title: Sialic acid (N-acetyl neuraminic acid) utilization by Bacteroides fragilis requires a novel N-acetyl mannosamine epimerase.
abstract: We characterized the nanLET operon in Bacteroides fragilis, whose products are required for the utilization of the sialic acid N-acetyl neuraminic acid (NANA) as a carbon and energy source. The first gene of the operon is nanL, which codes for an aldolase that cleaves NANA into N-acetyl mannosamine (manNAc) and pyruvate. The next gene, nanE, codes for a manNAc/N-acetylglucosamine (NAG) epimerase, which, intriguingly, possesses more similarity to eukaryotic renin binding proteins than to other bacterial NanE epimerase proteins. Unphosphorylated manNAc is the substrate of NanE, while ATP is a cofactor in the epimerase reaction. The third gene of the operon is nanT, which shows similarity to the major transporter facilitator superfamily and is most likely to be a NANA transporter. Deletion of any of these genes eliminates the ability of B. fragilis to grow on NANA. Although B. fragilis does not normally grow with manNAc as the sole carbon source, we isolated a B. fragilis mutant strain that can grow on this substrate, likely due to a mutation in a NAG transporter; both manNAc transport and NAG transport are affected in this strain. Deletion of the nanE epimerase gene or the rokA hexokinase gene, whose product phosphorylates NAG, in the manNAc-enabled strain abolishes growth on manNAc. Thus, B. fragilis possesses a new pathway of NANA utilization, which we show is also found in other Bacteroides species.
authors.affiliation: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA 02111, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Brigham
authors.forename: Ruth
authors.initials: R
authors.lastname: Caughlan
authors.forename: Rene
authors.initials: R
authors.lastname: Gallegos
authors.forename: Mary Beth
authors.initials: MB
authors.lastname: Dallas
authors.forename: Veronica G
authors.initials: VG
authors.lastname: Godoy
authors.forename: Michael H
authors.initials: MH
authors.lastname: Malamy
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001441:Bacteroides fragilis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D002238:Carbohydrate Epimerases / Q000235:genetics / Q000378:metabolism*; D006595:Hexosamines / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D006595:Hexosamines; C008322:mannosamine; D002238:Carbohydrate Epimerases; D019158:N-Acetylneuraminic Acid