Discovery of a Cryptic Depsipeptide from Streptomyces ghanaensis via MALDI‐MS‐Guided High‐Throughput Elicitor Screening

Abstract Microbial genomes harbor an abundance of biosynthetic gene clusters, but most are expressed at low levels and need to be activated for characterization of their cognate natural products. In this work, we report the combination of high‐throughput elicitor screening (HiTES) with matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) for the rapid identification of cryptic peptide natural products. Application to Streptomyces ghanaensis identified amygdalin as an elicitor of a novel non‐ribosomal peptide, which we term cinnapeptin. Complete structural elucidation revealed cinnapeptin as a cyclic depsipeptide with an unusual 2‐methyl‐cinnamoyl group. Insights into its biosynthesis were provided by whole genome sequencing and gene deletion studies, while bioactivity assays showed antimicrobial activity against Gram‐positive bacteria and fission yeast. MALDI‐HiTES is a broadly applicable tool for the discovery of cryptic peptides encoded in microbial genomes.