Human milk fat substitute (HMFS) is structured lipids with characteristics similar to human milk fat used in infant formulas. HMFS is designed to contain 60%-70% palmitic acid at the sn-2 position and unsaturated fatty acids at the sn-1,3 positions in the triacylglycerol structure. HMFS is synthesised by enzymatic interesterification of vegetable oils or animal fats. Generally, the type of HMFS synthesised is sn-2 palmitate, with the main triacylglycerol being 1,3-dioleoyl-2-palmitoylglycerol (OPO). Palm oil fractions are used as raw material to synthesise sn-2 palmitate because it contains high palmitic acid. This article reviews the synthesis of sn-2 palmitate based on palm oil fractions via enzymatic interesterification. It gives a detailed description of the potential of palm oil fractions as substrates, lipases as biocatalysts, methods and reactors synthesis, and fractionation to increase the triacylglycerol of OPO. The information presented in this review can be used to develop future strategies for the synthesis of palm-based HMFS.