CN106434815A - Method for preparing soybean antihypertensive peptides by membrane separation-electrodialysis technology - Google Patents
Method for preparing soybean antihypertensive peptides by membrane separation-electrodialysis technology Download PDFInfo
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- CN106434815A CN106434815A CN201611102584.9A CN201611102584A CN106434815A CN 106434815 A CN106434815 A CN 106434815A CN 201611102584 A CN201611102584 A CN 201611102584A CN 106434815 A CN106434815 A CN 106434815A
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- antihypertensive peptides
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- kcl
- antihypertensive
- peptides derived
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C07K1/24—Extraction; Separation; Purification by electrochemical means
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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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- 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
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
A method for preparing soybean antihypertensive peptides by membrane separation-electrodialysis technology includes the steps: (1) soybean protein hydrolysate preparation: placing soybean protein isolate into an enzyme reactor, adding an appropriate amount of distilled water, uniformly stirring the soybean protein isolate and the distilled water, performing enzymolysis after adjusting temperature and pH (potential of hydrogen), and denaturing enzyme and performing centrifugal separation after enzymolysis to obtain soybean protein hydrolysate; (2) soybean antihypertensive peptide separation: performing ultra-filtration for the soybean protein hydrolysate by a neutral ultra-filtration membrane to obtain coarse hydrolysate inhibiting peptide (ACE); (3) electrodialysis: collecting and separating KCl ions in KCl (1) to obtain the soybean antihypertensive peptides with negative charge, and collecting and separating KCl ions in KCl (2) to obtain the soybean antihypertensive peptides with positive charge. The concentration of the KCl ions in design application of an electrodialysis instrument is 0.5-1.0g/L, the antihypertensive peptides are separated, membrane separation and electrodialysis are combined by the technology, the soybean antihypertensive peptides with the positive charge are separated from the soybean antihypertensive peptides with the negative charge, and selectivity of target products is improved.
Description
Technical field
The invention belongs to foods processing technique, relates generally to a kind of using membrance separation-electrodialytic technique preparation Semen sojae atricolor blood pressure lowering
The method of peptide.
Background technology
In order to obtain the bioactive peptide of high-purity specific function, current the best way be using modern chromatographic techniques to hydrolysis
Thing is progressively separated, although and this separation method is finally obtained in that targeted activity peptide, obtained by yield often
It is all Gamma Magnitude, and requires a great deal of time and pay the Financial cost that purchase chromatographic column consumables associated therewith brings, it is difficult to
Realize the scale to biologically active peptide to separate.On the other hand, although be used alone pressure-actuated membrane separation technique can be big
Scale separation protein hydrolysate, but due to its relatively low selectivity, can only often separate the larger activity of molecular weight difference
Peptide and aminoacid, good for biological activity, and effective concentration effect of molecular weight bioactive peptide relatively is relatively poor.And
High-throughout membrance separation can bring the severe contamination of film bag, affect separating effect.
Compared with other isolation technics, membrance separation-electrodialytic technique can according to the isoelectric point, IP of target product and molecular weight,
Select, suitable extra electric field, the pH value of separating liquid and ultrafilter membrane is set, and the present invention is by designing variable concentrations gradient
KCl separating liquid isolates positively charged and negatively charged Antihypertensive Peptides while carrying out rough segmentation to antihypertensive peptides derived from soy protein, realize to mesh
The selectivity of mark product, extensive separation.
Content of the invention
The technical problem to be solved is to overcome above-mentioned the deficiencies in the prior art, provide a kind of using membrance separation-
The method that electrodialytic technique prepares antihypertensive peptides derived from soy protein.
The technical problem to be solved is achieved through the following technical solutions：
A kind of method that utilization membrance separation-electrodialytic technique prepares antihypertensive peptides derived from soy protein, the method is comprised the following steps：(1)
The preparation of soybean protein hydrolyate：Soybean protein isolate is placed in enzyme reactor, appropriate distilled water stirring and evenly mixing is added, is carried out
The pre-heat treatment, then temperature is transferred to 55 DEG C, it is 8.5 to adjust pH, adds the Alcalase protease hydrolysiss that concentration of substrate is 4%
4h, keeps system pH constant in course of reaction, after hydrolysis terminates, boiling water bath enzyme denaturing 10min, and it is cooled to after room temperature 3000g's
Under the conditions of be centrifuged 15min take supernatant, obtain soybean protein hydrolyate；(2) separation of antihypertensive peptides derived from soy protein：Protolysate is led to
Neutral ultrafilter membrane is crossed, after ultrafiltration, obtains thick hydrolysate-peptide for inhibiting (ACE)；(3) electrodialysis：The DC source of electric osmose analyzer is sent out
The voltage and current of raw device is respectively set to 20V, 0.17A, by anion exchange membrane (AXM) and cation exchange membrane (CXM)
Concurrently separate Antihypertensive Peptides of the acquisition with positive and negative charge.
The concentration that soybean protein isolate is placed in enzyme reactor is 8% (w/v), heat treatment under 90 DEG C of water bath condition
10min.
Hydrolysis temperature is 55 DEG C, and enzymolysis pH is 8.5, enzymolysis time 4h.
Protolysate is to lead to 1kDa, most Antihypertensive Peptides of the relative molecular mass less than 1kDa by neutral ultrafilter membrane
Cross.
Membrance separation-electrodialytic recovery pond is equipped with 15-20g/L NaCl solution, plays a part of buffer ions exchange,
Under electric field action, collected in KCl (1) and KCl (2) by the negatively charged and positively charged Antihypertensive Peptides of ultrafilter membrane respectively.
In the design application of electric osmose analyzer, KCl ion concentration is to collect isolated band in KCl (1)
The antihypertensive peptides derived from soy protein of negative charge, the scope of its relative molecular mass is collected mainly between 200-300, in KCl (2) and is separated
To positively charged antihypertensive peptides derived from soy protein, the scope of relative molecular mass is mainly between 400-500；Membrance separation-electrodialytic technique
(EDUF) can be used for separating-purifying soybean protein source antihypertensive peptide, with preferable selectivity and separating effect and easy to operate
Equipment is simple.
This method can be used for separating-purifying soybean protein source antihypertensive peptide, with preferable selectivity and separating effect, grasp
Make to facilitate the simple feature of equipment.
Description of the drawings
Accompanying drawing inventive film separation-electrodialysis plant schematic diagram.
Specific embodiment
Below in conjunction with the accompanying drawings the specific embodiment of the invention is described in detail.
A kind of method that utilization membrance separation-electrodialytic technique prepares antihypertensive peptides derived from soy protein, the method is comprised the following steps：(1)
The preparation of soybean protein hydrolyate：Soybean protein isolate is placed in enzyme reactor, the appropriate distilled water stirring and evenly mixing of addition, 90 DEG C
The pre-heat treatment 10min, then reduce the temperature to 55 DEG C stably, it is 8.5 to adjust pH, adds the Alcalase egg that concentration of substrate is 4%
White enzyme hydrolysiss 4h, keeps system pH constant in course of reaction, after hydrolysis terminates, boiling water bath enzyme denaturing 10min, and it is cooled to after room temperature
Under conditions of 3000g, centrifugation 15min takes supernatant, obtains soybean protein hydrolyate；(2) separation of antihypertensive peptides derived from soy protein：By albumen
Hydrolysate obtains thick hydrolysate peptide for inhibiting (ACE) by neutral ultrafilter membrane after ultrafiltration；(3) electrodialysis：The design of electric osmose analyzer should
It is 0.5-1.0g/L with middle KCl ion concentration, in KCl (1), isolated negatively charged antihypertensive peptides derived from soy protein is collected, which is relative
The scope of molecular mass collects isolated positively charged antihypertensive peptides derived from soy protein, phase mainly between 200-300, in KCl (2)
To the scope of molecular mass mainly between 400-500, Antihypertensive Peptides are made to reach detached purpose.
Embodiment 1
5g soybean protein isolate is placed in enzyme reactor, adds 100mL distilled water stirring and evenly mixing, 90 DEG C of the pre-heat treatment
10min, then reduce the temperature to 55 DEG C stably, it is 8.5 to adjust pH with 0.1M NaOH solution, and it is 4% to add concentration of substrate
Alcalase protease hydrolysiss 4h, keeps system pH constant in course of reaction, after hydrolysis terminates, boiling water bath enzyme denaturing 10min, and cooling
To after room temperature, under conditions of 3000g, centrifugation 15min takes supernatant, obtains soybean protein hydrolyate by membrance separation-electrodialysis
Device, in the design application of electric osmose analyzer, KCl ion concentration is 1.0g/L, collects isolated negatively charged in KCl (1)
Antihypertensive peptides derived from soy protein, the scope of its relative molecular mass collects isolated positively charged mainly between 200-300, in KCl (2)
The antihypertensive peptides derived from soy protein of lotus, the scope of relative molecular mass makes Antihypertensive Peptides reach detached purpose mainly between 400-500.
Claims (6)
1. a kind of method that utilization membrance separation-electrodialytic technique prepares antihypertensive peptides derived from soy protein, it is characterised in that the method includes following
Step：(1) preparation of soybean protein hydrolyate：Soybean protein isolate is placed in enzyme reactor, adds appropriate distilled water stirring
Mix, the pre-heat treatment is carried out, then temperature is transferred to 55 DEG C, it is 8.5 to adjust pH, adds the Alcalase egg that concentration of substrate is 4%
White enzyme hydrolysiss 4h, keeps system pH constant in course of reaction, after hydrolysis terminates, boiling water bath enzyme denaturing 10min, and it is cooled to after room temperature
Under conditions of 3000g, centrifugation 15min takes supernatant, obtains soybean protein hydrolyate；(2) separation of antihypertensive peptides derived from soy protein：By albumen
Hydrolysate obtains thick hydrolysate-peptide for inhibiting (ACE) by neutral ultrafilter membrane after ultrafiltration；(3) electrodialysis：By the straight of electric osmose analyzer
The voltage and current of stream power source generator is respectively set to 20V, 0.17A, by anion exchange membrane (AXM) and cation exchange
Film (CXM) concurrently separates Antihypertensive Peptides of the acquisition with positive and negative charge.
2. the method that a kind of utilization membrance separation-electrodialytic technique according to claim 1 prepares antihypertensive peptides derived from soy protein, its feature
It is, the concentration that soybean protein isolate is placed in enzyme reactor is 8% (w/v), heat treatment 10min under 90 DEG C of water bath condition.
3. the method that utilization membrance separation-electrodialytic technique according to claim 1 prepares antihypertensive peptides derived from soy protein, its feature exists
In hydrolysis temperature is 55 DEG C, and enzymolysis pH is 8.5, enzymolysis time 4h.
4. the method that utilization membrance separation-electrodialytic technique according to claim 1 prepares antihypertensive peptides derived from soy protein, its feature exists
In protolysate is to pass through 1kDa, most Antihypertensive Peptides of the relative molecular mass less than 1kDa by neutral ultrafilter membrane.
5. the method that utilization membrance separation-electrodialytic technique according to claim 1 prepares antihypertensive peptides derived from soy protein, its feature exists
In membrance separation-electrodialytic recovery pond is equipped with 15-20g/L NaCl solution, plays a part of buffer ions exchange, makees in electric field
With under, collected in the KCl (1) and KCl (2) by the negatively charged and positively charged Antihypertensive Peptides of ultrafilter membrane respectively.
6. the method that utilization membrance separation-electrodialytic technique according to claim 1 prepares antihypertensive peptides derived from soy protein, its feature exists
In in the design application of electric osmose analyzer, KCl ion concentration is 0.5-1.0g/L, collects isolated negatively charged in KCl (1)
Antihypertensive peptides derived from soy protein, the scope of its relative molecular mass just collecting isolated band in the KCl (2) mainly between 200-300
The antihypertensive peptides derived from soy protein of electric charge, the scope of relative molecular mass is mainly between 400-500；Membrance separation-electrodialytic technique (EDUF)
Can be used for separating-purifying soybean protein source antihypertensive peptide, with preferable selectivity and separating effect, and the letter of easy to operate equipment
Single.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611102584.9A CN106434815A (en) | 2016-12-05 | 2016-12-05 | Method for preparing soybean antihypertensive peptides by membrane separation-electrodialysis technology |
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| CN201611102584.9A CN106434815A (en) | 2016-12-05 | 2016-12-05 | Method for preparing soybean antihypertensive peptides by membrane separation-electrodialysis technology |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208309A (en) * | 2017-12-28 | 2018-06-29 | 保龄宝生物股份有限公司 | The production method of low ash content soybean peptide containing profitable probliotics |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1847260A (en) * | 2006-03-08 | 2006-10-18 | 江苏大学 | Wheat germ protein depressor peptide desalting process |
| CN102094060A (en) * | 2010-11-24 | 2011-06-15 | 河南工业大学 | Method for producing pressure reduction peptide from soybean as raw material |
-
2016
- 2016-12-05 CN CN201611102584.9A patent/CN106434815A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1847260A (en) * | 2006-03-08 | 2006-10-18 | 江苏大学 | Wheat germ protein depressor peptide desalting process |
| CN102094060A (en) * | 2010-11-24 | 2011-06-15 | 河南工业大学 | Method for producing pressure reduction peptide from soybean as raw material |
Non-Patent Citations (1)
| Title |
|---|
| RONG HE等: "Selective separation and concentration of antihypertensive peptides from rapeseed protein hydrolysate by electrodialysis with ultrafiltration membranes", 《FOOD CHEMISTRY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208309A (en) * | 2017-12-28 | 2018-06-29 | 保龄宝生物股份有限公司 | The production method of low ash content soybean peptide containing profitable probliotics |
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Application publication date: 20170222 |