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Isolation, biochemical and functional characterization of L - amino acid - oxidase from yellow venom of the snake Crotalus durissus terrificus

Grant number: 12/02839-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): April 01, 2012
Effective date (End): December 31, 2012
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Maurício Ventura Mazzi
Grantee:Ariana Aparecida Pianca
Home Institution: Pró-Reitoria de Pós-Graduação e Pesquisa. Centro Universitário Hermínio Ometto (UNIARARAS). Fundação Hermínio Ometto (FHO). Araras , SP, Brazil

Abstract

Toxins found in venomous and poisonous animals have been the focus of much research over the past 70 years, most of them directed toward understanding the physiological and biochemical mechanisms by which can trigger their dramatic pathological effects. The crotalidae venom consists abundantly by enzymes and peptides that have important effects on skeletal muscles, nervous system, kidneys and blood clotting. The L-amino acid oxidases (LAAOs, EC 1.4.3.2) are homodimers flavoproteins found in snake venoms and described as inducing a series of toxic effects, such as platelet aggregation, hemorrhage, edema, cytotoxicity and apoptosis inducers, among others effects. So the aim of this project is to isolate, characterize, biochemistry and functionally an L-amino acid oxidase of yellow venom of snake species Crotalus durissus terrificus. The enzyme will be isolated by chromatographic techniques (molecular exclusion, ion exchange and affinity) according to protocols to be standardized. The LAAOcdt will be isolated and characterized as: i)-biochemical and enzymatic properties (determination of molecular mass, kinetic characterization depending on pH, temperature and concentration of different substrates: L-Tyr, L-Asp, L-Phe, L-Glu, L-Trp, L-His, L-Gln, L-Ile, L-Met, L-Leu) and ii)-biological effects (myotoxicity, necrosis and pro and / or anticoagulant effects).

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