Present study addresses the fortification of mayonnaise with fish protein hydrolysate by partial replacement of egg yolk in the product. Red meat of yellowfin tuna (Thunnus albacares), a by-product from tuna canning industry was used as the source of protein hydrolysate (TPH). Substitution of egg yolk with hydrolysate imparted noticeable fish flavor to the product only at higher levels of replacement (50% and above). Emulsion stability of mayonnaise samples was not significantly affected at the given range of substitution. The preliminary product acceptability parameters indicated higher desirability with a sensory score of 7.6 ± 0.7 for a replacement ratio of 1:2::TPH:egg yolk. Hence, the same combination was further subjected to morphological, rheological characterization, and opted for storage stability studies. Fortified mayonnaise exhibited lower particle size, indicative of higher emulsion quality which was also evident in the rheological properties of the sample. Results indicated better oxidative and physicochemical stability for fortified samples compared to control under chilled conditions, suggesting the applicability of fish protein hydrolysate as fortifying and stabilizing agent in mayonnaise preparations.