The human gut hosts a dynamic microbial ecosystem essential for gut health and immune function. Influenced by delivery mode, diet, and antibiotics, the infant microbiota shapes health outcomes. Human-origin probiotics are gaining attention for their ability to restore microbial balance and offer therapeutic benefits. This study explores potential probiotic strains isolated from infant feces, focusing on safety, functional properties, and antioxidant capabilities. From fecal samples of 10 healthy infants aged 2–24 months, 47 isolates were screened. Comprehensive evaluations identified two promising strains: Lacticaseibacillus paracasei IF5YR and Enterococcus faecium IF5RW2. Both strains exhibited robust survival under simulated gastrointestinal conditions, including tolerance to low pH (1.5) and high bile concentrations (2%). These strains demonstrated strong autoaggregation abilities (>70%) and significant hydrophobicity (>90% for xylene), suggesting their potential to colonize the gut and inhibit pathogens. Functional assays confirmed bile salt hydrolase (BSH) activity, highlighting their role in cholesterol metabolism. Antioxidant evaluations revealed high reducing power (>75% in FRAP) and moderate free radical scavenging activity in ABTS and DPPH assays. Molecular characterization via 16S rRNA sequencing confirmed their identity, while safety was ensured by non-hemolytic activity and susceptibility to clinically relevant antibiotics. These findings position Lb. paracasei IF5YR and Ent. faecium IF5RW2 as potent candidates for probiotic applications, particularly in functional foods targeting gut health and oxidative stress management. Further, in vivo studies are warranted to substantiate their therapeutic potential.