This paper focused on the fermentation of conjugated linoleic acid(CLA) using immobilized lactic acid bacteria through embedding method.The results showed that the best diameter of immobilized ball was 2~3 mm.Furthermore,immobilized ball should be immersed in the mixture of calcium chloride and boric acid overnight.The best ferment conditions of immobilized ball were as follows: fermentation temperature of 37 ℃,initial pH of 6.5,fermentation time of 28 h,the substrate adding amount of 0.3 %,and maximum yield of CLA reached 92.061 μg/mL.7 batches of conjugated linoleic acid fermentation have been carried out using the immobilized lactic acid bacteria ball,and their outputs were higher than 80.476 μg/mL.And fermentation production of CLA using immobilized UV3-9 was 1.28 times of which using free UV3-9.