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RevistaMolecules
Año2017
Volumen22
Páginas91
Internacional

Desorption of lipases immobilized on octyl-agarose beads and coated with ionic polymers after thermal inactivation. Stronger adsorption of polymers/ unfolded proteins composites.

Autores:Cristina Otero Hernández, Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos, Biocatalisis y Bioenergía (GBB)
Jose J. Virgen-Ortíz+,1, Sara G. Pedrero+,2, Laura Fernandez-Lopez2, Nerea Lopez-Carrobles2, Beatriz C. Gorines2, Cristina Otero2, Roberto Fernandez-Lafuente2*
1   CONACYT -  Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD) - Centro de Innovación y Desarrollo Agroalimentario de Michoacán, A.C. (CIDAM), Km. 8 Antigua Carretera a Pátzcuaro s/n, C.P. 58341, Morelia, Michoacán, México.
2   Departamento de Biocatálisis, Instituto de Catálisis-CSIC, C/ Marie Curie 2, Campus UAM-CSIC. Cantoblanco, 28049 Madrid, Spain


Lipases from Candida antarctica (isoform B) and Rhizomucor miehei (CALB and RML) have been immobilized on octyl-agarose (OC) and further coated with polyethylenimine (PEI) and dextran sulfate (DS). The enzymes just immobilized on OC supports could be easily released from the support using 2% SDS at pH 7, both intact or after thermal inactivation (in fact, after inactivation most enzyme molecules were already desorbed). The coating with PEI and DS greatly reduced the enzyme release during thermal inactivation and improved enzyme stability. However, using OC-CALB/RML-PEI-DS, the full release of the immobilized enzyme to reuse the support required more drastic conditions: a pH value of 3, a buffer concentration over 2 M, and temperatures above 45°C. However, even these conditions were not able to fully release the thermally inactivated enzyme molecules from the support, being necessary to increase the buffer concentration to 4 M sodium phosphate and decrease the pH to 2.5. The formation of unfolded protein/polymers composites seems to be responsible for this strong interaction between the octyl and some anionic groups of OC supports. The support could be reused five cycles using these conditions with similar loading capacity of the support and stability of the immobilized enzyme.
Palabras clave:Lipase immobilization, octyl-agarose, Enzyme inactivation, enzyme desorption, support reuse, enzyme physical crosslinking with polymers
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