title: Production of α-galactosylceramide by a prominent member of the human gut microbiota.
abstract: While the human gut microbiota are suspected to produce diffusible small molecules that modulate host signaling pathways, few of these molecules have been identified. Species of Bacteroides and their relatives, which often comprise >50% of the gut community, are unusual among bacteria in that their membrane is rich in sphingolipids, a class of signaling molecules that play a key role in inducing apoptosis and modulating the host immune response. Although known for more than three decades, the full repertoire of Bacteroides sphingolipids has not been defined. Here, we use a combination of genetics and chemistry to identify the sphingolipids produced by Bacteroides fragilis NCTC 9343. We constructed a deletion mutant of BF2461, a putative serine palmitoyltransferase whose yeast homolog catalyzes the committed step in sphingolipid biosynthesis. We show that the Δ2461 mutant is sphingolipid deficient, enabling us to purify and solve the structures of three alkaline-stable lipids present in the wild-type strain but absent from the mutant. The first compound was the known sphingolipid ceramide phosphorylethanolamine, and the second was its corresponding dihydroceramide base. Unexpectedly, the third compound was the glycosphingolipid α-galactosylceramide (α-GalCer(Bf)), which is structurally related to a sponge-derived sphingolipid (α-GalCer, KRN7000) that is the prototypical agonist of CD1d-restricted natural killer T (iNKT) cells. We demonstrate that α-GalCer(Bf) has similar immunological properties to KRN7000: it binds to CD1d and activates both mouse and human iNKT cells both in vitro and in vivo. Thus, our study reveals BF2461 as the first known member of the Bacteroides sphingolipid pathway, and it indicates that the committed steps of the Bacteroides and eukaryotic sphingolipid pathways are identical. Moreover, our data suggest that some Bacteroides sphingolipids might influence host immune homeostasis.
authors.affiliation: Department of Bioengineering and Therapeutic Sciences and the California Institute for Quantitative Biosciences, University of California, San Francisco, California, United States of America.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Wieland Brown
authors.forename: Cristina
authors.initials: C
authors.lastname: Penaranda
authors.forename: Purna C
authors.initials: PC
authors.lastname: Kashyap
authors.forename: Brianna B
authors.initials: BB
authors.lastname: Williams
authors.forename: Jon
authors.initials: J
authors.lastname: Clardy
authors.forename: Mitchell
authors.initials: M
authors.lastname: Kronenberg
authors.forename: Justin L
authors.initials: JL
authors.lastname: Sonnenburg
authors.forename: Laurie E
authors.initials: LE
authors.lastname: Comstock
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Bluestone
authors.forename: Michael A
authors.initials: MA
authors.lastname: Fischbach
mesh_terms: D000818:Animals; D001441:Bacteroides fragilis / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D002478:Cells, Cultured; D005699:Galactosylceramides / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009154:Mutation; D055611:Natural Killer T-Cells / Q000378:metabolism
chemical_list: D005699:Galactosylceramides; C493154:alpha-galactosylceramide