Food proteins are important nutritional constituents, as they act as sources of peptides and amino acids required in the diet. In addition to the factor, these nutritional organic macromolecules have biotechnological applications as sources of bioactive substances, such as bioactive peptides. These protein derivatives can act through the regulation of physiological functions in living organisms, or even in vitro actions that are beneficial for applications such as nutraceutical products, cosmetics, and/or food preservatives. Traditionally, dairy proteins are explored as sources of prospecting for bioactive peptides, especially those from bovine milk. However, unusual animal products such as donkey milk have demonstrated a high potential for use as a basic product for prospecting numerous bioactive peptides. Despite the high potential for prospecting bioactives from food proteins, bioactive peptides face challenges in living organisms until their absorption, such as the low permeability and selectivity of the intestinal membrane, the possibility of new enzymatic hydrolysis, and their consequent inactivation, manipulation by action of intestinal microbiota metabolism, among others. Due to these factors, they can reach low serum concentration levels, especially due to the rapid rate of renal clearance that results in easy elimination of the substance from the body. Therefore, the encapsulation technique, such as liposome vesicle technology, is an important technological instrument capable of promoting stability, and functioning as a controlled delivery mechanism for bioactives. The present study evaluated the in vitro biological activity of peptides with molecular mass less than 3 kDa prospected from the enzymatic hydrolysis of whole donkey milk proteins through the action of commercial trypsin. In general, the peptides, free and encapsulated in liposome vesicles, exhibited antioxidant actions against the oxidative radicals ABTS^•+, DPPH^•, superoxide, and hydroxyl, broad-spectrum antibacterial, anti-inflammatory activity through the inhibition of protein denaturation and photoprotective activity.