The human gut microbiota is composed of more than 50% of Bacteroides species whose membranes are enriched in sphingolipids. In particular, Bacteroides fragilis produced glycosphingolipids termed as α-Galactosylceramide (BfaGCs) have the ability to activate mouse type I Natural Killer T (NKT) cells upon presentation by the antigen-presenting molecule CD1d. While they share key chemical similarities with the type I NKT cell marker antigen, α-Galactosylceramide (KRN7000), they possess distinctive structural features including short, terminal branching with the variations in their hydroxyl group positions. Here, using X-ray crystallography, we have determined two crystal structures of NKT TCR-mouse CD1d-BfaGC lipid ternary complexes revealing a parallel docking mode of the TCR atop the F’-pocket of CD1d in recognizing the presented bacterial sphingolipids. The measured binding affinities between the mouse CD1d presented BfaGCs and NKT TCR were observed to be in the nanomolar range. As translation of results from benchside to bedside with KRN7000 has proven challenging, our study of these physiologically relevant BfaGCs may open new avenues in designing novel immunomodulatory agents to achieve desired immune outcomes in various clinical conditions. Further, this study sheds light on a better understanding of the existing symbiotic relationship between the microbes producing these endogenous lipids and the host.