Preparation of diacylglycerols from Acer truncatum seed oil via enzymatic glycerolysis and evaluation of their neuroprotective effects

Acer truncatum seed oil (ATO), rich in nervonic acid, was enzymatically converted to diacylglycerols (DAG) via optimized glycerolysis (1:1 ATO:glycerol ratio, 2 % Novozym 435, 60 °C, 24 h), achieving a >50 % DAG yield. The process demonstrated scalability (50-fold) and enzyme reusability (6 cycles), yielding 50 % DAG content in scaled batches. Purification through molecular distillation and silica gel chromatography produced high-purity DAG (98.7 %) while preserving fatty acid composition. In H₂O₂-induced PC12 cells, ATO-derived DAG (40 μg mL⁻¹) enhanced cell viability by 32 % (vs. H₂O₂ controls), reduced reactive oxygen species (ROS) by 49 %, lowered malondialdehyde (MDA) levels to 20.8 μM mg⁻¹ protein, and restored the activities of antioxidant enzymes, including superoxide dismutase (SOD) and glutathione peroxidase (GPx). These results establish enzymatic glycerolysis as a scalable method for neuroprotective DAG production and position ATO-derived DAG as a promising therapeutic against oxidative neurodegeneration.