Lactobacillus reuteri is a resident of the human and animal intestinal tracts. The ability of L. reuteri to survive passage through the intestinal tract is a key point in its function as a probiotic. In order to examine the nature of bile salt tolerance by L. reuteri, its protein synthesis was analyzed in liquid cultures containing two different bile salt conditions. Significant cell growth inhibition was observed in the presence of 1.2g/L (higher concentration) bile salts. Two-dimensional gel electrophoresis allowed us to identify 28 proteins spots that were consistently and significantly altered in the presence of bile in the growth medium. Peptide mass fingerprinting was used to identify these 28 proteins, and functional annotation revealed their involvement in carbohydrate metabolism, transcription-translation, nucleotide metabolism, amino acid biosynthesis, pH homeostasis and stress responses, oxidation-reduction reactions, and unknown functions. These findings, which suggest that bile salts induce complex physiological responses in L. reuteri may provide early new insights into the inducible mechanisms underlying the capacity of intestinal L. reuteri to tolerate bile stress.