Conjugated Linoleic Acid（CLA） is a general term for a kind of octadecadienoic acid with different positions and geometric isomers. CLA have different physiological functions because of its various structures. Currently， c9， t11-CLA and t10， c12-CLA are the active isomers that are investigated much. Biosynthesis method is favored by researchers because of its uni-structure and higher safety. In this study， Ten strains are screened and identified from five traditional fermentation yogurts of Inner Mongolia and the fermentation performance of Lactobacillus plantarum HB01 with high yield of CLA was determined. The optimal fermentation conditions by uniform design are as follows： 4% inoculum， 0.7 mg/mL LA， fermentation for 32 h at pH 6.5 and 37 ℃， ripening for 12 h at 4 ℃. Provide a certain basis for further increasing CLA produce.