Metabolomics and Genomics Approach for the Discovery of Serrawettin W2 Lipopeptides from NP2.

A metabolomics/peptidomics and genomics approach, using UPLC-MS, molecular networking, and genome mining, was used to describe the serrawettin W2 lipopeptide family produced by NP2. Seven known serrawettin W2 analogues were structurally elucidated along with 17 new analogues, which varied based on the first (fatty acyl length of C, C, C, or C), fifth (Phe, Tyr, Trp, or Leu/Ile), and sixth (Leu, Ile, or Val) residues. Tandem MS results suggested that the previously classified serrawettin W3 may be an analogue of serrawettin W2, with a putative structure of cyclo(CHO-Leu-Ser-Thr-Leu/Ile-Val). Chiral phase amino acid analysis enabled the distinction between l/d-Leu and l-Ile residues within nine purified compounds. H and C NMR analyses confirmed the structures of four purified new analogues. Additionally, genome mining was conducted using genome sequences available on the NCBI database to identify the gene using the antiSMASH software. NRPSpredictor2 predicted the specificity score of the adenylation-domain within with 100% for the first, second, and third modules (Leu-Ser-Thr), 60-70% for the fourth module (Phe/Trp/Tyr/Val), and 70% for the fifth module (Val/Leu/Ile), confirming MS data. Finally, antibacterial activity was observed for compounds and against a clinical strain.