RT Journal Article
T1 Desorption of Lipases Immobilized on Octyl-Agarose Beads and Coated with Ionic Polymers after Thermal Inactivation. Stronger Adsorption of Polymers/Unfolded Protein Composites
A1 Virgen Ortíz, Jose
A1 Pedrero, Sara
A1 Fernández López, Laura
A1 López Carrobles, Nerea
A1 Gorines, Beatriz
A1 Otero, Cristina
A1 Fernandez Lafuente, Roberto
AB 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.
PB MDPI
SN 1420-3049
YR 2017
FD 2017-01-05
LK https://hdl.handle.net/20.500.14352/19239
UL https://hdl.handle.net/20.500.14352/19239
LA eng
NO Ministerio de Economía y Competitividad (MINECO)
DS Docta Complutense
RD 21 abr 2025