Abstract

VIVAS RUIZ, Dan et al. Barnetobin. Rev. Soc. Quím. Perú [online]. 2010, vol.76, n.3, pp.261-270. ISSN 1810-634X.

A thrombin- like enzyme, barnetobin, was purified from Bothrops barnetti, peruvian snake venom using CM Sephadex C-50 followed by Sephadex G-100, in both two cases with 0,05 M amonium acetate buffer pH 5,0. The enzyme was purified 45 fold with 14% of yield and the PAGE-SDS showed only protein band of 52 kDa under reducing condition with 2β- mercaptoetanol and 48 kDa under non reducing condition indicating that the enzyme has a single polypeptide chain with disulfide bond. The PNGase treatment showed that it is a basic glycoprotein containing 45% total carbohydrates. The enzyme has coagulant activity on fibrinogen and citrated plasma and amidolytic activity on BApNA and Chromozym TH. The coagulant potency was equivalent to 131 NIH thrombin Units/ mg. In addition the enzyme is inhibited by PMSF and soybean trypsin inhibitor suggesting that is a serine proteinase. The enzyme had optimal amidolytic activity pH was 8,0 and is stable until 40 ºC. The antigenicity and neutralization of enzyme was demonstrated by inmunodiffusion and inmunoelectrophoresis with the polyvalent antibothropic serum on citrated plasma (Effective Doses: 250 µl of antivenom/ mg of enzyme).

Keywords : Venom; snake; enzyme; thrombin like; Bothrops barnetti; coagulant.

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