Improved purification process of β- and α-trypsin isoforms by ion-exchange chromatography

The purpose of this work was to improve the separation and yield of pure β- and α-trypsin isoforms by ion-exchange chromatography and to characterize some physical-chemical properties of these isoforms. Purification of trypsin isoforms was performed by ion-exchange chromatography in 0.1 mol/L tris-HC buffer, pH 7.10 at 4ºC. The sample loading, salt concentration, flow rate and pH of mobile phase were varied to determine their effects on the resolution of the separation. The resolution was optimized mainly between β- and α-trypsin. Pure isoforms were obtained by chromatographying 100 mg of commercial trypsin during seven days, yielding 51 mg of high purity β-trypsin and 13 mg of α-trypsin partially pure, with small amounts of contaminating of ψ-trypsin. Thus, time and resolution of purification were optimized yielding large amounts of pure active enzymes that are useful for several research areas and biotechnology.

Bibliographic Details

Main Authors:	Santos,Alexandre Martins Costa, Oliveira,Jamil Silvano de, Bittar,Eustáquio Resende, Silva,Anderson Lourenço da, Guia,Marcos Luiz dos Mares, Bemquerer,Marcelo Porto, Santoro,Marcelo Matos
Format:	Digital revista
Language:	English
Published:	Instituto de Tecnologia do Paraná - Tecpar 2008
Online Access:	http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000400009
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