| Autor: |
Anderson Lourenço da Silva, Jamil S. Oliveira, Marcos Luiz dos Mares Guia, Marcelo M. Santoro, E.R. Bittar, Alexandre Martins Costa Santos, Marcelo P. Bemquerer |
| Rok vydání: |
2008 |
| Předmět: |
|
| Zdroj: |
Brazilian Archives of Biology and Technology. 51:511-521 |
| ISSN: |
1516-8913 |
| DOI: |
10.1590/s1516-89132008000400009 |
| Popis: |
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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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