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A kind of tea polypeptide with ACE inhibitory activity and its application

A technology of tea and inhibitors, applied in the field of biomedicine, can solve undiscovered problems and achieve good development value and application prospects

Active Publication Date: 2020-11-03
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the application of tea polypeptide in inhibiting ACE, lowering blood pressure and preventing cardiovascular diseases has not been found

Method used

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  • A kind of tea polypeptide with ACE inhibitory activity and its application
  • A kind of tea polypeptide with ACE inhibitory activity and its application
  • A kind of tea polypeptide with ACE inhibitory activity and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation and identification of peptide chain IQDKEGIPPDQQR:

[0024] 1. Weigh 1g (±0.01g) of tea (Tiaohong) powder into a conical flask, add 75ml of boiling ultrapure water, extract in a boiling water bath for 45min, shake once every 15min, suction filter after the extraction is completed, and use Dilute to 100ml with ultrapure water.

[0025] 2. Use 1kDa ultrafiltration membrane to roughly separate the peptide chain, and then use Tricine-SDS-PAGE electrophoresis technology to separate and purify (parameters are 4% stacking gel, 10% interlayer gel, 16% / 6M urea, constant pressure 30V to run to concentration At the end of the glue, change to 200V and run to the end). The band pattern after staining and decolorization is shown in figure 1 shown.

[0026] 3. Cut the gel and recover, and use nano-UPLC-MS / MS to identify the peptide sequence. The present invention identifies the IQDKEGIPPDQQR sequence from this process.

[0027] The specific steps are as follows: the pep...

Embodiment 2

[0031] 1. The sequence information of the polypeptide chain was obtained according to Example 1, and the polypeptide was artificially synthesized with a purity of 98%. Subsequent in vitro inhibitory activity and enzyme kinetic experiments were all carried out with the artificially synthesized peptide chain.

[0032] 2. Identification of ACE inhibitory activity of peptide chain IQDKEGIPPDQQR in vitro

[0033] 10 μl ACE enzyme solution (final system concentration 12.5U / L, dissolved in 50mM tris-HCl, containing 0.3M NaCl, pH7.5) was added to a 96-well plate and mixed with 40 μl peptide solution. 150 μl of 0.88 mM FAPPGG (dissolved in 50 mM tris-HCl with 0.3 M NaCl, pH 7.5, prewarmed at 37° C.) was added. Immediately transfer the 96-well plate to a microplate reader, record the absorbance value at 340 nm every 30 s, and record for 30 min. In the blank control, the inhibitor was replaced with water. ACE inhibitory activity is expressed by the following formula:

[0034] ACE inhi...

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Abstract

The invention discloses tealeaf polypeptide having ACE inhibitory activity and application thereof, and belongs to the technical field of bio-medicines. An amino acid sequence of the tealeaf polypeptide is Ile-Gln-Asp-Lys-Glu-Gly-Ile-Pro-Pro-Asp-Gln-Gln-Arg. ACE inhibitory peptide is obtained through separating the tealeaf polypeptide from striped red tealeaves; the study on enzyme kinetics determines that the tealeaf polypeptide has stronger ACE inhibitory activity, Lineweaver-Burk plot is drawn to define action ways; polypeptide is uncompetitive inhibition, and the inhibition type is very seldom seen in food source ACE inhibitory peptide, and is a good material which develops and researches multi-target point hypotensor medicines, and the tealeaf polypeptide can be used as an important component of a medicine or a health care product for treating and / or preventing hypertension diseases and cardiovascular diseases; therefore, the tealeaf polypeptide has a good development value and an application prospect on development and research of cardiovascular medicines.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a tea polypeptide with ACE inhibitory activity and its application. Background technique [0002] Cardiovascular disease and type II diabetes are the most serious global health problems, and high blood pressure is the most important risk factor for cardiovascular disease, which can easily cause atherosclerosis, coronary heart disease, myocardial infarction, heart failure and other problems. Angiotensin-converting enzyme (ACE) is the main component of the renin-angiotensin-aldosterol system (RAAS), which can convert the two carbon-terminal ends of the inactive precursor polypeptide angiotensin I into Amino acids are excised and transformed into angiotensin II, which has a strong vasoconstriction effect. At the same time, ACE can also decompose bradykinin, which has a vasodilation effect, to make it ineffective, eventually causing an increase in blood pressure. Therefore, inhi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K38/10A61P9/12A61P9/00A23L33/18
CPCA23L33/18A23V2002/00A61K38/00A61P9/00A61P9/12C07K7/08A23V2200/30A23V2200/326A23V2250/55
Inventor 王校常陆娅婷陆鹏范冬梅王羽黄旦益
Owner ZHEJIANG UNIV
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