title: Purification, characterization and antibacterial activity of L-amino acid oxidase from Cerastes cerastes.
abstract: Antibiotic resistance presents a real problem in which new antibacterial molecules from natural secretions could be beneficial in the development of new drugs. In this study, Cerastes cerastes venom was investigated for its antibacterial activity against Gram-positive and Gram-negative bacteria. The antibacterial activity was evaluated by measuring the halo inhibition and minimum inhibitory concentration (MIC). An L-amino acid oxidase (CcLAAO) was purified from this venom using three chromatographic steps; its homogeneity (60 kDa) was confirmed by SDS-PAGE. LC-MS/MS analysis of CcLAAO showed similarities with other LAAO enzymes from Echis ocellatus and Viridovipera stejnegeri venoms. CcLAAO presents an antibacterial activity against three bacterial strains (Staphylococcus aureus, Methicillin-resistant S. aureus, and Pseudomonas aeruginosa) with MIC values of 10, 10, and 20 μg/mL, respectively. However, no effect was observed against Escherichia coli and yeast strains. Kinetic parameters of CcLAAO evaluated on L-leucine at pH 8.0 and 20°C were Km = 0.06 mmol and Vmax = 164 mmol/min.
authors.affiliation: USTHB, Laboratory of Cellular and Molecular Biology, Faculty of Biological Sciences, BP 32, El-Alia, Bab Ezzouar, 16111, Algiers, Algeria.
authors.forename: Ziad-Meziane
authors.initials: ZM
authors.lastname: Hanane-Fadila
authors.forename: Laraba-Djebari
authors.initials: LD
authors.lastname: Fatima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004926:Escherichia coli / Q000187:drug effects; D050830:L-Amino Acid Oxidase / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D053719:Tandem Mass Spectrometry; D014757:Viper Venoms / Q000201:enzymology*; D017819:Viperidae
chemical_list: D014757:Viper Venoms; D050830:L-Amino Acid Oxidase
keywords: Antibacterial Activity; Cerastes cerastes; Kinetics; Viperidae Venom; l-Amino Acid Oxidase