2‐DE and MS analysis of interactions between Lactobacillus fermentum I5007 and intestinal epithelial cells

Abstract

Lactobacillus is a probiotic commonly used for supplementation to human and animal diets. In this study, we used 2-DE and MS to analyze changes in the proteomes of Lactobacillus and intestinal epithelial cells in two model systems. The in vivo and in vitro models were involved the inoculation of Lactobacillus fermentum I5007 into the rabbit jejunum for 4 h and the culture of the bacterium with Caco-2 cells for 1 h, respectively. Our results indicate that, after exposure to the intestinal environment, the bacterium exhibited decreases in key enzymes involved in energy metabolism (e.g., lactate dehydrogenase, dihydrolipoamide dehydrogenase, and nicotinate phosphoribosyltransferase) and amino acid metabolism (e.g., arginyl-tRNA synthetase and aspartate-semialdehyde dehydrogenase), but increases in glycoside hydrolase (an enzyme for mucin degradation) and fructose-6-phosphate phosphoketolase (an enzyme of the pentose phosphate pathway). In response to an interaction with L. fermentum I5007, Caco-2 cells showed changes in proteins that were beneficial for gut integrity, including voltage-dependent anion channel 1, glutathione transferase, and heat shock protein gp96. On the basis of their functions, we suggest that these proteins serve as useful biomarkers for metabolic changes in Lactobacillus and intestinal epithelial cells in response to their interactions.

Isolation and identification of bacteriocinogenic strain ofLactobacillus plantarumwith potential beneficial properties from donkey milk

“…To investigate in vitro interactions between probiotic bacteria and the intestinal mucosa, several cell lines and different culturing models have been used elsewhere (Boti c et al 2007;Yang et al 2007;Ewaschuk et al 2008;Zoumpopoulou et al 2009). The human colon tumorigenic cell lines CaCo2, T84 and HT-29 have been widely utilized for bacterial adhesion assays and mechanistic studies but are not an appropriate model for pathogenhost interactions, because they do not derive from the small intestine, have a tumorigenic phenotype distinct from that of normal gut epithelia, express modified surface glycoconjugates and are much more glycosylated than normal cells.…”

Effect of three lactobacilli with strain-specific activities on the growth performance, faecal microbiota and ileum mucosa proteomics of piglets

“…The protein spots of interest were confirmed in the Coomassie Brilliant Blue stained gels and manually excised for the treatment of digestion by trypsin. The matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was used for protein identification as described by early reports [24, 25]. The peptide fragments produced from each protein spot were employed to produce peptide-mass mapping (PMM) data.…”

Defatted Biomass of the Microalga, Desmodesmus sp., Can Replace Fishmeal in the Feeds for Atlantic salmon

“…Interestingly, the energy digestibility of feeds that contained the algal biomass was lower compared to that of the control feed, significantly in the case of feeds with 10% of the algal biomass. A decrease in Dld was suggested to help probiotic bacteria such as Lactobacillus fermentum to survive in the small intestine (Yang et al, 2007). The physiological relevance of the underexpression of the three proteins in the distal intestine needs to be verified through further studies.…”