Genomic characterization reveals probiotic potentials in Leuconostoc falkenbergense isolated from naturally fermented buffalo milk curd

Background The genus Leuconostoc has significant importance in food fermentations and is increasingly evaluated for probiotic applications. Although Leuconostoc falkenbergense is a recently identified species, and its in-depth genomic characterization and probiotic attributes remain largely unexplored, particularly for strains derived from diverse natural sources such as traditional dairy food matrices. This study therefore aimed to isolate and characterize a L. falkenbergense strain (BSMRAU-M1L5) from naturally fermented traditional buffalo milk curd and performed whole-genome sequencing with detailed genomic analyses to evaluate its probiotic potential and safety profile for future applications. Results The draft genome of L. falkenbergense BSMRAU-M1L5 spans 1.78 Mb, assembled into 96 contigs, and encodes 1,844 genes. Phylogenetic and average nucleotide analyses with 21 other L. falkenbergense strains confirmed that BSMRAU-M1L5 is most closely related to strain C, previously isolated from yogurt in China. Pan-genome analysis revealed a substantial core gene set and 124 strain-specific genes in BSMRAU-M1L5, including 99 hypothetical genes, while the remainder were linked to metabolic versatility, survivability, and functional adaptation. Functional annotation revealed genes involved in carbohydrate utilization, vitamin and acetate biosynthesis, acid tolerance, antioxidant defense, stress response, adhesion, AI-2 signaling, exopolysaccharide production, and antimicrobial activity, highlighting its strong probiotic potential. The genome also harbors CRISPR-Cas systems, insertion sequence elements, and prophages, while lacking virulence and biogenic amine–related genes, confirming its genomic safety. Despite harboring three vancomycin resistance genes, the strain was susceptible to most tested antibiotics and exhibited γ-hemolysis, supporting its non-pathogenic nature and overall safety profile. Conclusion The genome of L. falkenbergense BSMRAU‑M1L5 encodes diverse metabolic and probiotic properties, supported by both genomic and phenotypic evidence of safety, positioning it as a promising candidate for use in fermented food production and broader biotechnological applications.