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RevistaBiotechnology Progress
Año1970
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Internacional

Preparation of immobilized / stabilized biocatalysts of betaglucosidases from different sources: importance of the support active groups and the immobilization protocol

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Diandra de Andrades,1 Natália G. Graebin,1 Marco A. Z. Ayub,1 Roberto Fernandez-Lafuente*,2 and Rafael C. Rodrigues1,*
 
1.- Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institut, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil; 
2.-  Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049, Madrid, Spain
 

β‐glucosidases from two different commercial preparations, Pectinex Ultra SP‐L and Celluclast® 1.5 L, were immobilized on divinyl sulfone supports at pH 5.0, 7.0, 9.0, and 10. In addtion, the biocatalysts were also immobilized in agarose beads activated by glyoxyl, and epoxide as reagent groups. The best immobilization results were observed using higher pH values on divinyl sulfone‐agarose, and for Celluclast® 1.5 L, good results were also obtained using the glyoxil‐agarose immobilization. The biocatalyst obtained using Pectinex Ultra SP‐L showed the highest thermal stability, at 65°C, and an operational stability of 67% of activity after 10 reuses cycles when immobilized on DVS‐agarose immobilized at pH 10 and blocked with EDA. The β‐glucosidase from Celluclast® 1.5 L produced best results when immobilized on DVS‐agarose immobilized at pH 9 and blocked with glycine, reaching 7.76‐fold higher thermal stability compared to its free form and maintaining 76% of its activity after 10 successive cycles. The new biocatalysts obtained by these protocols showed reduction of glucose inhibition of enzymes, demonstrating the influence of immobilization protocols, pH, and blocking agent.

Palabras clave:covalent immobilization, Enzyme stabilization, divinyl sulfone, epoxide, glucose tolerance
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