Heterologous expression of a cryptic gene cluster from Streptomyces leeuwenhoekii C34T yields a novel lassopeptide, leepeptin

Analysis of the genome sequence of Streptomyces leeuwenhoekii C34T identified biosynthetic gene clusters (BGCs) for three different lasso peptides (Lp1, Lp2 and Lp3) which were not known to be made by the strain.  Lasso peptides are a relatively new member of the RiPP (ribosomally synthesized and post-translationally modified peptides) family of natural products and have not been extensively studied.  Lp3, whose production could be detected in culture supernatants from S. leeuwenhoekii C34T and after heterologous expression of its BGC in Streptomyces coelicolor, is identical to the previously characterised chaxapeptin.  Lp1, whose production could not be detected or achieved heterologously, appears to be identical to a recently identified member of the citrulassin family of lasso peptides.  Since production of Lp2 by S. leeuwenhoekii C34T was not observed, its BGC was also expressed in S. coelicolor.  The lasso peptide was isolated and its structure confirmed by mass spectrometry and nuclear magnetic resonance analyses revealing a novel structure that appears to represent a new family of lasso peptides.&nbsp