<jats:title>Abstract</jats:title><jats:p>Backbone N-methylation and macrocyclization improve the pharmacological properties of peptides by enhancing their proteolytic stability, membrane permeability and target selectivity. Borosins are backbone N-methylated peptide macrocycles derived from a precursor protein which contains a peptide α-N-methyltransferase domain autocatalytically modifying the core peptide located at its C-terminus. Founding members of borosins are the omphalotins from the mushroom <jats:italic>Omphalotus olearius</jats:italic> (omphalotins A-I) with nine out of 12 <jats:italic>L</jats:italic>-amino acids being backbone N-methylated. The omphalotin biosynthetic gene cluster codes for the precursor protein OphMA, the protease prolyloligopeptidase OphP and other proteins that are likely to be involved in other post-translational modifications of the peptide. Mining of available fungal genome sequences revealed the existence of highly homologous gene clusters in the basidiomycetes <jats:italic>Lentinula edodes</jats:italic> and <jats:italic>Dendrothele bispora</jats:italic>. The respective borosins, referred to as lentinulins and dendrothelins are naturally produced by <jats:italic>L. edodes</jats:italic> and <jats:italic>D. bispora</jats:italic> as shown by analysis of respective mycelial extracts. We produced all three homologous peptide natural products by coexpression of OphMA hybrid proteins and OphP in the yeast <jats:italic>Pichia pastoris</jats:italic>. The recombinant peptides differ in their nematotoxic activity against the plant pathogen <jats:italic>Meloidogyne incognita</jats:italic>. Our findings pave the way for the production of borosin peptide natural products and their potential application as novel biopharmaceuticals and biopesticides.