In this study, α-linolenic acid-enriched diacylglycerols (ALA-DAGs) were prepared via a two-step enzymatic way by combi-lipase using silkworm pupae oils as substrates. Firstly, several factors including temperature, mass ratio of water to oil, pH and enzyme loading were optimized for the hydrolysis of silkworm pupae oil. The maximum fatty acid content (96.51%) was obtained under the conditions: temperature 40 °C, water/oil 3:2 (w/w), pH 7, lipase TL100L loading 400 U/g, lipase PCL loading 30 U/g. Then, ALA was enriched by urea inclusion, with an increased ALA content of 82.50% being obtained. Secondly, the ALA-enriched silkworm pupae DAG oil (SPDO) was prepared by lipase PCL-catalyzed esterification reaction. After molecular distillation, the final SPDO product contained contents of DAGs (97.01%) and ALA (82.50%). This two-step enzymatic way for production of ALA–DAGs was successfully applied in a 100-fold scale-up reaction. Overall, our study provides a promising way for the preparation of ALA–DAGs.

在这项研究中，以蚕蛹油为底物，通过组合脂肪酶通过两步酶促方式制备了富含α-亚麻酸的二酰基甘油（ALA-DAGs）。首先，对蚕蛹油水解的温度、水油质量比、pH值和酶载量等几个因素进行了优化。最大脂肪酸含量 (96.51%) 是在以下条件下获得的：温度 40 °C，水/油 3:2 ( w / w)，pH 7，脂肪酶 TL100L 加载 400 U/g，脂肪酶 PCL 加载 30 U/g。然后，通过包含尿素来富集 ALA，获得了 82.50% 的增加的 ALA 含量。其次，通过脂肪酶PCL催化的酯化反应制备了富含ALA的蚕蛹DAG油(SPDO)。分子蒸馏后，最终的 SPDO 产品含有 DAGs (97.01%) 和 ALA (82.50%)。这种用于生产 ALA-DAG 的两步酶促方法已成功应用于 100 倍放大反应。总的来说，我们的研究为制备 ALA-DAGs 提供了一种有前景的方法。