Identification of a Novel Two-Peptide Lantibiotic from Vagococcus fluvialis

Rosenbergová, Zuzana; Oftedal, Thomas F.; Ovchinnikov, Kirill V.; Thiyagarajah, Thasanth; Rebroš, Martin; Diep, Dzung B.

doi:10.1128/spectrum.00954-22

“…Two-peptide lantibiotics are believed to use the dual mode of action only, where the α-peptide forms a complex with lipid II which recruits the β-peptide to form a pore ( 23 ). The propidium iodide pore formation assay has been used previously to examine the mode of action of bacteriocins, including two-peptide lantibiotics ( 55 , 56 ). The mode of action of the bacteriocin vagococcin T, with sequence homology to romsacin ( Fig.…”

Section: Discussionmentioning

confidence: 99%

The novel bacteriocin romsacin from Staphylococcus haemolyticus inhibits Gram-positive WHO priority pathogens

Wolden,

Ovchinnikov,

Venter

et al. 2023

Microbiol Spectr

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Staphylococcus haemolyticus is an increasingly relevant nosocomial pathogen. The combination of multi-drug resistance and ability to form biofilms makes S. haemolyticus infections difficult to treat. Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria to inhibit growth of often closely related bacteria. Due to differences in the modes of action between bacteriocins and antibiotics, bacteriocins are normally equally potent against antibiotic-resistant and antibiotic-sensitive strains. To find bacteriocins able to inhibit S. haemolyticus and related species, clinical and commensal S. haemolyticus isolates ( n = 174) were assayed for bacteriocin production. One commensal isolate produced an antimicrobial substance inhibiting S. haemolyticus and Staphylococcus aureus . The substance had physicochemical properties that are characteristic of bacteriocins. Purification, whole-genome sequencing, and mass spectrometry identified the antimicrobial as a novel two-peptide lantibiotic, hereafter named romsacin. The bacteriocin was active against a broad range of Gram-positive bacteria, such as the World Health Organization priority pathogens S. aureus [methicillin-resistant S. aureus (MRSA)] and Enterococcus faecium [vancomycin-resistant E. faecium (VRE)]. Importantly, the bacteriocin also eradicated S. haemolyticus , Staphylococcus epidermidis , MRSA, and VRE biofilms. IMPORTANCE Bacteria produce bacteriocins to inhibit growth of other bacterial species. We have studied the antimicrobial activity of a new bacteriocin produced by the skin bacterium S. haemolyticus . The bacteriocin is effective against several types of Gram-positive bacteria, including highly virulent and antibiotic-resistant strains such as Staphylococcus aureus and Enterococcus faecium . Effective antimicrobials are important for the treatment of infections and the success of major surgery and chemotherapy. Bacteriocins can be part of the solution to the global concern of antimicrobial resistance.

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“…On the other hand, V. fluvialis L21 isolated from the gut of gilthead sea bream [16] acted as a fish probiotic, effectively preventing infection with Vibrio anguillarum [17]. Moreover, ribosomally synthesized and post-translationally modified peptides of the lantibiotic class have been purified from V. fluvialis LMGT 4216 isolated from fermented maize [18]. A sewage isolate phylogenetically related to V. fluvialis was found to produce polysaccharides with flocculation properties [19], which could be of interest for water treatment.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, ribosomally synthesized and post-translationally modified peptides of the lantibiotic class have been purified from V. fluvialis LMGT 4216 isolated from fermented maize [18]. A sewage isolate phylogenetically related to V.…”

Section: Introductionmentioning

confidence: 99%

Vagococcus luciliae sp. nov., isolated from the common green bottle fly Lucilia sericata

Guzman

¹

,

Daniel

²

,

Kämpfer

³

et al. 2023

International Journal of Systematic and Evolutionary Microbiology

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The genus Vagococcus belongs to the family Enterococcaceae (order Lactobacillales) and is closely related to the genus Enterococcus . Currently, 19 species of the genus have been validly named. In this study, we isolated strain G314FT from the common green bottle fly Lucilia sericata collected in Germany. Sequencing of its almost-complete 16S rRNA gene revealed that the isolate belongs to the genus Vagococcus , being closely related to Vagococcus bubulae SS1994T with high sequence identity (99.50 %), followed by Vagococcus martis D7T301T (98.86 %), Vagococcus vulneris SS1995T (98.71 %), Vagococcus teuberi DSM 21459T (98.64 %), Vagococcus silagei 2B-2T (98.64 %) and Vagococcus penaei CD276T (98.64 %). Genome sequencing of strain G314FT was performed by a combination of Illumina and Oxford Nanopore technology, yielding a circular genome with a size of 2 139 468 bp and an 11 kb plasmid. Average nucleotide identity and digital DNA–DNA hybridization values were calculated between G314FT and its closest-related taxa, and found to be <91 % and <40 %, respectively, thus strongly supporting that strain G314FT represents a novel species of the genus Vagococcus . Phylogenetic and core protein-based phylogenomic trees revealed that G314FT was closely related to a group of three species, V. bubulae SS1994T, V. martis D7T301T and V. teuberi DSM 21459T. Comparatively, the genome of G314FT is the smallest in the group of the four related species, and the biochemical pathway comparison using BlastKOALA revealed that G314FT has lost some amino acid biosynthetic proteins; however, it has gained enzymes for carbohydrate metabolism. Phenotypically, G314FT was consistent with other species of the genus Vagococcus including a negative catalase reaction and non-motility. Using the polyphasic approach, our data supports that the isolate represents a new species, for which we propose the name Vagococcus luciliae G314FT (=DSM 112651T= CCM 9164T).

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“…However, the stereochemistry of the Lan and MeLan residues of the great majority of newly reported lanthipeptides is not determined. Therefore, as more and more lanthipeptides are discovered by genome mining, − a convenient method for determining their stereochemistry that is accessible to most laboratories would be valuable. We report here such a method including access to standards that use biochemical approaches that are complementary to chemical synthesis and methods available in most if not all laboratories studying lanthipeptides or RiPPs.…”

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confidence: 99%

Facile Method for Determining Lanthipeptide Stereochemistry

Luo,

Xu,

Frerk

et al. 2024

Anal. Chem.

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Lanthipeptides make up a large group of natural products that belong to the ribosomally synthesized and post-translationally modified peptides (RiPPs). Lanthipeptides contain lanthionine and methyllanthionine bis-amino acids that have varying stereochemistry. The stereochemistry of new lanthipeptides is often not determined because current methods require equipment that is not standard in most laboratories. In this study, we developed a facile, efficient, and user-friendly method for detecting lanthipeptide stereochemistry, utilizing advanced Marfey’s analysis with detection by liquid chromatography coupled with mass spectrometry (LC-MS). Under optimized conditions, 0.05 mg of peptide is sufficient to characterize the stereochemistry of five (methyl)lanthionines of different stereochemistry using a simple liquid chromatography setup, which is a much lower detection limit than current methods. In addition, we describe methods to readily access standards of the three different methyllanthionine stereoisomers and two different lanthionine stereoisomers that have been reported in known lanthipeptides. The developed workflow uses a commonly used nonchiral column system and offers a scalable platform to assist antimicrobial discovery. We illustrate its utility with an example of a lanthipeptide discovered by genome mining.

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Identification of a Novel Two-Peptide Lantibiotic from Vagococcus fluvialis

Cited by 10 publications

References 81 publications

The novel bacteriocin romsacin from Staphylococcus haemolyticus inhibits Gram-positive WHO priority pathogens

The novel bacteriocin romsacin from Staphylococcus haemolyticus inhibits Gram-positive WHO priority pathogens

Vagococcus luciliae sp. nov., isolated from the common green bottle fly Lucilia sericata

Facile Method for Determining Lanthipeptide Stereochemistry

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