title: Anchorless surface associated glycolytic enzymes from Lactobacillus plantarum 299v bind to epithelial cells and extracellular matrix proteins.
abstract: An important criterion for the selection of a probiotic bacterial strain is its ability to adhere to the mucosal surface. Adhesion is usually mediated by proteins or other components located on the outer cell surface of the bacterium. In the present study we characterized the adhesive properties of two classical intracellular enzymes glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and enolase (ENO) isolated from the outer cell surface of the probiotic bacterium Lactobacillus plantarum 299v. None of the genes encoded signal peptides or cell surface anchoring motifs that could explain their extracellular location on the bacterial surface. The presence of the glycolytic enzymes on the outer surface was verified by western blotting using polyclonal antibodies raised against the specific enzymes. GAPDH and ENO showed a highly specific binding to plasminogen and fibronectin whereas GAPDH but not ENO showed weak binding to mucin. Furthermore, a pH dependent and specific binding of GAPDH and ENO to intestinal epithelial Caco-2 cells at pH 5 but not at pH 7 was demonstrated. The results showed that these glycolytic enzymes could play a role in the adhesion of the probiotic bacterium L. plantarum 299v to the gastrointestinal tract of the host. Finally, a number of probiotic as well non-probiotic Lactobacillus strains were analyzed for the presence of GAPDH and ENO on the outer surface, but no correlation between the extracellular location of these enzymes and the probiotic status of the applied strains was demonstrated.
authors: Bioneer A/S, Kogle Allé 2, DK-2970 Hørsholm, Denmark. jacob.glenting@alk.net
authors: Jacob
authors: J
authors: Glenting
authors: Hans Christian
authors: HC
authors: Beck
authors: Astrid
authors: A
authors: Vrang
authors: Holger
authors: H
authors: Riemann
authors: Peter
authors: P
authors: Ravn
authors: Anne Maria
authors: AM
authors: Hansen
authors: Martin
authors: M
authors: Antonsson
authors: Siv
authors: S
authors: Ahrné
authors: Hans
authors: H
authors: Israelsen
authors: Søren
authors: S
authors: Madsen
mesh_terms: D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004847:Epithelial Cells / Q000382:microbiology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000378:metabolism*; D006801:Humans; D048191:Lactiplantibacillus plantarum / Q000201:enzymology* / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D011485:Protein Binding; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D016326:Extracellular Matrix Proteins; D008565:Membrane Proteins; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); D010751:Phosphopyruvate Hydratase