Many kinds ofLactobacillusare common occupants of humans’ digestive tract that support the preservation of a balanced microbial environment that benefits host health. In this study, the unique lactic acid bacterium strainLimosilactobacillus fermentumU-21, which was isolated from the feces of a healthy human, was examined for its metabolite profile in order to compare it to that of the strainL. fermentum279, which does not have antioxidant (AO) capabilities. By using GC × GC−MS, the metabolite fingerprint of each strain was identified, and the data were then subjected to multivariate bioinformatics analysis. TheL. fermentumU-21 strain has previously been shown to possess distinctive antioxidant properties in in vivo and in vitro studies, positioning it as a drug candidate for the treatment of Parkinsonism. The production of multiple distinct compounds is shown by the metabolite analysis, demonstrating the unique characteristics of theL. fermentumU-21 strain. According to reports, some of theL. fermentumU-21 metabolites found in this study have health-promoting properties. The GC × GC−MS-based metabolomic tests defined strainL. fermentumU-21 as a potential postbiotic with significant antioxidant potential.