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A Plasmid-Encoded Surface Polysaccharide Partly Blocks Ceduovirus Infection in Lactococci

delete2025-03-11
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OA
AI
C
Claudia Rendueles
J
Javier Nicolás Garay‐Novillo
M
Martin Holm Rau
P
Paula Gaspar
J
José Á. Ruiz-Masó
J
Jennifer Mahony
A
Ana Rodrı́guez
J
José Luis Barra
G
Gloria del Solar
DOI:10.3390/ijms26062508delete
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Abstract

Abstract

En 中文
Bacteriophages (or phages) remain the leading cause of failure in dairy fermentations. Thereby, phage-resistant Lactococcus lactis and Lactococcus cremoris dairy starters are in continuous demand. In this work, our goal was to identify phage defense mechanisms against ceduoviruses encoded by two wild isolates of dairy origin named L. lactis IPLA517 and IPLA1064. These strains were previously subjected to experimental evolution to select derivatives that are resistant to the bacteriocin Lcn972. It was observed that the Lcn972R derivatives became sensitive to phage infection; however, the underlying mechanism was not defined. The long-read sequencing technologies applied in this work reveal that all of the Lcn972R derivatives shared the loss of a 41 kb endogenous plasmid (p41) that harbors a putative exopolysaccharide (EPS) gene cluster with significant homology to one described in Lactococcus garvieae. Using a CRISPR-Cas9-based approach, p41 was selectively cured from L. lactis IPLA1064. Phage infection assays with three ceduoviruses demonstrated that curing p41 restored phage sensitivity at levels comparable to the Lcn972R-IPLA1064 derivatives. Phage adsorption to Delta p41 cells was also increased, consistent with the hypothesis of EPS production hindering access to the phage receptor protein Pip. Our results reinforce the role of EPSs in protecting Lactococcus against phage infection, a phenomenon that is rarely reported for ceduoviruses. Moreover, the results also exemplify the likely horizontal gene transfer that can occur between L. lactis and L. garvieae in a dairy environment.
Keywords:
CRISPR-Cas9
dairy starters
endogenous plasmids
nanopore sequencing
phage adsorption

Journal

International Journal of Molecular Sciences cover
International Journal of Molecular Sciences
IF:
4.9
Papers:
1.9W
Citations:
44.5W

Organization

No organization information available