One-step immobilization and stabilization of a recombinant Enterococcus faecium DBFIQ E36 L-Arabinose isomerase for D-Tagatose synthesis

• Autor(a) principal: Sousa, Marylane de
• Data de Publicação: 2018
• Outros Autores: 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.
• Tipo de documento: Artigo
• Idioma: por
• Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
• Texto Completo: http://www.repositorio.ufc.br/handle/riufc/62917
• topic: 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.