title: Characterization and cDNA sequence of Bothriechis schlegeliil-amino acid oxidase with antibacterial activity.
abstract: Snake venoms are complex mixtures of proteins including l-amino acid oxidase (lAAO). A lAAO (named BslAAO) with a mass of 56kDa and a theoretical Ip of 5.79, was purified from Bothriechis schlegelii venom through size-exclusion, ion exchange and affinity chromatography. The entire protein sequence of 498 amino acids, was determined from cDNA using reverse-transcribed mRNA isolated from venom gland. The enzyme showed dose-dependent inhibition of bacterial growth. BslAAO showed inhibitory effect against S. aureus with a MIC of 4μg/mL and a MBC of 8μg/mL. Against Acinetobacter baumannii, showed a MIC of 2μg/mL and MBC of 4μg/mL, No effect was observed in Escherichia coli. This antibacterial activity was inhibited by catalase, indicating that antimicrobial activity was due to H2O2 production. BslAAO did not show any cytotoxic activity toward mouse myoblast cell line C2C12 or peripheral blood mononuclear cells. The enzyme oxidated l-Leu, with a Km of 16.37μM and a Vmax of 0.39μM/min. Snake venoms lAAOs, are potential frames of different therapeutics molecules since these enzymes exhibit low MICs and MBCs and show to be harmless to human cells due to microorganisms being generally several fold more sensitive to reactive oxygen species than human tissues.
authors.affiliation: Programa de Ofidismo/Escorpionismo, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Calle 70 No. 52-2, A.A. 1226, Medellín, Colombia. Electronic address: johana2104@gmail.com.
authors.forename: Leidy Johana
authors.initials: LJ
authors.lastname: Vargas Muñoz
authors.affiliation: Programa de Ofidismo/Escorpionismo, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Calle 70 No. 52-2, A.A. 1226, Medellín, Colombia; Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Estrada-Gomez
authors.affiliation: Programa de Ofidismo/Escorpionismo, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Calle 70 No. 52-2, A.A. 1226, Medellín, Colombia; Escuela de microbiología, Universidad de Antioquia UdeA, Medellín, Colombia.
authors.forename: Vitelbina
authors.initials: V
authors.lastname: Núñez
authors.affiliation: Consejo Superior de Investigaciones Científicas, Jaume Roig 11, 46010 Valencia, Spain.
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.affiliation: Consejo Superior de Investigaciones Científicas, Jaume Roig 11, 46010 Valencia, Spain.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
mesh_terms: D040981:Acinetobacter baumannii / Q000187:drug effects; D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000201:enzymology; D018076:DNA, Complementary / Q000235:genetics*; D006801:Humans; D050830:L-Amino Acid Oxidase / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017819:Viperidae
chemical_list: D000900:Anti-Bacterial Agents; D003435:Crotalid Venoms; D018076:DNA, Complementary; D050830:L-Amino Acid Oxidase
keywords: Antibacterial activity; B. schlegelii; lAAO