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Functional and Genomic Analysis of Leuconostoc citreum DMLC16 Reveals Its Potential as a Probiotic and Antimicrobial Starter

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This study aimed to assess the probiotic potential of Leuconostoc citreum DMLC16 previously selected as a fermentation starter candidate based on safety and enzymatic activity evaluations by further examining its health-promoting properties and antimicrobial efficacy. Strain DMLC16 exhibited higher acid tolerance, bile salt resistance, and intestinal adhesion ability than its type strain L. citreum KACC 11860T . Moreover, it demonstrated antimicrobial activities against a range of foodborne pathogens and spoilage organisms, including Bacillus cereus, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, Alcaligenes xylosoxidans, Flavobacterium sp., and Salmonella enterica. Strain DMLC16 also showed antifungal activities against Clonostachys rosea, Epicoccum nigrum, and Penicillium citrinum. Whole-genome analysis confirmed the safety of DMLC16, revealing the absence of toxin-encoding and plasmids containing acquired antibiotic resistance genes. It also identified genes associated with enzymatic function, salt tolerance, and survival under extreme conditions such as gastric acidity. Although no specific bacteriocin genes were detected, genes implicated in the production of antimicrobial substances were present. The complete genome sequence of DMLC16 provides valuable insight into genetic determinants underlying its probiotic traits and supports its potential application as a functional starter culture or probiotic in the food industry.

Abstract

This study aimed to assess the probiotic potential of Leuconostoc citreum DMLC16 previously selected as a fermentation starter candidate based on safety and enzymatic activity evaluations by further examining its health-promoting properties and antimicrobial efficacy. Strain DMLC16 exhibited higher acid tolerance, bile salt resistance, and intestinal adhesion ability than its type strain L. citreum KACC 11860T . Moreover, it demonstrated antimicrobial activities against a range of foodborne pathogens and spoilage organisms, including Bacillus cereus, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, Alcaligenes xylosoxidans, Flavobacterium sp., and Salmonella enterica. Strain DMLC16 also showed antifungal activities against Clonostachys rosea, Epicoccum nigrum, and Penicillium citrinum. Whole-genome analysis confirmed the safety of DMLC16, revealing the absence of toxin-encoding and plasmids containing acquired antibiotic resistance genes. It also identified genes associated with enzymatic function, salt tolerance, and survival under extreme conditions such as gastric acidity. Although no specific bacteriocin genes were detected, genes implicated in the production of antimicrobial substances were present. The complete genome sequence of DMLC16 provides valuable insight into genetic determinants underlying its probiotic traits and supports its potential application as a functional starter culture or probiotic in the food industry.

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