---
title: "Biosynthesis and diversity of leaderless multipeptide bacteriocins: new genomic and functional ins"
id: "biorxiv-0-novel-insights-into-the-biosynthesis-and-diversity-of-leaderless-multipeptide"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-0-novel-insights-into-the-biosynthesis-and-diversity-of-leaderless-multipeptide"
content_type: "clinical_feed_article"
specialty: "Infectious Disease"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1?rss=1"
published_at: "2026-09-07T12:00:00.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Biosynthesis and diversity of leaderless multipeptide bacteriocins: new genomic and functional ins
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-0-novel-insights-into-the-biosynthesis-and-diversity-of-leaderless-multipeptide
- **Specialty:** [Infectious Disease](https://medichelpline.com/clinical-feed/infectious-disease.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1?rss=1)
- **Published At:** 2026-09-07T12:00:00.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- The study focuses on **Garvicin KS (GarKS)**, a three-peptide, **leaderless** broad-spectrum **multipeptide bacteriocin** active against many Gram-positive bacteria, including foodborne pathogens and antibiotic-resistant strains. - A hybrid sequencing approach produced a high-quality closed genome of the native producer strain originally labelled Lactococcus garvieae KS1546; the bacteriocin genes were located on a ~50 kb plasmid designated **pKS50**. - Comparative sequence analysis indicated the producer strain should be reclassified as **Lactococcus petauri**. - Two biosynthetic genes were functionally characterised by heterologous expression: a putative transcriptional regulator (**gakR**) and a putative immunity protein (**gakI**). - Deletion/removal of **gakR** reduced GarKS production approximately sixfold, indicating a positive role in biosynthesis regulation. - Expression of **gakI** in recipient cells lowered susceptibility to GarKS by about 250-fold and produced varying levels of cross-immunity to other multipeptide bacteriocins. - Spontaneous GarKS-resistant mutants were isolated in Lactococcus lactis and whole-genome sequenced; resistance levels were low overall. - Mutation analysis implicated **ythA**, a PspC-domain-containing protein linked to a phage stress response pathway, in the GarKS-susceptible phenotype. - Genome mining across databases identified 11 candidate loci for new leaderless multipeptide bacteriocins; four candidate peptides were synthesised and confirmed bioactive, inhibiting pathogens including **Listeria monocytogenes** and **enterococci**. - The work provides genomic, regulatory and functional data relevant to potential applications in food preservation and medical uses, while noting that the report is a preprint not yet peer reviewed.
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Telke, Tage Thorstensen, [ View ORCID Profile](http://orcid.org/0000-0002-4767-8666)Kirill V. Ovchinnikov, Dzung B. Diep, [ View ORCID Profile](http://orcid.org/0000-0003-4448-9082)Morten Kjos doi: https://doi.org/10.64898/2026.09.04.749385 This article is a preprint and has not been certified by peer review [[what does this mean?](https://www.biorxiv.org/about/FAQ#unrefereed)]. Thomas Førland Oftedal 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Thomas%2BF%C3%B8rland%2BOftedal%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Oftedal%20TF&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AThomas%2BF%25C3%25B8rland%2BOftedal%2B) * [ORCID record for Thomas Førland Oftedal](http://orcid.org/0000-0003-4182-3831 "Open in new tab") * For correspondence: thomas.forland.oftedal@nmbu.no Simen Hermansen 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Simen%2BHermansen%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Hermansen%20S&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ASimen%2BHermansen%2B) * [ORCID record for Simen Hermansen](http://orcid.org/0009-0006-2542-3693 "Open in new tab") Even Amandus Haslund 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Even%2BAmandus%2BHaslund%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Haslund%20EA&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AEven%2BAmandus%2BHaslund%2B) Amar A. Telke 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Amar%2BA.%2BTelke%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Telke%20AA&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AAmar%2BA.%2BTelke%2B) Tage Thorstensen 2 Agribiotix * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Tage%2BThorstensen%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Thorstensen%20T&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ATage%2BThorstensen%2B) Kirill V. Ovchinnikov 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Kirill%2BV.%2BOvchinnikov%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Ovchinnikov%20KV&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AKirill%2BV.%2BOvchinnikov%2B) * [ORCID record for Kirill V. Ovchinnikov](http://orcid.org/0000-0002-4767-8666 "Open in new tab") Dzung B. Diep 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Dzung%2BB.%2BDiep%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Diep%20DB&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ADzung%2BB.%2BDiep%2B) Morten Kjos 1 Norwegian University of Life Sciences; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Morten%2BKjos%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Kjos%20M&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AMorten%2BKjos%2B) * [ORCID record for Morten Kjos](http://orcid.org/0000-0003-4448-9082 "Open in new tab") * [Abstract](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_art/node:5762917/1) * [Info/History](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.article-info)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_info/node:5762917/1) * [Metrics](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.article-metrics)[](https://www.biorxiv.org/panels_ajax_tab/article_tab_metrics/node:5762917/1) * [Supplementary material](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.supplementary-material)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_data/node:5762917/1) * [ Preview PDF](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.full.pdf+html)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_pdf/node:5762917/1) ![Loading](https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif) ## Abstract Garvicin KS (GarKS) is a three-peptide, leaderless, broad-spectrum bacteriocin that is active against a wide range of Gram-positive bacteria, including several foodborne pathogens and antibiotic-resistant strains. This bacteriocin is considered a candidate for application in food preservation and medical treatments; however, key knowledge about the producer strain, regulation of production, and the mechanism of action is still lacking for leaderless, multipeptide bacteriocins, including GarKS. A hybrid sequencing strategy was used to obtain a high-quality closed genome assembly of the native GarKS producer _Lactococcus garvieae_ KS1546, which showed that the bacteriocin was encoded on a 50 kb plasmid (pKS50). Comparative analysis with updated sequence databases indicates that the producer strain should be reclassified as _Lactococcus petauri_. The roles of the biosynthetic genes, putatively encoding a transcriptional regulator (_gakR_) and immunity protein (_gakI_), were examined using heterologous expression. Removal of _gakR_ resulted in a 6-fold decrease in GarKS production, and expression of _gakI_ caused a 250-fold decrease in susceptibility to GarKS, with varying degrees of cross-immunity to other multipeptide bacteriocins. Isolation and whole-genome sequencing of spontaneous GarKS mutants in _L. lactis_ showed that resistance levels are low, but that _ythA_ , a PspC-domain-containing protein involved in a phage stress response pathway is involved in the GarKS susceptible phenotype. To decipher conserved features involved in production of these bacteriocins, we finally used genome mining to identify 11 candidates for new multipeptide bacteriocins; four of them were obtained synthetically and confirmed to be bioactive peptides inhibiting important pathogens including _Listeria monocytogenes_ and enterococci. ### Competing Interest Statement The authors have declared no competing interest. ## Funder Information Declared The Research Council of Norway, 341639 Norwegian University of Life Sciences, https://ror.org/04a1mvv97 Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a [CC-BY-NC-ND 4.0 International license](http://creativecommons.org/licenses/by-nc-nd/4.0/). bioRxiv and medRxiv thank the following for their generous financial support: > The Chan Zuckerberg Initiative, Cold Spring Harbor Laboratory, the Sergey Brin Family Foundation, California Institute of Technology, Centre National de la Recherche Scientifique, Fred Hutchinson Cancer Center, Imperial College London, Massachusetts Institute of Technology, Stanford University, The University of Edinburgh, University of Washington, and Vrije Universiteit Amsterdam. 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[ Download PDF](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.full.pdf) Print/Save Options [Download PDF](https://www.biorxiv.org/content/biorxiv/early/2026/09/07/2026.09.04.749385.full.pdf)Full Text & In-line FiguresXML [More Info](https://www.biorxiv.org/about/FAQ#PrintOptions "More Information on Print/Save Options") [Supplementary Material ](https://www.biorxiv.org/content/10.64898/2026.09.04.749385v1.supplementary-material) [ Email](https://www.biorxiv.org/ "Email this Article") [ Share](https://www.biorxiv.org/) Novel insights into the biosynthesis and diversity of leaderless multipeptide bacteriocins Thomas Førland Oftedal, Simen Hermansen, Even Amandus Haslund, Amar A. Telke, Tage Thorstensen, Kirill V. Ovchinnikov, Dzung B. Diep, Morten Kjos bioRxiv 2026.09.04.749385; doi: https://doi.org/10.64898/2026.09.04.749385 This article is a preprint and has not been certified by peer review [[what does this mean?](https://www.biorxiv.org/about/FAQ#unrefereed)]. 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