Walnut meal anti-inflammatory peptide FPY and application thereof

A technology of anti-inflammatory peptides and anti-inflammatory drugs is applied in the field of anti-inflammatory peptides isolated from enzymatic hydrolysis products of walnut meal, which can solve the problems of lengthy, increased operating costs, environmental safety threats, etc.

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 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 FPY and application thereof
  • Walnut meal anti-inflammatory peptide FPY and application thereof
  • Walnut meal anti-inflammatory peptide FPY 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 FPY (Phe-Pro-Tyr, molecular weight 425.2Da) was artificially synthesized to verify its anti-inflammatory activity.

[0073] Polypeptide FPY was entrusted to 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] First, the safety of the polypeptide FPY was explored, such as Image 6 As shown, it was found that in the concentration range of 0-500 μM, the four polypeptides all had different degrees of promoting cell proliferation, that is, they had no toxicity ( Image 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 the...

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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 thereof. The amino acid sequence of the anti-inflammatory peptide from the C terminal to the N terminal is Phe-Pro-Tyr, FPY 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 FPY has the anti-inflammatory activity. The compound has different degrees of inhibition rates on the generation amounts of four inflammatory mediators (NO, TNF-[alpha], IL-6 and IL-1[beta]). Molecular docking results show that FPY 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): C07K5/087A61K38/06A61P29/00
CPCC07K5/0812A61P29/00A61K38/00
Inventor 任迪峰尹玉茜曾宪坤周敏
Owner BEIJING FORESTRY UNIVERSITY
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