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Earthworm bioactive small peptide as well as preparation method and application thereof

A technology of biologically active peptides and earthworms, applied in the field of small peptides, can solve the problems that there are no reports of anti-oxidation of small peptides of earthworm protein hydrolyzate

Active Publication Date: 2021-12-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no anti-oxidation report of the small peptide of earthworm protein hydrolyzate

Method used

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  • Earthworm bioactive small peptide as well as preparation method and application thereof
  • Earthworm bioactive small peptide as well as preparation method and application thereof
  • Earthworm bioactive small peptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of embodiment 1 active polypeptide

[0033] (1) Extraction of Earthworm Crude Protein: Dried Earthworm is ground into powder for later use. Take 40g of Earthworm powder in a beaker, add 1L of deionized water, and extract for 2.5 hours at a temperature of 60°C and a pH of 10. , centrifuged at 3800r / min for 20min, adjusted the pH of the supernatant to 4.5 with 1mol / L HCl solution, centrifuged again after standing for a certain period of time, and freeze-dried the precipitate to obtain the earthworm crude protein.

[0034] (2) Preparation and grading of polypeptides: dissolve the crude protein into a solution with a concentration of 3% in water, and then add alkaline protease to the solution at 3% of the weight of the crude protein. Solution for 2 hours, heat inactivation, centrifuge, and take the supernatant. Finally, the supernatant is separated through an ultrafiltration membrane with a molecular weight cut-off of 5 kDa, and the polypeptide is obtained ...

Embodiment 2

[0041] Example 2: Screening of Active Peptides

[0042] (1) Structural Identification of Antioxidant Peptides from Earthworm

[0043]The structure of the dilongide prepared in Example 1 was identified by liquid chromatography-mass spectrometry (UPLC-ESI-MS / MS). UPLC conditions: chromatographic column: Accucore 100*2.1 mm, 2.6 μm; mobile phase A: 0.1% formic acid-acetonitrile; B: 0.1% formic acid aqueous solution. Mass spectrometry conditions: ion source ESI+MS, spray voltage 3800V, capillary temperature 275°C, see the total ion chromatogram of earthworm antioxidant peptide Figure 5 . The liquid mass results were analyzed with the help of Xcalibur software, and by comparing with the protein sequence of Dilong in the UniProt protein database, and sorted according to the Abundances value, it was predicted that the first 80 peptides might have good antioxidant activity.

[0044] (2) Molecular docking

[0045] Using the Autodock molecular docking software, 80 peptides were use...

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Abstract

The invention discloses a small peptide which is derived from earthworms and has antioxidant biological activity. The amino acid sequence of the small peptide is as shown in SEQ ID No. 1 or SEQ ID No. 2. The applicant firstly extracts and separates the bioactive peptide from the traditional Chinese medicine lumbricus, and then screens out the small peptide with strong antioxidant activity from the bioactive peptide through a large number of tests such as liquid chromatography-mass spectrometry, mass spectrometry analysis, sequence alignment, molecular docking, activity verification and the like. The small peptide is separated and identified from the traditional Chinese medicine lumbricus for the first time, a new way is provided for fully utilizing the lumbricus medicinal material, and meanwhile, a new resource is provided for the development of antioxidants.

Description

technical field [0001] The invention relates to a small peptide, especially a small peptide derived from earthworm and having antioxidant biological activity, and also relates to a preparation method and application of the small peptide. Background technique [0002] Human cells will produce a large amount of reactive oxygen species (ROS) during the metabolic process, and the body's antioxidant defense system can effectively remove these ROS, that is, under normal circumstances, the production and elimination of ROS are in a delicate balance. When ROS is excessively produced or the body's antioxidant defense system fails, this balance will be broken, and the body will be in a state of oxidative stress. Studies have confirmed that the damage of oxidative stress can lead to various chronic diseases such as cancer, diabetes, inflammation, cardiovascular disease, asthma and Alzheimer's disease. [0003] Antioxidant peptides are a class of biologically active peptides that have ...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K5/11C07K1/34C07K1/30C12P21/06A23L33/18A61K8/64A61Q19/08
CPCC07K7/06C07K5/1019C12P21/06A23L33/18A61K8/645A61Q19/08A61K2800/522A23V2002/00A23V2200/30A23V2250/55
Inventor 侯焘李泊远何慧马雪琪李凯凯
Owner HUAZHONG AGRI UNIV