Walnut meal anti-inflammatory peptide WSL and application thereof

A technology for anti-inflammatory peptides and anti-inflammatory drugs is applied in the field of anti-inflammatory peptides isolated from enzymatic hydrolysis products of walnut meal, and can solve the problems of expensive purification method chromatography technology, increased operating costs, and high purification costs.

Active Publication Date: 2021-08-27
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main problem faced in the production of peptides is the high cost of purification, and purification methods such as chromatography are usually expensive and lengthy, and a large amount of organic waste is generated in each operation during conventional separation, which poses a great potential threat to environmental safety. Increased operating costs, and the solubility and stability of the peptide during purification is also a challenge for the purification process

Method used

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  • Walnut meal anti-inflammatory peptide WSL and application thereof
  • Walnut meal anti-inflammatory peptide WSL and application thereof
  • Walnut meal anti-inflammatory peptide WSL and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation, structure identification and molecular docking of walnut meal anti-inflammatory polypeptide

[0052] 1. Preparation of polypeptides with different molecular weights

[0053] Enzymatic hydrolysis of walnut dregs: Crushed walnut dregs, passed through a 200-mesh sieve, extracted with n-hexane at a ratio of 1:4 for 14 hours, then filtered, collected residues, and placed in a fume hood to volatilize the remaining organic solvents to obtain degreasing Walnut meal, stored at 4°C for later use.

[0054] Weigh a certain amount of defatted walnut meal, add 200mL distilled water with a solid-liquid ratio of 1:15 (w / v), adjust the pH value to 10.0 with 1mol / L NaOH solution, and perform ultrasonic treatment after magnetic stirring for 5min. The ultrasonic conditions are 50°C, 20min, after the end of ultrasonication, let stand in the room for 2h, centrifuge at a speed of 6000r / min for 15min, adjust the pH value of the supernatant to 4.5 with 1mol / L HCL, contin...

Embodiment 2

[0071] Example 2 Anti-inflammatory activity of walnut meal anti-inflammatory polypeptide

[0072] In order to further obtain anti-inflammatory active peptides, the top-ranked peptide WSL (Trp-Ser-Leu, molecular weight 404.2Da) was artificially synthesized to verify its anti-inflammatory activity.

[0073] Polypeptide WSL entrusts Nanjing Yuanpeptide Biological Co., Ltd. to synthesize it through solid-phase synthesis, with a purity of >95%, which meets the requirements of related experiments for activity detection.

[0074] (1) Cytotoxicity

[0075] Firstly, the safety of peptide WSL was explored, such as Figure 6 As shown, it was found that in the concentration range of 0-500 μM, it has different degrees of promoting cell proliferation, that is, it has no toxicity ( Figure 6 The first column is the blank group, the second column is the induction group, and there is no positive control group in the safety experiment).

[0076] Among them, the effect of peptide WSL on cell ...

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Abstract

The invention belongs to the technical field of food deep processing, and particularly relates to an anti-inflammatory peptide separated from walnut meal enzymolysis products and application of the anti-inflammatory peptide. The amino acid sequence of the anti-inflammatory peptide from the C terminal to the N terminal is Trp-Ser-Leu, WSL for short. The anti-inflammatory polypeptide is prepared from walnut meal through enzymolysis, purifying is carried out by adopting ultrafiltration and sephadex chromatography, the polypeptide sequence is identified through LC-MS / MS, anti-inflammatory activity fragments are screened by utilizing a molecular docking technology, the polypeptide with relatively high score is subjected to artificial synthesis, the anti-inflammatory activity of the polypeptide is verified, and the invention finds that the WSL has the anti-inflammatory activity, and has different degrees of inhibition rates on the generation amounts of four inflammatory mediators (NO, TNF-alpha, IL-6 and IL-1beta). Molecular docking results show that the WSL and iNOS are connected mainly through pi-pi accumulation, alkyl bonds, pi-alkyl bonds and hydrogen bonds.

Description

Technical field: [0001] The invention belongs to the technical field of food deep processing, and in particular relates to an anti-inflammatory peptide isolated from an enzymatic hydrolysis product of walnut dregs and an application thereof. Background technique: [0002] In recent years, various chronic diseases caused by chronic inflammation have seriously threatened human health. Inflammation inducers trigger the production of various inflammatory mediators through various sensors present in immune cells, and then trigger inflammatory responses through several signaling pathways. Inflammatory mediators refer to chemical substances released by immune cells or produced by body fluids that can cause inflammatory reactions during inflammation. They can be divided into seven categories according to their biochemical characteristics: vasoactive amines, vasoactive peptides, lipid mediators, and complement components. fragments, cytokines, chemokines and proteolytic enzymes (Medz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/097A61K38/06A61P29/00
CPCC07K5/0821A61P29/00A61K38/00
Inventor 任迪峰任昊宇邓津常敏
Owner BEIJING FORESTRY UNIVERSITY
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