In the present study, Lactobacillus acidophilus LA-5 was microencapsulated in sodium alginate, followed by fish gelatin coating (0.5, 1.5, and 3%). The survival of L. acidophilus in bread before and after encapsulation in alginate/fish gelatin during the baking and 7-day storage was investigated. Moreover, the effect of alginate/fish gelatin-encapsulated L. acidophilus on the technological properties of bread (hardness, staling rate, water content, oven spring, specific volume, and internal texture structure) was evaluated. Compared with control (free bacteria), encapsulated L. acidophilus in alginate/fish gelatin showed an increase in the viability of bread until 2.49 and 3.07 log CFU/g during baking and storage, respectively. Good viability of (106 CFU/g) for probiotic in encapsulated L. acidophilus in alginate/fish gelatin (1.5 and 3%, respectively) after 4-day storage was achieved. Fish gelatin as a second-layer carrier of the bacteria had a positive effect on improving the technical quality of bread. Furthermore, the staling rate of bread containing encapsulated L. acidophilus alginate/fish gelatin 0.5, 1.5, and 3% decreased by 19.5, 25.8, and 31.7%, respectively. Overall, the findings suggested encapsulation of L. acidophilus in alginate/fish gelatin capsule had great potential to improve probiotic bacteria’s survival during baking and storage and to serve as an effective bread enhancer.