Actividad antibacteriana in vitro del veneno de Hadruroides sp. “escorpión”, frente a Acinetobacter baumannii, Klebsiella pneumoniae y Serratia marcescens
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Date
2023
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Universidad Nacional de Trujillo
Abstract
Se caracterizó el veneno crudo de Hadruroides sp. (escorpión) en su composición proteica y se evaluó su actividad antibacteriana frente a Acinetobacter baumannii ATCC19606, Klebsiella pneumoniae ATCC 700603 y Serratia marcescens. El estudio es experimental. Los 115 Hadruroides sp se muestrearon en la ciudad de Jaén y se trabajó con una cepa de A. baumannii, K. pneumoniae y S. marcescens. Para el ordeño se aplicó una descarga eléctrica entre 15 a 20 voltios y el contenido proteico se mensuró por espectrofotometría UV. Para la separación, identificación y cuantificación se usó Sephadex CM-25, después la determinación del peso molecular se llevó a cabo mediante electroforesis en geles de poliacrilamida en condiciones desnaturalizantes con dodecil sulfato de sodio y cromatografía líquida de alta resolución acoplado a espectrometría de masas en tándem (UHPLC-MS/MS). La inhibición del crecimiento de las tres especies bacterianas se ensayó por microdilución. A través del análisis por SDS-PAGE los péptidos tuvieron PM de10 kDa. Por Sephadex CM-25 se obtuvieron 11 picos agrupados en cinco fracciones, el análisis por UHPLC-MS/MS de las fracciones obtenidas por CM-25, determino que solo la fracción III presenta moléculas de la naturaleza catiónica a los tR (tiempo de retención) de 2.509 y 3.041 min con un rango de masa entre 628 y 699 Da. la fracción III presentó una CMI de 0,002 mg/mL para A. baumannii. Mientras que para las cepas K. pneumoniae y S. marcescens no presentó CMI a las concentraciones evaluadas. El veneno de Hadruroides sp. presentó péptidos catiónicos que inhiben el crecimiento de A. baumannii.
The raw venom of Hadruroides sp. (scorpion) in its protein composition and its antibacterial activity was evaluated against against Acinetobacter baumannii ATCC19606, Klebsiella pneumoniae ATCC 700603 and Serratia marcescens. The study is experimental. The 115 Hadruroides sp were sampled in Jaén and we worked with a strain of A. baumannii, K. pneumoniae and S. marcescens. For venom milking, an electric stimulation with 15 to 20 V was applied and the protein content was quantified by the absorption method. Sephadex CM-25 was used for separation, identification and quantification, then molecular weight determination was performed by electrophoresis in polyacrylamide gels under denaturing conditions with sodium dodecyl sulfate and high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Growth inhibition of the three bacterial species was tested by microdilution. Using SDS-PAGE analysis determined size was around peptides 10 kDa. By Sephadex CM-25, 11 peaks grouped into five fractions were obtained, the analysis by UHPLC-MS/MS of the fractions obtained by CM-25, determined that only fraction 3 presents molecules of a cationic nature at the tR (retention time) of 2,509 and 3,041 min with a mass range between 628 and 699 Da. The fraction III presented a MIC of 0,002 mg/mL for the strain A. baumannii. Meanwhile K. pneumoniae and S. marcescens did not presented MIC at the concentrations evaluated. The venom of Hadruroides sp. present cationic peptides that inhibit the growth of A. baumannii
The raw venom of Hadruroides sp. (scorpion) in its protein composition and its antibacterial activity was evaluated against against Acinetobacter baumannii ATCC19606, Klebsiella pneumoniae ATCC 700603 and Serratia marcescens. The study is experimental. The 115 Hadruroides sp were sampled in Jaén and we worked with a strain of A. baumannii, K. pneumoniae and S. marcescens. For venom milking, an electric stimulation with 15 to 20 V was applied and the protein content was quantified by the absorption method. Sephadex CM-25 was used for separation, identification and quantification, then molecular weight determination was performed by electrophoresis in polyacrylamide gels under denaturing conditions with sodium dodecyl sulfate and high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Growth inhibition of the three bacterial species was tested by microdilution. Using SDS-PAGE analysis determined size was around peptides 10 kDa. By Sephadex CM-25, 11 peaks grouped into five fractions were obtained, the analysis by UHPLC-MS/MS of the fractions obtained by CM-25, determined that only fraction 3 presents molecules of a cationic nature at the tR (retention time) of 2,509 and 3,041 min with a mass range between 628 and 699 Da. The fraction III presented a MIC of 0,002 mg/mL for the strain A. baumannii. Meanwhile K. pneumoniae and S. marcescens did not presented MIC at the concentrations evaluated. The venom of Hadruroides sp. present cationic peptides that inhibit the growth of A. baumannii
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Keywords
Hadruroides sp., Acinetbacter baumanni, Klebsiella pneumoniae, Serratia marcescens, Antibacteriano, Veneno de escorpión