Abstract: The breastfeeding rate is generally low in China and some other developing countries due to economic, social and personal factors. Human milk fat has a high content of palmitic acid at the sn-2 position of the glycerol backbone compared with ruminant milk fat and common vegetable oils. Such a unique structure helps the simultaneous absorption of fatty acids in the gut lumen of the infant and the loss of calcium through the feces. As an important ingredient in infant formula, human milk fat substitutes with high similarity to human milk fat have great potential in industrial applications. Human milk fat substitutes can be synthesized using sn-1, 3specific lipases due to mild reaction conditions, high sn-1, 3-regioselectivity and fatty acid specificity of these lipases. In this article, the definition and enzymatic synthesis of human milk fat substitutes were reviewed, and study on human milk fat substitutes in the future was pointed out. Particular attention was paid to choice of natural oils and acyl donors, lipase screening, synthesis methods and techniques, analytical procedures, similarity evaluation and oxidation stability.