The β-carotene emulsion system using high-acyl gellan gum (HA) as an emulsifier was fabricated and systematically studied. The stability and stabilizing mechanism of the emulsion using medium-chain triglyceride as oil phase with a water-oil mass ratio of 9:1 under different physicochemical conditions of heat, pH, and ions were investigated by analyzing mean particle size (MPS), emulsion yield (EY), and dynamic stability. The effects of the HA-β-carotene emulsion system on the bioaccessibility of β-carotene in vitro were conducted. During the simulated oral digestion stage (SODP) and simulated gastric digestion stage (SGDP), the emulsion systems stabilized with different HA contents showed good stability, and the changes of MPS and zeta potential (ZP) were within 2.5 μm and 3.0 mV, respectively. After entering the simulated intestinal digestion phase (SIDP), β-carotene was released from oil droplets and formed micelles with bile salts, phospholipids, etc. HA-β-carotene emulsion can enhance the release rate of free fatty acid (FFA), which ultimately affects the β-carotene bioaccessibility. These results indicate that HA can be used to prepare carotene emulsion and improve its bioavailability. The study provides a reference for the application of HA as a natural emulsifier and the delivery of β-carotene.