Techno‐Functional Characterization and Safety Evaluation of Human‐Derived Enterococcus faecium as Potential Probiotics (In Vitro Study)

ABSTRACT Probiotics are considered live microorganisms that contribute to health benefits by modulating gut microbiota and improving gastrointestinal function. The present study aimed to isolate and characterize indigenous Enterococcus faecium ( E. faecium ) strains from healthy human fecal samples to assess their probiotic potential. Eleven isolates were selected based on Gram‐positive, catalase‐negative, nonmotile, and nonhemolytic characteristics, and were identified using MALDI‐TOF MS and 16S rRNA gene sequencing. The bacterial isolates were assessed for acid and bile tolerance, antimicrobial activity, cell surface hydrophobicity, tolerance to osmotic (NaCl) and phenolic stress, exopolysaccharide (EPS) production, bile salt hydrolase and DNase activity, cholesterol‐lowering capacity, and antioxidant activity. Findings demonstrated strong acid and bile tolerance, high NaCl and phenol resilience, positive BSH and EPS activity, and absence of DNase. Antimicrobial activity was moderate to high against gastrointestinal pathogens, antioxidant activity and surface hydrophobicity ranged from moderate to high, while cholesterol‐lowering efficiency varied among strains. Cumulative probiotic potential scores were determined that highlighting strain‐specific strengths, signifying that these E. faecium isolates possess multifunctional probiotic attributes suitable for functional foods and nutraceutical applications. Further in vivo studies are necessary to confirm their efficacy and safety for human consumption.