Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides

Abstract Exopolysaccharides (EPS) play critical roles in microbial survival, stress adaptation, and biofilm formation across diverse environments. In food-associated bacteria such as Leuconostoc mesenteroides, understanding the regulation of EPS production under environmental stress is important for...

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Main Authors: Miguel Fernandez de Ullivarri, Colin Buttimer, Janneke Wijman, Eelco Heintz, Paul Ross, Matthew P. McCusker, Colin Hill
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-04141-x
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author Miguel Fernandez de Ullivarri
Colin Buttimer
Janneke Wijman
Eelco Heintz
Paul Ross
Matthew P. McCusker
Colin Hill
author_facet Miguel Fernandez de Ullivarri
Colin Buttimer
Janneke Wijman
Eelco Heintz
Paul Ross
Matthew P. McCusker
Colin Hill
author_sort Miguel Fernandez de Ullivarri
collection DOAJ
description Abstract Exopolysaccharides (EPS) play critical roles in microbial survival, stress adaptation, and biofilm formation across diverse environments. In food-associated bacteria such as Leuconostoc mesenteroides, understanding the regulation of EPS production under environmental stress is important for both spoilage control and industrial applications. However, the mechanisms linking cold stress to EPS biosynthesis remain poorly understood. Here, we show that sucrose and low temperature (8 °C) trigger a metabolic shift from dextran-only to combined dextran and levan biosynthesis in four meat-borne Leuc. mesenteroides strains. Two high EPS-producing strains (HEPRs) possess the sacB_1 gene, which encodes a previously uncharacterized levansucrase absent from low EPS-producing strains (LEPRs) that only carry the levS gene. This is the first study to describe the role of sacB_1 in cold-induced EPS production. Notably, sacB_1 was also identified in Leuc. mesenteroides strains isolated from plant-based fermentations such as kimchi and birch sap, but the HEPR strains analyzed here are the only known meat-derived isolates to carry this gene. Genomic analyses revealed highly conserved biosynthetic clusters for dextran, heteropolysaccharide, and levan. Gene expression profiling showed that levS and sacB_1 were upregulated at 8 °C, while dsrD expression was favoured at 25 °C. Cold-induced sucrose metabolism, characterized by high expression of levS, sacB_1, and dsrD, enhanced cell viability under oxidative stress. Furthermore, heterologous expression of sacB_1 in Leuc. mesenteroides and Lactococcus lactis improved resilience under cold and high-aeration conditions, confirming the protective role of levan. These findings advance the understanding of temperature-dependent EPS regulation in LAB and highlight sucrase diversity as a key factor in microbial adaptation to environmental stress.
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spelling doaj-art-a7b51e5490fe4179881d5f6ec78f0b162025-08-20T04:01:26ZengNature PortfolioScientific Reports2045-23222025-07-0115111710.1038/s41598-025-04141-xCold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroidesMiguel Fernandez de Ullivarri0Colin Buttimer1Janneke Wijman2Eelco Heintz3Paul Ross4Matthew P. McCusker5Colin Hill6APC Microbiome Ireland, School of Microbiology, University College CorkAPC Microbiome Ireland, School of Microbiology, University College CorkNiacet, A Kerry® CompanyNiacet, A Kerry® CompanyAPC Microbiome Ireland, School of Microbiology, University College CorkKerry Taste & Nutrition, Global Technology & Innovation CentreAPC Microbiome Ireland, School of Microbiology, University College CorkAbstract Exopolysaccharides (EPS) play critical roles in microbial survival, stress adaptation, and biofilm formation across diverse environments. In food-associated bacteria such as Leuconostoc mesenteroides, understanding the regulation of EPS production under environmental stress is important for both spoilage control and industrial applications. However, the mechanisms linking cold stress to EPS biosynthesis remain poorly understood. Here, we show that sucrose and low temperature (8 °C) trigger a metabolic shift from dextran-only to combined dextran and levan biosynthesis in four meat-borne Leuc. mesenteroides strains. Two high EPS-producing strains (HEPRs) possess the sacB_1 gene, which encodes a previously uncharacterized levansucrase absent from low EPS-producing strains (LEPRs) that only carry the levS gene. This is the first study to describe the role of sacB_1 in cold-induced EPS production. Notably, sacB_1 was also identified in Leuc. mesenteroides strains isolated from plant-based fermentations such as kimchi and birch sap, but the HEPR strains analyzed here are the only known meat-derived isolates to carry this gene. Genomic analyses revealed highly conserved biosynthetic clusters for dextran, heteropolysaccharide, and levan. Gene expression profiling showed that levS and sacB_1 were upregulated at 8 °C, while dsrD expression was favoured at 25 °C. Cold-induced sucrose metabolism, characterized by high expression of levS, sacB_1, and dsrD, enhanced cell viability under oxidative stress. Furthermore, heterologous expression of sacB_1 in Leuc. mesenteroides and Lactococcus lactis improved resilience under cold and high-aeration conditions, confirming the protective role of levan. These findings advance the understanding of temperature-dependent EPS regulation in LAB and highlight sucrase diversity as a key factor in microbial adaptation to environmental stress.https://doi.org/10.1038/s41598-025-04141-xLeuconostoc mesenteroidesExopolysaccharideLevansucraseCold stressSucrose-induced metabolism
spellingShingle Miguel Fernandez de Ullivarri
Colin Buttimer
Janneke Wijman
Eelco Heintz
Paul Ross
Matthew P. McCusker
Colin Hill
Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
Scientific Reports
Leuconostoc mesenteroides
Exopolysaccharide
Levansucrase
Cold stress
Sucrose-induced metabolism
title Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
title_full Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
title_fullStr Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
title_full_unstemmed Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
title_short Cold induced expression of a novel levansucrase gene sacB1 enhances exopolysaccharide production and stress resilience in Leuconostoc mesenteroides
title_sort cold induced expression of a novel levansucrase gene sacb1 enhances exopolysaccharide production and stress resilience in leuconostoc mesenteroides
topic Leuconostoc mesenteroides
Exopolysaccharide
Levansucrase
Cold stress
Sucrose-induced metabolism
url https://doi.org/10.1038/s41598-025-04141-x
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