Genome-assisted Identification, Purification, and Characterization of Bacteriocins

Ovchinnikov, Kirill V.; Oftedal, Thomas F.; Reich, Sebastian J.; Bar, Nadav; Holo, Helge; Skaugen, Morten; Riedel, Christian U.; Diep, Dzung B.

doi:10.21769/bioprotoc.4477

Cited by 4 publications

(2 citation statements)

References 28 publications

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“…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%

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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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Section: Discussionmentioning

confidence: 99%

“…Bacteriocin purification was performed similarly as described by Ovchinnikov et al ( 56 ), with some modifications. One liter of BHI was inoculated with 2% (vol/vol) of an overnight culture of S. haemolyticus 57-27.…”

Section: Methodsmentioning

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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A bacteriocin-based coating strategy to prevent vancomycin-resistant Enterococcus faecium biofilm formation on materials of interest for indwelling medical devices

Kranjec,

Mathew,

Ovchinnikov

et al. 2024

Biofilm

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Antimicrobial peptide production with Corynebacterium glutamicum on lignocellulosic side streams

Waldschitz,

Neudert,

Kitzmueller

et al. 2024

Preprint

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Background:Biorefineries usually focus on the production of low-value commodities, such as bioethanol, platform chemicals or single cell protein.Shifting production to bioactive compounds, such as antimicrobial peptides, could provide an opportunity to increase the economic viability of biorefineries. Results: Recombinant production of the antimicrobial peptide pediocin PA-1 in Corynebacterium glutamicum was transferred from yeast extract based media to minimal media based on lignocellulosic spent sulfite liquor.Induced batch, fed batch, and extended batch process modes were compared for highest pediocin PA-1 production. Conclusion: For pediocin PA-1 production on lignocellulosic residues, extended batch cultivation was identified as the optimal process mode, producing up to $\simeq$ 104 mg/L active pediocin PA-1.Moreover, the production of pediocin PA-1 on this sustainable second generation resource exceeded its state-of-the-art production on yeast extract based media $\simeq$ 1.5-fold.

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Genome-assisted Identification, Purification, and Characterization of Bacteriocins

Cited by 4 publications

References 28 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

A bacteriocin-based coating strategy to prevent vancomycin-resistant Enterococcus faecium biofilm formation on materials of interest for indwelling medical devices

Antimicrobial peptide production with Corynebacterium glutamicum on lignocellulosic side streams

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