PT - JOURNAL ARTICLE AU - Efthimia Pashou AU - Sebastian J. Reich AU - Dominik Weixler AU - Bernhard J. Eikmanns AU - Christian U. Riedel AU - Oliver Goldbeck TI - Identification, Characterization and Mode of Action of Corynaridin, a Novel Linaridin from <em>Corynebacterium lactis</em> AID - 10.1101/2022.05.11.491181 DP - 2022 Jan 01 TA - bioRxiv PG - 2022.05.11.491181 4099 - http://biorxiv.org/content/early/2022/05/11/2022.05.11.491181.short 4100 - http://biorxiv.org/content/early/2022/05/11/2022.05.11.491181.full AB - Genome analysis of Corynebacterium lactis revealed a bacteriocin gene cluster encoding a putative bacteriocin of the linaridin family of ribosomally synthesized and posttranslationally modified peptides (RIPPs). The locus harbors typical linaridin modification enzymes but lacks genes for a decarboxylase and methyltransferase, which is unusual for type B linaridins. Supernatants of Corynebacterium lactis RW3-42 showed antimicrobial activity against Corynebacterium glutamicum. Deletion of the precursor gene crdA clearly linked the antimicrobial activity of the producer strain to the identified gene cluster. Following purification, we observed potent activity of the peptide against Actinobacteria, mainly other members of the genus Corynebacterium including the pathogenic species Corynebacterium striatum and Corynebacterium amycolatum. Also, low activity against some Firmicutes was observed, but no activity against Gram-negative species. The peptide is resilient towards heat but sensitive to proteolytic degradation by trypsin and proteinase K. Notably, time-kill kinetics and experiments using live biosensors to monitor membrane integrity suggest bactericidal activity that does not involve formation of pores in the cytoplasmic membrane. As Corynebacterium species are ubiquitous in nature and include important commensal and pathogens of mammalian organisms, secretion of bacteriocins by species of this genus could be a hitherto neglected trait with high relevance for intra- and interspecies competition and infection.Importance Bacteriocins are antimicrobial peptides produced by bacteria to fend off competitors in ecological niches and are considered to be important factors influencing the composition of microbial communities. However, bacteriocin production by bacteria of the genus Corynebacterium has been a hitherto neglected trait, although its species are ubiquitous in nature and make up large parts of the microbiome of humans and animals. In this study, we describe and characterize a novel linaridin-family bacteriocin from Corynebacterium lactis and show its narrow spectrum activity, mainly against other actinobacteria. Moreover, we were able to extend the limited knowledge on linaridin bioactivity in general and for the first time describe the mode of action of such a bacteriocin. Interestingly, the peptide, which was named corynaridin, appears bactericidal, but without formation of pores in the bacterial membrane.Competing Interest StatementThe authors have declared no competing interest.