Reuse of anion exchangers as supports for enzyme immobilization: Reinforcement of the enzyme-support multiinteraction after enzyme inactivation
| Autor: | Vicente Cortes-Corberan, Maria Russo, Antonio Marzocchella, Roberto Fernandez-Lafuente, Sara Peirce, Veymar G. Tacias-Pascacio, Jose J. Virgen-Ortíz |
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| Přispěvatelé: | Virgen Ortíz, Jose J., Peirce, Sara, Tacias Pascacio, Veymar G., Cortes Corberan, Vicente, Marzocchella, Antonio, Russo, Maria Elena, Fernandez Lafuente, Roberto |
| Rok vydání: | 2016 |
| Předmět: |
Reversibly immobilization
Immobilized enzyme Sodium chemistry.chemical_element Bioengineering 02 engineering and technology 01 natural sciences Applied Microbiology and Biotechnology Biochemistry chemistry.chemical_compound Enzyme desorption Desorption Incubation chemistry.chemical_classification Chromatography Ion exchange 010405 organic chemistry Support reuse 021001 nanoscience & nanotechnology Phosphate 0104 chemical sciences Support-enzyme multi-interaction Enzyme chemistry Ion exchange Reversibly immobilization Enzyme unfolding Support-enzyme multi-interaction Enzyme desorption Support reus Enzyme unfolding Agarose 0210 nano-technology |
| Zdroj: | Process biochemistry (1991) 51 (2016): 1391–1396. doi:10.1016/j.procbio.2016.06.020 info:cnr-pdr/source/autori:Virgen-Ortiz, Jose J.; Peirce, Sara; Tacias-Pascacio, Veymar G.; Cortes-Corberan, Vicente; Marzocchella, Antonio; Russo, Maria Elena; Fernandez-Lafuente, Roberto/titolo:Reuse of anion exchangers as supports for enzyme immobilization: Reinforcement of the enzyme-support multiinteraction after enzyme inactivation/doi:10.1016%2Fj.procbio.2016.06.020/rivista:Process biochemistry (1991)/anno:2016/pagina_da:1391/pagina_a:1396/intervallo_pagine:1391–1396/volume:51 |
| ISSN: | 1359-5113 |
| DOI: | 10.1016/j.procbio.2016.06.020 |
| Popis: | beta-Galactosidase from Aspergillus oryze has been immobilized on agarose beads coated with polyethyleneimine. The fresh enzyme was released from the support using 500 mM NaCl at pH 7. After thermal inactivation or inactivation in the presence of organic solvents, the active enzyme still could be easily released from the support using similar conditions. However, SDS-PAGE analysis of the enzyme contained in the support after enzyme desorption showed that enzyme molecules remained in the support (inactivated enzyme molecules). This effect was stronger on enzyme preparations inactivated in an organic medium. Now the conditions should be greatly strengthen to permit the full enzyme desorption: only after incubation in 2 M sodium phosphate at pH 2 and 50 degrees C full release of the enzyme molecules was achieved. This could be repeated several cycles with any difference neither in the immobilization performance nor on the SDS-PAGE analysis. Therefore, the reversibility of the immobilization is a real fact, but recovery of a support fully free of protein molecules is not an easy objective after enzyme inactivation, because the inactivated enzymes seemed to unfold increasing in a great way the interaction with the support, driving to a very strong enzyme-support multi-interaction that difficulty its desorption. (C) 2016 Elsevier Ltd. All rights reserved. |
| Databáze: | OpenAIRE |
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