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Silkworm pupa protein peptide HPP capable of reducing blood fat, and application of silkworm pupa protein peptide HPP

A silkworm chrysalis protein and a blood lipid-lowering technology are applied to the silkworm chrysalis protein peptide HPP and its application fields to achieve the effect of improving the comprehensive utilization level

Active Publication Date: 2020-01-31
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports of silkworm chrysalis-specific protein peptides that regulate blood lipids at home and abroad.

Method used

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  • Silkworm pupa protein peptide HPP capable of reducing blood fat, and application of silkworm pupa protein peptide HPP
  • Silkworm pupa protein peptide HPP capable of reducing blood fat, and application of silkworm pupa protein peptide HPP
  • Silkworm pupa protein peptide HPP capable of reducing blood fat, and application of silkworm pupa protein peptide HPP

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The acquisition route of silkworm chrysalis protein peptide HPP

[0028] (1) Neutral protease degrades silkworm chrysalis protein

[0029] The enzymatic hydrolysis conditions of silkworm chrysalis protein were studied by using the dual function index of hydrolysis degree and HMGCR inhibition rate. Through response surface optimization experiments, the optimal enzymatic hydrolysis conditions for the preparation of silkworm chrysalis protein lipid-lowering (cholesterol) peptides were obtained: the amount of neutral protease enzyme was 5.1% (w / w), the enzymatic hydrolysis time was 5 hours, the enzymatic hydrolysis temperature was 52°C, and the enzyme The solution pH was 7.0 and the bottom / water ratio (w / v) was 3.9%. The enzymatic hydrolysis solution of silkworm chrysalis protein is obtained.

[0030] (2) Separation and purification of silkworm chrysalis protein enzymatic hydrolyzate

[0031]Peptides with a molecular weight less than 3kDa in the enzymatic hydrolyzate of ...

Embodiment 2

[0049] (1) The molecular docking of HPP and HMGCR was carried out using the Docking module in the MOE software. The HMGCR structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 1HW8).

[0050] (2) The molecular docking of HPP and LDLR was carried out by using the Docking module in the MOE software. The HMGCR structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 2MG9).

[0051] (3) The molecular docking of HPP and SQS was carried out by using the Docking module in the MOE software. The HMGCR structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 3WC9).

[0052] The molecular docking technique was used to analyze the interaction mechanism of HPP with HMGCR, LDLR and SQS. HPP can combine with HMMCR through three hydrogen bonds, which are the three amino acids Arg D595, Arg D641 and Ala A783 in the active pocket of HM...

Embodiment 3

[0054] (1) HepG2 cells were inoculated into 6-well plates, and after 24 hours of culture, they were cultured overnight in the starvation solution. According to different groups, 500ng / mL HPP was not added or added, and the cells were cultured for 8 hours.

[0055] (2) Add 1.0ml Trizol to the cells in each well to lyse the cells, then transfer to a centrifuge tube, add 200μl chloroform; shake vigorously for 15Sec (avoid shaking with an oscillator), let stand at room temperature for 2min, then centrifuge at 12000g for 10min at 4°C; Use a pipette to draw the upper aqueous phase into another clean centrifuge tube, add 600 μL of isopropanol and mix it upside down, centrifuge at 12,000 g at 4°C for 15 min; discard the aqueous phase, add 1 mL of 75% ethanol to wash the precipitate, centrifuge at 12,000 g at 4°C for 5 min, discard The aqueous phase was dried at room temperature for 10 minutes, and the RNA was dissolved in 50 μl of DEPC water, and stored in a -80°C refrigerator for late...

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Abstract

The invention provides a silkworm pupa protein peptide HPP capable of reducing blood fat, and application of the silkworm pupa protein peptide HPP, and belongs to the biotechnology field. The amino acid sequence of the silkworm pupa protein peptide HPP capable of reducing the blood fat is His-Pro-Pro. The invention also discloses the application of the silkworm pupa protein peptide HPP for preparing a drug capable of reducing the blood fat. Blood fat reduction is realized through reduction of the synthetic amount of cholesterol and / or removal of a low density lipoprotein. An experiment indicates that the silkworm pupa protein peptide HPP has the functions of down-regulating the expression level of HMGCCR and SQS genes and / or proteins and / or up-regulating the expression level of LDLR genes and / or proteins. An enzyme activity inhibition experiment indicates that the silkworm pupa protein peptide HPP also can inhibit the activity of 3-hydroxy-3-methyl glutaryl coenzyme A reductase.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a blood lipid-lowering silkworm chrysalis protein peptide HPP and an application thereof. Background technique [0002] Hyperlipidemia is a disease caused by abnormal blood lipid metabolism in the body. Excessive blood lipid levels, especially blood cholesterol levels, can induce atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular and cerebrovascular diseases, which have seriously threatened human health. Therefore, the screening of peptides with hypolipidemic efficacy and related research on the development of hypolipidemic drugs have attracted the attention of the academic community. [0003] A large number of animal experiments and clinical experiments have shown that blood lipid-lowering drugs have some side effects, such as gastrointestinal discomfort, muscle pain, myositis and so on. Therefore, food-borne substances have attracte...

Claims

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Application Information

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IPC IPC(8): C07K5/097A61K38/06A61P3/06
CPCA61K38/00A61P3/06C07K5/0821
Inventor 王君虹孙素玲王伟张玉朱作艺李雪
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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