A coacervation method with double emulsion strategy (w/o/w) was used to prepare immobilized Lactobacillus E1. Diatomite was chosen as the carrier for bacteria. Sodium alginate, dextrin and gelatin were used as protective solutes for the preservation of Lactobacillus E1 and their effects on the storage viability during storage were discussed. The influence of storage temperature on the storage viability was also examined. The results show that high bacteria viable count over 109 cfu/g for an extended shelf life of 37 d can be achieved with 2% sodium alginate, 5% dextrin and 4% gelatin as protective solutes, at 10 ℃ of the storage temperature. This immobilized Lactobacillus E1 has potential use as functional food ingredient for both human dairy food and animal feedstuff. A coacervation method with double emulsion strategy (w / o / w) was used to prepare immobilized Lactobacillus E1. Diatomite was chosen as the carrier for bacteria. Sodium alginate, dextrin and gelatin were used as protective solutes for the preservation of Lactobacillus E1 and their effects on the storage viability during storage were discussed. The influence of storage temperature on the storage viability was also examined. The results show that high bacteria viable count over 109 cfu / g for an extended shelf life of 37 d can be achieved with 2% Sodium alginate, 5% dextrin and 4% gelatin as protective solutes, at 10 ° C of the storage temperature. This immobilized Lactobacillus E1 has potential use as functional food ingredients for both human dairy food and animal feedstuff.