New Heterofunctional Supports Based on Glutaraldehyde-Activation: A Tool for Enzyme Immobilization at Neutral pH

Autor(es)

Rodrigues de Melo, Ricardo ; Carlos Alnoch, Robson ; Lopes Vilela, Adriana Ferreira ; de Souza, Emanuel Maltempi ; Krieger, Nadia ; Ruller, Roberto ; Sato, Helia Harumi ; Mateo, Cesar

• Tipo de documento: Artigo Científico
• Fonte: Molecules; v. 22, n. 7 JUL 2017.
• Citações Web of Science: 15

Resumo

Immobilization is an exciting alternative to improve the stability of enzymatic processes. However, part of the applied covalent strategies for immobilization uses specific conditions, generally alkaline pH, where some enzymes are not stable. Here, a new generation of heterofunctional supports with application at neutral pH conditions was proposed. New supports were developed with different bifunctional groups (i.e., hydrophobic or carboxylic/metal) capable of adsorbing biocatalysts at different regions (hydrophobic or histidine richest place), together with a glutaraldehyde group that promotes an irreversible immobilization at neutral conditions. To verify these supports, a multi-protein model system (E. coli extract) and four enzymes (Candida rugosa lipase, metagenomic lipase, beta-galactosidase and beta-glucosidase) were used. The immobilization mechanism was tested and indicated that moderate ionic strength should be applied to avoid possible unspecific adsorption. The use of different supports allowed the immobilization of most of the proteins contained in a crude protein extract. In addition, different supports yielded catalysts of the tested enzymes with different catalytic properties. At neutral pH, the new supports were able to adsorb and covalently immobilize the four enzymes tested with different recovered activity values. Notably, the use of these supports proved to be an efficient alternative tool for enzyme immobilization at neutral pH. (AU)

• Processo FAPESP: 13/01710-1 - Ligantes enzimáticos: novos modelos de triagem
• Beneficiário: Quezia Bezerra Cass
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 16/15037-5 - Co-imobilização das enzimas acetilcolinesterase e beta-secretase 1: estudo de condições para triagem de ligantes
• Beneficiário: Adriana Ferreira Lopes Vilela
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado