Candida albicans has been recently added to the WHO critical priority group of fungal pathogens based on its impact on global public health. C. albicans is a mucosal commensal yeast in humans that can cause severe gastrointestinal-borne disseminated candidiasis in immunocompromised patients. C. albicans interaction with enterocytes is thus a key step in the pathophysiology of disseminated candidiasis. Here, we show that, during the first steps of the infection process, C. albicans releases a peptide from the Repressed by Tup1 protein 1 (Rbt1), which disorganizes the cell‒cell adhesion junctions in Caco-2 intestinal epithelial cells by notably downregulating constitutive proteins of the tight junction complex (i.e. ZO-1). This is the first report to show that a peptide of a human pathogenic fungus promotes fungal invasion into the gut epithelium by disorganizing the protective epithelial barrier.