Biosynthetic diversification of non-ribosomal peptides through activity-based protein profiling of adenylation domains

We describe activity-based protein profiling for analyzing the adenylation domains of non-ribosomal peptide synthetases (ABPP-NRPS) in bacterial proteomes. Using a range of non-proteoinogenic amino acid sulfamoyladenosines, the competitive format of ABPP-NRPS provided substrate tolerance toward non-proteinogenic amino acids. When coupled with precursor-directed biosynthesis, a non-proteinogenic amino acid (O-allyl-l-serine) was successfully incorporated into gramicidin S.

Automated summary

We present a method called activity-based protein profiling for studying the adenylation domains of non-ribosomal peptide synthetases (ABPP-NRPS) in bacterial proteomes. This method allows substrate tolerance towards non-proteinogenic amino acids by using a range of non-proteinogenic amino acid sulfamoyladenosines in a competitive format. Additionally, we demonstrate successful incorporation of a non-proteinogenic amino acid (O-allyl-L-serine) into gramicidin S using precursor-directed biosynthesis.