Fosfolipase de Bothrops alternatus: purificação e avaliação de potencial antimicrobiano

Abstract

Introduction: The increasing bacterial resistance to conventional antibiotics is a public health concern due to the high resistance rates of microorganisms. This scenario highlights the need for new sources of antimicrobial agents. Snake venoms from the genus Bothrops are complex mixtures of bioactive molecules, including enzymes such as phospholipases A₂ (PLA₂) with potential therapeutic activity. These enzymes stand out for their action on membrane phospholipids and for their relevant biological activity, including antimicrobial potential against Gram-positive and Gram-negative bacteria. Materials and Methods: Crude venom from B. alternatus was subjected to sequential chromatographic fractionation (ion exchange, molecular exclusion, and hydrophobic interaction) for the purification of phospholipases. The fractions were analyzed for protein profile by SDS-PAGE, phospholipase activity in agarose gel emulsified with egg yolk, and antibacterial effect against reference strains of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, using disk diffusion method. Results: Fractionation of B. alternatus venom yielded a phospholipase with an approximate molecular mass of 17 kDa and 70% recovery in the final chromatographic step. The isolated phospholipase retained enzymatic activity but did not show antibacterial activity against the tested strains. However, antibacterial potential against S. aureus was identified in fraction A5B2. Conclusion: The methods applied in this study were effective in isolating from B. alternatus venom a phospholipase with preserved enzymatic activity. Although the isolated enzyme did not display antibacterial activity, antimicrobial potential was observed in intermediate fractions of the venom, suggesting the possible involvement of other proteins or synergistic action between different components. These findings reinforce the relevance of this research and indicate the need for further investigations focused on the structural characterization and therapeutic evaluation of snake venom toxins.