Autor(es): Sousa, Marylane de ; Melo, Vânia M. M. ; Hissa, Denise Cavalcante ; Manzo, Ricardo M. ; Mammarella, Enrique J. ; Antunes, André Saraiva Leão Marcelo ; García, José L. ; Pessela, Benevides C. ; Gonçalves, Luciana R. B.
Data: 2021
Origem: Oasisbr
Assunto(s): L-Arabinose isomerase; Chelate-agarose; D-Tagatose; Enterococcus faecium; Immobilization; Enzyme activity
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.
