In conclusion, our study demonstrates that 2’FL supplementation in the context of HFD-feeding can counteract obesity and metabolic alterations and it is associated with alterations in the intestinal mucus layer through increased expression of secreted and transmembrane mucins, glycosyltransferases and alterations in mucin glycans composition. The gene discussed is MUC5AC; the disease is obesity due to melanocortin 4 receptor deficiency.