Engineering sortase A by screening a second‐generation library using phage display

Abstract

Sortase‐mediated ligation is one of the most commonly used chemo‐enzymatic techniques for the site‐specific modification of proteins. We have established a new library of sortase mutants for directed evolution of sortase substrate selectivity. Phage display screens of this second‐generation library yielded sortase mutants that ligate substrate proteins containing an APxTG or FPxTG recognition sequence instead of the canonical LPxTG sorting motif. These findings indicate that the second‐generation sortase library is well suited for sortase engineering in order to increase the versatility of sortase‐mediated ligation. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.