# Enzymatic synthesis of phosphatidyl-EPA/DHA using Candida antarctica lipase B immobilized on mesoporous MIL-88 A

## TL;DR

This paper presents a new method for making health-benefiting phosphatidyl-EPA/DHA using an immobilized enzyme on a mesoporous material, achieving high efficiency and eco-friendly production.

## Contribution

A novel green synthesis of Meso-MIL-88A and its use for immobilizing lipase to efficiently produce phosphatidyl-EPA/DHA.

## Key findings

- Meso-MIL-88A synthesized via water-washing showed larger pores and less sulfur residue than ethanol-based methods.
- CalB@Meso-MIL-88A achieved 86.8% sn-1 positional incorporation of EPA/DHA with high donor efficiency.
- Molecular docking showed higher DHA-EE substrate affinity due to improved transfer efficiency in solvents.

## Abstract

Mesoporous MIL-88A (Meso-MIL-88A) shows significant potential as an effective carrier for immobilizing large molecules such as lipases. This study investigates Meso-MIL-88A immobilized lipase for the catalysis of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) incorporation into soybean phosphatidylcholine. Phosphatidyl-EPA and phosphatidyl-DHA are known for their superior health benefits and have garnered significant global attention. Firstly, we developed a novel and green water-washing strategy to synthesize Meso-MIL-88A, demonstrating its significant potential as an effective carrier for immobilizing large molecules such as lipases. Nitrogen adsorption/desorption and XPS analyses revealed that water as the eluent yielded larger average pore diameters and lower sulfur residue content compared to ethanol. This optimized Meso-MIL-88A carrier was used to immobilize Candida antarctica lipase B (CalB@Meso-MIL-88A), which was applied to the production of phosphatidyl-EPA and phosphatidyl-DHA. The immobilized CalB@Meso-MIL-88A exhibited exceptional catalytic efficiency, achieving an unprecedented sn-1 positional incorporation rate of 86.8% (44.2% EPA, 42.7% DHA) with 90% EPA/DHA-ethyl ester (EE) donors, while 97% DHA-EE resulted in a record-high 90.1% DHA incorporation at the sn-1 position. Kinetic studies and molecular docking simulations indicated a higher substrate affinity for DHA-EE, attributed to enhanced transfer efficiency of EPA/DHA-EE in organic solvent system. This study presents the first demonstration of the potential of Meso-MIL-88A for industrial lipase immobilization via this optimized route, offering an eco-friendly and highly efficient catalytic application for nutraceutical synthesis.

The online version contains supplementary material available at 10.1186/s40643-025-00959-5.