In this study, we investigated the spray-drying microencapsulation of β-carotene in oil co-stabilized by soy protein isolate-epigallocatechin-3-gallate conjugate (SPE) and small molecule surfactants [sodium dodecyl sulfate (SDS), hexadecyl trimethyl ammonium bromide (CTAB), and tea saponin (TS)] of different concentrations [0.1, 0.5, and 1.0% (w/v)], as a prospective approach to stabilize β-carotene. The results show that different surfactant types and concentrations significantly affect the encapsulation efficiency, water dispersibility, microstructure, and digestion of the microcapsules. Interactions between the surfactants and the SPE at the interface were found to include both synergistic and competitive effects, and they depended on the surfactant type and concentration. Moreover, the addition of SDS and TS before spray drying significantly improved the microencapsulation performance of the microcapsules and the water dispersion behavior of the corresponding spray-dried powders. The highest encapsulation efficiency was achieved for the SPE-0.1TS-encapsulated β-carotene microcapsules. In contrast, the addition of CTAB was not conducive to microcapsule formation, resulting in poor encapsulation efficiency, water dispersibility, thermal stability, β-carotene retention rate, and oxidation stability. In vitro gastrointestinal digestion results revealed that the addition of CTAB promotes the release of β-carotene and improves the bioaccessibility of β-carotene. In contrast, except for SPE-1.0SDS, the addition of SDS and TS inhibited β-carotene release and reduced β-carotene bioaccessibility. This study demonstrated that this novel β-carotene encapsulation formulation can overcome stability limitations for the development of β-carotene supplements with a high bioaccessibility.