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The use of probiotics has been considered as a new therapy option for ulcerative colitis (UC), and yeast has recently received widespread recommendation for human health. In this study, the probiotic characteristics of four yeast strains, <i>Saccharomyces boulardii</i> CNCMI-745, <i>Kluyveromyces marxianus</i> QHBYC4L2, <i>Saccharomyces cerevisiae</i> QHNLD8L1, and <i>Debaryomyces hansenii</i> QSCLS6L3, were evaluated in vitro; their ability to ameliorate dextran sulfate sodium (DSS)-induced colitis was investigated. Among these, <i>S. cerevisiae</i> QHNLD8L1 protected against colitis, which was reflected by increased body weight, colon length, histological injury relief, decreased gut inflammation markers, and intestinal barrier restoration. The abundance of the pathogenic bacteria <i>Escherichia–Shigella</i> and <i>Enterococcaceae</i> in mice with colitis decreased after <i>S. cerevisiae</i> QHNLD8L1 treatment. Moreover, <i>S. cerevisiae</i> QHNLD8L1 enriched beneficial bacteria <i>Lactobacillus</i>, <i>Faecalibaculum,</i> and <i>Butyricimonas,</i> enhanced carbon metabolism and fatty acid biosynthesis function, and increased short chain fatty acid (SCFAs) production. Taken together, our results indicate the great potential of <i>S. cerevisiae</i> QHNLD8L1 supplementation for the prevention and alleviation of UC.