Combining phospholipase C hydrolysis of soybean concentrated phospholipids with column chromatography to produce of high-purity phosphatidylinositol

Phosphatidylinositol (PI) is vital for studying metabolic diseases and cancer; however, its purification is challenging. An enzymatic-chromatographic approach for producing high-purity PI from concentrated soybean phospholipids was developed in this study. This method employs phospholipase C (PLC), which selectively hydrolyses phosphatidylcholine (PC) and phosphatidylethanolamine (PE) but leaves PI intact. This selective reaction serves as a critical pretreatment step for enriching PI content in mixtures. The optimal reaction conditions included a SCPs to water ratio of 1:1.57 (g/mL), temperature of 39.6 °C, PLC dosage of 22 mg/g, and reaction time of 3.3 h, complete hydrolysis of PE and PC was achieved. Subsequently, defatting and deglycosylation the hydrolysed phospholipids concentrated the PI from 152.90 to 513.30 mg/g. This step markedly facilitated the downstream PI purification process. Chloroform-methanol volume ratio (1.8:1) chromatography at a flow rate of 8 mL/min was used to purify the PI further, yielding a high-purity product with a purity of 91.86 g/hg and yield of 77.85 g/hg. High-purity PI is suitable for medical applications owing to its lipid properties. This efficient purification strategy paves the way for the development of new therapeutics. • New enzymatic-chromatographic method for high-purity PI from SCPs was established. • PE/PC fully hydrolyze without damaging PI at optimized PLC hydrolysis conditions. • Defatting and deglycosylation raised PI in PLC hydrolysis SCPs from 15.29 to 51.33 g/100 g. • Novel method produced high-purity PI at 91.86% purity with 77.85% yield.