Inflammatory bowel disease (IBD) is an intestinal chronic inflammatory disease, and its incidence is steadily increasing. IBD is closely related to the intestinal microbiota, and probiotics are known to be a potential therapeutic agent for IBD. In our study, we evaluated the protective effect ofLactobacillus sakeiCVL-001, isolated fromBaechukimchi, on dextran sulfated sodium (DSS)-induced colitis in mice. The oral administration ofL. sakeiCVL-001 according to the experimental schedule alleviated weight loss and disease activity in the mice with colitis. Furthermore, the length and histopathology of the colon improved. The expression of tumor necrosis factor (TNF)-αand interleukin(IL)-1βgenes decreased in the colons of mice that were administeredL. sakeiCVL-001, whereas that ofIL-10increased. The expressions of genes coding for E-cadherin, claudin3, occludin, and mucin were also restored. In co-housed conditions,L. sakeiCVL-001 administration did not improve disease activity, colon length, and histopathology. Microbiota analysis revealed thatL. sakeiCVL-001 administration increased the abundance of microbiota and alteredFirmicutes/Bacteroidetesratio, and decreasedProteobacteria. In conclusion,L. sakeiCVL-001 administration protects mice from DSS-induced colitis by regulating immune response and intestinal integrity via gut microbiota modulation.