To develop new functional foods for the prevention of atherosclerosis, we evaluated the cholesterol-lowering effects of several strains of Lactobacillus (L.) brevis. These strains included the γ-amino butyric acid-producing strains L. brevis 119-2 and L. brevis 119-6, isolated from turnip “tsuda kabu” and pickled turnip “tsuda kabu zuke”, respectively. To evaluate the cholesterol-lowering effects of L. brevis in vitro and to examine potential mechanisms of this property, we evaluated changes in levels of cholesterol, sodium taurocholate, and cholic acid in growth medium containing cholesterol and sodium taurocholate. Moreover, to assess the potential of L. brevis 119-2 and L. brevis 119-6 for probiotic use, we examined the tolerance of L. brevis to artificial digestive fluid and its adherence to epithelial cells. Our results indicate that L. brevis 119-2 incorporates cholesterol from the medium into the cell, and that L. brevis 119-2 and L. brevis 119-6 are probiotic bacteria. Therefore, we propose that viable cells of L. brevis 119-2 can reach the small intestine, where they can inhibit intestinal absorption of cholesterol in vivo by incorporation of cholesterol into the cell.