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Walnut meal anti-inflammatory peptide WPL 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: 2021-08-03
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 oper

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

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

[0052] 1, preparation of different molecular weight polypeptides

[0053] Walnut mealase solution: The cold squeezed walnut passes after pulverizing the 200 mesh sieve, which is filtered after 1: 4, and is filtered, collect residue, and collect the residue in the fume hood to remove the remaining organic solvent, degreasing Walnut meal, stored at 4 ° C.

[0054] It is weighed with a certain amount of degreasing walnut, and 200 mL of distilled water was added to the material of 1: 15 (w / v), and the pH is adjusted to 10.0 with 1 mol / L of NaOH solution, and the magnetic force is stirred for 5 min to perform ultrasonic treatment, ultrasound conditions After 50 ° C, 20 min, the ultrasonic end is allowed to stand for 2 h, and the pH is adjusted by 1 mol / L of the HCl of 1 mol / L by the rotational speed of 6000 r / min, and the obtained precipitated water washed t...

Example Embodiment

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

[0072] In order to further obtain an anti-inflammatory active polypeptide, artificial synthesis was carried out against the ranked polypeptide WPL (TRP-PRO-LEU, molecular weight of 414.2 Da) to verify its anti-inflammatory activity.

[0073] Polypeptide WPL Entrusted Nanjing Source Protection Biochemical Co., Ltd., Synthesized, purity> 95% by solid phase synthesis, to meet the relevant experiments of activity detection.

[0074] (1) Cell toxicity

[0075] First, the safety of polypeptide WPL is explored, such as Image 6 As shown, it is found that the WPL has different degrees of promoting cell proliferation in the range of 0 to 500 μm, that is, does not have toxicity. Image 6 The first list is a blank group, and the second list is induced as an induction group, and the safety experiment does not have a positive control group).

[0076] The effect of polypeptide WPL on cell viability As the concentrati...

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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-Pro-Leu, WPL for short. The preparation method comprises the following steps: preparing anti-inflammatory polypeptide from walnut meal through enzymolysis, purifying by adopting ultrafiltration and sephadex chromatography, identifying a polypeptide sequence through LC-MS/MS, screening anti-inflammatory activity fragments by utilizing a molecular docking technology, artificially synthesizing the polypeptide with relatively high score, verifying the anti-inflammatory activity of the polypeptide, and finding that the WPL has the anti-inflammatory activity. The compound has different degrees of inhibition rates on the generation amounts of three inflammatory media (NO, TNF-[alpha] and IL-1[beta]). Molecular docking results show that WPL 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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IPC IPC(8): C07K5/097A61K38/06A61P29/00
CPCC07K5/0821A61P29/00A61K38/00
Inventor 任迪峰梁雅馨程乐凌予希
Owner BEIJING FORESTRY UNIVERSITY
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