biostudies-literatureQi SNational Natural Science Foundation of China174https://www.ebi.ac.uk/biostudies/studies/S-EPMC8877149biostudies-literatureUnknown8(2)Defensins are a class of cationic disulfide-bridged antimicrobial peptides (AMPs) present in many eukaryotic organisms and even in bacteria. They primarily include two distinct but evolutionarily related superfamilies (<i>cis</i> and <i>trans</i>). Defensins in fungi belong to the members of the <i>cis</i>-superfamily with the cysteine-stabilized α-helical and β-sheet fold. To date, many fungal defensin-like peptides (fDLPs) have been found through gene mining of the genome resource, but only a few have been experimentally characterized. Here, we report the structural and functional characterization of Pyronesin4 (abbreviated as Py4), a fDLP previously identified by genomic sequencing of the basal filamentous ascomycete <i>Pyronema confluens</i>. Chemically, synthetic Py4 adopts a native-lJournal of fungi (Basel, Switzerland)A Fungal Defensin Inhibiting Bacterial Cell-Wall Biosynthesis with Non-Hemolysis and Serum Stability.PMC887714931870766Gao BQi SZhu SfalseA Fungal Defensin Inhibiting Bacterial Cell-Wall Biosynthesis with Non-Hemolysis and Serum Stability.Defensins are a class of cationic disulfide-bridged antimicrobial peptides (AMPs) present in many eukaryotic organisms and even in bacteria. They primarily include two distinct but evolutionarily related superfamilies (<i>cis</i> and <i>trans</i>). Defensins in fungi belong to the members of the <i>cis</i>-superfamily with the cysteine-stabilized α-helical and β-sheet fold. To date, many fungal defensin-like peptides (fDLPs) have been found through gene mining of the genome resource, but only a few have been experimentally characterized. Here, we report the structural and functional characterization of Pyronesin4 (abbreviated as Py4), a fDLP previously identified by genomic sequencing of the basal filamentous ascomycete <i>Pyronema confluens</i>. Chemically, synthetic Py4 adopts a native-l2022-01-01T00:00:00Z2022 Feb2026-04-08T17:21:32.531Z2025-02-19T05:04:49.243ZS-EPMC88771493520592810.3390/jof8020174