title: Purification and characterization of a novel protein produced by Bifidobacterium longum SBT2928 that inhibits the binding of enterotoxigenic Escherichia coli Pb176 (CFA/II) to gangliotetraosylceramide.
abstract: A novel protein (BIF) which shows inhibitory activity on the binding of enterotoxigenic Escherichia coli Pb176 (ETEC with colonization factor antigen (CFA) II, which consists of coli surface-associated antigens CS1 and CS3) to gangliotetraosylceramide (asialo GM1 or GA1) was isolated from the culture supernatant fluid of Bifidobacterium longum SBT2928 (BL2928) at its stationary phase. The homogeneity of the final preparation of BIF was demonstrated by SDS-PAGE, polyacrylamide gel electrofocusing and N-terminal amino acid sequencing. The BIF was characterized as (i) a protein with an M(r) of approximately 104 kDa when chromatographed on a gel filtration column, and 52 kDa when separated on SDS-PAGE, and (ii) having an isoelectric point of 5.9. No change in size was produced by thiol reduction. These results suggest that BIF is a homodimer consisting of identical 52 kDa monomers. The purified BIF at the concentration of 25 micrograms protein ml-1 caused a 50% reduction in binding of the ETEC strain to GA1.
authors.affiliation: Technology and Research Institute, Snow Brand Milk Products Co., Ltd, Saitama, Japan. ht4s-fjwr@asahi-net.or.jp
authors.forename: S
authors.initials: S
authors.lastname: Fujiwara
authors.forename: H
authors.initials: H
authors.lastname: Hashiba
authors.forename: T
authors.initials: T
authors.lastname: Hirota
authors.forename: J F
authors.initials: JF
authors.lastname: Forstner
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D001644:Bifidobacterium / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000096:biosynthesis; D004926:Escherichia coli / Q000187:drug effects* / Q000378:metabolism; D005677:G(M1) Ganglioside / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data
chemical_list: C119733:BIF protein, Bifidobacterium longum; D001426:Bacterial Proteins; D004768:Enterotoxins; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside