Artículo
Analysis of the interactions between Eudragit® L100 and porcine pancreatic trypsin by calorimetric techniques
Fecha de publicación:
01/2012
Editorial:
Elsevier Science
Revista:
International Journal of Biological Macromolecules
ISSN:
0141-8130
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Flexible-chain polymers with charge (polyelectrolytes) can interact with globular proteins with a net charge opposite to the charge of the polymers forming insoluble complexes polymer?protein. In this work, the interaction between the basic protein trypsin and the anionic polyelectrolyte Eudragit® L100 was studied by using isothermal calorimetric titrations and differential scanning calorimetry. Turbidimetric assays allowed determining that protein?polymer complex was insoluble at pH below 5 and the trypsin and Eudragit® L100 concentrations required forming the insoluble complex. DSC measurements showed that the Tm and denaturalization heat of trypsin increased in the polymer presence and the complex unfolded according to a two-state model. DH y DS binding parameters obtained by ITC were positives agree with hydrophobic interaction between trypsin and polymer. However, ionic strength of 1.0 M modified the insoluble complex formation. We propose a mechanism of interaction between Eudragit® L100 and trypsin molecules that involves both hydrophobic and electrostatic interactions. Kinetic studies of complex formation showed that the interaction requires less than 1 min achieving the maximum quantity of complex. Finally, a high percentage of active trypsin was precipitated (approximately 76% of the total mass of protein). These findings could be useful in different protocols such as a protein isolation strategy, immobilization or purification of a target protein.
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Articulos(CCT - ROSARIO)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - ROSARIO
Citación
Braia, Mauricio Javier; Tubio, Gisela; Nerli, Bibiana Beatriz; Loh, Watson; Romanini, Diana; Analysis of the interactions between Eudragit® L100 and porcine pancreatic trypsin by calorimetric techniques; Elsevier Science; International Journal of Biological Macromolecules; 50; 1; 1-2012; 180-186
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