Purification and antibacterial activities of an L-amino acid oxidase from king cobra (Ophiophagus hannah) venom

Abstract

Some constituents of snake venom have been found to display a variety of biological activities. The antibacterial property of snake venom, in particular, has gathered increasing scientific interest due to antibiotic resistance. In the present study, king cobra venom was screened against three strains of Staphylococcus aureus [including methicillin-resistant Staphylococcus aureus (MRSA)], three other species of gram-positive bacteria and six gram-negative bacteria. King cobra venom was active against all the 12 bacteria tested, and was most effective against Staphylococcus spp. (S. aureus and S. epidermidis). Subsequently, an antibacterial protein from king cobra venom was purified by gel filtration, anion exchange and heparin chromatography. Mass spectrometry analysis confirmed that the protein was king cobra L-amino acid oxidase (Oh-LAAO). SDS-PAGE showed that the protein has an estimated molecular weight of 68 kDa and 70 kDa under reducing and non-reducing conditions, respectively. The minimum inhibitory concentrations (MIC) of Oh-LAAO for all the 12 bacteria were obtained using radial diffusion assay method. Oh-LAAO had the lowest MIC value of 7.5 µg/mL against S. aureus ATCC 25923 and ATCC 29213, MRSA ATCC 43300, and S. epidermidis ATCC 12228. Therefore, the LAAO enzyme from king cobra venom may be useful as an antimicrobial agent.

Synergistic antibacterial effect of co-administering adipose-derived mesenchymal stromal cells and Ophiophagus hannah l-amino acid oxidase in a mouse model of methicillin-resistant Staphylococcus aureus-infected wounds

“…SDS-PAGE conducted on purified Oh-LAAO revealed a single band with a molecular weight of approximately 68 kDa (see Additional file 1: Figure S3) which corresponds to data previously reported to be the expected size of Oh-LAAO [4]. …”

“…Characterization of the protein was performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, and liquid chromatography-mass spectrometry/mass spectrometry (LCMS/MS). Enzymatic analyses including LAAO activity, cytotoxicity, and antibacterial assays confirmed functionality and a minimum inhibitory concentration (MIC) value similar to published data [4]. Purity of the enzyme was reconfirmed by SDS-PAGE prior to in vivo studies.…”

“…l -amino acid oxidase isolated from Ophiophagus hannah (Oh-LAAO) has been demonstrated to be more efficacious in its antimicrobial activity against many strains of Gram-positive bacteria commonly associated with cutaneous wounds compared to routinely used antibiotics [3]. The antimicrobial activity of Oh-LAAO on MRSA has also been previously demonstrated [4], as has properties of thermal stability and retention of activity under alkaline conditions [5]. …”

Therapeutic Uses of Amino Acids

“…In another study of the heat stable l-amino acid oxidase isolated from the king cobra (O. hannah) venom, Phua et al [33] Therapeutic Uses of Amino Acids http://dx.doi.org/10.5772/intechopen.68932…”

Predicting antibacterial activity from snake venom proteomes

“…Crude venoms and their constituents have previously been determined to be effective against bacteria. For example, venom extracted from Ophiophagus hannah was shown to hinder Pseudomonas aeruginosa, Salmonella enteritidis, Escherichia coli, and Staphylococcus aureus growth [6,7], including resistant strains such as Methicillin-resistant S. aureus (MRSA). Numerous studies focused on the antibacterial properties of snake venom with over 58% of those studies reporting effective results [8].…”