Milk fat globules (MFG) are the primary source of energy and functional factor in dairy-based products. To evaluate the suitability of commercial MFG substitutes for infant and adult consumers, the lipolysis properties and regulatory factors in digesting bovine, caprine, and human MFG were compared. The lipolysis kinetic models of bovine and caprine MFG are comparable, with bovine MFG displaying the highest degree of lipolysis (50.21%) during gastric digestion in infants, resulting in the production short-chain saturated fatty acids. Caprine MFG has a high rate of lipolysis due to its weak phospholipid-glycoprotein layer, which is prominent in both infants’ and adults’ digestion. The effects of consuming bovine and caprine MFG for adults are comparable in terms of lipolysis properties and nutritional value of fatty acid. After gastrointestinal digestion, human MFG has the highest digestibility (93.98%). The advantages of human MFG include its phospholipid structure, exogenous lipase, and lipolysis-regulating proteins. This study provides recommendation for the selection of milk fat substitutes and improvements in commercial dairy.