Immobilization of lipases on glyoxyl-octyl supports: Improved stability and reactivation strategies

• Published in: Process Biochemistry
• Main Authors: Suescun, Angélica, Rueda, Nazzoly, Sousa dos Santos, José Cleiton, Castillo, John J., Ortiz, Claudia, Torres Sáez, Rodrigo, Barbosa, Oveimar, Fernández-Lafuente, Roberto
• Other Authors: Ministerio de Economía y Competitividad (España), Colciencias (Colombia), Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil)
• Format: Article in Journal/Newspaper
• Language: unknown
• Published: Elsevier 2015
• Subjects: Enfolding/refolding; Covalent immobilization; Enzyme reactivation; Enzyme stabilization; Immobilization of lipases via interfacial activation; Rugosa; Antarc*; Antarctica
• Online Access: http://hdl.handle.net/10261/190168; https://doi.org/10.1016/j.procbio.2015.05.010; https://doi.org/10.13039/501100003593; https://doi.org/10.13039/501100003329

Lipases from Candida rugosa (CRL) and from Candida antarctica (isoform A) (CALA) have been successfully immobilized on octyl–glyoxyl agarose (OCGLX) beads and compared to the octyl–agarose (OC) or glyoxyl (GLX) beads immobilized counterparts. Immobilization on OCGLX gave similar hyperactivations than those found for the immobilization on OC supports, although the incubation at pH 10.0 for 4 h decreased the activity of both enzymes by 25%. After reduction, more than 95% of the enzyme activity was covalently attached to the support. The fraction not covalently attached was desorbed by washing with detergent. These biocatalysts were more stable than the octyl counterparts in thermal or organic solvent inactivation. More interestingly, the irreversible immobilization permitted the reactivation of CALA biocatalysts inactivated by incubation in organic solvent, after unfolding in the presence of guanidine and refolding in aqueous buffer (around 55% of the activity could be recovered during 3 successive cycles of inactivation/reactivation). GLX–CALA permitted to recover 75% of the activity, but the thermal stability and activity was much lower, and this strategy could not be applied to CRL. Neither the enzyme immobilized on cyanogen bromide nor the enzyme immobilized on OCGLX exhibited significant activity after the unfolding/refolding strategy. We gratefully recognize the support from the MINECO of Spanish Government, CTQ2013-41507-R. The predoctoral fellowships for Ms. Rueda (Colciencia, Colombian Goberment) and Mr dos Santos (CNPq, Brazil) are also recognized. Peer Reviewed