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

Author(s)

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

• Document type: Journal article
• Source: Molecules; v. 22, n. 7 JUL 2017.
• Web of Science Citations: 15

Abstract

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)

• FAPESP's process: 13/01710-1 - Enzyme ligand: new models of screening
• Grantee: Quezia Bezerra Cass
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/15037-5 - Co-immobilization of enzymes acetylcholinesterase and beta-secretase 1: study of conditions for screening of ligands
• Grantee: Adriana Ferreira Lopes Vilela
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate