CN113508828B - Preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread - Google Patents
Preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread Download PDFInfo
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- CN113508828B CN113508828B CN202110773291.8A CN202110773291A CN113508828B CN 113508828 B CN113508828 B CN 113508828B CN 202110773291 A CN202110773291 A CN 202110773291A CN 113508828 B CN113508828 B CN 113508828B
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- mackerel
- active peptide
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- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
- A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
- A21D2/24—Organic nitrogen compounds
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- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
- A21D13/06—Products with modified nutritive value, e.g. with modified starch content
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
Abstract
The invention discloses a preparation method of mackerel antihypertensive active peptide and application thereof in bread. The preparation method comprises the following steps: adding a proper amount of deionized water into the mackerel meat emulsion to obtain a meat emulsion solution, heating in a boiling water bath for 15-20 min, and cooling to room temperature; then adding alkaline protease for enzymolysis for 1.5-3.5 h; and (3) after the enzymolysis is finished, carrying out water bath for 10min at 100 ℃, centrifuging to remove sediment, and freeze-drying supernatant fluid to obtain the mackerel antihypertensive active peptide. The mackerel antihypertensive active peptide prepared by the invention has good inhibition effect on ACE and good blood pressure control function, and when the mackerel antihypertensive active peptide is applied to bread, patients with hypertension can control blood pressure through daily diet therapy, the intake of medicines is reduced, and the requirements of people on health foods are met.
Description
Technical Field
The invention belongs to the technical field of food processing, and particularly relates to a preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread.
Background
In recent years, bioactive peptides have been attracting attention because of their various biological activities such as antihypertensive, anticoagulant and antitumor. The marine protein source active peptide has the advantages of complex structure, multiple types, strong activity, low side effect and the like, and therefore has wide application prospect in the fields of health foods, medicines, cosmetics and the like. With the continuous improvement of research technology and the expansion of research range of bioactive substances, more and more marine fish bioactive peptides are being explored. Mackerel contains a considerable proportion of muscle protein, which has high nutritive value and balanced amino acid composition, and also contains substances such as bioactive peptides with various physiological functions.
Along with the improvement of living standard, the hypertension population in China is more and more. Hypertension patients generally control blood pressure by taking captopril and other medicaments, and the medicaments generally have certain toxic and side effects. Therefore, finding safer ways to lower blood pressure without side effects is an urgent need for every hypertensive patient. The polypeptide with the activity of inhibiting Angiotensin Converting Enzyme (ACE) in food sources can effectively reduce blood pressure, has no toxic or side effect, and can be used as functional food for resisting hypertension to replace antihypertensive drugs.
Toast bread is western bread, has rich and varied taste, is convenient and quick to eat, is easy to digest and absorb, and is popular with people in fast-paced life. Traditional bread products on the market are mainly in the aspects of convenient carrying and eating, delicious taste and the like, and have less attention on the functional aspects of the products. In recent years, the living standard of people is gradually improved, and consumers are not limited to pursuing mouthfeel and flavor for bread snack foods, but pay more attention to whether the nutrition of the food is balanced and comprehensive and whether the food has a health care function.
Disclosure of Invention
The invention aims to provide a preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the activity of the polypeptide is determined by the molecular size, amino acid types and sequence, and the polypeptides with different effects can be obtained by different enzymolysis methods from different sources. The invention takes mackerel muscle as a raw material, prepares active peptide with good blood pressure reducing function through enzymolysis, and applies the active peptide to food to develop toast bread with the blood pressure reducing function. The mackerel antihypertensive active peptide is added into the bread, so that the edible nutritive value of the bread can be improved to a great extent, the hypertension patient can control the blood pressure through diet therapy, the intake of medicines is reduced, and the requirements of people on health foods are met.
The preparation method of mackerel antihypertensive active peptide provided by the invention comprises the following steps:
1) Adding deionized water into mackerel meat emulsion, and stirring and uniformly mixing to obtain meat emulsion solution;
2) Heating the meat emulsion solution in boiling water bath for 15-20 min, cooling to room temperature, then adjusting the pH of the solution, and adding protease for enzymolysis to obtain enzymolysis solution;
3) And (3) carrying out water bath on the enzymolysis liquid at 100 ℃ for 10-15min, centrifuging to remove sediment, and collecting supernatant for freeze drying to obtain the mackerel antihypertensive active peptide.
The ratio of the mackerel meat emulsion to the water in the step 1) is 1mg to 3-5 mL.
And 2) the pH value of the solution in the enzymolysis process is 8.0-9.0.
The protease in the step 2) is alkaline protease, and the addition amount of the protease is 3000U/g mackerel meat emulsion.
The enzymolysis conditions of the step 2) are as follows: the water bath at 50 ℃ is carried out for 1.5 to 3.5 hours, the enzymolysis process is accompanied by stirring, and the stirring speed is 80 to 100r/min.
The rotational speed of the centrifugation in the step 3) is 7000-9000 r/min, and the centrifugation time is 20-30min.
The mackerel antihypertensive active peptide is used for preparing bread and comprises the following specific steps:
(1) Preparing materials: weighing 0.5-2 parts of mackerel antihypertensive active peptide, 200-300 parts of high gluten flour, 30-50 parts of sugar, 2-4 parts of salt, 15-25 parts of butter, 4-6 parts of yeast, 40-60 parts of eggs and 50-150 parts of milk according to parts by weight;
(2) Dough kneading: placing milk and egg at the bottom of bread machine, sequentially placing high gluten flour, mackerel antihypertensive active peptide, sugar and salt into opposite corners of bread machine, preventing yeast from contacting sugar and salt, burying in flour, and starting bread machine; stirring for 10min, adding softened butter, and stirring for 10min;
(3) Primary fermentation: kneading dough, putting the smooth bone surface upwards into a fermentation basin for oiling, covering a preservative film for primary fermentation for 1h, and removing the dough when the area of the dough is twice as large for pressing and exhausting;
(4) Rolling: dividing the dough obtained in the step (3) into three equal parts, rounding and standing for 15 minutes, and covering a preservative film to prevent air drying; rolling the dough twice, and relaxing for 10min after the first rolling;
(5) Secondary fermentation: putting the rolled bread embryo in the step (4) into a toast box along the same spiral direction for secondary fermentation for 1h;
(6) Baking: baking with upper and lower fire at 180 deg.C for 30min.
The invention has the beneficial effects that: (1) The mackerel antihypertensive active peptide prepared by the invention has good inhibition effect on Angiotensin Converting Enzyme (ACE), and has a half inhibition concentration (IC 50) of only 39.3 mu mol/L and good blood pressure control function. (2) Under the condition of ensuring the taste, the food-originated mackerel antihypertensive active peptide is applied to bread to prepare the functional meal replacement bread with antihypertensive effect. The hypertension patients can control the blood pressure through daily diet therapy, reduce the intake of medicines and meet the current requirements of people on health foods.
Drawings
FIG. 1 shows breads prepared in examples 3-6 and comparative examples;
fig. 2 shows the test results of the blood pressure lowering effect (ACE inhibition ratio) of example 7.
Detailed Description
EXAMPLE 1 preparation of mackerel antihypertensive active peptide
Taking mackerel meat emulsion, and adding deionized water into the mackerel meat emulsion according to the solid-to-liquid ratio of 1mg to 3-5 mL to obtain a meat emulsion solution; heating the meat emulsion solution in a boiling water bath for 15-20 minutes, and naturally cooling to room temperature. Then, the pH of the solution is regulated to 8.5, alkaline protease is added according to the standard of 3000U/g meat emulsion, water bath is carried out at 50 ℃, and stirring (80-100 r/min) is carried out for enzymolysis for 2h. And after enzymolysis is finished, carrying out water bath for 10min at 100 ℃, centrifuging the solution (8000 r/min for 20 min), removing precipitates, and freeze-drying supernatant fluid to obtain the antihypertensive active peptide dry powder product.
The molecular weight distribution of the obtained mackerel antihypertensive active peptide is shown in Table 1. The average relative molecular weight of the active peptide prepared by the method is 727. Wherein the molecular weight is lower than 2000Da and is 94.5%, which belongs to small molecular polypeptide and is easy to be absorbed.
TABLE 1 molecular weight distribution of blood pressure-lowering active peptides from mackerel
EXAMPLE 2 antihypertensive Effect of mackerel active polypeptide (ACE inhibitory Activity)
Angiotensin Converting Enzyme (ACE) is a dipeptide carboxypeptidase that plays a critical role in the cardiovascular system of the human body and ACE increases blood pressure by converting angiotensin I to angiotensin ii. Angiotensin converting enzyme inhibitory peptides, also known as ACE inhibitory peptides, inhibit the conversion of angiotensin i to angiotensin ii, thereby lowering hypertension. The mackerel antihypertensive active peptide is prepared into polypeptide solutions with different concentrations by deionized water, the inhibition capability of the mackerel antihypertensive active peptide on ACE is analyzed to evaluate the antihypertensive effect of the polypeptide, and the result shows that the half inhibition concentration (IC 50) of the mackerel polypeptide prepared by the invention on ACE is 39.3 mu mol/L. Other food-derived bioactive peptides, such as marine chlorella-derived ACE inhibitory peptide IC50 of 219 μmol/L and egg protein-derived tripeptide ACE inhibitory peptide IC50 of 250 μmol/L. In contrast, the mackerel active polypeptide prepared by the invention has lower ACE half-inhibition concentration, which indicates that the mackerel active polypeptide has better blood pressure reducing effect, and the mackerel active polypeptide has small molecular weight and is easier to be absorbed, thus being suitable for developing functional food for reducing hypertension.
Example 3 preparation of antihypertensive active peptide bread
A method for preparing bread containing mackerel antihypertensive active peptide, comprising the following steps:
(1) Preparing materials: weighing 0.5 part of mackerel antihypertensive active peptide, 250 parts of high-gluten flour, 40 parts of sugar powder, 3 parts of salt, 20 parts of butter, 5 parts of yeast, 50 parts of eggs and 100 parts of milk according to parts by weight;
(2) Dough kneading: placing milk and egg at the bottom of bread machine, sequentially placing high gluten flour, mackerel antihypertensive active peptide, sugar powder and salt into opposite corners of bread machine, preventing yeast from contacting sugar and salt, and burying into flour to start bread machine; stirring for 10min, adding softened butter, and stirring for 10min;
(3) Primary fermentation: kneading dough, putting the smooth bone surface upwards into a fermentation basin for oiling, covering a preservative film for primary fermentation for 1h, and removing the dough when the area of the dough is twice as large for pressing and exhausting;
(4) Rolling: dividing the dough obtained in the step (3) into three equal parts, rounding and standing for 15 minutes, and covering a preservative film to prevent air drying; rolling the dough twice, and relaxing for 10min after the first rolling;
(4) Secondary fermentation: putting the rolled bread embryo in the step (4) into a toast box along the same spiral direction for secondary fermentation for 1h;
(6) Baking: baking with upper and lower fire at 180 deg.C for 30min.
Example 4 preparation of antihypertensive active peptide bread
The amount of the polypeptide in example 1 was adjusted to 1 part, and the other was kept the same as in example 3, to obtain a blood pressure-lowering active peptide bread.
Example 5 preparation of antihypertensive active peptide bread
The amount of the polypeptide in example 1 was adjusted to 1.5 parts, and the other ingredients were kept the same as in example 3, to obtain a blood pressure-lowering active peptide bread.
EXAMPLE 6 preparation of antihypertensive active peptide bread
The amount of the polypeptide in example 1 was adjusted to 2 parts, and the other was kept the same as in example 3, to obtain a blood pressure-lowering active peptide bread.
Comparative example
The addition of mackerel antihypertensive active peptide in example 3 was omitted, and the other ingredients were the same as in example 3, to obtain bread.
Example 7 ACE inhibitory Activity of breads prepared in examples 3-6
The sample added in the research is high-quality ACE inhibitory peptide with ACE inhibitory activity of 39.3 mu mol/L which is screened out in the front stage of a key laboratory of the university of Fujian province of offshore resource biotechnology, and has good angiotensin converting enzyme inhibitory activity, thereby playing a role in reducing blood pressure. The effect of different amounts of mackerel antihypertensive active peptide on the ACE inhibition of the bread extract is shown in fig. 2, and the ACE inhibition rate of the bread extract is remarkably increased with the increase of the amount of mackerel antihypertensive active peptide. When the addition amount is 2 parts (namely example 6), the ACE inhibition rate reaches 82.39 percent, and the antihypertensive food has good antihypertensive activity and wide application prospect as antihypertensive functional food.
Example 8 organoleptic evaluation of breads prepared in examples 3-6
Sensory evaluation test method: sensory evaluation is seen in table 2. The sensory evaluation results of the breads prepared in examples 3 to 6 are shown in Table 3. The sensory scores of the toast bread are higher than those of the control group by adding a proper amount of mackerel antihypertensive active polypeptide (examples 3-4), which shows that the proper amount of mackerel antihypertensive active polypeptide can not only achieve the effect of reducing blood pressure, but also improve the quality of the bread. However, too high an amount of polypeptide would affect the sensory scores, such as example 6, which is significantly lower than the control.
Table 2 sensory evaluation table
Table 3 examples 3-6 bread sensory evaluation scores
Claims (3)
1. A preparation method of mackerel antihypertensive active peptide is characterized by comprising the following steps:
1) Adding water into mackerel meat emulsion, and stirring and uniformly mixing to obtain meat emulsion solution;
2) Heating the meat emulsion solution in a boiling water bath for 15-20 min, cooling to room temperature, then adjusting the pH of the solution to 8.0-9.0, and adding protease for enzymolysis to obtain an enzymolysis solution;
the protease is alkaline protease, the addition amount of the protease is 3000U/g mackerel meat emulsion, and the enzymolysis conditions are as follows: 50. water bath at the temperature of between 1.5 and 3.5 hours, stirring during enzymolysis, wherein the stirring speed is 80 to 100 r/min;
3) And (3) carrying out water bath on the enzymolysis liquid at 100 ℃ for 10-15min, centrifuging to remove sediment, and collecting supernatant for freeze drying to obtain the mackerel antihypertensive active peptide.
2. The method for preparing the mackerel antihypertensive active peptide of claim 1, wherein the ratio of mackerel meat emulsion to water in the step 1) is 1 mg/3-5 mL.
3. The method for preparing the mackerel antihypertensive active peptide of claim 1, wherein the rotational speed of centrifugation in the step 3) is 7000-9000 r/min and the centrifugation time is 20-30min.
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| CN101130800A (en) * | 2007-09-01 | 2008-02-27 | 浙江大学 | Method for preparing antihypertensive peptide by enzymolysis of hairtail chine |
| CN101168762B (en) * | 2007-10-23 | 2011-05-11 | 浙江大学 | Method for preparing antihypertensive peptide from hairtail backbone by fermentation method |
| CN101240312A (en) * | 2008-01-21 | 2008-08-13 | 南昌大学 | Method for preparing ACE inhibition peptide originate from fish skin |
| JP5416964B2 (en) * | 2008-12-26 | 2014-02-12 | ヤマキ株式会社 | Novel tripeptides, methods for producing these tripeptides, and methods for producing angiotensin converting enzyme inhibitors |
| CN105146045B (en) * | 2015-09-22 | 2018-09-14 | 华南理工大学 | Sea-fish protein prepares the method for combining blood pressure lowering peptide with fresh water fish protein |
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