# Enzymatic Hydrolysis-Assisted Separation and Purification of High F-Value Oligopeptides from Sea Cucumbers and Their Anti-Fatigue Mechanism

## TL;DR

This paper explores a new method to purify sea cucumber peptides that reduce fatigue and explains how they work.

## Contribution

The study introduces an optimized enzymatic hydrolysis and membrane separation process to produce high F-value oligopeptides with anti-fatigue properties.

## Key findings

- A two-step enzymatic hydrolysis and membrane separation yielded 45.00% sea cucumber peptides and 51.28% polysaccharides.
- High-F-value oligopeptides increased swimming duration and grip strength in mice, indicating anti-fatigue effects.
- The peptides reduce exercise-induced metabolites, enhance antioxidant activity, and protect muscles from fatigue.

## Abstract

Sea cucumber peptides have been shown to possess a number of functions, including antioxidant, anti-inflammatory, anti-tumor, and anti-fatigue effects, as well as immune regulation and promotion of collagen synthesis. Among these, high F-value oligopeptides are a promising natural active ingredient demonstrating excellent anti-fatigue effects. This study utilized fresh sea cucumbers as the primary raw material, employing membrane separation technology to investigate the simultaneous separation of sea cucumber polysaccharides and peptides. The process for removing aromatic amino acids during the preparation of high F-value oligopeptides from sea cucumbers was optimized, and the mechanism underlying their anti-fatigue effects was explored. A two-step enzymatic hydrolysis method using neutral protease and composite flavor protease was employed, followed by membrane separation using a 10,000 Da molecular weight ultrafiltration membrane, yielding a sea cucumber peptide yield of 45.00 ± 0.12% and a sea cucumber polysaccharide yield of 51.28 ± 0.63%. Following the removal of aromatic amino acids by means of activated carbon adsorption, the F-value of the high-F-value oligopeptides attained 23.82, with a yield of 24.56%. The experimental findings demonstrated that high-F-value oligopeptides exhibited a substantial increase in the swimming duration of mice and a notable enhancement in their grip strength. These observations signified their substantial anti-fatigue potential. Furthermore, studies have indicated that sea cucumber high-F-value oligopeptides reduce metabolites produced by exercise, enhance muscle protection, increase the activity of antioxidant enzymes in the body, and alleviate fatigue, thereby achieving an anti-fatigue effect.