Chapter 10 Using Phosphopantetheinyl Transferases for Enzyme Posttranslational Activation, Site Specific Protein Labeling and Identification of Natural Product Biosynthetic Gene Clusters from Bacterial Genomes

摘要: Abstract Phosphopantetheinyl transferases (PPTases) covalently attach the phosphopantetheinyl group derived from coenzyme A (CoA) to acyl carrier proteins or peptidyl as part of enzymatic assembly lines fatty acid synthases (FAS), polyketide (PKS), and nonribosomal peptide synthetases (NRPS). PPTases have demonstrated broad substrate specificities for cross‐species modification embedded in PKS NRPS modules. PPTase Sfp Bacillus subtilis AcpS Escherichia coli also transfer small molecules diverse structures their CoA conjugates proteins. Short tags thus been developed efficient substrates site‐specific labeling peptide‐tagged fusion with biotin organic fluorophores. This chapter discusses use vivo vitro enzymes protein labeling. We describe a phage selection method based on PPTase‐catalyzed identification genes bacterial genomes.