Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis
ABSTRACT 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...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-02-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01756-22 |
| _version_ | 1811164373611184128 |
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| author | Efthimia Pashou Sebastian J. Reich Alexander Reiter Dominik Weixler Bernhard J. Eikmanns Marco Oldiges Christian U. Riedel Oliver Goldbeck |
| author_facet | Efthimia Pashou Sebastian J. Reich Alexander Reiter Dominik Weixler Bernhard J. Eikmanns Marco Oldiges Christian U. Riedel Oliver Goldbeck |
| author_sort | Efthimia Pashou |
| collection | DOAJ |
| description | ABSTRACT 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 there was no activity against Gram-negative species. The peptide is resilient towards heat but sensitive to proteolytic degradation by trypsin and proteinase K. Analysis by mass spectrometry indicates that corynaridin is processed by cleaving off the leader sequence at a conserved motif and posttranslationally modified by dehydration of all threonine and serin residues, resulting in a monoisotopic mass of 3,961.19 Da. 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 commensals 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 bactericidal activity of such a bacteriocin. Interestingly, the peptide, which was named corynaridin, appears bactericidal, but without formation of pores in the bacterial membrane. |
| first_indexed | 2024-04-10T15:21:32Z |
| format | Article |
| id | doaj.art-6266ea92a4484e6395b0ad1c5cfdf626 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-10T15:21:32Z |
| publishDate | 2023-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-6266ea92a4484e6395b0ad1c5cfdf6262023-02-14T14:15:49ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-02-0111110.1128/spectrum.01756-22Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactisEfthimia Pashou0Sebastian J. Reich1Alexander Reiter2Dominik Weixler3Bernhard J. Eikmanns4Marco Oldiges5Christian U. Riedel6Oliver Goldbeck7Institute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyInstitute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyInstitute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Jülich, GermanyInstitute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyInstitute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyInstitute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Jülich, GermanyInstitute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyInstitute of Microbiology and Biotechnology, University of Ulm, Ulm, GermanyABSTRACT 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 there was no activity against Gram-negative species. The peptide is resilient towards heat but sensitive to proteolytic degradation by trypsin and proteinase K. Analysis by mass spectrometry indicates that corynaridin is processed by cleaving off the leader sequence at a conserved motif and posttranslationally modified by dehydration of all threonine and serin residues, resulting in a monoisotopic mass of 3,961.19 Da. 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 commensals 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 bactericidal activity of such a bacteriocin. Interestingly, the peptide, which was named corynaridin, appears bactericidal, but without formation of pores in the bacterial membrane.https://journals.asm.org/doi/10.1128/spectrum.01756-22Corynebacteriumlinaridinantimicrobial peptidesbacteriocins |
| spellingShingle | Efthimia Pashou Sebastian J. Reich Alexander Reiter Dominik Weixler Bernhard J. Eikmanns Marco Oldiges Christian U. Riedel Oliver Goldbeck Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis Microbiology Spectrum Corynebacterium linaridin antimicrobial peptides bacteriocins |
| title | Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis |
| title_full | Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis |
| title_fullStr | Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis |
| title_full_unstemmed | Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis |
| title_short | Identification and Characterization of Corynaridin, a Novel Linaridin from Corynebacterium lactis |
| title_sort | identification and characterization of corynaridin a novel linaridin from corynebacterium lactis |
| topic | Corynebacterium linaridin antimicrobial peptides bacteriocins |
| url | https://journals.asm.org/doi/10.1128/spectrum.01756-22 |
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