阅读说明：本技术 一种大蒜降压肽的制备方法 (Preparation method of garlic antihypertensive peptide ) 是由 李宇馨 王婷婷 于 2021-02-06 设计创作，主要内容包括：本发明公开了一种大蒜降压肽的制备方法,本发明涉及大蒜降压肽制备技术领域,包括：A1．原料选择：取新鲜大蒜为原料,清洗干净,用刀片剁碎成渣,并放入冷冻保鲜；再选择一颗新鲜大蒜,将大蒜清洗干净,将大蒜碾压成蒜泥；A2．蒜泥处理：蒜泥加入玻璃容器中,A3．酶解液提取；将冷冻的大蒜渣取出,大蒜渣放入玻璃容器中,向玻璃容器中加入重量为大蒜渣重量十倍的蒸馏水和95wt％乙醇以及蒜泥液,在本发明中,将装有冷冻后的大蒜渣的玻璃容器置于水浴盘上,有利于大蒜渣快速进行回流反应。提高提取效率,利用蒸馏水与蒜泥搅拌得到蒜泥的溶液和蒸馏水以及乙醇充分融合,提高降压肽产量,便于对降压肽进行分组份研究。(The invention discloses a preparation method of garlic antihypertensive peptide, relating to the technical field of garlic antihypertensive peptide preparation, and comprising the following steps: A1. selecting raw materials: taking fresh garlic as a raw material, cleaning, chopping into residues by using a blade, and freezing for preservation; selecting a fresh garlic, cleaning the garlic, and grinding the garlic into mashed garlic; A2. and (3) mashed garlic treatment: adding the mashed garlic into a glass container, and A3, extracting enzymatic hydrolysate; taking out the frozen garlic residues, putting the garlic residues into a glass container, and adding distilled water and 95 wt% ethanol which are ten times of the weight of the garlic residues and garlic paste liquid into the glass container. The extraction efficiency is improved, the solution of mashed garlic obtained by stirring the distilled water and the mashed garlic is fully fused with the distilled water and the ethanol, the yield of the antihypertensive peptide is improved, and the research on the antihypertensive peptide in groups is facilitated.)

1. A preparation method of garlic antihypertensive peptide is characterized by comprising the following steps:

A1. selecting raw materials: taking fresh garlic as a raw material, cleaning, chopping into residues by using a blade, and freezing for preservation; selecting a fresh garlic, cleaning the garlic, and grinding the garlic into mashed garlic;

A2. and (3) mashed garlic treatment: adding mashed garlic into a glass container, adding distilled water with the weight four times that of the mashed garlic into the glass container, and fully stirring;

A3. extracting enzymolysis liquid; taking out the frozen garlic residues, putting the garlic residues into a glass container, adding distilled water with the weight ten times that of the garlic residues, 95 wt% ethanol and garlic paste liquid into the glass container, and carrying out reflux reaction for 2 hours at 95 ℃;

and (3) carrying out enzymolysis on the filtrate: cooling the filtrate to 45-60 deg.C, adjusting pH to 9-11, adding alkaline protease based on 3-5% of the weight of Bulbus Allii residue raw material, stirring for enzymolysis for 3 hr to obtain enzymolysis solution;

A4. keeping the enzymolysis liquid at 100 ℃ for 15min to inactivate enzyme;

A5. filtering the enzymolysis solution by an ultra-membrane to remove protease;

A6. adding the protease with the volume of 1/2 enzymolysis liquid into the solution after removing the protease again, and stirring for enzymolysis for 0.5 h.

A7. Separating and filtering the obtained solution through mass spectrometry of a mass spectrometer, and concentrating the obtained solution after separating the solution through the mass spectrometry;

A8. and drying the concentrated solution in vacuum at low temperature.

2. The method of claim 1, wherein the volume of the 95 wt% ethanol and the volume of the mashed garlic are the same, and the volume of the 95 wt% ethanol and the mashed garlic is 2ml in the step A3.

3. The method according to claim 1, wherein in the step A3, the frozen garlic clove residues are placed in a glass, and the glass is placed in a water bath tray with a temperature of 30 °.

4. The method for preparing garlic antihypertensive peptide according to claim 1, wherein in the filtration treatment of step a5, the ultrafiltration membrane is a cellulose acetate membrane, and the filtration temperature is 30 ° to 35 °.

5. The method for preparing garlic antihypertensive peptide according to claim 1, wherein 1ml of protease is added in the treatment of step a6, and the temperature during the treatment is 40 ° to 45 °.

6. The method for preparing garlic antihypertensive peptide according to claim 1, wherein in the mass spectrometric separation of step a7, the separation environment temperature is 40 ° to 50 ° and the gas pressure is 23 MPa.

Technical Field

The invention belongs to the technical field of preparation of garlic antihypertensive peptides, and particularly relates to a preparation method of garlic antihypertensive peptides.

Background

Antihypertensive peptides derived from food are a general name of micromolecular polypeptides capable of reducing human blood pressure, generally the molecular weight of the active antihypertensive peptides is below 1000, the antihypertensive peptides are generally obtained by hydrolyzing protein under a relatively mild condition by protease, the antihypertensive peptides have obvious antihypertensive effect, have no influence on normal blood pressure and no side effect, garlic contains three active peptides, the garlic has high yield in life and is easy to take materials, and therefore, the preparation of the antihypertensive peptides by using the active peptides of the garlic becomes a research hotspot;

at present, researchers use an enzymatic hydrolysis method to carry out enzymolysis on garlic residues, the garlic residues are unfrozen at a normal state at room temperature after being frozen, the unfreezing speed is low, the antihypertensive peptides in one garlic are extracted at one time, the yield of the antihypertensive peptides is low, and the research on the obtained antihypertensive peptides in parts is inconvenient.

Disclosure of Invention

In view of this, the present invention aims to provide a method for preparing garlic antihypertensive peptides, so as to effectively solve the problems of slow thawing speed when thawing is performed at normal room temperature, extraction of antihypertensive peptides from garlic at one time, and low yield of antihypertensive peptides in the garlic.

In order to achieve the purpose, the invention provides the following technical scheme:

a preparation method of garlic antihypertensive peptide comprises the following steps:

A1. selecting raw materials: taking fresh garlic as a raw material, cleaning, chopping into residues by using a blade, and freezing for preservation; selecting a fresh garlic, cleaning the garlic, and grinding the garlic into mashed garlic;

A2. and (3) mashed garlic treatment: adding mashed garlic into a glass container, adding distilled water with the weight four times that of the mashed garlic into the glass container, and fully stirring;

A3. extracting enzymolysis liquid; taking out the frozen garlic residues, putting the garlic residues into a glass container, adding distilled water with the weight ten times that of the garlic residues, 95 wt% ethanol and garlic paste liquid into the glass container, and carrying out reflux reaction for 2 hours at 95 ℃;

and (3) carrying out enzymolysis on the filtrate: cooling the filtrate to 45-60 deg.C, adjusting pH to 9-11, adding alkaline protease based on 3-5% of the weight of Bulbus Allii residue raw material, stirring for enzymolysis for 3 hr to obtain enzymolysis solution;

A4. keeping the enzymolysis liquid at 100 ℃ for 15min to inactivate enzyme;

A5. filtering the enzymolysis solution by an ultra-membrane to remove protease;

A6. adding the protease with the volume of 1/2 enzymolysis liquid into the solution after removing the protease again, and stirring for enzymolysis for 0.5 h.

A7. Separating and filtering the obtained solution through mass spectrometry of a mass spectrometer, and concentrating the obtained solution after separating the solution through the mass spectrometry;

A8. drying the concentrated solution in vacuum at low temperature;

preferably, in the step A3, the added 95 wt% ethanol and garlic clove juice are the same volume, and the volume of the 95 wt% ethanol and garlic clove juice is 2 ml.

Preferably, in the process of step a3, the frozen garlic clove residues are placed in a glass, and the glass is placed in a water bath tray with a temperature of 30 °.

Preferably, in the filtration treatment of the step A5, the ultrafiltration membrane adopts a cellulose acetate membrane, and the temperature of the filtration environment is 30-35 degrees.

Preferably, in the step A6 treatment, 1ml of protease is added, and the temperature during the treatment is 40-45 deg.

Preferably, in the mass spectrum separation of the step A7, the separation environment temperature is 40-50 ℃ and the air pressure is 23 MPa.

Compared with the prior art, the invention has the beneficial effects that:

(1) according to the invention, the frozen garlic residues are placed in the heat-conducting glass container, the glass container is placed on the water bath tray filled with warm water, the temperature of the warm water is absorbed by the garlic residues by utilizing the heat conduction principle, the thawing speed of the garlic residues is improved, the garlic residues are thawed without a heating mode, the activity of polypeptides in the garlic residues is protected, the thawing speed of the garlic residues is improved, and the quick reflux reaction of the garlic residues is facilitated. The extraction efficiency is improved.

(2) According to the invention, distilled water and mashed garlic are stirred to obtain a mashed garlic solution, the mashed garlic solution contains a large amount of polypeptide, the mashed garlic solution obtained by stirring the distilled water and the mashed garlic is fully fused with the distilled water and ethanol, the yield of the antihypertensive peptide is improved, the research on the antihypertensive peptide in groups is facilitated, the antihypertensive peptide does not need to be extracted for many times, and the cost and the time are saved.

(3) In the invention, the solution of the garlic residues is decomposed by protease twice, so that the smell in the solution of the garlic residues is effectively eliminated, the pungent smell is reduced, the smell is prevented from remaining on the inner wall of the extraction equipment, the cleaning of the extraction equipment is facilitated, and the quality of the solution after enzymolysis is improved.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The first embodiment is as follows:

a preparation method of garlic antihypertensive peptide comprises the following steps:

A1. selecting raw materials: taking fresh garlic as a raw material, cleaning, chopping into residues by using a blade, and freezing for preservation; selecting a fresh garlic, cleaning the garlic, and grinding the garlic into mashed garlic;

A2. and (3) mashed garlic treatment: adding mashed garlic into a glass container, adding distilled water with the weight four times that of the mashed garlic into the glass container, and fully stirring;

A3. extracting enzymolysis liquid; taking out the frozen garlic residues, placing the frozen garlic residues in a glass cup, placing the glass cup in a water bath tray at the temperature of 30 ℃, placing the garlic residues in a glass container, adding distilled water and 95 wt% ethanol which are ten times of the weight of the garlic residues and garlic paste liquid into the glass container, and carrying out reflux reaction for 2 hours at the temperature of 95 ℃;

and (3) carrying out enzymolysis on the filtrate: cooling the filtrate to 45-60 deg.C, adjusting pH to 9-11, adding alkaline protease based on 3-5% of the weight of Bulbus Allii residue raw material, stirring for enzymolysis for 3 hr to obtain enzymolysis solution;

A4. keeping the enzymolysis liquid at 100 ℃ for 15min to inactivate enzyme;

A5. filtering the enzymolysis solution by an ultra-membrane to remove protease;

A6. adding the protease with the volume of 1/2 enzymolysis liquid into the solution after removing the protease again, and stirring for enzymolysis for 0.5 h.

A7. Separating and filtering the obtained solution through mass spectrometry of a mass spectrometer, and concentrating the obtained solution after separating the solution through the mass spectrometry;

A8. drying the concentrated solution in vacuum at low temperature;

example two:

a preparation method of garlic antihypertensive peptide comprises the following steps:

A1. selecting raw materials: taking fresh garlic as a raw material, cleaning, chopping into residues by using a blade, and freezing for preservation; selecting a fresh garlic, cleaning the garlic, and grinding the garlic into mashed garlic;

A2. and (3) mashed garlic treatment: adding mashed garlic into a glass container, adding distilled water with the weight twice that of the mashed garlic into the glass container, and fully stirring;

A3. extracting enzymolysis liquid; taking out the frozen garlic residues, placing the frozen garlic residues in a glass cup, placing the glass cup in a water bath tray at the temperature of 50 ℃, placing the garlic residues in a glass container, adding distilled water and 95 wt% ethanol which are ten times of the weight of the garlic residues and garlic paste liquid into the glass container, and carrying out reflux reaction for 1h at the temperature of 95 ℃;

and (3) carrying out enzymolysis on the filtrate: cooling the filtrate to 45-60 deg.C, adjusting pH to 9-11, adding alkaline protease based on 3-5% of the weight of Bulbus Allii residue raw material, stirring for enzymolysis for 1.5 hr to obtain enzymolysis solution;

A4. keeping the enzymolysis liquid at 100 ℃ for 7.5min to inactivate enzyme;

A5. filtering the enzymolysis solution by an ultra-membrane to remove protease;

A6. adding the protease with the volume of 1/2 enzymolysis liquid into the solution after removing the protease again, and stirring for enzymolysis for 25 min.

A7. Separating and filtering the obtained solution through mass spectrometry of a mass spectrometer, and concentrating the obtained solution after separating the solution through the mass spectrometry;

A8. drying the concentrated solution in vacuum at low temperature;

for the 2 embodiments described above:

comparing the thawing time of the frozen garlic residue to obtain the following table:

	temperature of	Time
			Example 1	30°	20min
Example 2	50°	11min

According to the method, the thawing speed of the garlic residues is obviously accelerated by cooling through the method provided by the invention, and the speed of the garlic residues entering the reflux reaction can be increased.

Comparing the content of the antihypertensive peptide obtained by adding the garlic paste liquid into the frozen garlic residue solution to obtain the following table:

	amount of mashed garlic	Antihypertensive peptide content
			Example 1	2ml	5％
Example 2	1ml	2.5％

As can be seen from the above table, after the thawing by the method provided by the invention, the thawing time of the garlic residue is accelerated, the extraction speed of the antihypertensive peptide is effectively improved, and the extraction amount of the antihypertensive peptide is effectively improved after the garlic puree is added.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.