Substrate preference of an ABC importer corresponds to selective growth on β-(1,6)-galactosides in Bifidobacterium animalis subsp. lactis

Mia Christine Theilmann, Folmer Fredslund, Birte Svensson, Leila Lo Leggio, Maher Abou Hachem*

*Corresponding author for this work

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Abstract

Bifidobacteria are exposed to substantial amounts of dietary β-galactosides. Distinctive preferences for growth on different β-galactosides are observed within Bifidobacterium members, but the basis of these preferences remains unclear. We previously described the first β-(1,6)/(1,3)-galactosidase from Bifidobacterium animalis subsp. lactis Bl-04. This enzyme is relatively promiscuous, exhibiting only 5-fold higher efficiency on the preferred β-(1,6)-galactobiose than the β-(1,4) isomer. Here, we characterize the solute-binding protein (Bal6GBP) that governs the specificity of the ABC transporter encoded by the same β-galactoside-utilization locus. We observed that although Bal6GBP recognizes both β-(1,6)- and β-(1,4)-galactobiose, Bal6GBP has a 1630-fold higher selectivity for the former, reflected in dramatic differences in growth, with several hours lag on less preferred β-(1,4)- and β-(1,3)-galactobiose. Experiments performed in the presence of varying proportions of β-(1,4)/ β-(1,6)-galactobioses indicated that the preferred substrate was preferentially depleted from the culture supernatant. This established that the poor growth on the non-preferred β-(1,4) was due to inefficient uptake. We solved the structure of Bal6GBP in complex with β-(1,6)-galactobiose at 1.39 Å resolution, revealing the structural basis of this strict selectivity. Moreover, we observed a close evolutionary relationship with the human milk disaccharide lacto-N-biose-binding protein from Bifidobacterium longum, indicating that the recognition of the non-reducing galactosyl is essentially conserved, whereas the adjacent position is diversified to fit different glycosidic linkages and monosaccharide residues. These findings indicate that oligosaccharide uptake has a pivotal role in governing selectivity for distinct growth substrates and have uncovered evolutionary trajectories that shape the diversification of sugar-uptake proteins within Bifidobacterium.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume294
Issue number31
Pages (from-to)11701-11711
Number of pages12
ISSN0021-9258
DOIs
Publication statusPublished - 2019

Keywords

  • ABC transport
  • Actinobacteria
  • Bifidobacteria
  • Crystal structure
  • Enzyme kinetics
  • Galactoogliosaccharides (GOS)
  • Human milk ogliosaccharides (HMO)
  • Human gut microbiota
  • Isothermal titration calorimetry (ITC)
  • Microbiome
  • Prebitoic
  • Probiotic
  • Protein evolution
  • Surface plasmon resonance (SPR)

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