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Method for high-throughput screening of ACE inhibitory peptide

A peptide-inhibiting, high-throughput technology, applied in the field of preparation and screening of biological peptides, can solve the problems of time-consuming and laborious, and achieve the effects of improving screening efficiency, shortening time-consuming, and reducing consumption.

Inactive Publication Date: 2021-07-02
JIANGXI NORMAL UNIV
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Problems solved by technology

However, these traditional separation and purification processes are time-consuming and labor-intensive

Method used

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  • Method for high-throughput screening of ACE inhibitory peptide
  • Method for high-throughput screening of ACE inhibitory peptide
  • Method for high-throughput screening of ACE inhibitory peptide

Examples

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Embodiment 1

[0056] 1) Preparation of protein hydrolyzate: take 1g of snakehead fish meat protein, dissolve it in 20mL of water, adjust the pH to 9, add 5% alkaline protease (Beijing Suo Laibao Technology Co., Ltd.), hydrolyze at 55°C, and hydrolyze for 3 hours .

[0057] 2) Determination of the optimal screening concentration of the hydrolyzate: set the concentration of the hydrolyzate to 1, 2, 4, 5, 8, 10 mg / mL, evaluate its ACE inhibitory activity, and obtain the optimal binding concentration of 5 mg / mL ( Figure 2A ).

[0058] 3) Bio-affinity ultrafiltration: Mix ACE (100mU / mL) and hydrolyzate (optimum concentration) according to the volume of 2:1 and incubate at 37°C for 1h. And the same experiment was carried out with inactivated ACE as a control.

[0059] Select an ultrafiltration centrifuge tube with a molecular weight of 10kDa, perform ultrafiltration centrifugation 3 times (4000rpm each time, 20min), use a pipette after each centrifugation to prevent the filter membrane from be...

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Abstract

The invention provides a method for high-throughput screening of ACE inhibitory peptides, and relates to the technical field of preparation and screening of biological peptides. The method comprises the following steps: 1) preparing a protein hydrolysate; 2) determining the optimal screening concentration of hydrolysate; (3) carrying out biological affinity ultrafiltration; and 4) performing high-throughput identification on peptide composition and performing molecular docking secondary screening by taking binding energy as an index. According to the method, biological affinity ultrafiltration is combined with a high-throughput identification technology, so that the screening efficiency is greatly improved; the screening accuracy is improved to the maximum extent on the basis of the biocompatibility of interaction of the micromolecule ligand and the enzyme; the consumption of enzymes and samples in the whole screening process is greatly reduced, and the ACE inhibitory activity of the peptide can be predicted and the binding site of the peptide and ACE can be analyzed by molecular docking simulation screening taking binding energy as an index; and the method disclosed by the invention has the advantages of high screening efficiency, short time consumption and high accuracy, and can be widely applied to screening of the ACE inhibitory peptide.

Description

technical field [0001] The invention belongs to the technical field of preparation and screening of biological peptides, in particular to a method for high-throughput screening of ACE inhibitory peptides. Background technique [0002] Hypertension is a clinical syndrome characterized by elevated arterial systolic or diastolic blood pressure and is a risk factor for cardiovascular disease (arteriosclerosis, stroke, and myocardial infarction) and end-stage renal disease. Globally, hypertension Topped the list with a death rate of 12.8%. Angiotensin-converting enzyme plays an important role in maintaining the stability of the body's blood pressure. It has dual functions of hydrolyzing the inactive decapeptide Ang I into the vasoconstriction active substance Ang II and inactivating bradykinin, causing blood pressure to rise. Inhibition of angiotensin-converting enzyme activity can effectively lower blood pressure. Currently, clinically used angiotensin-converting enzyme inhibit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/22C07K1/16C07K1/34G01N30/02
CPCC12P21/06C07K1/22C07K1/16C07K1/34G01N30/02G01N2030/027
Inventor 涂宗财马天新张露李金林
Owner JIANGXI NORMAL UNIV
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