The purpose of this study was to develop a structured lipid (named DCAO) from amaranth oil to resemble breast milk fat in milk-based infant formula. We increased the content of palmitic acid in amaranth oil using Novozym 435 lipase, specifically at the sn-2 position from 2.1 to 20.8%. Docosahexaenoic acid (DHA) was incorporated at the sn-1,3 positions using specific Lipozyme RM IM lipase. DCAO contained 33.0% palmitic acid and 0.9% DHA. Sterol content increased 19% compared to amaranth oil, however interesterification and purification procedures reduced the content of tocopherols (94.5%) and squalene (99.8%). Antioxidants are recommended even though we observed good oxidative stability. DCAO showed higher melting temperatures, iodine and saponification values than amaranth oil. The infant formula containing DCAO closely resembled breast milk fat composition, meeting the recommended DHA (0.2%) content, and supplying more palmitic acid at the sn-2 position (33.0%), for proper development and better fat absorption, respectively.