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Angiotensin-converting enzyme inhibitory peptide, and preparation method and application thereof

An angiotensin and inhibitory peptide technology, which is applied in the field of biomedicine to achieve the effects of reasonable process design, good application prospects and strong operability

Active Publication Date: 2013-08-14
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the ACE inhibitory peptides derived from the four-cornered clam.

Method used

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  • Angiotensin-converting enzyme inhibitory peptide, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation method of angiotensin-converting enzyme inhibitory peptide, which comprises the following steps:

[0024] (1) Preparation of four-horned clam enzymolysate:

[0025] Wash the soft body of the four-cornered clams, add 3 times the amount of water and cook for 2 times, 45 minutes each time, separate the decoction liquid and the meat residue, and drain; take the meat residue, add 3 volumes of water to homogenize, and add enzyme activity It is 30000U / g trypsin hydrolysis, the weight of trypsin is 1.0% of the weight of meat residue, the temperature of enzymatic hydrolysis is 48 ℃, the pH of enzymatic hydrolysis is 8.50, and the reaction time of enzymatic hydrolysis is 2 h; Inactivated in a boiling water bath for 15 min, then centrifuged at 10,000 rpm and 4 °C for 20 min, and the supernatant was taken;

[0026] (2) Ultrafiltration purification:

[0027] Take the supernatant obtained by enzymatic hydrolysis in step (1), filter it with a 0.45 μm microporous memb...

Embodiment 2

[0030] Example 2 Sequence analysis of angiotensin converting enzyme inhibitory peptide

[0031] Take the angiotensin-converting enzyme inhibitory peptide prepared in Example 1, and use ESI-Q TOF MS / MS to analyze the amino acid sequence of the polypeptide. The mass spectrometry conditions are: ESI source, scanning mode: positive ion mode, mass scanning range: 50-1000 m / z; the molecular weight of the active polypeptide obtained by analysis is 619.42 Da, and the mass spectrometry detection chart is as follows figure 1 As shown, the amino acid sequence obtained by determination is: Leu-Ala-Ser-Pro-Thr-Met.

Embodiment 3

[0032] Example 3 Synthesis of Angiotensin Converting Enzyme Inhibitory Peptide

[0033] According to the amino acid sequence obtained in Example 2, the method of solid-phase synthesis was used to synthesize the angiotensin converting enzyme inhibitory peptide Leu-Ala-Ser-Pro-Thr-Met, the synthesized polypeptide was analyzed by HPLC and the purity was 98%, and the mass spectrometry was determined. The molecular weight is 619.46 Da, which is consistent with the molecular weight of the purified polypeptide, and the fragments of the secondary mass spectrometry are consistent with the purified polypeptide fragments.

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Abstract

The invention discloses an angiotensin-converting enzyme inhibitory peptide, and a preparation method and an application thereof, wherein the amino acid sequence of the inhibitory peptide is Leu-Ala-Ser-Pro-Thr-Met. The angiotensin-converting enzyme inhibitory peptide is prepared by tracking, screening and estimating an active ingredient for inhibiting the angiotensin-converting enzyme in mactra veneriformis by virtue of modern biochemical technical means, and purifying the active ingredient by methods of enzymolysis, ultrafiltration, reversed-phase high-performance liquid chromatography and the like; and experiment results indicate that the inhibitory peptide has an excellent effect of inhibiting the angiotensin-converting enzyme and a good anti-hypertension effect, and is good in safety performance, and thus has a bright application prospect.

Description

technical field [0001] The invention relates to an active polypeptide, in particular to a polypeptide with ACE-inhibiting activity isolated from an enzymatic hydrolyzate of clam molluscs, and belongs to the technical field of biomedicine. Background technique [0002] Angiotensin Converting Enzyme (ACE) is a zinc ion-dependent dipeptide carboxypeptidase that is widely present in mammalian tissues and can convert angiotensin Ⅰ (Angiotensin Ⅰ) into angiotensin Ⅱ ( Angiotensin Ⅱ), the latter is a strong vasoconstrictor, which promotes the increase of blood pressure; on the other hand, ACE can convert bradykinin, which has vasodilation effect, into an inactive fragment, inhibiting the antihypertensive system-activation Peptide release enzyme - kinin system (Kallikrein-Kinin System, KKS), also lead to increased blood pressure. Therefore, inhibiting the activity of ACE can relieve the increase of blood pressure and has the effect of treating hypertension. [0003] Modern researc...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/20C12P21/06A61K38/08A61P9/12
Inventor 刘睿吴皓朱蕴菡程建明卞慧敏王令充王欣之
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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