Biochemical characterisation of non-ribosomal peptide synthetases from glycopeptide antibiotic biosynthesis

Abstract

In this study, the entire glycopeptide antibiotic teicoplanin biosynthesis non-ribosomal peptide synthetase (NRPS) assembly line was analysed. Our findings provided deeper mechanistic understanding how peptide biosynthesis assembly lines function. Here, for the first time using A-domain amino acid activation assays and assembly line reassembly, we have reconstituted the activity of all NRPS modules from the biosynthesis of the antibiotic teicoplanin and demonstrated the formation of the teicoplanin heptapeptide precursor biosynthesis in vitro. In addition, several novel biosynthetic pathway reengineering strategies, were designed and experimentally demonstrated, which allow the biosynthesis of peptides with altered sequences.