSF) suggesting the presence of serine protease. The serine protease(s) fraction exhibiting a promising fibrinogenolytic activity at 25 μg and 50 μg concentration by hydrolyzing Aα and Bβ subunit of fibrinogen, whereas γ-subunit undergo partial degradation. The stability of protease activity against various parameters viz., temperature, pH has been evaluated and its optimum activity was found to be 60 °C & pH 7.0 respectively. The AqCDF shows a promising recalcification time up to 80% against trypsin. More interestingly AqCDF (300 μg) did not induce hemolysis even at higher concentration. These findings suggest the possible role of the dialyzed fraction of Cucumis dipsaceus in hemostasis and the wound healing process.