Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (original) (raw)
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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.
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.
Assessment of adhesion properties of novel probiotic strains to human intestinal mucus
International Journal of Food Microbiology, 2001
Potential new probiotic strains Lactobacillus brevis PELI, L. reuteri ING1, L. rhamnosus VTT E-800 and L. rhamnosus LC-705 were assessed for their adhesion properties using the human intestinal mucus model. The effect on the adhesion of exposure to acid and pepsin and to milk were tested to simulate gastric and food processing conditions, and the effect of different growth media on adhesion was tested. The properties of the four strains were compared to the well-investigated probiotic L. rhamnosus strain GG. Three of the tested strains showed significant adhesion properties in the mucus model, while L. brevis PELI had intermediate adhesion and L. rhamnosus LC-705 adhered poorly. Pretreatment with different milks decreased the adhesion and low pH and pepsin treatment reduced the adhesion of all tested strains except L. rhamnosus LC-705. No competitive exclusion of pathogenic Salmonella typhimurium or Escherichia coli SfaII was observed. The results indicate that major differences exist between tested proposed probiotic strains. The growth media and the food matrix significantly affect the adhesive ability of the tested strains. This has previously not been taken into account when selecting novel probiotic strains.
Indonesian Journal of Biotechnology, 2017
One of the primary selection criteria for potential probiotics is the ability to adhere to the host gastrointestinal tract. This study evaluated the in vitro adhesion ability on gastric mucin of two Lactobacillus casei strains (AP and AG) and two Pediococcus acidilactici strains (BE and BK), and identifed the corresponding genes responsible for adherence. Adhesion assays were performed in 96-well polystyrene microtiter plates using gastric mucin from porcine stomach as the matrix. An in vitro study on gastric mucin revealed that lactobacilli had a greater adherence ability compared with pediococci strains. The potential adherence genes were investigated using polymerase chain reaction (PCR) technology. Using specifc primers, PCR studies amplifed 150 base pairs of a potential mub gene and 161 base pairs of a potential ef-Tu gene, but no amplifed bands for potential map and bac genes were obtained. Sequence comparisons showed that the 150 and 161 amplifed base pairs were respectively ...
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.
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...
Journal of Applied Microbiology, 2007
Aims: To isolate lactobacilli from the mucus layer of the human intestine and evaluate their adhesion abilities using a BIACORE assay. Methods and Results: Thirty strains of lactobacilli were isolated from the mucus layer of normal human intestinal tissues using conventional plate culture. The strains were identified using homology comparisons of the 16S rDNA sequence to databases as Lactobacillus salivarius (26%), Lactobacillus fermentum (13%), Lactobacillus gasseri (10%), Lactobacillus paracasei (7%), Lactobacillus casei (3%), Lactobacillus mucosae (3%) and Lactobacillus plantarum (3%). Lactobacillus plantarum LA 318 shows the highest adhesion to human colonic mucin (HCM) using the BIACORE assay at 115AE30 ± 12AE37 resonance unit (RU). The adhesion of cell wall surface proteins from strain LA 318 was significantly higher to HCM than to bovine serum albumin (BSA; P < 0AE05). Conclusions: We isolated 30 strains of lactobacilli. Lactobacillus salivarius was the predominant species of lactobacilli isolated in this study. The adhesion of strain LA 318 isolated from human transverse colon to its mucin was shown. The adhesion could be mediated by lectin-like components on the bacterial cell surface. Significance and Impact of the Study: This is the first study where lactobacilli were isolated from human intestinal tissues and shown to adhere to HCM.
Prebiotic Treatment Influence the Adhesion Properties of Three Lactobacillus strains
International Journal of Current Microbiology and Applied Sciences, 2017
Different species from the genus Lactobacillus are commonly used as probiotics (Zago et al., 2011). By definition, probiotics are live microorganisms that confer a health benefit on the host when administered in adequate amounts (FAO/WHO, 2001). Their efficacy has been demonstrated for the treatment of gastrointestinal disorders, respiratory infections and allergic symptoms (Wohlgemuth et al., 2010). The general mechanisms of action that induce these beneficial results are: production of antibacterial substances; induction of defensin production by intestinal epithelial cells; competitive exclusion of pathogenic bacteria; influence on host microbiota and pathogenic bacteria; improved intestinal barrier function; modulation of host immune (Wohlgemuth et al., 2010). The predominant number of species applied as probiotics belong to the group of lactic acid bacteria (LAB), named after the main end product of their carbohydrate metabolism. LAB are GramInternational Journal of Current Mic...
Food research international (Ottawa, Ont.), 2018
Gastrointestinal conditions along the digestive tract are the main stress to which probiotics administrated orally are exposed because they must survive these adverse conditions and arrive alive to the intestine. Adhesion to epithelium has been considered one of the key criteria for the characterization of probiotics because it extends their residence time in the intestine and as a consequence, can influence the health of the host by modifying the local microbiota or modulating the immune response. Nevertheless, there are very few reports on the adhesion properties to epithelium and mucus of microorganisms after passing through the gastrointestinal tract. In the present work, we evaluate the adhesion ability in vitro of L. paracasei strains isolated from kefir grains after acid and bile stress and we observed that they survive simulated gastrointestinal passage in different levels depending on the strain. L. paracasei CIDCA 8339, 83120 and 83123 were more resistant than L. paracasei...
The ability of probiotic bacteria to bind to human intestinal mucus
FEMS Microbiology …, 1998
Human mucus was isolated from faecal samples of newborns, two and six month old infants and adults. The adhesion to this mucus by the bacteria mentioned below was assessed in vitro. Depending on the age group: 44–46% of the applied Lactobacillus GG, 23–30% of Bifidobacterium lactis Bb-12, 9–14% of Lactobacillus johnsonii LJ-1, 3–10% of Lactobacillus salivarius LM2-118, Lactobacillus crispatus M247, Lactobacillus paracasei F19 and 2% of L. crispatus Mu5 adhered. All the strains adhered better to the mucus of adults than to that of infants. With some of the strains significant differences between the infant age groups were also observed. In conclusion, the age of the target group may be worthy of consideration when planning a schedule for probiotic or functional food therapy.