Prospecção de peptídeos antimicrobianos na peçonha do escorpião Tityus obscurus
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Data
2022-12-01
Autores
Vilela, Daniela Araújo
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Universidade Estadual Paulista (Unesp)
Resumo
Os venenos escorpiônicos são misturas complexas de componentes, como, nucleotídeos, mucopolissacarídeos, oligopeptídeos, moléculas de baixa massa molecular, proteínas e peptídeos. Além disso, os peptídeos podem ser divididos em duas categorias, os peptídeos de ponte dissulfeto (DBPs) e peptídeos sem ponte dissulfeto (NDBPs). Este estudo tem como objetivo caracterizar bioquimicamente os peptídeos no veneno do escorpião preto amazônico Tityus obscurus, após fracionamento cromatográfico, a fim de avaliar a presença de atividade antimicrobiana. A amostra do veneno foi separada por Cromatografia Líquida de Alta Eficiência – Fase Reversa (CLAE ou em inglês “HPLC - RP”), equipado com uma coluna analítica C18 para obtenção de seis frações, que foram testadas sobre os microrganismos Staphylococcus aureus, Escherichia coli, Candida albicans e C. tropicalis. O percentual de inibição de crescimento microbiano efetivo das frações ocorreu em C. albicans (F5 – 18,36% e F6 – 38,95%) e S. aureus (F5 – 16,52%). No entanto, na maioria dos casos houve leve aumento da proliferação microbiana. As análises por espectrometria de massas da fração com maior índice de inibição de crescimento microbiano indicaram a presença de três moléculas: uma fosfolipase, uma metalopeptidase e uma neurotoxina. Atividades antibióticas derivadas de hidrolases já são conhecidas, inclusive para Tityus sp., restando investigar o possível papel desta neurotoxina na atividade antibiótica aqui reportada para T. obscurus. O uso de peptídeos antimicrobianos advindos de fontes alternativas, como visto em venenos escorpiônicos, geraram interesse e notoriedade devido a sua vasta atividade biológica. Este conhecimento pode auxiliar tratamentos adjuntos e/ou alternativos para as infecções resistentes aos antibióticos convencionais.
Scorpion venoms are complex mixtures of components, such as nucleotides, mucopolysaccharides, oligopeptides, low molar mass molecules, proteins and peptides. Furthermore, peptides can be divided into two categories, disulfide-bridged peptides (DBPs) and non-disulfide-bridged peptides (NDBPs). This study aims to characterize biochemically the peptides in the venom of the Amazonian black scorpion Tityus obscurus after chromatographic fractionation in order to assess antimicrobial activity presence. The venom sample was injected into a High Performance Liquid Chromatography - Reverse Phase (HPLC - RP), equipped with a C18 analytical column to obtain the six fractions, which were tested for the microorganisms Staphylococcus aureus, Escherichia coli, Candida albicans and C. tropicalis. The percentage of effective microbial growth inhibition of the fractions occurred in C. albicans (F5 – 18.36% and F6 – 38.95%) and S. aureus (F5 – 16.52%). Nevertheless, there was an slight increase in microbial proliferation. The mass spectrometry analysis of the fraction with the highest microbial growth inhibition rate indicated the presence of three molecules: a phospholipase, a metallopeptidase and a neurotoxin. Antibiotic activities derived from hydrolases are already known, including for Tityus sp, and the possible role of this neurotoxin in the antibiotic activity reported here for T. obscurus remains to be investigated. The use of antimicrobial from alternative sources, as seen in scorpion venoms, has generated interest and notoriety due to their broad biological activity. This knowledge can aid adjunctive and/or alternative treatments for infections resistant to conventional antibiotics.
Scorpion venoms are complex mixtures of components, such as nucleotides, mucopolysaccharides, oligopeptides, low molar mass molecules, proteins and peptides. Furthermore, peptides can be divided into two categories, disulfide-bridged peptides (DBPs) and non-disulfide-bridged peptides (NDBPs). This study aims to characterize biochemically the peptides in the venom of the Amazonian black scorpion Tityus obscurus after chromatographic fractionation in order to assess antimicrobial activity presence. The venom sample was injected into a High Performance Liquid Chromatography - Reverse Phase (HPLC - RP), equipped with a C18 analytical column to obtain the six fractions, which were tested for the microorganisms Staphylococcus aureus, Escherichia coli, Candida albicans and C. tropicalis. The percentage of effective microbial growth inhibition of the fractions occurred in C. albicans (F5 – 18.36% and F6 – 38.95%) and S. aureus (F5 – 16.52%). Nevertheless, there was an slight increase in microbial proliferation. The mass spectrometry analysis of the fraction with the highest microbial growth inhibition rate indicated the presence of three molecules: a phospholipase, a metallopeptidase and a neurotoxin. Antibiotic activities derived from hydrolases are already known, including for Tityus sp, and the possible role of this neurotoxin in the antibiotic activity reported here for T. obscurus remains to be investigated. The use of antimicrobial from alternative sources, as seen in scorpion venoms, has generated interest and notoriety due to their broad biological activity. This knowledge can aid adjunctive and/or alternative treatments for infections resistant to conventional antibiotics.
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Palavras-chave
Tityus obscurus, Peptidômica, Antimicrobiano, Bioprospecção, AMPs, Metalopeptidase, Fosfolipase, Actina, Beta toxina, Escorpionismo, Peptidomics, Bioprospecting, Antimicrobial