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• Synergistic interaction between sodium cholate and Tween 80 were explored. • The effect of the crowding media on the digestion varies with oil compositions. • The lipolysis rate and transformation increased when using binary emulsifiers. • The denaturation temperature of lipase changed in the crowding media. • The lipolysis in crowding media was affected by the stability of lipase. Control of lipid digestibility by various food components has received great attention in recent decades. However, there is limited literature on investigating the synergistic effect of exogenous emulsifiers and endogenous sodium cholate (SC) on lipid digestion in a simulated physiological crowded medium. In this work, the synergistic interaction of Tween80 and SC according to the regular solution theory, and the hydrolysis of lipid emulsions containing tricaprylin, glyceryltrioleate or soybean oil in crowding medium was studied. The results show that emulsions stabilized by a combination of Tween80 and SC showed higher digestion rate and transformation than those with Tween80 or SC. The digestion rate could be increased by polyethylene glycols (PEGn) with varying crowding degree. The denaturation temperature of the lipase was increased in macromolecular crowded medium. This work allows for better understanding of the interaction between the amphiphiles and the macromolecular crowding effect on lipase digestion in the physiological environment. [ABSTRACT FROM AUTHOR]