Walnut meal acetylcholine esterase inhibiting peptide as well as preparation method and application thereof

A technology for acetylcholinesterase and inhibitory peptides, which is applied in the field of acetylcholinesterase inhibitory peptides and its preparation, can solve the problems of wasting walnut protein resources and hindering the development of walnut industry, and achieve low production costs, obvious enzyme inhibition, and high economic benefits Effect

Active Publication Date: 2021-11-26
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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  • Walnut meal acetylcholine esterase inhibiting peptide as well as preparation method and application thereof
  • Walnut meal acetylcholine esterase inhibiting peptide as well as preparation method and application thereof
  • Walnut meal acetylcholine esterase inhibiting peptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: the extraction method of walnut meal polypeptide

[0042] (1) Degreasing walnut meal:

[0043] The walnut pulp was crushed through a 40-mesh sieve, extracted with petroleum ether at a solid-liquid ratio (w / v) of 1:5 for 2 hours, filtered, and the residue was collected, extracted continuously for 3 times, and the residue was placed in a fume hood to evaporate the organic solvent. The degreased walnut meal powder was obtained and stored at 4°C for future use.

[0044] (2) Alkali-soluble acid precipitation to extract walnut protein

[0045] Accurately weigh 10g of degreased walnut meal powder into a 400mL beaker, add 200mL of distilled water, adjust the pH to 10.0, stir on a magnetic stirrer for 5min, then ultrasonicate for 20min, take it out immediately after the ultrasonication is over, and let it stand for 1h, 6000r / min Centrifuge for 20 min, and take the supernatant for later use. Use 1mol / L HCl to adjust the pH to 4.5, let it stand for 1h, and centrif...

Embodiment 2

[0055] The inhibitory ability of walnut meal polypeptides with different molecular weights to acetylcholinesterase after the ultrafiltration of embodiment 2

[0056] The enzymolysis solution of walnut meal obtained by combined enzymolysis with alkaline protease and pepsin contains free amino acids, polypeptides of different lengths, and proteins with a small degree of hydrolysis, etc. purification.

[0057] In this example, walnut meal polypeptides were purified by means of ultrafiltration, using acetylcholinesterase inhibitory ability as an indicator, and the polypeptide sequences of the purified components were identified by LC-MS / MS. Through PyRx virtual screening combined with Autodock Vina molecular docking and literature research, the peptides that may have strong acetylcholinesterase inhibitory ability were screened out.

[0058] (1) ultrafiltration

[0059] The enzymatic hydrolysis product of walnut meal prepared in Example 1 was subjected to ultrafiltration to purif...

Embodiment 3

[0084] Example 3 Verification of polypeptide activity

[0085] (1) Artificial synthesis of peptides

[0086] The synthesis of peptides adopts the Fmoc solid-phase synthesis method, and peptides are synthesized according to the amino acid sequence, and powdered peptides are obtained after cleavage, precipitation, and purification. The synthesis process was entrusted to Nanjing Yuanpeptide Biotechnology Co., Ltd. The purity of peptides YVPHW, LAPF and FYRR were all greater than 95%.

[0087] (2) Activity verification of artificially synthesized peptides

[0088] According to the method described in Example 1, the inhibitory rate of different concentrations of peptides to acetylcholinesterase was measured,

[0089] According to the fitting calculation of the experimental results, the half-inhibitory concentration (IC) of the peptide to acetylcholinesterase was calculated. 50 ). IC of peptide YVPHW 50 The value is 139.10±1.34 μg / mL, the IC of LAPF 50 The value is 318.80±1.9...

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Abstract

The invention belongs to the technical field of deep processing of foods and particularly relates to an acetylcholine esterase inhibiting peptide extracted from walnut meal as well as a preparation method and application of the acetylcholine esterase inhibiting peptide. An amino acid sequence of the acetylcholine esterase inhibiting peptide from an N terminal to a C terminal is as follows: Tyr-Val-Pro-His-Trp (YVPHW), Phe-Tyr-Arg-Arg (FYRR) or Leu-Ala-Pro-Phe (LAPF), and a half inhibitory concentration IC50 of the acetylcholine esterase inhibiting peptide is 139.10+/-1.34 micrograms per millimeter, 304.20+/-2.57 micrograms per millimeter and 318.80+/-1.98 micrograms per millimeter separately. The oligopeptidase provided by the invention is obvious in inhibiting effect, the walnut meal is adopted as a raw material, the production cost is low, and the economic benefit is high. According to the invention, a double-nano emulsion loaded with an oligopeptide YVPHW is also prepared by a two-step emulsification method, and the encapsulation efficiency can reach 92.65%+/-0.77%. The double-nano emulsion is relatively uniform in distribution and relatively good in environmental stability, the defect that the oligopeptide YVPHW is quickly digested in gastrointestinal tracts can be overcome, a slow release effect is achieved, and a theoretical basis for deep processing of the walnut meal is provided.

Description

Technical field: [0001] The invention belongs to the technical field of food deep processing, and in particular relates to an acetylcholinesterase inhibitory peptide isolated from walnut dregs, a preparation method and application thereof. Background technique: [0002] Alzheimer's disease (AD) is the leading cause of neurodegenerative dementia and one of the main sources of morbidity and mortality in the elderly population. Its main manifestations are memory loss, degeneration of language ability, abnormal behavior and cognitive dysfunction. AD patients have brought a heavy burden to the family and society due to lack of daily living ability. Therefore, Alzheimer's disease is an urgent public health problem, and it is also the focus of research in recent years. The pathological features of AD mainly include loss of acetylcholine (ACh), amyloid deposition, hyperphosphorylation of tau protein and so on. The cholinergic nerves in the central nervous system constitute an imp...

Claims

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Application Information

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Patent Type & Authority Applications(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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