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Angiotensin-converting enzyme inhibition peptide sourcing from marine microalgae

An angiotensin and inhibitory peptide technology, applied in the field of active peptide screening, to achieve the effect of novel structure, small molecular weight and easy absorption

Active Publication Date: 2014-07-16
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies on ACE inhibitory peptides from marine microalgae, especially bait microalgae, have not been reported yet.

Method used

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  • Angiotensin-converting enzyme inhibition peptide sourcing from marine microalgae
  • Angiotensin-converting enzyme inhibition peptide sourcing from marine microalgae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Separation and purification of angiotensin-converting enzyme inhibitory peptide

[0021] (1) Preparation of Isochrysis globosa protein:

[0022] Add Isoflagellates to 1.5% (w / v) sodium chloride solution at a ratio of 1:15 (1 gram of algae powder is added to 15 ml of sodium chloride solution), adjust the pH to 7.0, extract with ultrasonic waves, and crush at high speed Refrigerate and centrifuge at 4°C and 5000r / min for 15min, centrifuge to separate the supernatant, add sodium chloride solution to the precipitated algal mud and ultrasonically crush it twice, with an interval of 1s, combine all the supernatant obtained after the above crushing, and freeze-dry That is crude protein.

[0023] (2) Enzymolysis:

[0024] With the crude protein in step (1), according to the enzyme and crude protein ratio of 6:100, add trypsin with an enzyme activity of 10000U / g, adjust the pH of the solution to 8.0, and the concentration of the substrate protein to be 5.5g / 100ml. T...

Embodiment 2

[0033] Example 2 Sequence Analysis of Angiotensin Converting Enzyme Inhibitor Peptide

[0034] Taking the angiotensin-converting enzyme inhibitory peptide prepared in Example 1, the amino acid composition of the active polypeptide was first analyzed, and the results showed that its main amino acids were Met, Lys, and Leu, and it also contained Tyr, Gly, and Asp. Using MALDI-TOF / TOF-MS / MS to analyze the amino acid sequence of the polypeptide, the molecular weight of the active polypeptide was analyzed to be 839.4061Da, and the determined amino acid sequence was: Tyr-Met-Gly-Leu-Asp-Leu-Lys. De novo sequencing analysis was performed on the structure, and the results were consistent with the above data, confirming that the polypeptide is Tyr-Met-Gly-Leu-Asp-Leu-Lys (SEQ ID NO: 1), which is a new type of active polypeptide structure.

Embodiment 3

[0035] Example 3 Effect of angiotensin-converting enzyme inhibitory peptide

[0036] The antihypertensive peptide derived from marine microalgae obtained in Example 1 was directly administered into essential hypertensive rats (SHR), and its effect on the blood pressure of SHR was detected. The experiment selected 24 rats with a body weight of 240 ± 20 grams. The rats were randomly divided into 3 groups, 8 in each group, including a blank control group (injection of the same volume of normal saline), an ACE enzyme inhibition group and a synthetic antihypertensive drug card. Positive control group for Topril. The experimental temperature was 20°C, the relative humidity was 70%, and the pre-feeding was carried out for 1 week. The administration dose was 10 mg / kg, and the time was 24 days. The systolic pressure of the tail artery was measured once at 0, 2, 4, 6, and 8 hours. The results showed that the ACE inhibitory peptide group had a significant effect compared with the blank...

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Abstract

The invention aims at providing a novel angiotensin-converting enzyme inhibition peptide, namely an ACE inhibition peptide prepared form isochrysis galbana, and with the amino acid sequence of SEQ ID NO:1. The invention provides a preparation method of a high-activity angiotensin-converting enzyme inhibition peptide sourcing from marine microalgae. The current angiotensin-converting enzyme inhibiter mainly comes from food and large macroalgae of laver and the like, and coming from bait microalgae is not reported. The structure of the obtained active peptide with a unique structure is Tyr-Met-Gly-Leu-Asp-Leu-Lys. The structure is novel and has the characteristics of being small in molecular weight and easily absorbed by a human body.

Description

technical field [0001] The invention belongs to the technical field of active polypeptide screening, and specifically relates to an angiotensin-converting enzyme inhibitory peptide, that is, an angiotensin-converting enzyme (ACE) inhibitory peptide isolated from marine microalgae dinoflagellates. Background technique [0002] Angiotensin converting enzyme (ACE) is a zinc metalloprotease, a carboxyl dipeptidase, and one of the important proteases in the renin-angiotensin system. It plays an important role in the regulation of human blood pressure. By acting on the end of angiotensin I to remove His-Leu to generate angiotensin II, it can contract arterial smooth muscle and rapidly cause blood pressure to rise. Inhibition of ACE activity is an effective way to lower blood pressure. Most of the current drugs for the treatment of hypertension are chemical synthetic products, which have some adverse reactions, such as cough, taste disturbance and skin rash. ACE-inhibiting peptid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12P21/02A61K38/08A61P9/12C07K1/16
Inventor 徐年军吴昊孙雪余虹
Owner NINGBO UNIV
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