Artículo
Interaction between trypsin and alginate: An ITC and DLS approach to the formation of insoluble complexes
Braia, Mauricio Javier
; Loureiro, Dana Belen
; Tubio, Gisela
; Lienqueo, María Elena; Romanini, Diana
Fecha de publicación:
07/2017
Editorial:
Elsevier Science
Revista:
Colloids and Surfaces B: Biointerfaces
ISSN:
0927-7765
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Trypsin is a protease widely used in several industrial areas for leather and meat softening and to produce enzymatic detergents, among others applications. The high demand for this enzyme has motivated the development of purification, stabilization and immobilization methods Formation of insoluble complexes between proteins and polyelectrolytes is a methodology that may include these features. The aim of this paper is to give evidence for a novel methodology that combines precipitation of the insoluble trypsin-alginate complex and hydrophobic interaction chromatography. This methodology allows the interaction between trypsin and alginate and their separation when necessary. It could be applied to isolation, stabilization and/or immobilization of trypsin. Isothermal titration calorimetry experiments showed that 232 μmol of trypsin interacts electrostatically with 1 g of alginate to form an insoluble complex that can be separated from soluble contaminants by decantation. Dynamic light scattering experiments confirmed the calorimetric results and allowed measuring the Rh of the soluble complex at pH 3.5 (185 nm). When the optimal conditions were applied to precipitate commercially available trypsin, the recovery of the precipitation was around 92%. Finally, hydrophobic interaction chromatography allowed separating alginate from trypsin in order to obtain a polymer-free enzyme.
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Articulos(CCT - ROSARIO)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Articulos(IPROBYQ)
Articulos de INST. DE PROCESOS BIOTECNOLOGICOS Y QUIMICOS ROSARIO
Articulos de INST. DE PROCESOS BIOTECNOLOGICOS Y QUIMICOS ROSARIO
Articulos(SEDE CENTRAL)
Articulos de SEDE CENTRAL
Articulos de SEDE CENTRAL
Citación
Braia, Mauricio Javier; Loureiro, Dana Belen; Tubio, Gisela; Lienqueo, María Elena; Romanini, Diana; Interaction between trypsin and alginate: An ITC and DLS approach to the formation of insoluble complexes; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 155; 7-2017; 507-511
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