2‐DE and MS analysis of key proteins in the adhesion of Lactobacillus plantarum, a first step toward early selection of probiotics based on bacterial biomarkers

Abstract

The identification of cell components involved in probiotic activities is a challenge in current probiotic research. In this work, a new approach based on proteomics as an analytical tool for the identification of characteristic protein profiles related to adhesion to mucin as a model probiotic property was used. Three Lactobacillus plantarum strains with different adhesion rates were used for proteomic analysis: L. plantarum WHE 92 (15.9%), L. plantarum 299 v (9.1%) and L. plantarum CECT 4185 (1.4%). Cell wall extracts were subjected to proteomic analysis of differential protein expression using 2-DE, tryptic digestion, chip-LC-QTOF mass analysis and protein identification using database search. Several proteins, previously reported to be involved in bacterial adhesion: elongation factor EF-Tu, GroEL chaperonin, molecular chaperone DnaK and glyceraldehyde-3-phosphate dehydrogenase were found to be overexpressed in the cell wall proteome of the highly adhesive strain L. plantarum WHE 92. The overexpression of two spots containing GroES co-chaperonin in the most adhesive strain also suggested the involvement of this protein in the adhesion process. The association of proteomic profiles and proteins with particular probiotic properties opens the way for the use of such profiles and proteins as bacterial biomarkers for the properties of bacteria but probably also for their potential health effects.

Decrease in Penicillin Susceptibility Due to Heat Shock Protein ClpL in Streptococcus pneumoniae

“…Similarly, the Coxiella burnetii DnaK homologue is localized to the cell wall and the cytoplasm (35). Consistent with this, in Gram-positive bacteria, DnaK is associated with the cell wall in Lactobacillus salivarius (24), Lactobacillus plantarum (23), and Listeria monocytogenes (55). In Staphylococcus aureus, DnaK is a member of the cell wall stress stimulon and induced by inhibition of peptidoglycan biosynthesis (47).…”

“…The dnaK mutant of S. aureus is highly susceptible to cell wall-active antibiotic stress conditions (56). Moreover, both GroEL and DnaK are found on the bacterial surface in Mycobacterium tuberculosis (21) and L. plantarum (23). Thus, GroEL or DnaK facilitate protein refolding on the cell wall or cell surface.…”

Diminished Exoproteome of Frankia spp. in Culture and Symbiosis

“…A surface-localized glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in streptococci binds to host extracellular matrix glycoproteins and cytoskeletal proteins, and a GAPDH abundant in the cell wall proteome of Lactobacillus plantarum is implicated in adhesion to mucin (17,28). CcI3 also has a second GAPDH that is homologous to that located on the surface in other gram-positive bacteria.…”

Cell surface proteins play an important role in probiotic activities of Lactobacillus reuteri

“…It has been proposed that cell surface proteins are involved in cell protection and surface recognition and that they could be potential mediators in the initial steps involved in autoaggregation and adhesion [8][9][10]. In cell envelope proteome studies of potentially probiotic bacteria, the cell wall protein fraction has typically been extracted by lysozyme-containing buffer [11][12][13][14][15][16], lithium chloride [16,17], or some other cell surface molecule or proteinsolubilizing agent [15][16][17][18].…”