Desorption of Lipases Immobilized on Octyl-Agarose Beads and Coated with Ionic Polymers after Thermal Inactivation. Stronger Adsorption of Polymers/Unfolded Protein Composites

• Published in: Molecules
• Main Authors: Virgen-Ortíz, José J., Pedrero, Sara G., Fernández-López, Laura, López-Carrobles, Nerea, Gorines, Beatriz C., Otero Hernández, Cristina, Fernández-Lafuente, Roberto
• Other Authors: Consejo Superior de Investigaciones Científicas (España), Ministerio de Economía y Competitividad (España)
• Format: Article in Journal/Newspaper
• Language: unknown
• Published: Multidisciplinary Digital Publishing Institute 2017
• Subjects: Martínez; Antarc*; Antarctica
• Online Access: http://hdl.handle.net/10261/149339; https://doi.org/10.3390/molecules22010091; https://doi.org/10.13039/501100003329; https://doi.org/10.13039/501100003339

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. We gratefully recognize the support from the MINECO from Spanish Government, (project numbers CTQ2013-41507-R and CTQ2016-78587-R). The authors wish to thank Ramiro Martínez (Novozymes, Spain) for kindly supplying some of the enzymes used in this research. The help and comments from Ángel Berenguer (Instituto de Materiales, Universidad de Alicante) are gratefully recognized. We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).