Anchorless surface associated glycolytic enzymes from Lactobacillus plantarum 299v bind to epithelial cells and extracellular matrix proteins (original) (raw)
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Animal Science Journal, 2012
Adhesion tests are complex, time-consuming and expensive, while the most important criterion for a probiotic lactobacilli is the ability to adhere to the human intestine. Thirty lactobacilli isolates from human intestinal tissues were measured for cell surface glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity using a microtiter plate screening method. GAPDH activities were detected in 21 out of 30 samples from 12 h cultures and in all samples from 18 h cultures. This suggests GAPDH is universally expressed on the bacterial cell surfaces from many lactobacilli. A statistically significant positive correlation was shown between GAPDH activity and adhesion using the BIACORE adhesion assay (P < 0.01). The new screening method using GAPDH enzymatic activity without an adhesion test may be possible due to the significant positive correlation of GAPDH activity with adhesion of lactobacilli derived from the human intestine.
Journal of Applied Microbiology, 2008
Aims: To characterize the adhesion molecule of Lactobacillus plantarum LA 318 that shows high adhesion to human colonic mucin (HCM). Methods and Results: The adhesion test used the BIACORE assay where PBS-washed bacterial cells showed a significant decrease in adherence to HCM than distilled water-washed cells. A component in the PBS wash fraction adhered to the HCM and a main protein was detected as a c. 40-kDa band using SDS-PAGE. Using homology comparisons of the N-terminal amino acid sequences compared with sequence databases, this protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The DNA sequence of LA 318 GAPDH was 100% identical to the GAPDH (gapB) of L. plantarum WCFS1. The purified GAPDH adhered to HCM. Conclusions: We found the adhesin of L. plantarum LA 318 to HCM in its culture PBS wash fraction. The molecule was identified as GAPDH. Because LA 318 possesses the same adhesin as many pathogens, the lactobacilli GAPDH may compete with pathogens infecting the intestine. Significance and Impact of the Study: This is the first report showing GAPDH expressed on the cell surface of lactobacilli adheres to mucin suggesting L. plantarum LA 318 adheres to HCM using GAPDH binding activity to colonize the human intestinal mucosa.
ELECTROPHORESIS, 2009
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-3phosphate 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.
Evaluation of adhesion properties of lactobacilli probiotic candidates
Monatshefte für Chemie - Chemical Monthly
Bacterial adhesion is a complex phenomenon implicated in the host-bacterial interaction that is pivotal for probiotic activity. Eight probiotic lactobacilli candidates (Lactobacillus reuteri, L. plantarum, L. mucosae, L. murinus) were screened for their ability to adhere to abiotic and biotic surfaces in vitro. Adhesion to hydrocarbons was used for hydrophobicity assessment. Three strains of L. reuteri and L. murinus C were evaluated as hydrophobic, others as intermediate. All tested strains were able to form the biofilm on polystyrene. L. mucosae D and L. reuteri E were tested for adhesion to epithelial cell lines (HeLa and Caco-2). Both were more adherent to HeLa than to Caco-2. The adhesivity degree in HeLa reached the highest value after 8 h of co-cultivation in both lactobacilli tested, then decreased. In Caco-2, adhesion was increased within 24 h from the beginning of the co-cultivation. Mucus-binding protein gene, implicated in adhesion, was detected in L. mucosae D. Therefore, the involvement of proteinaceous substances in binding process was investigated. Cells of L. mucosae D were digested by three proteolytic enzymes (proteinase K, pronase E, trypsin) and evaluated for time-dependent adhesivity changes to HeLa, Caco-2, and L929 cell lines. Results confirmed that proteins are most likely to play an important role in binding of lactobacilli to eukaryotic cells. One hour after treatment, L. mucosae D was able to overcome the effect of proteolytic cleavage. We assume that it was due to the restoration of its cell-surface binding structures. Co-cultivation of HeLa and L. mucosae D led to protuberance and communication channels formation in eukaryotic cells.
The Effect of Digestive Enzymes on the Adhesion of Probiotic Bacteria In Vitro
Journal of Food Science, 2001
Specific probiotics have several clinically proven health effects. Adhesion to the intestinal mucosa is considered important for many of these effects. In the current study, the effect of digestive enzymes and bile on the adhesion to intestinal mucus of 5 selected probiotics was studied. All of the digestive enzymes affected the adhesion of at least some of the tested strains. Bile was observed to reduce the adhesion of all strains tested. When the strains were sequentially exposed to 3 enzyme preparations and bile, the level of adhesion was reduced for all strains. The results suggest that the selection criterion "adhesion" for probiotics should be further refined.
Archives of Microbiology, 2009
Adhesion to intestinal epithelium is an outcome property for the selection of probiotic lactic acid bacteria strains. We have analyzed the adhesion properties of a collection of Lactobacillus casei strains from different origins, ranging from cheese isolates to commercial probiotics. Analysis of the surface characteristics of the strains by measuring adhesion to solvents (MATS test) showed that most of the strains have a basic and hydrophobic surface. The strains were able to bind ex vivo to human colon fragments at different levels and, in most cases, this adhesion correlated with the ability to in vitro binding of mucin. Attachment to this later substrate was not enhanced by growing the cells in the presence of mucin and was independent of proteinaceous factors. On the contrary, adhesion to other extracellular matrix components, such as collagen, fibronectin, or fibrinogen was partially or totally dependent on the presence of surface proteins. These results show that most of L. casei strains have in their surfaces factors that promote binding to intestinal epithelium, however, no clear correlation appears to exist between the origin of the strains and their adhesion capacities.
2014
Background: Adhesiveness to intestinal epithelium, beneficial immunomodulating effects and the production of pathogen-inhibitory compounds are generally considered as beneficial characteristics of probiotic organisms. We showed the potential health-promoting properties and the mechanisms of probiotic action of seven swine intestinal Lactobacillus amylovorus isolates plus the type strain (DSM 20531 T ) by investigating their adherence to porcine intestinal epithelial cells (IPEC-1) and mucus as well as the capacities of the strains to i) inhibit the adherence of Escherichia coli to IPEC-1 cells, ii) to produce soluble inhibitors against intestinal pathogens and iii) to induce immune signaling in dendritic cells (DCs). Moreover, the role of the L. amylovorus surface (S) -layers -symmetric, porous arrays of identical protein subunits present as the outermost layer of the cell envelope -in adherence to IPEC-1 cells was assessed using a novel approach which utilized purified cell wall fragments of the strains as carriers for the recombinantly produced S-layer proteins.
Study on the Adhesion of Lactobacillus Plantarum Strains with Probiotic Properties to MDCK
2016
One of the requirements for probiotic strains is to adhere to epithelial cells or cell lines. The presence of S-layer proteins in three Lactobacillus plantarum strains with probiotic properties was examined as well as their ability to adhere to the epithelial monolayer model non-cancerous cell line MDCK. The three strains lacked S-layer proteins, but Lactobacillus plantarum X2 and Lactobacillus plantarum LBRZ12 adhered to the cells of MDCK, while Lactobacillus plantarum F3 didn’t. Along with their other probiotic properties these make them suitable for inclusion in the composition of probiotics and probiotic foods.
FEMS Microbiology Letters, 2005
Lactobacillus salivarius subsp. salivarius UCC118 is a probiotic bacterium that was originally isolated from human intestinal tissues and was subsequently shown in a pilot study to alleviate symptoms associated with mild-moderate CrohnÕs disease. Strain UCC118 can adhere to animal and human intestinal tissue, and to both healthy and inflamed ulcerative colitis mucosa, irrespective of location in the gut. In this study, an enzymatic technique has been combined with proteomic analysis to correlate bacterial growth phase with the presence of factors present in the cell wall of the bacterium. Using PAGE electrophoresis, it was determined that progression from lag to log to stationary growth phases in vitro correlated with increasing prominence of an 84 kD protein associated with in vitro adherence ability. Isolated proteins from the 84 kD band region were further separated by two-dimensional electrophoresis, resolving this band into 20 individual protein spots at differing isoelectric points. The protein moieties were excised, trypsin digested and subjected to tandem mass spectrometry. The observed proteins are analogous to those reported to be associated with the Listeria monocytogenes cell-wall proteome, and include DnaK, Ef-Ts and pyruvate kinase. These data suggest that at least some of the beneficial attributes of probiotic lactobacilli, and in particular this strain, may be due to nonpathogenic mimicry of pathogens and potentially be mediated through a form of attenuated virulence.
Microorganisms, 2021
Adhesion capacity is considered one of the selection criteria for probiotic strains. The purpose of this study was to determine the adhesion properties of two candidate probiotics, Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7. The evaluation included the hydrophobicity of the cell surface using microbial adhesion to hydrocarbons (MATH), autoaggregation, and the adhesion of L. plantarum Dad-13 and L. plantarum Mut-7 to the intestinal mucosa of Sprague Dawley rat, followed by genomic analysis of the two L. plantarum strains. L. plantarum Dad-13 and L. plantarum Mut-7 showed a high surface hydrophobicity (78.9% and 83.5%) and medium autoaggregation ability (40.9% and 57.5%, respectively). The exposure of both isolates to the surface of the rat intestine increased the total number of lactic acid bacteria on the colon compartment, from 2.9 log CFU/cm2 to 4.4 log CFU/cm2 in L. plantarum Dad-13 treatment and to 3.86 log CFU/cm2 in L. plantarum Mut-7 treatment. The resul...