A kind of walnut meal acetylcholinesterase inhibitory peptide and its preparation method and application

A technology for acetylcholinesterase and inhibitory peptide, applied in the field of acetylcholinesterase inhibitory peptide and preparation thereof, can solve the problems of waste of walnut protein resources, hinder the development of walnut industry, etc., and achieves low production cost, obvious enzyme inhibitory effect and high economic benefit. Effect

Active Publication Date: 2022-05-24
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After walnut oil extraction, a large amount of by-product walnut meal is produced, which is usually sold at a low price as feed, which not only causes a waste of walnut protein resources, but also seriously hinders the development of the walnut industry

Method used

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  • A kind of walnut meal acetylcholinesterase inhibitory peptide and its preparation method and application
  • A kind of walnut meal acetylcholinesterase inhibitory peptide and its preparation method and application
  • A kind of walnut meal acetylcholinesterase inhibitory peptide and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Extraction method of walnut meal polypeptide

[0042] (1) Walnut meal degreasing:

[0043] Walnut meal crushed through 40 mesh sieve, at 1:5 feed-to-liquid ratio (w / v) added petroleum ether extraction for 2h, filtering, collecting residue, continuous extraction 3 times, the residue in a fume hood to evaporate the organic solvent, that is, walnut meal degreasing powder, stored at 4 °C conditions.

[0044] (2) Alkali-soluble acid sink extracts walnut protein

[0045] Accurately weigh 10g of walnut meal degreasing powder in a beaker of 400mL, add 200mL of distilled water, adjust the pH to 10.0, stir on a magnetic stirrer for 5min, then sonicate for 20min, take out immediately after sonication, let stand for 1h, 6000r / min centrifugation for 20min, take the supernatant for later. Adjust the pH to 4.5 with 1mol / L HCl, stand for 1h, centrifuge at 6000r / min for 20min to obtain walnut meal protein precipitation, adjust the pH value to 7.0 with 1mol / L NaOH, and obtain wal...

Embodiment 2

[0055] Example 2 After ultrafiltration, walnut meal polypeptide of different molecular weight inhibition capacity of acetylcholinesterase

[0056] The walnut meal enzyme solution obtained by the basic protease and pepsin complex enzyme solution contains free amino acids, polypeptides of different lengths and proteins with a small degree of hydrolysis, etc., and the composition is complex, and it is necessary to purify the polypeptide with high activity and purity.

[0057] In the present embodiment, the means of ultrafiltration, the acetylcholinesterase inhibition capacity as the index to purify the walnut meal polypeptide, and by LC-MS / MS identification of the purified component of the polypeptide sequence. Through PyRx virtual screening combined with Autodock Vina molecular docking and literature research, peptides with strong acetylcholinesterase inhibition ability were screened.

[0058] (1) Ultrafiltration

[0059] The walnut meal lysate prepared in Example 1 is ultrafiltra...

Embodiment 3

[0084] Example 3 polypeptide activity validation

[0085] (1) Peptide artificial synthesis

[0086] The synthesis of peptides adopts Fmoc solid-phase synthesis method, which synthesizes peptides according to the amino acid sequence, and obtains powdered polypeptides after cutting, precipitation and purification. The synthesis process is entrusted to Nanjing Source Peptide Biotechnology Co., Ltd. The purity of peptides YVPHW, LAPF, and FYRR is greater than 95%.

[0087] (2) Verification of artificial synthesis of peptide activity

[0088]Determination of inhibition of acetylcholinesterase by different concentrations of peptides described in Example 1,

[0089] Calculate the semi-inhibitory concentration (IC) of peptides to acetylcholinesterase based on the experimental results fit 50 )。 IC of peptide YVPHW 50 The value is 139.10± 1.34 μg / mL, LAPF IC 50 ICs with values of 318.80± 1.98 μg / mL, FYRR 50 Values of 304.20± 2.57 μg / mL have good acetylcholinesterase inhibition activity, whi...

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Abstract

The invention belongs to the technical field of food deep processing, and in particular relates to an acetylcholinesterase inhibitory peptide extracted from walnut dregs, a preparation method and application thereof. The amino acid sequence of the acetylcholinesterase inhibitory peptide from N-terminal to C-terminal is: Tyr-Val-Pro-His-Trp (YVPHW), Phe-Tyr-Arg-Arg (FYRR) or Leu-Ala-Pro-Phe (LAPF ), the half-inhibitory concentration IC 50 They were 139.10±1.34 μg / mL, 304.20±2.57 μg / mL and 318.80±1.98 μg / mL, respectively. The short peptidase provided by the invention has obvious inhibitory effect, uses walnut meal as raw material, has low production cost and high economic benefit. The present invention also prepares double nanoemulsions loaded with short peptide YVPHW through a two-step emulsification method, and the encapsulation efficiency can reach 92.65±0.77%. The double nanoemulsion is relatively uniform in distribution and has good environmental stability, which can improve the shortcoming of the short peptide YVPHW being quickly digested in the gastrointestinal tract, achieve the effect of slow release, and provide a theoretical basis for the deep processing of walnut meal.

Description

Technical fields: [0001] The present invention belongs to the field of food deep processing technology, specifically relates to an acetylcholinesterase inhibitory peptide isolated from walnut meal and preparation method and application thereof. Background: [0002] Alzheimer's disease (AD) is the leading cause of neurodegenerative dementia and one of the leading sources of morbidity and mortality in the elderly population. It is characterized by memory loss, language impairment, behavioral abnormalities, and cognitive dysfunction. People with AD place a heavy burden on families and society due to their lack of ability to perform their daily lives. Therefore, Alzheimer's disease is a public health problem that needs to be solved urgently, and it is also the focus of research in recent years. The pathological features of AD mainly include acetylcholine (ACh) deletion, amyloid deposition, tau protein superphosphorylation, etc. The cholinergic nerves in the central nervous system are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K5/107C07K5/103C12P21/06C07K1/36C07K1/34C07K1/16A23L33/18A23L29/00A61K38/07A61K38/08A61K9/107A61K47/12A61K47/24A61P25/28
CPCC07K7/06C07K5/1016C07K5/101C12P21/06A23L33/18A23L29/05A23L29/04A61K9/1075A61K47/12A61K47/24A61P25/28A23V2002/00A61K38/00A23V2200/322A23V2250/55A23V2250/1842A23V2250/1872
Inventor 任迪峰王子纯翟鑫禹尹玉茜
Owner BEIJING FORESTRY UNIVERSITY
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