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Angiotensin converting enzyme inhibitory peptide and preparation method thereof

An angiotensin and inhibitory peptide technology, applied in the field of inhibitory peptides and their preparation, can solve the problems of low content of anti-ACE peptides, no reports of amino acid sequences, and high production costs, so as to improve the ACE inhibition rate, avoid hydrolysis inactivation, and reduce The effect of production costs

Inactive Publication Date: 2010-09-29
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Angiotensinase (ACE) inhibitors (captopril, arapril, etc.) currently used for the treatment of hypertension, due to their fast absorption and excretion, can cause side effects such as coughing, kidney damage, and angioneurotic edema. The ACE inhibitory peptide derived from whey protein has the characteristics of strong physiological functions, no side effects, and high safety. Compared with other drugs, it has great advantages. However, the amino acid sequence of the ACE inhibitory peptide derived from whey protein is currently unknown no report
[0005] The existing methods for preparing whey protein-derived anti-ACE peptides are mainly prepared by enzymatic hydrolysis, and the proteolytic enzymes used are relatively single, mainly alkaline protease, pepsin, trypsin or neutral protease, etc., so that the anti-ACE peptides in milk The content in the albumin hydrolyzate is low, the separation and purification are difficult, and the production cost is high. At the same time, the anti-ACE peptide produced in the enzymatic hydrolysis reaction solution is further hydrolyzed and inactivated due to failure to separate in time

Method used

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  • Angiotensin converting enzyme inhibitory peptide and preparation method thereof
  • Angiotensin converting enzyme inhibitory peptide and preparation method thereof
  • Angiotensin converting enzyme inhibitory peptide and preparation method thereof

Examples

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Embodiment 1

[0030] One, the preparation of whey protein source anti-ACE peptide, comprises the following steps:

[0031] 1) Preparation of whey protein concentrate: 1.0 kg of commercially available whey powder is selected and dissolved in 9 kg of warm water at a temperature of 40° C. to obtain a whey solution. The pressure is 1.0MPa, the temperature is ultrafiltration at 38°C, and the filtrate is then ultrafiltered with a polysulfone membrane with a molecular weight cut-off of 10KDa at a pressure of 1.0MPa and a temperature of 38°C, and the resulting 4.5kg whey protein is ultrafiltered The pH value of the filtered concentrate is adjusted to 8.0;

[0032] 2) Enzymolysis reaction of whey protein concentrate: add whey protein concentrate to enzymolysis reaction vessel 1, and add Lactobacillus helveticus wall protease with an activity of 500U / mg, pepsin at 2000U / mg and 1500U / mg 20 grams of mixed protease composed of trypsin, in which the mass ratio of Lactobacillus helveticus cell wall prote...

Embodiment 2

[0044] The same as Example 1, the difference is: take 12kg of fresh milk and centrifuge at 6000rpm×20min to remove the fat to obtain skim milk, then adjust the pH value of the obtained skim milk to 4.6 with 0.10N dilute hydrochloric acid, and centrifuge at 6000rpm×20min to remove the casein precipitate Obtain whey solution. Adjust the pH of the obtained whey to 7.8, and use a polysulfone membrane with a molecular weight cut-off of 10-15KDa to perform ultrafiltration at a pressure of 1.5MPa and a temperature of 38°C to remove small molecular substances such as lactose, salts, and water, and then Add the resulting 4kg whey protein ultrafiltration concentrate to enzymolysis reaction vessel 1, and add a mixture of Lactobacillus helveticus wall protease with an activity of 700U / mg, pepsin at 2500U / mg, and trypsin at 1600U / mg 15 grams of protease were hydrolyzed at 41° C. for 2.5 hours through the insulation jacket 2 and the temperature control system 3, and continuously stirred by ...

Embodiment 3

[0047] The same as in Example 1, the difference is that 1.2kg of milk powder is dissolved in 9kg of warm water at a temperature of 45°C, and the fat is removed by centrifugation at 6000rpm×20min to obtain skim milk, and the resulting skim milk is then adjusted to pH 4.6 with dilute hydrochloric acid. Centrifuge at 6000rpm×20min to remove casein precipitates to obtain whey solution. Adjust the pH of the obtained whey to 7.8, and use a polysulfone membrane with a molecular weight cut-off of 10-15KDa to perform ultrafiltration at a pressure of 1.5MPa and a temperature of 39°C to remove small molecular substances such as lactose, salts, and water, and then Put the obtained 0.5kg whey protein ultrafiltration concentrate into the enzymolysis reaction vessel 1, and add a mixture consisting of Lactobacillus helveticus wall protease with an activity of 800U / mg, pepsin at 3000U / mg and trypsin at 1800U / mg 2 grams of protease were hydrolyzed at 41.5° C. for 2.8 hours through the insulatio...

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Abstract

The invention discloses an angiotensin converting enzyme inhibitory peptide, which is characterized by comprising the following amino acid sequences: SEQ NO1: Arg-Leu-Ser-Phe-Asn-Pro, SEQ NO2: Leu-Leu, and SEQ NO3: Ala-Leu-Pro-Met-His-Ile-Arg. The method for preparing the inhibitory peptides comprises the following steps of: (1) preparing lactalbumin concentrated solution; (2) preparing lactalbumin hydrolysis liquid; (3) preparing angiotensin converting enzyme-containing inhibitory peptide concentrated solution; (4) performing separation and purification on the inhibitory peptide-containing concentrated solution through gel chromatography and reverse-phase high-pressure liquid chromatography; and (5) obtaining the product by freeze drying. The invention has the advantages that: three amino acid sequences of the angiotensin converting enzyme inhibitory peptide derived from the lactalbumin are disclosed for the first time, the preparation cost is low and the extraction ratio is high.

Description

technical field [0001] The invention relates to an inhibitory peptide and a preparation method thereof, in particular to an angiotensin-converting enzyme inhibitory peptide and a preparation method thereof. Background technique [0002] Angiotensin-converting enzyme (ACE) plays an important role in the process of blood pressure regulation in the human body, and it exists in blood vessel endothelial cells or epithelial cells of various tissues and body fluids such as plasma. It plays an important role in blood pressure regulation in the human renin-angiotensin system and kininase-kinin system, and can simultaneously act on angiotensin I (Asp-Arg-Val-Tyr-Ile-His Leu-Val-Ile-His) and bradykinin to convert angiotensin I into angiotensin II (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe), while hydrolyzing bradykinin to make it inactivation, leading to an increase in blood pressure. Anti-ACE peptides refer to a class of polypeptide substances with ACE-inhibiting activity. They reduce the pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K5/062C12P21/06C07K1/36A61P9/12
Inventor 潘道东
Owner NINGBO UNIV
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