Identification and purification of the three toxins from Odotobuthus bidentatus, Lourenco and Pezier 2002, venom (scorpions, buthidae)

Abstract

Odonthobuthus belongs to Buthidae family from which 2 species O.doriea and O. bidentatus have been collected and reported from Iran. This study investigated the venom of O. bidentatus to identify mammal toxic fractions. After collecting scorpions, they were milked with electroshock technique and then the venoms was lyophilized. The LD50 of the venom was determined by injecting the venom to 18-20 g mice via., tail vein. In order to separate different fractions, the freeze dried venom was solubilized in distilled water and centrifuged at 15000 rpm for 20 min, to separate insoluble material and the clear supernatant containing soluble venom was loaded on a chromatography column packed with sephaidex G50 gel and the fractions were collected according to UV absorption at 280 nm wavelength. Second fraction was toxic in Lab animal and in order to study sub fractions, fraction II (O2) was loaded first on an anionic ion exchange resin (DEAE sephacel) out of which we obtained 2 toxic fractions (O21 and O23). Then these 2 fraction were loaded on a cationic resin (CM sephadex C25) out of which, from fraction 21, we got 2 toxic fractions (O211, O213) and from fraction O23 we got one toxic fraction (O232). SDS PAGE electrophoretic studies on these toxins showed single band appearance that indicates purified toxins with molecular weight of O211 9.2, O213 8.6 and O233 6.8 kD. © 2009 Asian Network for Scientific Information.