Purificação e caracterização bioquímica de peptídeos antimicrobianos dos venenos das serpentes Bothrops atrox e Bothrops jararacussu

Abstract

The emergence of pathogenic agents resistant to conventional drugs has stimulated the search for new classes of antimicrobial and antiparasitic agents from natural sources. Among these, it highlights antimicrobial peptides (AMPs), which have mechanisms not dependent on the interaction with a specific receptor, providing new possibilities for the development of drugs against resistant organisms. The present study aimed to purify and characterize biochemically the peptides of B. atrox and B. jararacussu snakes venoms, as well as to evaluate the antimicrobial and antiparasitic activities of the peptides against the strains: Staphylococcus aureus (ATCC 29213), Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Klebsiella pneumoniae (ATCC 13883); Leishmania amazonensis (IFLA/BR/97/PH8), and Plasmodium falciparum (clone W2, resistant to chloroquine). In this work, we carried out a comprehensive analysis of peptides presents in the venom of B. atrox and B. jararacussu using the combination of Amicon ultrafiltration of 3kDa, high performance liquid chromatography in reverse phase and mass spectrometry with electrospray ionization (Electrospray-Ion Trap-Time of Flight). Sixteen peptides were identified in both venoms, with mass range 444.17 to 1356.73 Da and primary structure between 3 to 13 residues. Among them 13 are unique sequences, including 7 peptides and 6 bradykinin-potentiating peptides (BBPs). The other peptides refer to 2 metallopeptidases inhibitor found in both peptidome venoms, but not described for these species, besides 1 BPP identified in the venom of B. atrox, the BPP-Bax12 with peptide sequence described. Some of the new peptides identified showed antibacterial activity against K. pneumoniae, P. aeruginosa and S. aureus, and low hemolytic activity, but did not demonstrate antiparasitic activity. Therefore, in this study 13 new peptides were identified from the B. atrox and B. jararacussu snake venoms, and some with relevant bacterial activities, which can assist in the development of new drugs against multiresistant microorganisms.