RT Journal Article
T1 Enzyme loading in the support and medium composition during immobilization alter activity, specificity and stability of octyl agarose-immobilized Eversa Transform
A1 Sabi, Guilherme J.
A1 De Souza, Leonardo
A1 Abellanas Perez, Pedro
A1 Tardioli, Paulo W.
A1 Mendes, Adriano A.
A1 Rocha Martín, Javier
A1 Fernandez Lafuente, Roberto
AB Eversa Transform (ETL) was immobilized on octyl agarose beads at two different enzymes loadings (1 mg/g and 15 mg/g) under 18 different conditions, including different pH values, buffers, additives (different solvents, Ca2+, NaCl). Their activity was analyzed at pH 5 and 7 with p-nitrophenyl butyrate and at pH 5 with triacetin, determining also its stability at pH 5 and 7 (in different media). Ca2+ stabilized ETL biocatalysts while phosphate destabilized them. The overloaded biocatalysts were generally less stable and with a lower specific activity than the lowly loaded biocatalyst. Results show that enzyme activity (even by a 3 fold factor) and stability of the immobilized enzyme may be tailored by controlling the immobilization conditions, but the effects of the immobilization conditions on activity depend on the substrate and conditions of activity determination, the effects on stability depend on the inactivation conditions. Moreover, the enzyme loading of the biocatalysts defines the effects of the immobilization conditions, and there are clear interactions between immobilization conditions (e.g., immobilization pH determines the effect of the presence of NaCl). These suggest that the extrapolation of the results obtained with one substrate under one condition to other conditions can lead to wrong decisions.
PB Elsevier
SN 0141-8130
YR 2025
FD 2025
LK https://hdl.handle.net/20.500.14352/118416
UL https://hdl.handle.net/20.500.14352/118416
LA eng
NO Sabi, G. J., De Souza, L., Abellanas-Perez, P., Tardioli, P. W., Mendes, A. A., Rocha-Martin, J., & Fernandez-Lafuente, R. (2025). Enzyme loading in the support and medium composition during immobilization alter activity, specificity and stability of octyl agarose-immobilized Eversa Transform. International Journal of Biological Macromolecules, 295, 139667. https://doi.org/10.1016/j.ijbiomac.2025.139667
NO RF-L gratefully recognizes the financial support from Ministerio de Ciencia e Innovación and Agencia Estatal de Investigación (Spanish Government) (PID2022-136535OB-I00). AAM recognizes the support from CNPq (project 306253/2023-2). Guilherme J. Sabi and Leonardo de Sousa gratefully recognice Coordenação de Aperfeiçoamento de Pessoal de Nível Superior–Brasil (CAPES, Finance Code 001; CAPES-PRINT Process Numbers 88887.936648/2024-00; CAPES-PDSE Process Numbers 88881.933605/2024-01) for supporting the stay at ICP-CSIC.
NO Ministerio de Ciencia e Innovación (España)
NO Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil)
NO Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Brasil)
DS Docta Complutense
RD 19 ene 2026