A general strategy for the evolution of bond-forming enzymes using yeast display

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 108; no. 28; pp. 11399 - 11404
• Main Authors: Chen, Irwin, Dorr, Brent M., Liu, David R.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 12.07.2011; National Acad Sciences
• Subjects: Amino Acid Sequence; Aminoacyltransferases - chemistry; Aminoacyltransferases - genetics; Aminoacyltransferases - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Base Sequence; Biological Sciences; Catalysis; Catalysts; catalytic activity; CD154 antigen; chemical bonding; Chemical reactions; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - genetics; Cysteine Endopeptidases - metabolism; directed evolution; Directed molecular evolution; Directed Molecular Evolution - methods; DNA, Recombinant - genetics; Enzyme substrates; Enzymes; Enzymes - chemistry; Enzymes - genetics; Enzymes - metabolism; Escherichia coli - enzymology; Escherichia coli - genetics; Evolution; Flow cytometry; Fluorescence; Gene Library; Genetic mutation; HeLa Cells; Humans; Kinetics; Libraries; Models, Molecular; Molecular Sequence Data; Molecules; Mutant Proteins - chemistry; Mutant Proteins - genetics; Mutant Proteins - metabolism; Peptide Library; Protein Engineering; proteins; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; screening; sortase; sortase A; Staphylococcus aureus; Staphylococcus aureus - enzymology; Staphylococcus aureus - genetics; Substrate Specificity; Yeast; Yeasts

The ability to routinely generate efficient protein catalysts of bond-forming reactions chosen by researchers, rather than nature, is a long-standing goal of the molecular life sciences. Here, we describe a directed evolution strategy for enzymes that catalyze, in principle, any bond-forming reactio...