Mucosal adhesion properties of the probiotic Lactobacillus rhamnosus GG SpaCBA and SpaFED pilin subunits

I. von Ossowski, J. Reunanen, R.M. Satokari, S. Vesterlund, M. Kankainen, H. Huhtinen, S. Tynkkynen, S. Salminen, W.M. de Vos, A. Palva

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139 Citations (Scopus)

Abstract

Lactobacillus rhamnosus GG is a well-established Gram-positive probiotic strain, whose health-benefiting properties are in part dependent upon a prolonged residency in the gastrointestinal tract and likely dictated by an adherence to the intestinal mucosa. Previously, we identified two pilus gene clusters (spaCBA and spaFED) in the genome of this probiotic, each of which contained the predicted genes for three pilin subunits and a single sortase. We also confirmed the presence of SpaCBA pili on the cell surface and attributed an intestinal mucus-binding capacity to one of the pilin subunits (SpaC). Herein, we now report the cloning of the remaining pilin genes (spaA, spaB, spaD, spaE, and spaF) in Escherichia coli, production and purification of the recombinant proteins, and an assessment of their adherence to human intestinal mucus. Our findings indicate that the SpaB and SpaF pilin subunits also exhibit substantial mucus binding, which can be inhibited competitively in a dose-related manner. Moreover, the binding between SpaB pilin subunit and the mucosal substrate appears to operate through electrostatic contacts and is not related to a recognized mucus-binding domain. We conclude from these results that it is conceivable two pilin subunits (SpaB and SpaC) in the SpaCBA pilus fiber play a role in binding to intestinal mucus, but for the uncharacterized and putative SpaFED pilus fiber only a single pilin subunit (SpaF) is potentially responsible for mucus adhesion
Original languageEnglish
Pages (from-to)2049-2057
JournalApplied and Environmental Microbiology
Volume76
Issue number7
DOIs
Publication statusPublished - 2010

Keywords

  • human intestinal mucus
  • gram-positive bacteria
  • lactic-acid bacteria
  • cell-surface
  • streptococcus-agalactiae
  • gastrointestinal-tract
  • oral consumption
  • dodecyl-sulfate
  • protein
  • colonization

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