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Natural oligopeptide with hepatic cell oxidative damage protecting effect

An oligopeptide and effective concentration technology, applied in the field of natural oligopeptides, can solve the problems of carcinogenicity threats, limit the use range and dosage, etc., achieve the effect of improving activity, increasing oligopeptide and stability and biological activity, and weakening hydrolysis activity

Active Publication Date: 2019-09-17
长生无极(北京)生物科学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to reduce or inhibit the generation of free radicals, common artificial antioxidants tert-butylhydroquinone, di-tert-butyl-p-cresol and butyl anisole have been widely used in food preservation, but their liver damage and carcinogenicity defects have Potentially huge threats to human health, thus limiting the scope and amount of their use

Method used

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  • Natural oligopeptide with hepatic cell oxidative damage protecting effect
  • Natural oligopeptide with hepatic cell oxidative damage protecting effect
  • Natural oligopeptide with hepatic cell oxidative damage protecting effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A method for preparing a natural oligopeptide with liver cell oxidative damage protection, comprising:

[0042] 1) Preparation of protein hydrolyzate of Moringa oleifera: firstly mix crushed moringa oleifera seeds with ethyl acetate at a mass volume ratio of 1:6 for 2 days; dry moringa oleifera seeds naturally in the air, and dry the Mix Moringa oleifera seeds with pH 8.8, 1.5mol / L Tris-HCL at a mass volume ratio of 1:38 at 42°C, centrifuge after 100min, and take the supernatant; add 4.25mol / L to the supernatant Ammonium sulfate, precipitate protein at 25°C, desalt the precipitated Moringa seed protein with a dialysis bag, and then freeze-dry; prepare a suspension of Moringa seed protein with a mass volume ratio of 5%, take 100mL of the protein suspension, and then adjust the pH to 6.7, add the modified flavor protease, the total enzyme amount is 5%, and enzymatically hydrolyze at 50°C for 300min.

[0043] 2) separation and purification:

[0044] Ultrafiltration: The ...

Embodiment 2

[0052] Flavor protease should be modified before use. The selected flavor protease exopeptidase includes aminopeptidase and carboxypeptidase. Dissolve 23mg flavor protease in 4.5mL pH7.5 phosphate buffer, add 3mL35mmol / L glucose acetic acid The ester solution was mixed thoroughly, left at room temperature for 22 hours, dialyzed to remove the solvent, and freeze-dried.

[0053] All the other parts are completely consistent with Example 1. The yield of the oligopeptide whose sequence is Ser-Phe was determined to be 2.03%.

[0054] The yield of the target oligopeptide in Example 2 is higher than that in Example 1, which shows that after the flavor protease is modified, the characteristics of aminopeptidase and carboxypeptidase are changed, and the effect of aminopeptidase on the hydrophobic amino acid produced by the carboxyl terminal is weakened. The hydrolysis activity of the formed peptide bond and the hydrolysis activity of carboxypeptidase to the carboxyl terminus formed by...

Embodiment 3

[0056] In order to further improve the protective effect of moringa seed oligopeptides on liver cell oxidative damage, the following measures are also taken in this embodiment:

[0057] Moringa seed oligopeptide obtained in Example 1 was modified with strontium citrate. After FOXO3a is activated, it can induce the expression of downstream target genes. The target genes of FOXO3a include antioxidant genes, apoptosis and cell cycle arrest genes, etc. The PI3K-Ak / PKB signaling pathway increases the expression of peroxidase by regulating the activity of FOXO3a. Peroxidase has a higher affinity than catalase and can also produce the same at low levels of hydrogen peroxide. Effect. Akt / PKB consists of four parts: the N-terminal regulatory region, the intermediate enzyme active region, the C-terminal regulatory region, and the hinge region connecting the PH region and the kinase active region, and the C-terminal has a proline-rich hydrophobic domain (HM ), which contains the second...

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Abstract

The invention provides a natural oligopeptide with a hepatic cell oxidative damage protecting effect, and belongs to the field of deep processing of moringa seeds. The oligopeptide is separated from a protein hydrolysate of the moringa seeds, the amino acid sequence is Ser-Phe, and the molecular weight is 252.17 Da. The preparation method of the oligopeptide comprises the steps that protein of the moringa seeds is extracted and hydrolyzed with a flavourzyme, ultrafiltration, anion-exchange chromatography, gel filtration chromatography and reversed phase high-performance liquid chromatography are conducted, and the amino acid sequence and the molecular weight are determined. The invention further discloses application of the oligopeptide in research on a drug for treating diabetes and heart disease. The oligopeptide is high in hepatic cell oxidative damage protecting effect, and the preparation method is high in yield of the target peptide.

Description

technical field [0001] The invention belongs to the field of deep processing of Moringa oleifera seeds, and in particular relates to a natural oligopeptide capable of protecting liver cells from oxidative damage. Background technique [0002] Reactive oxygen species (ROS), including hydroxyl radicals, superoxide anion radicals, singlet oxygen, and hydrogen peroxide, are byproducts of normal metabolism in human cells and play important roles in cell signal transduction and proliferation. Excessive ROS will lead to an imbalance of oxidation and anti-oxidation, and when it exceeds the effective anti-oxidative response capacity of cells in the body, it will trigger oxidative stress in the body. Oxidative stress can cause oxidative damage such as protein cross-linking, lipid peroxidation, DNA and RNA damage, and then lead to a series of diseases such as diabetes, cardiovascular disease, rheumatoid arthritis, cancer, and Parkinson's disease. In order to reduce or inhibit the gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/062C07K1/36C07K1/34C07K1/20C07K1/18C07K1/16C12P21/06A61K38/05A61P3/10A61P9/00
CPCA61K38/00A61P3/10A61P9/00C07K5/06069C12P21/06
Inventor 孙坤来梁丽丽王斌陈荫赵玉勤蔡诗颖初雪梅
Owner 长生无极(北京)生物科学研究院有限公司
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