# Deep Eutectic Solvent Ultrasonic-Assisted Extraction of Polysaccharides from Red Alga Asparagopsis taxiformis: Optimization, Characterization, Mechanism, and Immunological Activity in RAW264.7 Cells

## TL;DR

A new method using deep eutectic solvents and ultrasound improves the extraction of polysaccharides from a red alga, but the extracted material has lower immunological activity than traditional methods.

## Contribution

The study introduces an optimized DES-ultrasonic extraction method for A. taxiformis polysaccharides and reveals its impact on structure and bioactivity.

## Key findings

- DES extraction achieved a 1.5 times higher yield than water extraction.
- The DES-extracted polysaccharide showed significantly lower immunological activity in macrophages.
- Molecular simulations suggest DES enhances galactose solubility through stronger hydrogen bonding.

## Abstract

Traditional polysaccharide extraction suffers from low efficiency and high energy consumption, while deep eutectic solvents (DESs) are promising sustainable solvents. This study used DES ChCl-LA (1:2) with ultrasonic assistance to extract polysaccharides from red alga A.
taxiformis. Optimized via single-factor experiments and response surface methodology (350 W, 1:30 g/mL, 75 °C), the yield reached 11.28% ± 0.50% (1.5 times higher than that obtained by water extraction). Structural characterization revealed that the DES extract was an acidic polysaccharide, mainly composed of galactose (89.2%), glucose (4.9%), xylose (4.9%), and glucuronic acid (1.0%), with a weight-average molecular weight of 99.88 kDa. Density functional theory and molecular dynamics simulations showed that ChCl-LA enhanced galactose solubility via stronger hydrogen bonding (−25.33 vs. −5.06 kcal/mol for water). Notably, the immunological activity of the DES-extracted polysaccharide was significantly compromised compared to the water-extracted counterpart (p < 0.05). At a concentration of 0.25 mg/mL, the water-extracted polysaccharide-treated group exhibited a 33.98% higher neutral red phagocytosis rate in macrophages, a nitric oxide (NO) secretion level of 34.14 μmol/L (94.98% higher) compared with the DES-extracted polysaccharide group, as well as significantly higher secretion levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). The observed disparity in bioactivity is likely due to the distinct chemical profiles resulting from the two extraction methods, including the significantly reduced molecular weight and potential alterations of sulfation degree, monosaccharide composition, and protein content in the DES-extracted polysaccharide. This mechanistic perspective is supported by the relevant literature on the structure–activity relationships of polysaccharides. This study demonstrates the potential of ChCl-LA and elucidates the complex effects of extraction methods on polysaccharide’s structure and function, thereby informing the high-value utilization of A. taxiformis in functional foods.