A cost-effective one-step enzymatic synthesis of 1-oleoyl-2-palmitoyl-3-linoleoylglycerol and its application in the preparation of high-similarity human milk fat substitutes with different sources

This study developed a cost-effective one-step enzymatic process for synthesizing 1-oleoyl-2-palmitoyl-3-linoleoylglycerol (OPL) and applied it to prepare human milk fat substitute (HMFS) with different sources. Fractionated tripalmitin and free fatty acids from a camellia/sunflower oil mixture were used as substrates. Reaction conditions were optimized using response surface methodology, yielding an OPL product containing 70.36% palmitic acid at the sn -2 position with 40.06% oleic acid and 37.53% linoleic acid at the sn -1,3 positions. The obtained OPL was subsequently used as a core ingredient to formulate three HMFSs: goat milk fat-based (HMFS-G), cow milk fat-based (HMFS-C), and plant oils-based (HMFS-P). Similarity evaluation demonstrated that HMFS-C achieved the highest overall similarity score (84.53) to human milk fat (HMF), followed by HMFS-G (81.66) and HMFS-P (77.35). The superior performance of HMFS-C was attributed to its optimized fatty acid and sn -2 fatty acid distributions, whereas the lower similarity of HMFS-P was due to its less optimal sn -2 fatty acid and triacylglycerol profiles. Compared with previously reported HMFSs and infant formulas, all three HMFSs exhibited improved structural resemblance to HMF. This study provides a practical and economically viable strategy for producing OPL and high-similarity HMFSs with different sources, demonstrating good potential for application in infant formula. • A cost-effective one-step enzymatic process was developed to synthesize OPL. • The optimized OPL showed 70.36% palmitic acid at the sn -2 position. • Three HMFSs were formulated based on goat milk fat, cow milk fat, and plant oils. • HMFS-C exhibited the highest overall similarity (84.53%) to human milk fat.