RT Journal Article
T1 Synergy of ion exchange and covalent Reaction: immobilization of penicillin G acylase on herofunctional amino-vinyl sulfone agarose
A1 da Rocha, Thays
A1 Morellon-Sterling, Roberto
A1 Gonçalves, Luciana
A1 Bolívar Bolívar, Juan Manuel
A1 Alcántara León, Andrés Rafael
A1 Rocha Martín, Javier
A1 Fernández-Lafuente, Roberto
AB Agarose-vinyl sulfone (VS) beads have proven to be a good support to immobilize several enzymes. However, some enzymes are hardly immobilized on it. This is the case of penicillin G acylase (PGA) from Escherichia coli, which is immobilized very slowly on this support (less than 10% in 24 h). This enzyme is also not significantly adsorbed in aminated MANAE-agarose beads, an anionic exchanger. In this study, MANAE-agarose beads were modified with divinyl sulfone (DVS) to produce MANAE-vinyl sulfone (VS) agarose beads. When PGA was immobilized on this support, the enzyme was fully immobilized in less than 1.5 h. PGA cannot be released from the support by incubation at high ionic strength, suggesting that the enzyme was rapidly immobilized in a covalent fashion. Considering that the amount of reactive VS groups was only marginally increased, the results indicated some cooperative effect between the anion exchange on the amine groups of the support, probably as the first step of the process, and the covalent attachment of the previously adsorbed PGA molecules. The covalent reaction of the previously adsorbed enzyme molecules proceeds much more efficiently than that of the free enzyme, due to the proximity of the reactive groups of the support and the enzyme. Finally, the steps of immobilization, incubation, and blocking with different agents were studied to determine the effects on final activity/stability. The stability of PGA immobilized on this new catalyst was improved with respect to the VS-agarose prepared at low ionic strength
PB MDPI
SN 2073-4344
YR 2023
FD 2023
LK https://hdl.handle.net/20.500.14352/101579
UL https://hdl.handle.net/20.500.14352/101579
LA eng
NO da Rocha, T. N., Morellon-Sterling, R., Gonçalves, L. R. B., Bolivar, J. M., Alcántara, A. R., Rocha-Martin, J., & Fernández-Lafuente, R. (2023). Synergy of Ion Exchange and Covalent Reaction: Immobilization of Penicillin G Acylase on Heterofunctional Amino-Vinyl Sulfone Agarose. Catalysts, 13(1). https://doi.org/10.3390/CATAL13010151
NO National Council for Scientific and Technological Development (Brazil)
NO Ministerio de Ciencia, Innovación y Universidades (España)
DS Docta Complutense
RD 4 abr 2025