AbstractSiphonaxanthin is a carotenoid found in certain green algae, and its promising beneficial properties, such as its anti‐obesity effect, have recently been demonstrated. However, there is little information about the molecular mechanisms underlying intestinal absorption of siphonaxanthin. In this study, we aimed to elucidate how siphonaxanthin is transported across the intestinal epithelium using differentiated Caco‐2 cells (dCaco‐2 cells), recombinant proteins, and an animal model. Siphonaxanthin was taken up by dCaco‐2 cells, a model of intestinal epithelial cells, and its uptake linearly increased up to at least 6 h. Pharmacological inhibition of Nieman‐Pick C1‐like 1 (NPC1L1), but not that of scavenger receptor class B type 1 (SR‐B1), significantly suppressed siphonaxanthin uptake by dCaco‐2 cells. Results from an in vitro binding assay suggested that the N‐terminal domain of NPC1L1, which is an extracellular domain of NPC1L1, binds with siphonaxanthin. Moreover, pretreatment with ezetimibe, an inhibitor of NPC1L1, significantly decreased the plasma level of siphonaxanthin following oral administration in mice. Considered together, we concluded that NPC1L1 promotes siphonaxanthin transport across the intestinal epithelium.