We used LC-MS to identify the metabolic changes in soybean fermented by Bacillus subtilis DC-15 to comprehend the biochemical alterations. Total 650 distinct metabolites were observed, with amino acids, dipeptides, fatty acids, small molecule sugars, and isoflavones having the largest abundances. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the primary metabolic pathways involved during the fermentation of soybeans were related to amino acid metabolism, including beta-alanine metabolism, phenylalanine metabolism, glycine, serine, and threonine metabolism. In the gastric phase, the maximum amount of soy isoflavones was successfully converted from its glycoside form (50.04 μg/mL) to the aglycone form (104.07 μg/mL). The distribution of protein and lipid particles in natto during various stages of digestion showed changes from aggregated particles to more scattered particles. For the future development and enhancement of fermented soybean products, our metabolomic data can shed light on activated biochemical pathways and changes in metabolic pattern.