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Layer-by-layer technology (LBL) and enzymatic cross-linking technology were used to make modified β-carotene emulsion. Two kinds of modified β-carotene emulsion were prepared by using whey protein isolate as emulsifier, and then 0.15% sugar beet pectin was added to cover the surface of particles under the action of electrostatic adsorption and enzymatic cross-linking. The results showed that compared with original emulsions, particle size of modified emulsions increases; however, the bioavailability of β-carotene in modified emulsions is obviously lower than that in original emulsion, and it suggests that β-carotene in modified emulsions, especially in emulsion modified through enzymatic cross-linking, has sustained-release function.