Abellanas Perez, PedroCarballares Navarro, DiegoFernandez Lafuente, RobertoRocha Martín, Javier2025-06-182025-06-182023Abellanas-Perez, Pedro, et al. «Glutaraldehyde Modification of Lipases Immobilized on Octyl Agarose Beads: Roles of the Support Enzyme Loading and Chemical Amination of the Enzyme on the Final Enzyme Features». International Journal of Biological Macromolecules, vol. 248, septiembre de 2023, p. 125853. DOI.org (Crossref), https://doi.org/10.1016/j.ijbiomac.2023.125853.0141-813010.1016/j.ijbiomac.2023.125853https://hdl.handle.net/20.500.14352/121485DC thank to Ministerio de Ciencia e Innovación-Spanish Government for a FPI. This research was funded by Ministerio de Ciencia e Innovación and Agencia Estatal de Investigación (Spanish Government) (PID2022-136535OB-I00 and TED2021-131462B-I00).Lipase B from Candida antarctica (CALB) and lipase from Thermomyces lanuginosus (TLL) have been immobilized on octyl agarose at low loading and at a loading exceeding the maximum support capacity. Then, the enzymes have been treated with glutaraldehyde and inactivated at pH 7.0 in Tris-HCl, sodium phosphate and HEPES, giving different stabilities. Stabilization (depending on the buffer) of the highly loaded biocatalysts was found, very likely as a consequence of the detected intermolecular crosslinkings. This did not occur for the lowly loaded biocatalysts. Next, the enzymes were chemically aminated and then treated with glutaraldehyde. In the case of TLL, the intramolecular crosslinkings (visible by the apparent reduction of the protein size) increased enzyme stability of the lowly loaded biocatalysts, an effect that was further increased for the highly loaded biocatalysts due to intermolecular crosslinkings. Using CALB, the intramolecular crosslinkings were less intense, and the stabilization was lower, even though the intermolecular crosslinkings were quite intense for the highly loaded biocatalyst. The stabilization detected depended on the inactivation buffer. The interactions between enzyme loading and inactivating buffer on the effects of the chemical modifications suggest that the modification and inactivation studies must be performed under the target biocatalysts and conditions.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal article1879-0003https://doi.org/10.1016/j.ijbiomac.2023.125853https://www.sciencedirect.com/science/article/pii/S0141813023027484?via%3Dihubopen access577.1566.095.8547.2Inter and intramolecular enzyme crosslinkingEnzyme stabilizationEffect of enzyme loadingEffect of the inactivating buffer on enzyme stabilityBioquímica (Biología)Química orgánica (Química)MaterialesBiotecnología2302.09 Enzimología2306 Química Orgánica2302 Bioquímica