In order to develop a new type of probiotics, a yeast strain was isolated from intestine of mice, and it was identified using the methods of morphology, physiology, biochemistry, gene sequencing and phylogenetic tree. The results showed that the bacterial colony presented in white milky color and oval shape. Through microscopic observation, we found its cell presented in oval shape, and the asexual reproduction was gemmation. The yeast could ferment glucose, sucrose and D–mannitol, as well as assimilate peptone and potassium nitrate, while, it could not tolerate high concentration of alcohol, and the urease test was negative. The results from 18S rDNA sequencing analysis showed that DNA homology of the strain (Accession No. KC534843) and Debaryomyces hansenii (Accession No. AB013567) reached up to 99%, moreover, 26S rDNA sequencing analysis showed that the homology between the strain (Accession No. KC534842) and Debaryomyces hansenii (Accession No. EU131182) was also 99%. From the result of phylogenetic tree showed that 18s rDNA gene sequence of the strain was closest to Debaryomyces hansenii (Accession No. AB013590), and 26S rDNA gene sequence of the strain was closest to Debaryomyces hansenii (Accession No. EU131182). We could draw the conclusion that the yeast strain was Debaryomyces hansenii.