Discovery and Biosynthesis of Nyuzenamides D and E by Genome Mining in Streptomyces hygroscopicus

Nyuzenamides belong to a class of bioactive cinnamoyl moieties containing nonribosomal peptides. Genome mining revealed a putative nyu BGC in Streptomyces hygroscopicus DSM 40578. Two new analogues, nyuzenamides D (1) and E (2), were subsequently isolated, and their structures were elucidated by detailed nuclear magnetic resonance spectroscopic and mass spectrometric data analyses. The absolute configuration of 1 was determined by a single-crystal X-ray diffraction study. Through retrobiosynthesis and CRISPR genome editing, the nonribosomal peptide synthetase biosynthetic gene cluster for nyuzenamides was confirmed. Nyuzenamides D and E are likely derived from nyuzenamide C through nonenzymatic hydrolysis. Our discovery enriches the diversity of cinnamoyl-containing nonribosomal peptides and validates the biosynthetic gene clusters for future genome mining research.