Puede utilizar el filtro de búsqueda del panel izquierdo para acotar los resultados
Tipo de publicación
Todos Libros Revistas
Palabras clave
Acceso DOI
Acceso digital CSIC
Datos técnicos

Using the versatility of glutaraldehyde chemistry to immobilize and stabilize different β-glucosidases: protocol optimization depends only on the enzyme

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Diandra de Andrades,1 Natália G. Graebin,1 Marina Kimiko Kadowaki2 Marco A. Z.
Ayub,1 Roberto Fernandez-Lafuente,3 Rafael C. Rodrigues1,*
1Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology
Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box
15090, ZC 91501-970, Porto Alegre, RS, Brazil;
2Center of Medical Sciences and Pharmaceutical, Western Paraná State University, Rua
Universitária 2069, ZC 85819-110, Cascavel, PR, Brazil.
3Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049,
Madrid, Spain
The β-glucosidases from Aspergillus niger and contained in Pectinex Ultra SP-L and Pectinex Ultra Clear, were
immobilized using different strategies: ionic adsorption on aminated (MANAE)-agarose beads at pH 5, 7, and 9,
followed by biocatalysts modification with glutaraldehyde, or on glutaraldehyde pre-activated supports. The pH of the
immobilization was altered to allow different enzyme molecule orientations on the support surface. The biocatalysts
from Pectinex Ultra SP-L showed the highest thermal and operational stabilities when immobilized on MANAEagarose-
glutaraldehyde at pH 7. The β-glucosidase from Pectinex Ultra Clear and from A. niger produced best results
when immobilized on MANAE-agarose beads at pH 5 and 7, respectively, which was later treated with glutaraldehyde.
The best immobilization results using pre-activated supports were observed for the enzyme present in Pectinex Ultra
SP-L, to which the highest thermal stabilities were obtained. Remarkably, the enzyme from A. niger, immobilized on
MANAE-agarose at pH 9 and subsequently treated with glutaraldehyde, produced the highest stabilization
(approximately 560 times more stable than soluble enzyme at 60 ºC). Results showed that optimal protocol for β-
glucosidases immobilizations using the glutaraldehyde chemistry must be individually tested and tailored to each type
of enzyme.
Palabras clave:MANAE-agarose; Glutaraldehyde; Enzyme stabilization; Ionic adsorption; Covalent attachment.
logo de CSIC