Characterization and cDNA sequence of Bothriechis schlegeliil-amino acid oxidase with antibacterial activity

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 o...

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Bibliographic Details
Published in:International journal of biological macromolecules 2014-08, Vol.69, p.200-207
Main Authors: Vargas Muñoz, Leidy Johana, Estrada-Gomez, Sebastian, Núñez, Vitelbina, Sanz, Libia, Calvete, Juan J.
Format: Article
Language:English
Subjects:
Acinetobacter baumannii
Acinetobacter baumannii - drug effects
Amino Acid Sequence
Animals
Anti-Bacterial Agents - chemistry
Anti-Bacterial Agents - pharmacology
Antibacterial activity
B. schlegelii
Cell Line
Cloning, Molecular
Crotalid Venoms - enzymology
DNA, Complementary - genetics
Escherichia coli
Humans
L-Amino Acid Oxidase - chemistry
L-Amino Acid Oxidase - genetics
L-Amino Acid Oxidase - pharmacology
lAAO
Mice
Models, Molecular
Molecular Sequence Data
Protein Structure, Secondary
Staphylococcus aureus
Viperidae
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Summary: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.
ISSN:0141-8130
1879-0003
DOI:10.1016/j.ijbiomac.2014.05.039