Co-encapsulation of acylglycerols and probiotics may improve the resistance of probiotics to adverse conditions. In this study, three probiotic microcapsule models were constructed using gelatin (GE)-gum arabic (GA) complex coacervate as wall material: microcapsules containing only probiotics (GE-GA), microcapsules containing triacylglycerol (TAG) oil and probiotics (GE-T-GA) and microcapsules containing diacylglycerol (DAG) oil and probiotics (GE-D-GA). The protective effects of three microcapsules on probiotic cells under environmental stresses (freeze-drying, heat treatment, simulated digestive fluid and storage) were evaluated. The results of cell membrane fatty acid composition and Fourier transform infrared (FTIR) spectroscopy revealed that GE-D-GA could improve the fluidity of cell membrane, maintain the stability of protein and nucleic acid structure, and decrease the damage of cell membrane. These characteristics supported the high freeze-dried survival rate (96.24 %) of GE-D-GA. Furthermore, regardless of thermotolerance or storage, GE-D-GA showed the best cell viability retention. More importantly, GE-D-GA provided the best protection for probiotics under simulated gastrointestinal conditions, as the presence of DAG reduced cell damage during freeze-drying and the degree of contact between probiotics and digestive fluids. Therefore, co-microencapsulation of DAG oil and probiotics is a promising strategy to resist adverse conditions.