The proteolytic activity of the venom of Bothrops jararaca

Abstract

The hydrolysis of casein and gelatin by the venom of B. jararaca is enhanced by Ca++ ions and inhibited by sulfide, cyanide, cysteine, reduced glutathione, ethylenediaminetetraacetic acid, 8-hydroxyquinoline and histamine, whereas the hydrolysis of benzolarginine amide is moderately inhibited only by reduced glutathione and histamine. The venom fraction precipitated between 0.50 and 0.55 saturation with (NH4)2 SO4 proved to be activated by Ca++ ions when hydrolyzing casein and had a specific activity about twice as high as that of the crude venom when hydrolyzing casein or gelatin; its benzoylarginine amidase specific activity was about the same as that of the crude venom. The venom fraction precipitated between 0.7 and 0.8 saturation with (NH4)2 SO4 had a specific activity about 4 times higher than the crude venom when hydrolyzing benzoylarginine amide; it was insensitive to Ca++ ions when hydrolyzing casein and its caseinase and gelatinase activities were lower than those of the crude venom. An isolation technique is described which leads to the preparation from the venom of a heat-resistant proteolytic enzyme which is precipitated between 0.7 and 0.8 saturation with (NH4)2 SO4 and has a benzoylarginine amidase specific activity 52 times higher than the crude venom.