Hyaluronidase and phospholypase activity in the venom of Mesobuthus Epeus epeus Scorpion

The Scorpion Mesobuthus Eupeus belongs to the Buthidae family. It is one of the most abundant scorpions, present in Khuzestan which is responsible for about forty five percent of stinges in this area. The venom of scorpions is composed mainly of a variety of neurotoxins. These neurotoxins specifically interact with ionic channels of excitable cell. Because of the importance of scorpion in this geographic area and since there was not much work done on the enzyme activity on Mesobuthus eupeus eupeus, we decided to do fractionation of Mesobuthus Eupeus eupeus venom with gel filtration chromatography and to study enzyme activity in all collected fractions. In this study the venom of Mesobuthus Epeus epeus was prepared in lyophilized form.One gram of crude venom was dissolved in 10 ml distilled water and centrifuged for 12 minute in 18000xg. The supernatant containing protein was applied to the sephadex G-50 gel filtration column chromatography, equilibrated with 20 mM ammonium acetate pH 4.7 with flow rate of 30 ml/h. Eluant was collected in 3 ml fractions by fraction collector system. Six fractions were separated. Absorbance of eluants was measured continuously in 280 nm. All fractions were tested for protein content, phospholipase A2 and hyaluronidase activity. In crude venom there was 816 mg protein. The activity of phospholipase A2 with indirect hemolysate in crude venom and all fractions showed that crude cvenom, Fraction I and Fraction II had 53.12, 76.30 and 2.43% hemolysis activity respectively. Hyaluronidase specific activity was 740 and 4574.5 TRU/mg (Turbidity Reducing Unit) in crude venom and in fraction I the fold of purification of hyaluronidase was 6.18 by gel filtration chromatography. It was found that phospholipase and hyaluronidase activities are present in Mesobuthus eupeus eupeus venom and both are separated in fraction I using gel filtration chromatography.