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ACE inhibitory peptide from snakehead protein source and preparation method of ACE inhibitory peptide

A protein source and inhibitory peptide technology, applied in the biological field, can solve problems such as the scarcity of snakeheads, and achieve the effects of low cost, wide source of raw materials, and deep utilization.

Active Publication Date: 2020-01-17
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few studies on the preparation of ACE inhibitory peptides using freshwater fish, and there are even fewer studies on snakehead

Method used

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  • ACE inhibitory peptide from snakehead protein source and preparation method of ACE inhibitory peptide
  • ACE inhibitory peptide from snakehead protein source and preparation method of ACE inhibitory peptide
  • ACE inhibitory peptide from snakehead protein source and preparation method of ACE inhibitory peptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Raw material preparation: keep the fresh snakehead only the back muscles, remove the thorns and skin, rinse 1-2 times in cooling water, mince, freeze-dry, grind, pass through a 40-mesh sieve, and collect the sifted fish meal at -20 ℃ refrigerated for later use;

[0026] (2) Preparation of snakehead protein enzymatic hydrolyzate: weigh 1% (w / v) fish meal and dissolve it in ultrapure water, stir overnight, centrifuge and take supernatant to obtain snakehead protein. Adjust the snakehead protein to pH = 3, add 4% pepsin, hydrolyze at 37°C for 6 hours, stir continuously during the hydrolysis process, and inactivate the enzyme in a water bath at 90°C for 10 minutes after the hydrolysis is completed. The enzymolyzed solution and unenzymolyzed protein were tested for ACE inhibitory activity, and the half-inhibitory concentration (IC 50 ) is 38.5mg / mL, the IC of enzymolysis solution 50 0.179 mg / mL;

[0027] (3) Separation, purification and peptide sequence identification...

Embodiment 2

[0029] (1) Raw material preparation: keep the fresh snakehead only the back muscles, remove the thorns and skin, rinse 1-2 times in cooling water, mince, freeze-dry, grind, pass through a 40-mesh sieve, and collect the sifted fish meal at -20 ℃ refrigerated for later use;

[0030] (2) Preparation of snakehead protein enzymatic hydrolyzate: weigh 1% (w / v) fish meal and dissolve it in ultrapure water, stir overnight, centrifuge and take supernatant to obtain snakehead protein. Adjust the snakehead protein to pH = 3, add 4% pepsin, hydrolyze at 37°C for 6 hours, stir continuously during the hydrolysis process, and inactivate the enzyme in a water bath at 90°C for 10 minutes after the hydrolysis is completed. The enzymolyzed solution and unenzymolyzed protein were tested for ACE inhibitory activity, and the half-inhibitory concentration (IC 50 ) is 38.5mg / mL, the IC of enzymolysis solution 50 0.179 mg / mL;

[0031] (3) Separation, purification and peptide sequence identification...

Embodiment 3

[0033] (1) Raw material preparation: keep the fresh snakehead only the back muscles, remove the thorns and skin, rinse 1-2 times in cooling water, mince, freeze-dry, grind, pass through a 40-mesh sieve, and collect the sifted fish meal at -20 ℃ refrigerated for later use;

[0034] (2) Preparation of snakehead protein enzymatic hydrolyzate: weigh 1% (w / v) fish meal and dissolve it in ultrapure water, stir overnight, centrifuge and take supernatant to obtain snakehead protein. Adjust the snakehead protein to pH = 3, add 4% pepsin, hydrolyze at 37°C for 6 hours, stir continuously during the hydrolysis process, and inactivate the enzyme in a water bath at 90°C for 10 minutes after the hydrolysis is completed. The enzymolyzed solution and unenzymolyzed protein were tested for ACE inhibitory activity, and the half-inhibitory concentration (IC 50 ) is 38.5mg / mL, the IC of enzymolysis solution 50 0.179 mg / mL;

[0035](3) Separation, purification and peptide sequence identification:...

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Abstract

The invention discloses ACE inhibitory peptide from a snakehead protein source and a preparation method of the ACE inhibitory peptide. Pepsin is used for hydrolyzing snakehead protein, ACE inhibitorypeptide enzymatic hydrolysate high in activity is prepared, an ultrafiltration pipe is used for rough separation, liquid chromatography is used for further separation, and identification is performedto obtain three corresponding polypeptide sequences FRVPTPNVS, HVNKDIAPKL and KIKIIAPPERKYS. Snakeheads are broad in source and low in cost, so that the preparation cost of the ACE inhibitory peptidecan be reduced, the utilization value of the snakeheads can be increased, and the ACE inhibitory peptide is hopeful to be applied to business preparation of blood pressure drugs. Deep utilization of freshwater fish sources is promoted.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an ACE inhibitory peptide derived from snakehead protein and a preparation method thereof. Background technique [0002] According to the World Health Organization, about 30% of the deaths in the world are caused by cardiovascular diseases. By 2020, stroke and heart disease will be the leading causes of death worldwide. Antihypertensive peptides lower blood pressure by inhibiting the activity of angiotensin converting enzyme (ACE), so they are also called ACE inhibitory peptides. ACE can catalyze the conversion of angiotensin I to angiotensin II and inactivate the vasodilator (bradykinin), thus leading to an increase in blood pressure (Food Chem, 2017, 228:506–517). Therefore, inhibition of ACE activity has become a major goal in the treatment of hypertension. Although some synthetic ACE inhibitors, such as enalapril, alacipril or lisinopril, are effective for hyperten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/08C12P21/06
CPCC07K1/20C07K1/34C07K7/06C07K7/08C12P21/06
Inventor 王远鹏唐碧灵
Owner XIAMEN UNIV
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