Biochemical Characterisation and Kinetic Analysis of Phospholipase A2 from Naja melanoleuca Venom

Abstract

This study aimed to isolate and partially purify the major basic phospholipase  A₂ (PLA₂) from the venom of Naja melanoleuca using Resource™ S cation-exchange chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as well as the determination of its characteristics and kinetic parameters. The venom was reconstituted in a buffer solution and subjected to chromatography. PLA₂-containing fractions were eluted and collected. SDS-PAGE analysis revealed a heterogeneous mixture of proteins with molecular weights of approximately 35, 26, 20, 13, and 7 kDa. The kinetic properties assessments revealed optimal activity at pH 8.0 and temperature 60℃ with 2.2mM and 0.00017µmol/min respectively for Km and Vmax where 6152.36 J/mol was the activation energy deduced for the PLA2.The relatively low intensity of the PLA₂ bands suggested a low yield, indicating the need for further optimization using reverse-phase HPLC. The results provide key insights into the composition of Naja melanoleuca venom and lay the groundwork for future venom research and therapeutic applications. The research work was conducted between February 2024 to June 2024 at the Department of Biochemistry, Faculty of Life Science, University of Maiduguri, Borno State, Nigeria.