1-油酸-2-棕榈酸-3-亚油酸甘油三酯（OPL）是人乳中主要的脂质组成，为满足婴幼儿配方食品母乳化需求，该试验探究了新型固定化脂肪酶ANL-MARE催化制备OPL的方法。试验成功制备和表征了固定化酶ANL-MARE，并以ANL-MARE为生物催化剂，建立了一种用三棕榈酸甘油三酯（PPP）、油酸（OA）和亚油酸（LA）高效酶催化制备富含OPL结构脂的方法。经单因素和响应面试验优化，获得OPL最优合成工艺为：PPP与总脂肪酸的摩尔比1:14.27，OA与LA摩尔比为1:0.76，脂肪酶添加量12.70%，反应温度50 ℃，反应4 h，此条件下产物中OPL相对含量为47.93%，2位棕榈酸（sn-2PA）占总棕榈酸（PA）的质量分数（sn-2PA相对含量）为71.69%。此外，固定化酶ANL-MARE与商业脂肪酶相比，表现出较好的催化合成OPL的活性。综上所述，固定化酶ANL-MARE具有催化制备OPL结构脂的重大潜力，为人乳脂替代脂的高效制备提供了新策略和理论基础。

The demand for infant formula similar to breast milk is growing. To address this, this study investigated the catalytic preparation of 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride (OPL), the main lipid component in human milk, using a new immobilized lipase, ANL-MARE. The immobilized lipase ANL-MARE was successfully prepared and characterized. Next, an efficient enzyme-catalyzed preparation of lipids rich in OPL structures from tripalmitin, oleic acid, and linoleic acid was established using ANL-MARE as the biocatalyst. After optimization via single-factor and response surface experiments, the optimal OPL synthesis process was obtained, with the following characteristics: a tripalmitin: total fatty acids molar ratio of 1:14.27; an oleic acid:linoleic acid molar ratio of 1:0.76; the addition of 12.70% ANL-MARE; a reaction temperature of 50 ℃ , and a reaction time of 4 h. Under these conditions, the relative OPL content reached 47.93%, and sn-2 palmitic acid accounted for 71.69% of the total palmitic acid content (reflecting the relative sn-2 palmitic acid content). In addition, the immobilized lipase ANL-MARE showed better catalytic activity than commercial lipases in OPL synthesis. In conclusion, the immobilized lipase ANL-MARE has significant potential to catalyze the preparation of lipids with OPL structures. This study provides a novel strategical and theoretical basis for the efficient preparation of human milk fat substitutes.

广东省科技厅重点实验室项目-广东省功能食品活性物重点实验室（2018B030322010）；广州市重点研发计划项目（SL2022B03J01001）；中 山市科技局项目-广式酱油风味控制与智能制造研究创新团队项目（CXTD2020006）