In vitro assays, probiogenomic analysis, safety assessment and probiotic potential of Enterococcus faecium F25

The probiogenomic approach has gained considerable attention for elucidating the genetic basis of probiotic functionality. In this study, the probiotic potential of Enterococcus faecium F25 was comprehensively evaluated using an integrated strategy combining traditional biochemical characterization, in vitro functional assays, and whole-genome-based analyses. The in vitro assessments were conducted using plate-based assays, while genomic investigations were performed using publicly available bioinformatic tools and platforms. Both approaches consistently confirmed the strain's typical enterococcal characteristics and its promising probiotic potential. Comprehensive genomic profiling revealed the absence of transmissible virulence-associated determinants and transferable antibiotic resistance genes, together with a comparatively low abundance of mobile genetic elements, supporting the strain's biosafety for potential industrial and health-related applications. Functional genome annotation further identified multiple probiotic-associated determinants involved in stress tolerance, metabolic adaptability, antimicrobial activity, and bioactive compound production. In particular, genes encoding enterocins A, B, and enterolysin A, together with stress response elements and enhanced metabolic capabilities, may contribute to improved environmental resilience, gastrointestinal survival, and antagonistic activity against pathogens. Collectively, these findings indicate that E. faecium F25 possesses a promising combination of safety, functional, and metabolic traits, supporting its potential as a versatile probiotic candidate. Further in vivo investigations are required to validate its probiotic efficacy and functional performance under host-associated conditions.