Adsorption features of reduced aminated supports modified with glutaraldehyde: Understanding the heterofunctional features of these supports

Immobilization of enzymes on aminated supports using the glutaraldehyde chemistry may involve three different interactions, cationic, hydrophobic, and covalent interactions. To try to understand the impact this heterofunctionality, we study the physical adsorption of the beta-galactosidase from Aspe...

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Detalles Bibliográficos
Autores: Andrades, Diandra de, Abellanas, Pedro, Carballares Navarro, Diego, Alcántara León, Andrés Rafael, Polizeli, Maria de Lourdes Teixeira de Moraes, Rocha Martín, Javier, Fernandez-Lafuente, Roberto
Tipo de recurso: artículo
Fecha de publicación:2024
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/119650
Acceso en línea:https://hdl.handle.net/20.500.14352/119650
Access Level:acceso abierto
Palabra clave:577.15
66.081
66.092
Heterofunctional supports
Mixed adsorption
Enzyme release
Biología molecular (Biología)
Bioquímica (Biología)
Ingeniería química
Biotecnología
2403 Bioquímica
3302 Tecnología Bioquímica
3313 Tecnología E Ingeniería Mecánicas
2302.09 Enzimología
2304 Química Macromolecular
Descripción
Sumario:Immobilization of enzymes on aminated supports using the glutaraldehyde chemistry may involve three different interactions, cationic, hydrophobic, and covalent interactions. To try to understand the impact this heterofunctionality, we study the physical adsorption of the beta-galactosidase from Aspergillus niger, on aminated supports (MANAE) and aminated supports with one (MANAE-GLU) or two molecules of glutaraldehyde (MANAE-GLU-GLU). To eliminate the chemical reactivity of the glutaraldehyde, the supports were reduced using sodium borohydride. After enzyme adsorption, the release of the enzyme from the supports using different NaCl concentrations, Triton X100, ionic detergents (SDS and CTAB), or different temperatures (4 °C to 55 °C) was studied. Using MANAE support, at 0.3 M NaCl almost all the immobilized enzyme was released. Using MANAE-GLU, 0.3 M, and 0.6 M NaCl similar results were obtained. However, incubation at 1 M or 2 M NaCl, many enzyme molecules were not released from the support. For the MANAE-GLU-GLU support, none of the tested concentrations of NaCl was sufficient to release all enzyme bound to the support. Only using high temperatures, 0.6 M NaCl, and 1 % CTAB or SDS, could the totality of the proteins be released from the support. The results shown in this paper confirm the heterofunctional character of aminated supports modified with glutaraldehyde.