The present study investigated the deconjugation of bile salts and co-precipitation of cholesterol with deconjugated bile salts by seven Propionibacterium spp. strains in vitro. Propionibacterium spp. could deconjugate sodium glycocholate (1.53-5.31 mM) and sodium taurocholate (0.08-0.25 mM) bile salts. The highest cholesterol precipitation (47.8 mu g/mL) was determined with Propionibacterium freudenreichii subsp. freudenreichii SP3 strain in a 0.15% oxgall-containing medium. Significant (P < 0.05) correlations were observed between cholesterol co-precipitation and deconjugation of sodium glycocholate among the strains. In vitro bile salt deconjugation activity studies of P. freudenreichii subsp. freudenreichii SP3 strain revealed that this strain may have potential as a probiotic strain for deconjugation of bile salts in vivo studies.