A novel helper phage for HaloTag-mediated co-display of enzyme and substrate on phage

Phage display is an established technique for the molecular evolution of peptides and proteins. For the selection of enzymes based on catalytic activity however, simultaneous coupling of an enzyme and its substrate to the phage surface is required. To facilitate this process of co-display, we developed a new helper phage displaying HaloTag, a modified haloalkane dehalogenase that binds specifically and covalently to functionalized haloalkane ligands. The display of functional HaloTag was demonstrated by capture on streptavidin-coated magnetic beads, after coupling a biotinylated haloalkane ligand, or after on-phage extension of a DNA oligonucleotide primer with a biotinylated nucleotide by phi29 DNA polymerase. We also achieved co-display of HaloTag and phi29 DNA polymerase, thereby opening perspectives for the molecular evolution of this enzyme (and others) towards new substrate specificities. publisher: Elsevier articletitle: A novel helper phage for HaloTag-mediated co-display of enzyme and substrate on phage journaltitle: Biochemical and Biophysical Research Communications articlelink: http://dx.doi.org/10.1016/j.bbrc.2015.03.019 content_type: article copyright: Copyright © 2015 Elsevier Inc. All rights reserved. ispartof: Biochemical and Biophysical Research Communications vol:460 issue:2 pages:245-249 ispartof: location:United States status: published