CN109355344A - A kind of preparation method of antihypertensive activity peptide - Google Patents
A kind of preparation method of antihypertensive activity peptide Download PDFInfo
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- CN109355344A CN109355344A CN201910014771.9A CN201910014771A CN109355344A CN 109355344 A CN109355344 A CN 109355344A CN 201910014771 A CN201910014771 A CN 201910014771A CN 109355344 A CN109355344 A CN 109355344A
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The present invention relates to active peptide preparation field, specifically a kind of preparation method of antihypertensive activity peptide.Digested using Alaska pollock as raw material through enzyme, after enzymolysis liquid enzyme deactivation, be centrifuged, filter 500-3000D Antihypertensive Peptides.The Alaska pollock blood pressure lowering peptide ACE inhibitory activity with higher that the present invention is prepared, blood pressure lowering effect is good, has no toxic side effect and the utilization that is easily absorbed by the body.Meanwhile the present invention will provide foundation for the comprehensive utilization of gadus resource, and provide foundation further to develop natural blood-pressure reducing health care product.
Description
Technical field
The present invention relates to active peptide preparation field, specifically a kind of preparation method of antihypertensive activity peptide.
Background technique
Hypertension is currently a popular most commonly used cardiovascular disease, be the major chronic illnesses that are detrimental to health it
One.In regulating blood pressure system, angiotensin converting enzyme (Angiotensin-converting enzyme, ACE) can be incited somebody to action
Angiotensin I hydrolysis, generating has the Angiotensin II for increasing blood pressure effect, it can also hydrolyze bradykinin, make it
Lose the effect to reduce blood pressure.Food-borne protein blood pressure-reducing peptide, i.e. inhibiting peptide of tonin (Angiotensin
I-conberting enzyme inhibitorypeptides, ACEI) with high security, Small side effects the advantages that, it is more next
More favored.Currently, people from vegetable protein, animal protein, aquatic product protein, lactoprotein zymolyte in had
The peptide fragment of ACE inhibitory activity.Marine product such as gadus has good protective effect to cardio-cerebrovascular, is conducive to pre-
The cardiovascular and cerebrovascular diseases such as anti-high blood, myocardial infarction.Wang Haiying etc. is drop with the low-temperature alkaline protease from marine microorganism
Instrumental purpose is solved in skin deep-sea cod, preparation has the active blood pressure lowering peptide of angiotensin converting enzyme inhibitor (Wang Haiying, Song
Phase forever, research [J] the modern food that the quiet enzymatic hydrolysis cod skin of grandson prepares angiotensin converting enzyme inhibitor ACEI is scientific and technological, and 2008
(10): 987-991. it) but the not anti-digestion of the Antihypertensive Peptides of above-mentioned most food sources, is easy to be digested enzyme decomposition in vivo, lose
To the inhibiting effect of ACE.Thus, using being subject to certain restrictions.
Summary of the invention
It is an object of that present invention to provide a kind of preparation methods of antihypertensive activity peptide.
To achieve the above object the invention adopts a technical scheme as:
A kind of preparation method of antihypertensive activity peptide, is digested using Alaska pollock as raw material through enzyme, after enzymolysis liquid enzyme deactivation, from
The heart, filter 500-3000D Antihypertensive Peptides.
Further, Alaska pollock oligopeptide or its protein peptides are dissolved through water, and enzyme is added after dissolution and is digested,
Enzymolysis liquid be heated to enzyme deactivation in boiling water, centrifugation, filter 500-3000D Antihypertensive Peptides；The additive amount of the enzyme is every 100ml former
0.15-4g is added in material lysate, wherein enzyme is bromelain, papain, compound protease, trypsase, alkalinity
Protease or acid protease.
Trypsase, alkali protease or acid protease are added in the lysate, is digested at 37 DEG C；Wherein, pancreas
The enzymatic hydrolysis system pH of protease and alkali protease is 8.0, and the enzymatic hydrolysis system pH of acid protease is 3.0.
Bromelain, papain or compound protease are added in the lysate, is digested at 55 DEG C, wherein
Above-mentioned enzymatic hydrolysis system pH is 7.0.
The enzymatic hydrolysis system constant temperature digests 2-5h, enzymolysis liquid is transferred in boiling water after enzymatic hydrolysis, 10-15min of enzyme deactivation, with
The speed of 3000-5000r/min is centrifuged 10-20min, collects clear liquid through 2500D ultrafiltration membrane ultrafiltration, filtered solution is obtained, to filtration
Liquid carries out reverse osmosis with 250D anti-seepage membrane to get the Antihypertensive Peptides of 500-3000D are arrived.
The oligomeric peptide molecular weight of gadus is 500-1000Da；Gadus polypeptide molecular weight is 1000-3000Da.
The Alaska pollock oligopeptide or its protein peptides are dissolved through water, acquisition concentration be 4%(w/v) solution.
Advantage for present invention:
The Alaska pollock blood pressure lowering peptide ACE inhibitory activity with higher that the present invention is prepared, blood pressure lowering effect is good, nothing
Toxic side effect and the utilization that is easily absorbed by the body.Meanwhile the present invention provides foundation for the comprehensive utilization of gadus resource, and is into one
Step develops natural blood-pressure reducing health care product and provides foundation.It is in particular in:
Oligopeptide and the polypeptide ACE inhibitory activity with higher that the present invention obtains, are difficult further to be digested in alimentary canal,
It can be directly absorbed and utilized, there is preferable antihypertensive activity, be laid the foundation for the exploitation of antihypertensive drugs.
Detailed description of the invention
Fig. 1 is that different enzymes provided in an embodiment of the present invention can try hard to the enzymatic hydrolysis of Alaska pollock oligopeptide.
Fig. 2 is that different enzymes provided in an embodiment of the present invention can try hard to the enzymatic hydrolysis of Alaska pollock protein peptides.
Specific embodiment
A specific embodiment of the invention is described further below in conjunction with example, it is noted that retouch in this place
The specific embodiment stated is simply to illustrate that with the present invention is explained, it is not limited to the present invention.
Embodiment 1
Precision weighs the gadus albumen that the gadus oligopeptide that 24g molecular weight is 500-1000Da or molecular weight are 1000-3000Da
Polypeptide is dissolved in respectively in the deionized water of 600 mL, and stirring, which makes it completely dissolved, obtains concentration as oligopeptide/egg of 4%(W/V)
White peptide lysate.Take the type and concentration (6 kinds of different protease, 3 use of every kind of enzyme of 18 50 mL centrifuge tube marker enzymes
Amount).500 mL large beakers are separately taken, oligopeptide/protein peptides lysate that the above-mentioned configured good concentration of 300 mL is 4% is added, makes
With sodium hydroxide or hydrochloric acid solution using the pH value of pH meter adjusting polypeptide liquid to 7.0, precision measures the polypeptide liquid each 30 of pH7.0
ML is transferred in 1-6 centrifuge tube and 10-12 centrifuge tube；500 mL large beakers are taken again, and it is above-mentioned configured good that 200 mL are added
Oligopeptide/albumen peptide solution, adjust the pH value of polypeptide liquid to 8.0 using pH meter using sodium hydroxide or hydrochloric acid solution, it is accurate
Each 30 mL of oligopeptide/albumen peptide solution for measuring pH8.0 is transferred to 1-6 centrifuge tube 7-9 centrifuge tube and No. 13-15 centrifugation
Guan Zhong；Take 100 mL beakers again, the above-mentioned configured good oligopeptide/albumen peptide solution of 100 mL be added, using sodium hydroxide or
Using the pH value of pH meter adjusting polypeptide liquid to 3.0, each 30 mL of oligopeptide solution that precision measures pH3.0 is transferred to hydrochloric acid solution
In 1-6 centrifuge tube 16-18 centrifuge tube.
According to the above-mentioned enzyme marked in advance type (bromelain, papain, compound protease, trypsase,
Alkali protease, acid protease) and relative to liquid volume enzyme dosage (g/100mL), the different enzyme of precision weighing is added
Into different centrifuge tubes.Then (bromelain, papain, compound egg are digested in the optimum temperature constant temperature of respective enzyme
The optimum temperature of white enzyme is 55 DEG C；Trypsase, alkali protease, acid protease optimum temperature be 37 DEG C).Wait digest
After the completion, hydrolyzate is heated to 10 min of enzyme deactivation in boiling water, is then centrifuged (3000-5000r/min) 15min, is obtained
Supernatant.The relative molecular mass of the ace inhibitory peptide generated after digesting according to food protein generally in 1500D hereinafter, with
2500D ultrafiltration membrane ultrafiltration, obtains filtered solution, carries out reverse osmosis (using 250D anti-seepage membrane) to filtered solution, obtained trapped fluid a part
By filling, high pressure sterilization, the Antihypertensive Peptides of the 500-3000D under different PH enzymatic hydrolysis systems are obtained；
Antihypertensive Peptides under above-mentioned difference PH enzymatic hydrolysis system digest Alaska pollock oligopeptide using ultraviolet spectrophotometry
Liquid supernatant carries out ACE inhibitory activity detection, measures the absorbance value (A under 228 nm228), it obtains each protease and most preferably digests
Condition (referring to table 1 and Fig. 1 and Fig. 2).
Its inhibiting rate is calculated as follows:
Wherein,
I:ACE maximum inhibition
A0: the absorbance value of control group
A1: the absorbance value of test sample
1 protease optimum enzymolysis condition of table
It would know that different protease under respective optimum hydrolysising condition to Alaska pollock oligopeptide or albumen peptase by above-mentioned
The activity that solution obtains Antihypertensive Peptides is different, wherein for Alaska pollock oligopeptide, the enzymatic hydrolysis capacity of water of 6 kinds of different enzymes
Are as follows: trypsase > acid protease > alkali protease > papain > bromelain > compound protease；For Arras
Add gadus polypeptide, the enzymatic hydrolysis capacity of water of 6 kinds of different enzymes are as follows: acid protease > trypsase > papain > compound egg
White enzyme > alkali protease > bromelain.Further by Fig. 1 and 2 as it can be seen that trypsase is to Alaska pollock oligopeptide
Enzymolysis activity it is best, inhibiting rate reaches as high as 93.7%；Acid protease to the enzymolysis activities of Alaska pollock protein peptides most
Good, inhibiting rate reaches as high as 94%, and trypsase is preferable to its hydrolysis ability, and inhibiting rate reaches as high as 90%.
Claims (6)
1. a kind of preparation method of antihypertensive activity peptide, it is characterised in that: digested using Alaska pollock as raw material through enzyme, enzyme
Solution liquid enzyme deactivation after, be centrifuged, filter 500-3000D Antihypertensive Peptides.
2. the preparation method of antihypertensive activity peptide according to claim 1, it is characterised in that: Alaska pollock oligopeptide or its
Protein peptides are dissolved through water, and enzyme is added after dissolution and is digested, enzymolysis liquid is heated to enzyme deactivation in boiling water, centrifugation, filters to obtain 500-
The Antihypertensive Peptides of 3000D；The additive amount of the enzyme is that 0.15-4g is added in every milliliter of dissolution of raw material liquid, wherein enzyme is bromelain
Enzyme, papain, compound protease, trypsase, alkali protease or acid protease.
3. the preparation method of antihypertensive activity peptide as described in claim 2, it is characterised in that:
Trypsase, alkali protease or acid protease are added in the lysate, is digested at 37 DEG C；Wherein, tryptose
The enzymatic hydrolysis system pH of enzyme and alkali protease is 8.0, and the enzymatic hydrolysis system pH of acid protease is 3.0.
4. the preparation method of antihypertensive activity peptide as described in claim 2, it is characterised in that: pineapple egg is added in the lysate
White enzyme, papain or compound protease digest at 55 DEG C, wherein above-mentioned enzymatic hydrolysis system pH is 7.0.
5. by the preparation method of antihypertensive activity peptide described in claim 1-4 any one, it is characterised in that: the enzymatic hydrolysis system
Constant temperature digests 2-5h, enzymolysis liquid is transferred in boiling water after enzymatic hydrolysis, 10-15min of enzyme deactivation, with the speed of 3000-5000r/min
It is centrifuged 10-20min, clear liquid is collected through 2500D ultrafiltration membrane ultrafiltration, obtains filtered solution, filtered solution is carried out instead with 250D anti-seepage membrane
It seeps to get the Antihypertensive Peptides of 500-3000D are arrived.
6. the preparation method of antihypertensive activity peptide as described in claim 2, it is characterised in that: the oligomeric peptide molecular weight of gadus is
500-1000Da；Gadus polypeptide molecular weight is 1000-3000Da.
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| CN201910014771.9A CN109355344A (en) | 2019-01-08 | 2019-01-08 | A kind of preparation method of antihypertensive activity peptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114214384A (en) * | 2021-12-22 | 2022-03-22 | 海南三元星生物科技股份有限公司 | Marine organism-derived collagen peptide, and extraction method and application thereof |
| CN114657229A (en) * | 2022-05-17 | 2022-06-24 | 中国科学院烟台海岸带研究所 | Method for extracting ACE inhibitory peptide from viscera of sea cucumber and application thereof |
Citations (4)
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|---|---|---|---|---|
| CN104975059A (en) * | 2014-04-08 | 2015-10-14 | 广州世优生物科技有限公司 | Method for preparing ACE (angiotensin converting enzyme) inhibitory peptide by using cod meat |
| CN105906709A (en) * | 2016-05-17 | 2016-08-31 | 青岛海博瑞克生物科技有限公司 | Alaska Pollock fish skin active oligopeptides as well as synthesis method and application thereof |
| CN107319401A (en) * | 2017-06-22 | 2017-11-07 | 新昌县瑞天食品科技有限公司 | The preparation method of cod activity extract |
| CN107937468A (en) * | 2018-01-15 | 2018-04-20 | 中国海洋大学 | Low bitter taste oligopeptides of a kind of gadus source function and preparation method thereof |
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2019
- 2019-01-08 CN CN201910014771.9A patent/CN109355344A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104975059A (en) * | 2014-04-08 | 2015-10-14 | 广州世优生物科技有限公司 | Method for preparing ACE (angiotensin converting enzyme) inhibitory peptide by using cod meat |
| CN105906709A (en) * | 2016-05-17 | 2016-08-31 | 青岛海博瑞克生物科技有限公司 | Alaska Pollock fish skin active oligopeptides as well as synthesis method and application thereof |
| CN107319401A (en) * | 2017-06-22 | 2017-11-07 | 新昌县瑞天食品科技有限公司 | The preparation method of cod activity extract |
| CN107937468A (en) * | 2018-01-15 | 2018-04-20 | 中国海洋大学 | Low bitter taste oligopeptides of a kind of gadus source function and preparation method thereof |
Non-Patent Citations (1)
| Title |
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| 王海英等: "酶解鳕鱼皮制备血管紧张素转换酶抑制剂ACEI 的研究", 《现代食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114214384A (en) * | 2021-12-22 | 2022-03-22 | 海南三元星生物科技股份有限公司 | Marine organism-derived collagen peptide, and extraction method and application thereof |
| CN114657229A (en) * | 2022-05-17 | 2022-06-24 | 中国科学院烟台海岸带研究所 | Method for extracting ACE inhibitory peptide from viscera of sea cucumber and application thereof |
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