Structure of macroamphiphiles from several Bifidobacterium strains

Abstract

Lipoteichoic acid-like substances, macroamphiphiles, were isolated from cell homogenates ofBifidobacterium bifidum YIT 4007 and YIT 4013, Bifidobacterium breve YIT 4010 and YIT 4014, and Bfidobacterium longum YIT 4021 by phenol extraction followed by nuclease digestion, gel chromatography, ion-exchange chromatography, and hydrophobic interaction chromatography. The macroamphiphile preparations from these five strains contained D-glucose, D-galactose, glycerol, phosphorus, L-alanine, and fatty acids in molar ratios of 1.00, 1.57 to 1.95, 1.02 to 1.99, 0.97 to 1.72, 0.15 to 0.46, and 0.16 to 0.43. Data from structural analyses including methylation, 'H nuclear magnetic resonance measurement, alkaline hydrolysis, mild acid hydrolysis, and hydrogen fluoride treatment led to the most likely common structure for the macroamphiphiles of the examined strains,L-Ala-Gro-P (m = 11 to 18; n = 8 to 12) where Gro-P is glycerophosphate, m is the number of repeating units of galactofuranan, and n is the number of repeating units of glucan. Whereas the polymers from the respective strains differed in the numbers of repeating units of the galactofuranan and glucan moieties and in the number of fatty acid residues, the proposed structure is essentially the same as that reported previously for the macroamphiphile of B. bifidum subsp. pennsylvanicum DSM 20239 by W. Fischer (Eur. J. Biochem. 165:639-46, 1987). H--5Galf1l)m(6Gcp,3l)-n-6Galp,Bl-3-diacylglyceroI 16 L-Ala-Gro-P (m = 7 to 10; n = 8 to 15) where Gro-p is glycerophosphate, m is the numbering of repeating units of galactofuranan, and n is the number of repeating units of glucan.Thus, it is of interest to establish the structure for the macroamphiphile of Bifidobacterium and to confirm the common occurrence of such amphipathic polymers in a variety of Bifidobacterium species. This report describes the * Corresponding author. results of studies on the isolation and structure of the macroamphiphiles from five Bifidobacterium strains. MATERIALS AND METHODSMacroorganisms and isolation of macroamphiphiles. Cells of B. bifidum YIT 4007 and YIT 4013 (ATCC 15696), Bifidobacterium breve YIT 4010 and YIT 4014 (ATCC 15700), and Bifidobacterium longum YIT 4021 (ATCC 15707), selected from the culture collection of Yakuruto Central Institute for Microbiological Research, were grown and harvested as described previously (8). The extraction and purification of macroamphiphiles were essentially as described previously for the preparation of lipoteichoic acids (10). Sonic disintegration of the cells, extraction with hot phenol, nuclease treatment, and purification by column chromatography of Sepharose CL-6B, DEAE-Sephacel, and Octyl-Sepharose CL-4B were included.Alkaline treatment. Mild alkaline hydrolysis for deacylation was carried out in 0.2 M NaOH at 30°C for 2 h, and alkaline hydrolysis for removal of glycerophosphate residues from macroamphiphiles was done in 0.3 M NaOH at 100°C for 4 h. After neutralization with 2 M HCI, the alkaline hydrolysis products were fractionated by chromatograp...

Atypical lipoteichoic acids of gram-positive bacteria

“…Antigenic cross-reactivities suggested that similar lipoglycans were present in other bifidobacteria, since extracts from seven of eight bifidobacterial serological groups cross-reacted with antisera to the B. bifidum antigen (49). Lipoglycans with essentially the same structure as that proposed by Fischer (16) have been confirmed to be present in Bifidobacterium breve and Bifidobacterium longum strains (30).…”

Bifidobacterial Lipoglycan as a New Cause for False-Positive Platelia Aspergillus Enzyme-Linked Immunosorbent Assay Reactivity

“…ELISA reactivity was clearly shown to be associated with bacterial lipoglycans containing a ␤-1,5-galactofuranosyl chain. Only a few bifidobacterial lipoglycans have been structurally analyzed, but also other bifidobacterial species seem to contain reactive residues in their cell wall (7,18,30). This specific epitope can be detected as a surface antigen, shown by the reactivity of the suspended cell pellet, but also as an exoantigen, shown by the reactivity of the cellfree supernatant of a liquid culture (Table 3).…”

The Sweet Tooth of Bacteria: Common Themes in Bacterial Glycoconjugates

“…These glycerophosphates are substituted with L-alanine residues. This basic structure is common to all bifidobacteria, but the numbers of polymers and repeating units vary among strains (216).…”