Characterization of Partially Purified Phospholipase A2 Enzyme of Naja Katiensis Venom

Abstract

Hassan AU, Egbe NE, Ugochukwu OO, Onuh K, Kereakede E and Umar SI

Snake bite issues has been for long a neglected public health concern in Nigeria and other African countries, causing high number of human fatalities annually. It however remains a serious medical and economic problem. This research is aimed at the characterization of partially purified Phospholipase A2 (PLA2 ) enzyme of Naja katiensis venom. The enzyme is extracted using a two way purification step (Gel filitration using G-75 and ion exchange chromatography on CM- Sephadex). The molecular weight of the enzyme is determine using Sodium Dodecyl-Sulphate Electrophoresis (SDS- Page). The extracted enzyme kinetic parameters were also determine, after which its relative optimum affinity/activity were also determine with related to different ranges of temperature, pH, metal ions and salts. The partially purified PLA2 gave a total enzyme activity of 3.11 μmol/min and an estimated extrapolated molecular weight of 17.5 KDa. Initial velocity data of the enzyme was used to compute the kinetic parameters of the enzyme, where the Km and Vmax of the enzyme was estimated to be 12.6 mg/ml and 3.32 μmoles/min respectivily. The enzyme optimum temperature and pH was found to be 35oC and 7.0. Ca2+ ions was revealed to increase the enzyme activity and affinity. This revealed data could provide an alternative natural way of producing a more friendly pharmaceutical formulation for the management and treatment of snake envenomation.

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