The aim of the study is to investigate the possible usage of oleaster fruit flour (OFF) (Elaeagnus angustifolia) in mayonnaise formulation by reducing the fat and egg yolk ratios. Mayonnaise samples with four different formulations as control, low-fat low-cholesterol mayonnaise-I (LFCM-I), low-fat low-cholesterol mayonnaise-II (LFCM-II) and low-fat low-cholesterol mayonnaise-III (LFCM-III) were produced. The oil, egg yolk and sugar ratios were decreased with the incremental percentage of OFF. The substitution of egg yolk and fat in the mayonnaise formula by the addition of OFF caused an increase in pH value, viscosity, firmness, consistency and work of shear, whereas a decrease in the acidity, total cholesterol content, peroxide value, TBARs value and conjugated dienes ratio. Considering all of the physical and chemical analysis results, the OFF can be used as a fat and egg yolk substitute in the mayonnaise formula; however, it was concluded that the mayonnaise-like sauce, which has higher emulsion stability and better physical characteristics, can be obtained by combining OFF with various stabilizers.