title: Chemical modification of enzymes for enhanced functionality.
abstract: The explosion in commercial and synthetic applications of enzymes has stimulated much of the interest in enhancing enzyme functionality and stability. Covalent chemical modification, the original method available for altering protein properties, has now re-emerged as a powerful complementary approach to site-directed mutagenesis and directed evolution for tailoring proteins and enzymes. Glutaraldehyde crosslinking of enzyme crystals and polyethylene glycol (PEG) modification of enzyme surface amino groups are practical methods to enhance biocatalyst stability. Whereas crosslinking of enzyme crystals generates easily recoverable insoluble biocatalysts, PEGylation increases solubility in organic solvents. Chemical modification has been exploited for the incorporation of cofactors onto protein templates and for atom replacement in order to generate new functionality, such as the conversion of a hydrolase into a peroxidase. Despite the breadth of applicability of chemically modified enzymes, a difficulty that has previously impeded their implementation is the lack of chemo- or regio-specificity of chemical modifications, which can yield heterogeneous and irreproducible product mixtures. This challenge has recently been addressed by the introduction of a unique position for modification by a site-directed mutation that can subsequently be chemically modified to introduce an unnatural amino acid sidechain in a highly chemo- and regio-specific manner.
authors.affiliation: Department of Chemistry, University of Toronto, 80 St George Street, Toronto, Ontario, M5S 3H6, Canada.
authors.forename: G
authors.initials: G
authors.lastname: DeSantis
authors.forename: J B
authors.initials: JB
authors.lastname: Jones
mesh_terms: D001709:Biotechnology; D003432:Cross-Linking Reagents; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering
chemical_list: D003432:Cross-Linking Reagents; D004798:Enzymes