One-step immobilization and stabilization of a recombinant enterococcus faecium DBFIQ E36 L-arabinose isomerase for D-tagatose synthesis

A recombinant L-arabinose isomerase from Enterococcus faecium DBFIQ E36 was immobilized onto multifunctional epoxide supports by chemical adsorption and onto a chelate-activated support via polyhistidine-tag, located on the N-terminal (N-His-L-AI) or on the C-terminal (C-His-L-AI) sequence, followed by covalent bonding between the enzyme and the support. The results were compared to reversible L-AI immobilization by adsorption onto charged agarose supports with improved stability. All the derivatives presented immobilization yields of above 75%. The ionic interaction established between agarose gels containing monoaminoethyl-N-aminoethyl structures (MANAE) and the enzyme was the most suitable strategy for L-AI immobilization in comparison to the chelate-activated agarose. In addition, the immobilized biocatalysts by ionic interaction in MANAE showed to be the most stable, retaining up to 100% of enzyme activity for 60 min at 60 °C and with Km values of 28 and 218 mM for MANAE-N-His-L-AI and MANAE-C-His-L-AI, respectively.

Bibliographic Details

Main Authors:	Sousa, Marylane de, Melo, Vânia M. M., Hissa, Denise C., Manzo, Ricardo M., Mammarella, Enrique J., Antunes, André Saraiva Leão Marcelo, García, José Luis, Pessela, Benevides C., Gonçalves, Luciana R. B.
Other Authors:	Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil)
Format:	artículo biblioteca
Language:	English
Published:	Springer 2019-06
Subjects:	Chelate-agarose, D-tagatose, Enterococcus faecium, Enzyme activity, Immobilization, L-Arabinose isomerase,
Online Access:	http://hdl.handle.net/10261/183587 http://dx.doi.org/10.13039/501100003033 http://dx.doi.org/10.13039/501100003593 http://dx.doi.org/10.13039/501100005283
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