Microbiota-derived tryptophan metabolites are essential signals for maintaining gut homeostasis, yet the potential contribution to modulating gut microbiota has been rarely investigated. In this study, ZJ316 (CCTCC No. M 208077) with a high production (43.14 μg/mL) of indole-3-lactic acid (ILA) was screened. ILA with 99.00% purity was prepared by macroporous resin, Sephadex G-25 and reversed-phase high-performance liquid chromatography. Purified ILA can effectively inhibit foodborne pathogens such as spp., spp., and . In an in vitro model of the human gut microbiota, a medium-dose ILA (172 mg/L) intervention increased the average relative abundance of phyla Firmicutes and Bacteroidota by 9.27% and 15.38%, respectively, while Proteobacteria decreased by 14.36% after 24 h fermentation. At the genus level, the relative abundance of and significantly increased to 5.36 ± 2.31% and 2.19 ± 0.77% ( < 0.01), respectively. and decreased to 16.41 ± 4.81% ( < 0.05) and 2.84 ± 1.02% ( < 0.05), respectively. Intestinal short-chain fatty acids, especially butyric acid, were significantly increased (2.98 ± 0.72 µmol/mL, < 0.05) and positively correlated with and . Overall, ILA has the potential to regulate the gut microbiota, and an in-depth understanding of the relationship between tryptophan metabolites and gut microbiota is needed in the future.
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