Development of an enzymatic method for efficient production of DHA-enriched phospholipids through immobilized phospholipase A1 in AOT-water reverse micelles

• Published in: Enzyme and microbial technology 2025-04, Vol.185, p.110600, Article 110600
• Main Authors: Gao, Qin, Yu, Xiaolin, Wei, Jianming, Hu, Xuechao, Ren, Lujing
• Format: Article
• Language: English
• Subjects: Immobilization; Phospholipase A1; Phospholipid-DHA; Reverse micelle system
• ISSN: 0141-0229; 1879-0909; 1879-0909
• DOI: 10.1016/j.enzmictec.2025.110600

The demand for omega-3 polyunsaturated fatty acids (PUFAs), particularly docosahexaenoic acid (DHA), has been steadily increasing due to their significant health benefits. Traditional methods for producing DHA-enriched phospholipids often suffer from low efficiency and high costs. In this study, we developed an efficient enzymatic process to prepare phospholipid-DHA, which used immobilized phospholipase A1 to catalyze transesterification in AOT-water reverse micelle systems. Initially, high concentrations of free fatty acids were produced via acid hydrolysis of algae oil followed by crystallization. Among six evaluated reverse micelle systems, one was selected for further optimization. The substrate/enzyme ratio, temperature, reaction time, and water content were optimized using single-factor experiments and response surface methodology. To enhance cost-efficiency and eco-friendly practices, substrate recycling was implemented to maximize substrate utilization. This study established a comprehensive process chain for the preparation of phospholipid-DHA, promoting its industrial production and providing a reference for the production of other phospholipid products. [Display omitted] •AOT-water reverse micelle systems was used in enzymatic production of PL-DHA.•Process optimization coupled with AOT system enhanced the DHA incorporation to 63.71 %.•Substrate recycling strategy was implemented to maximize substrate utilization.•An integrated process from fatty acid preparation to PL-DHA synthesis was established.