Hydrophobic interaction chromatography as separation method of alkaline proteases from viscera of Scomberomorus sierra

This study focused on recovering alkaline proteases from the viscera of Scomberomorus sierra through hydrophobic interaction chromatography. Three alkaline proteases were partially separated using this chromatographic technique; two of them, with molecular weights of 19 and 31 kDa, were identified a...

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Detalles Bibliográficos
Autores: Osuna-Amarillas, Pablo Sergio, Rouzaud-Sandez, Ofelia, Higuera-Barraza, Odilia Azucena, Arias-Moscoso, Joe Luis, López-Mata, Marco Antonio, Campos-García, Julio César, Valdez-Melchor, Ramón Gertrudis
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:México
Institución:UNIVERSIDAD NACIONAL AUTÓNOMA DE MÉXICO
Repositorio:TIP Revista especializada en ciencias químico-biológicas
Idioma:español
OAI Identifier:oai:ojs.ojs.escire.net:article/183
Acceso en línea:http://tip.zaragoza.unam.mx/index.php/tip/article/view/183
Access Level:acceso abierto
Palabra clave:chymotrypsin; hydrophobic interaction chromatography; Scomberomorus sierra; trypsin
quimiotripsina; cromatografía de interacción hidrofóbica; Scomberomorus sierra; tripsina
Descripción
Sumario:This study focused on recovering alkaline proteases from the viscera of Scomberomorus sierra through hydrophobic interaction chromatography. Three alkaline proteases were partially separated using this chromatographic technique; two of them, with molecular weights of 19 and 31 kDa, were identified as trypsin-like enzymes according to inhibition assays. The 31 kDa alkaline protease, the only isolated enzyme, was purified under following chromatographic conditions: ammonium sulfate 13% (w/v) and ethylene glycol 27% (w/v); this enzyme showed maximum activity at pH 9 – 10 and 50 – 60 °C and was strongly inhibited by soybean trypsin inhibitor (SBTI) and porcine trypsin inhibitor (TPI). A third alkaline protease with molecular weight of 20 kDa was partially separated and inhibited by tosyl phenylalanyl chloromethyl ketone (TPCK), showing optimum activity at pH 9 – 11 and 60 °C. These results show that the viscera of Scomberomorus sierra may be useful as source of proteases.