Making and Breaking Peptide Bonds: Protein Engineering Using Sortase

Abstract Sortases are a class of bacterial enzymes that possess transpeptidase activity. It is their ability to site‐specifically break a peptide bond and then reform a new bond with an incoming nucleophile that makes sortase an attractive tool for protein engineering. This technique has been adopted for a range of applications, from chemistry‐based to cell biology and technology. In this Minireview we provide a brief overview of the biology of sortase enzymes and current applications in protein engineering. We identify areas that lend themselves to further innovation and that suggest new applications.