Asparagus antihypertensive peptide extract and asparagus antihypertensive peptide and application thereof

A technology of antihypertensive peptides and dragon whiskers, applied in the field of bioactive peptides

Active Publication Date: 2021-10-29
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report of ACE inhibitory peptide isolated from asparagus

Method used

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  • Asparagus antihypertensive peptide extract and asparagus antihypertensive peptide and application thereof
  • Asparagus antihypertensive peptide extract and asparagus antihypertensive peptide and application thereof
  • Asparagus antihypertensive peptide extract and asparagus antihypertensive peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation method of asparagus antihypertensive peptide extract comprises the following steps:

[0025] (1) Asparagus powder 20g, add 500ml of distilled water and stir to swell for 2h.

[0026] (2) Add 500mg of trypsin (EC 3.4.23.4, ≥250.N.F.U / mg) for enzymolysis, the enzymolysis temperature is 42°C, the enzymolysis pH is controlled at 8.0, the enzymolysis time is 3h, and the enzyme is extinguished at 100°C after the enzymolysis is completed 10min, filter to remove impurities.

[0027] (3) The supernatant was precipitated with ethanol with a final volume concentration of 70% to remove the precipitate, and the supernatant was ultrafiltered to collect components <3 kDa, concentrated to 20 ml and freeze-dried to obtain the antihypertensive peptide extract.

[0028] (4) Determination of IC50 value of antihypertensive peptide by high performance liquid chromatography.

[0029] The medicines and instruments used in this example can be obtained commercially unless other...

Embodiment 2

[0037] The preparation method of asparagus antihypertensive peptide extract comprises the following steps:

[0038] (1) Asparagus was dried and crushed, 20g of dry powder was added to 400ml of distilled water, stirred, and swelled at room temperature for 4h.

[0039] (2) Add 400mg of trypsin (EC 3.4.23.4, ≥250.N.F.U / mg) and stir for enzymolysis. The enzymolysis temperature is controlled at 42°C, the enzymolysis pH is controlled at 7.8, and the enzymolysis time is 2 hours. After the enzymolysis is completed, boil water Inactivate the enzyme for 10 min, filter to remove impurities.

[0040] (3) The supernatant is precipitated with ethanol with a final volume concentration of 80% to remove the precipitate, and the supernatant is ultrafiltered to collect <3kDa components, and concentrated to 20ml, and freeze-dried to obtain the Asparagus antihypertensive peptide extract .

[0041] (4) Evaluate the ACE inhibitory activity of the Asparagus antihypertensive peptide extract by using...

Embodiment 3

[0046] The UPLC-ESI-Q-Exactive Focus-MS / MS mass spectrometry information of the above-mentioned asparagus <3kDa antihypertensive peptide extract of asparagus was used Mascot2.2 and asparagus transcriptome (Accession: SRX258772, downloaded from NCBI) database For comparison, the top 20 polypeptides were selected for selective synthesis, and then the polypeptides with ACE inhibitory activity were screened. It was identified that the polypeptide FQINMCILR had better ACE inhibitory activity. Methionine and cysteine ​​are easily oxidized in the body because they contain S atoms, and the S atoms are modified into a sulfoxide structure to enhance their stability in the body. FQINMCILR commissioned Shanghai Qiangyao Biotechnology Co., Ltd. to synthesize and selectively modify methionine to obtain a peptide with the sequence FQIN[M(O)]CILR, a purity of ≥98%, and a molecular weight of 1153.43Da identified by mass spectrometry.

[0047] (1) Determination of ACE inhibitory activity of pep...

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Abstract

The invention discloses asparagus antihypertensive peptide and its application in the preparation of antihypertensive drugs. The antihypertensive extract of asparagus is obtained by enzymatic hydrolysis, and the extract is further separated and purified to obtain a polypeptide with a specific sequence. The amino acid sequence of the antihypertensive peptide is Phe‑Gln‑Ile‑Asn‑[Met(O)]‑Cys ‑Ile‑Leu‑Arg (FQIN[M(O)]CILR), with a molecular weight of 1153.43 Da identified by mass spectrometry. The asparagus antihypertensive peptide screened by the invention has significant angiotensin-converting enzyme inhibitory activity. In vitro performance of temperature, pH, simulated gastrointestinal digestion and angiotensin converting enzyme (ACE) degradation stability. Enzyme kinetics studies show that the polypeptide is a non-competitive inhibitor of ACE. Both in vitro and in non-spontaneously hypertensive rats (SHRs) have shown good antihypertensive effects, and can be used as medicines related to the treatment of hypertension.

Description

technical field [0001] The invention relates to the technical field of bioactive peptides, and relates to the antihypertensive peptide extract of asparagus asparagus, the antihypertensive peptide with a specific sequence and its application in antihypertensive drugs. Background technique [0002] Hypertension is mainly characterized by elevated blood pressure in the human circulation arteries (high pressure ≥ 140mmHg, low pressure ≥ 90mmHg), and hypertension is associated with increased mortality from cardiovascular diseases and diabetes. Renin-angiotensin system (RAS) and kinin-bradykinin system (kallikreo-bradykinin system, KKS) can play an important role in blood pressure regulation. ACE is a Zn that can regulate the renin-angiotensin system (RAS) and kinin-bradykinin system (KKS) ubiquitously in mammalian tissues. 2+ dependent carboxypeptidase. Therefore, ACE is considered to be an important target for the treatment of hypertension. [0003] Most of the ACE inhibitory...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12P21/06C07K1/14A61K38/08A61P9/12A23L33/18
CPCA23V2002/00A61K38/00A23L33/18A61P9/12C07K7/06C12P21/06A23V2200/326A23V2250/55
Inventor 张全斌邓真真刘英娟王晶耿丽华岳洋
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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