PURIFICATION OF THE 28KDA CATHEPSIN L FROM FASCIOLA HEPATICA

• Autores: Arias, Luis, López, G. Miryam, Marcelo, Alvaro
• Formato: artículo
• Fecha de Publicación: 2015
• Institución: Universidad Nacional Federico Villarreal
• Repositorio: Revistas - Universidad Nacional Federico Villarreal
• Lenguaje: español
• OAI Identifier: oai:ojs2.revistas.unfv.edu.pe:article/1554
• Enlace del recurso: https://revistas.unfv.edu.pe/rtb/article/view/1554
• Nivel de acceso: acceso abierto
• Materia: cathepsin L; chromatography; electrophoresis; Fasciola hepatica; proteinase; catepsina L; cromatografía; electroforesis; proteinasa

This work aims to isolate the 28KDa proteinase from the parasite Fasciola hepatica (Linnaeus, 1758). This cysteinyl protease was purified by classical methods such as the ammonium sulphate precipitation, G-25 and G-75 gel filtration chromatographies, affinity chromatography with thiol sepharose 4B and SDS-PAGE 12% electrophoresis. The enzymatic activity of Cathepsin L was monitored in each step of purification using a specific substrate. The cysteine proteinase was observed as a pure and homogenous band in the SDS-PAGE gel. The recovery percentage was 12.7%, with a specific activity of 337,500 umol, min-1 mg-1 and a purification factor of the enzyme of 260. Using this simple scheme of purification we purified to homogeneity the cathepsin L of 28KDa from the regurgitate in an effective way compared with other alternative methods. The amount of protein obtained was 0.032 mg·mL-¹. The enzyme showed as a homogenous band during the electrophoresis in polyacrylamide gel with 12.5% SDS-PAGE and the molecular weight was 28 kDa according to its relative mobility.