CN110499351A - A kind of soybean peptide and the preparation method and application thereof with high ACE inhibitory activity - Google Patents
A kind of soybean peptide and the preparation method and application thereof with high ACE inhibitory activity Download PDFInfo
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- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 83
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 83
- 244000068988 Glycine max Species 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 230000002401 inhibitory effect Effects 0.000 title abstract description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 65
- 239000000843 powder Substances 0.000 claims abstract description 43
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 35
- 108010073771 Soybean Proteins Proteins 0.000 claims abstract description 27
- 235000019710 soybean protein Nutrition 0.000 claims abstract description 27
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 19
- 239000012528 membrane Substances 0.000 claims abstract description 15
- 108091005804 Peptidases Proteins 0.000 claims abstract description 8
- 239000004365 Protease Substances 0.000 claims abstract description 8
- 239000003513 alkali Substances 0.000 claims abstract description 7
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 230000036541 health Effects 0.000 claims abstract description 4
- 230000036772 blood pressure Effects 0.000 claims abstract description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 35
- 239000006228 supernatant Substances 0.000 claims description 21
- 239000000047 product Substances 0.000 claims description 17
- 230000009514 concussion Effects 0.000 claims description 16
- 238000004108 freeze drying Methods 0.000 claims description 15
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 238000010521 absorption reaction Methods 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 230000001376 precipitating effect Effects 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 238000005119 centrifugation Methods 0.000 claims description 6
- 235000018102 proteins Nutrition 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 4
- 229940079593 drug Drugs 0.000 claims description 4
- 238000001179 sorption measurement Methods 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 claims description 3
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 3
- 230000009849 deactivation Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 claims description 3
- 239000000654 additive Substances 0.000 claims description 2
- 230000000996 additive effect Effects 0.000 claims description 2
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- 238000005194 fractionation Methods 0.000 abstract description 6
- 238000005377 adsorption chromatography Methods 0.000 abstract description 5
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- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 36
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 36
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 36
- 235000019441 ethanol Nutrition 0.000 description 20
- 239000000758 substrate Substances 0.000 description 11
- 230000001476 alcoholic effect Effects 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 206010020772 Hypertension Diseases 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 102000035195 Peptidases Human genes 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 101000984728 Chiropsoides quadrigatus Angiotensin-converting enzyme inhibitory peptide Proteins 0.000 description 3
- 108010009736 Protein Hydrolysates Proteins 0.000 description 3
- 230000004531 blood pressure lowering effect Effects 0.000 description 3
- 239000003531 protein hydrolysate Substances 0.000 description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 239000000413 hydrolysate Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 208000010470 Ageusia Diseases 0.000 description 1
- 208000028185 Angioedema Diseases 0.000 description 1
- 206010003658 Atrial Fibrillation Diseases 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000019666 ageusia Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003276 anti-hypertensive effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000030613 peripheral artery disease Diseases 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004393 visual impairment Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- 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
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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
- C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof
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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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention belongs to technical field of food biotechnology, disclose a kind of soybean peptide and the preparation method and application thereof with high ACE inhibitory activity.The soybean peptide has high ACE inhibitory activity, and preparation method includes the following steps: to digest soybean protein isolate addition alkali protease to obtain enzymolysis product；Isolated soy peptide powder I is carried out to enzymolysis product using membrane ultrafiltration process；Soy peptide powder I is further purified using ethanol fractionation, obtains soy peptide powder II；Soy peptide powder II is further isolated and purified using activated carbon adsorption chromatography, obtains the soybean peptide with high ACE inhibitory activity.Soybean peptide of the invention has excellent ACE inhibitory activity and good blood pressure reduction effect, can be applied to food, field of health care products, and preparation method is simply controllable, is suitable for factory's volume production.
Description
Technical field
The invention belongs to biologically active peptide fields, and in particular to a kind of soybean peptide and its preparation with ACE inhibitory activity
Method and purposes.
Background technique
Hypertension is a kind of major global public health problem, and long-term hypertension is to cause coronary artery disease, apoplexy, mental and physical efforts
Failure, auricular fibrillation, peripheral artery disease, the Major Risk Factors of visual loss, while being still the maximum list of the global death rate
One factor.In China, the large sample cohort study that Chinese chronic disease perspective study (CKB research) is carried out is the result shows that every ten
Resident just has three with hypertension, and illness rate is up to 32.5%, and the control rate of hypertension is down to 4.2%.Currently, treatment
Hypertension mainly or by drug effect is realized, and is taken side effect caused by drug for hypertension for a long time and also do not allowed to neglect
Depending on, including dry cough, potassemia, fatigue is dizzy, headache, ageusia and angioedema etc..
Angiotensin converting enzyme (ACE) peptide for inhibiting is a kind of by inhibiting ACE activity to reach blood pressure lowering effect
Small molecule bioactive polypeptide general name.It is usually obtained by the food-borne albumen of protease hydrolytic, because its blood pressure lowering effect is obvious, nothing
Toxic side effect and receive significant attention.
The biologically active peptide with ACE inhibitory activity is obtained from food-borne albumen at present, wherein being using casein
12 peptide FFVAPFPGVFGK of raw material preparation show preferable ACE inhibitory activity, and include the caseic hydrolysate of this peptide
Patent of invention (Patent No.: US6514941B1) is applied for, and has been realized as antihypertensive product and commercially produce and sell
It sells.
Chinese patent application CN 105524966A also discloses a kind of enzymatic hydrolysis dregs of beans method for preparing ace inhibitory peptide, but by
In the raw material used for the dregs of beans (containing albumen, polysaccharide, minerals etc.) of complicated component, and only with simple separating technology
(acid sinks and ultrafiltration), resulting ace inhibitory peptide purity is lower, and whole ACE inhibitory activity is not high.
Summary of the invention
The primary purpose of the present invention is that providing a kind of preparation method of soybean peptide with high ACE inhibitory activity, the party
Method, which is characterized by, efficiently to be discharged and collects in conjunction with biological enzymolysis technology, Ultrafiltration Membrane and active carbon chromatographic techniques
With the soybean peptide of high ACE inhibitory activity in soybean protein isolate.
Another object of the present invention is to provide soybean peptides made from the above method.
A further object of the present invention is to provide the purposes of above-mentioned soybean peptide.
The purpose of the invention is achieved by the following technical solution:
A kind of preparation method of soybean peptide, includes the following steps:
(1) add water to mix soybean protein isolate, be added alkali protease, temperature be 45~55 DEG C, pH value be 7.0~
6~10h is digested under the conditions of 8.0, enzyme deactivation is collected by centrifugation supernatant, obtains soybean protein isolate enzymolysis product；
(2) by the ultrafiltration membrane of 3kDa after being diluted with water soybean protein isolate enzymolysis product, < 3kDa component is collected, it is cold
It is lyophilized dry, obtains soy peptide powder I；
(3) soy peptide powder I is dissolved in 61~80% ethanol solutions, is stood after stirring and evenly mixing, is centrifuged, collect supernatant,
Freeze-drying, obtains soy peptide powder II；
(4) plus water dissolves soy peptide powder II, and activated carbon adsorption is added, then filters, and collects precipitating, is rushed with deionized water
It is washed till neutrality, 61~80% ethanol solutions are added, concussion parsing, collected by suction supernatant obtain after freeze-drying at room temperature
A kind of soybean peptide with high ACE inhibitory activity；
In step (1), alkali protease is preferably letter (China) Bioisystech Co., Ltd, Novi
2.4L；
In step (1), the additive amount of alkali protease is the 0.5~1.0% of soybean protein isolate quality；
Enzyme deactivation described in step (1) is that reactant is kept the temperature to 10~20min at 85~100 DEG C；
Centrifugation described in step (1) refers to 15~30min of centrifugation at 1500~4000g；
In step (1), after adding water to mix, concentration of substrate is 5~10% (w/w)；
Step is diluted with water described in (2), is to be diluted to solid content as 0.5~1.0% (w/w)；
Its separation parameter of ultrafiltration described in step (2) are as follows: applied sample amount 5L, 4~8 DEG C of temperature, pressure 0.5MPa；
In step (3), soy peptide powder I is dissolved in 61~80% ethanol solutions, makes its final concentration of 5~10mg/ml；
Centrifugation described in step (3) refers to 15~30min of centrifugation at 1500~4000g；
The additional amount of step (4) described active carbon is 1~3 times of II mass of soy peptide powder；
Absorption described in step (4), system pH is 2.0~4.0,1~3h is adsorbed in concussion at room temperature；
The parsing of concussion described in step (4), preferably 1~3h of parsing.
There is the present invention soybean peptide of high ACE inhibitory activity can be used for preparing blood-pressure drug, health care product and food.
Soybean protein isolate (soybean protein isolate, SPI) is that soybean passes through shelling, degreasing, carbon elimination aquation
The protein component separated after the processing such as object is closed, protein content is up to 90% or more.Using soybean protein isolate as raw material, utilize
Modern biotechnology controlled enzymatic hydrolysis technology and separating and purifying technology produce the soybean peptide with high ACE inhibitory activity, have high society
It can meaning and economic value.
The present invention has the following advantages and effects with respect to the prior art:
(1) present invention uses soybean protein isolate to digest raw material, because the protein content of soybean protein isolate is up to 90%
More than, it is relatively single to digest prepared hydrolysate ingredient；And hydrophobic amino acid rich content in soybean protein isolate, it is right
ACE inhibitory activity contribution is big.
(2) present invention combines biological enzymolysis technology, film hyperfiltration technique, alcohol grading purification technique and adsorption separation technology system
Standby soybean peptide ACE inhibitory activity out is high.
(3) preparation method that the present invention uses is simply controllable, and short preparation period is suitable for factory's volume production.
Detailed description of the invention
Fig. 1 is the ACE inhibitory activity of different film ultrafiltration components in embodiment 1.
Fig. 2 is the ACE inhibitory activity of different concentration ethanol classification component in embodiment 1.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Embodiment 1
A kind of soybean peptide preparation method with high ACE inhibitory activity, includes the following steps:
(1) add water to mix soybean protein isolate, make its concentration of substrate 7.5% (w/w), In2.4L adding
Dosage is 0.75% (w/w), and temperature is 55 DEG C, and pH value keeps the temperature 10min in 100 DEG C of water after digesting 8h under conditions of being 7.5,
It is centrifuged 15min under the centrifugal force of 4000g, collects supernatant, obtains soybean protein isolate enzymolysis product (SPIH), obtains soybean
Protein isolate enzymolysis product (SPIH), enzymolysis liquid degree of hydrolysis are 6.07 ± 0.08%.
(2) SPIH is isolated and purified using membrane ultrafiltration process, SPIH is diluted with water to will after solid content 0.8%
Then it will transmit through the ultrafiltration membrane that liquid passes sequentially through 5kDa and 3kDa by 10kDa ultrafiltration membrane, collect each component, and freezing is dry
It is dry, obtain soy peptide powder I.
(3) soy peptide powder I is further purified using ethanol fractionation, soy peptide powder I is dissolved in 15% respectively,
35%, in 55%, 75%, 95% ethanol solution, make its final concentration of 10mg/ml, 30min, In are stood after stirring and evenly mixing
It is centrifuged 15min under the centrifugal force of 4000g, collects each concentration ethanol component supernatant, is freeze-dried, obtains soy peptide powder II.
(4) soy peptide powder II is further purified using Activated Carbon Adsorption Separation method, soy peptide powder II is dissolved in water, is made
Its final concentration of 10mg/ml, by substrate protein: quality of activated carbon ratio is that active carbon is added in 2:1 mass ratio, and regulation system pH value is extremely
3.0 filter after concussion absorption 2h at room temperature, collect precipitating, and rinsed to neutrality with deionized water, isometric 75% second is added
Alcoholic solution, at room temperature after concussion parsing 2h, collected by suction supernatant, one kind is obtained after freeze-drying, and there is high ACE to inhibit to live
The soybean peptide of property.
The ACE of 4 components of gained in enzymolysis product (SPIH) obtained by the present embodiment step (1) and step (2) is inhibited to live
Property is measured, as a result as shown in Figure 1, the results showed that the ACE inhibitory activity highest of < 3kDa component.
The ACE inhibitory activity of different ethanol concentration component in the present embodiment step (3) is measured, as a result such as Fig. 2 institute
Show, the results showed that the ACE inhibitory activity highest of 61~80% ethanol components.
The ACE inhibitory activity of 1 different component of table
Note: detectable concentration 0.2mg/mL.
Have a large amount of document and shows that ace inhibitory peptide has good blood pressure lowering effect, and safe without toxic side effect.By
Table 1 has ACE inhibitory activity, with going deep into for preparative separation process, soybean peptide it is found that inventing the soybean peptide of preparation herein
ACE inhibitory activity steps up, and the ACE inhibitory activity of final soybean peptide finished product is the 10.4 of former soybean separation proteolysis object
Times, show that soybean peptide prepared by the present invention has high ACE inhibitory activity, can be used in the industries such as drug, health care product or food.
Embodiment 2
A kind of soybean peptide preparation method with high ACE inhibitory activity, includes the following steps:
(1) add water to mix soybean protein isolate, make its concentration of substrate 5% (w/w), In2.4L addition
Amount is 0.5% (w/w), and temperature is 45 DEG C, and pH value keeps the temperature 20min, In after digesting 10h under conditions of being 7.0 in 85 DEG C of water
It is centrifuged 30min under the centrifugal force of 2000g, collects supernatant, obtains soybean protein isolate enzymolysis product (SPIH), obtains soybean point
From protein hydrolysate (SPIH), enzymolysis liquid degree of hydrolysis is 5.11 ± 0.03%.
(2) SPIH is isolated and purified using membrane ultrafiltration process, SPIH is diluted with water to will after solid content 0.5%
It collects < 3kDa component by the ultrafiltration membrane of 3kDa, and freeze-drying obtains soy peptide powder I.
(3) soy peptide powder I is further purified using ethanol fractionation, it is molten that soy peptide powder I is dissolved in 61% ethyl alcohol
In liquid, make its final concentration of 5mg/ml, stand 30min after stirring and evenly mixing, 30min is centrifuged under the centrifugal force of 2000g, collects
61% ethanol component supernatant, freeze-drying, obtains soy peptide powder II.
(4) soy peptide powder II is further purified using activated carbon adsorption chromatography, soy peptide powder II is dissolved in water, is made
Its final concentration of 5mg/ml, by substrate protein: quality of activated carbon ratio is that active carbon is added in 1:1 mass ratio, and regulation system pH value is extremely
2.0 filter after concussion absorption 1h at room temperature, collect precipitating, and rinsed to neutrality with deionized water, isometric 61% second is added
Alcoholic solution, at room temperature after concussion parsing 1h, collected by suction supernatant, one kind is obtained after freeze-drying, and there is high ACE to inhibit to live
The soybean peptide of property.
Embodiment 3
A kind of soybean peptide preparation method with high ACE inhibitory activity, includes the following steps:
(1) add water to mix soybean protein isolate, make its concentration of substrate 7.5% (w/w), In2.4L adding
Dosage is 0.5% (w/w), and temperature is 50 DEG C, and pH value keeps the temperature 10min in 100 DEG C of water after digesting 8h under conditions of being 7.5,
It is centrifuged 15min under the centrifugal force of 4000g, collects supernatant, obtains soybean protein isolate enzymolysis product (SPIH), obtains soybean
Protein isolate enzymolysis product (SPIH), enzymolysis liquid degree of hydrolysis are 5.61 ± 0.05%.
(2) SPIH is isolated and purified using membrane ultrafiltration process, SPIH is diluted with water to will after solid content 1.0%
It collects < 3kDa component by the ultrafiltration membrane of 3kDa, and freeze-drying obtains soy peptide powder I.
(3) soy peptide powder I is further purified using ethanol fractionation, soy peptide powder I is dissolved in 80% second respectively
In alcoholic solution, makes its final concentration of 10mg/ml, stand 15min after stirring and evenly mixing, be centrifuged under the centrifugal force of 4000g
15min, collects 80% ethanol component supernatant, and freeze-drying obtains soy peptide powder II.
(4) soy peptide powder II is further purified using activated carbon adsorption chromatography, soy peptide powder II is dissolved in water, is made
Its final concentration of 10mg/ml, by substrate protein: quality of activated carbon ratio is that active carbon is added in 1:1 mass ratio, and regulation system pH value is extremely
4.0 filter after concussion absorption 3h at room temperature, collect precipitating, and rinsed to neutrality with deionized water, isometric 80% second is added
Alcoholic solution, at room temperature after concussion parsing 3h, collected by suction supernatant, one kind is obtained after freeze-drying, and there is high ACE to inhibit to live
The soybean peptide of property.
Embodiment 4
A kind of soybean peptide preparation method with high ACE inhibitory activity, includes the following steps:
(1) add water to mix soybean protein isolate, make its concentration of substrate 10% (w/w), In2.4L adding
Dosage is 1.0% (w/w), and temperature is 55 DEG C, and pH value keeps the temperature 15min, In after digesting 6h under conditions of being 7.0 in 90 DEG C of water
It is centrifuged 20min under the centrifugal force of 3000g, collects supernatant, obtains soybean protein isolate enzymolysis product (SPIH), obtains soybean point
From protein hydrolysate (SPIH), enzymolysis liquid degree of hydrolysis is 5.73 ± 0.07%.
(2) SPIH is isolated and purified using membrane ultrafiltration process, SPIH is diluted with water to will after solid content 0.8%
It collects < 3kDa component by the ultrafiltration membrane of 3kDa, and freeze-drying obtains soy peptide powder I.
(3) soy peptide powder I is further purified using ethanol fractionation, soy peptide powder I is dissolved in 70% second respectively
In alcoholic solution, makes its final concentration of 8mg/ml, stands 20min after stirring and evenly mixing, be centrifuged 20min under the centrifugal force of 3000g,
70% ethanol component supernatant is collected, is freeze-dried, obtains soy peptide powder II.
(4) soy peptide powder II is further purified using activated carbon adsorption chromatography, soy peptide powder II is dissolved in water, is made
Its final concentration of 8mg/ml, by substrate protein: quality of activated carbon ratio is that active carbon is added in 3:1 mass ratio, and regulation system pH value is extremely
3.0 filter after concussion absorption 2h at room temperature, collect precipitating, and rinsed to neutrality with deionized water, isometric 70% second is added
Alcoholic solution, at room temperature after concussion parsing 2h, collected by suction supernatant, one kind is obtained after freeze-drying, and there is high ACE to inhibit to live
The soybean peptide of property.
Embodiment 5
A kind of soybean peptide preparation method with high ACE inhibitory activity, includes the following steps:
(1) add water to mix soybean protein isolate, make its concentration of substrate 5% (w/w), In2.4L addition
Amount is 0.75% (w/w), and temperature is 50 DEG C, and pH value keeps the temperature 15min, In after digesting 10h under conditions of being 8.0 in 90 DEG C of water
It is centrifuged 20min under the centrifugal force of 3000g, collects supernatant, obtains soybean protein isolate enzymolysis product (SPIH), obtains soybean point
From protein hydrolysate (SPIH), enzymolysis liquid degree of hydrolysis is 6.70 ± 0.07%.
(2) SPIH is isolated and purified using membrane ultrafiltration process, SPIH is diluted with water to will after solid content 0.5%
It collects < 3kDa component by the ultrafiltration membrane of 3kDa, and freeze-drying obtains soy peptide powder I.
(3) Gly-His-Lys I is further purified using ethanol fractionation, Gly-His-Lys I is dissolved in respectively in 75% ethanol solution,
It makes its final concentration of 5mg/ml, stands 20min after stirring and evenly mixing, and 20min is centrifuged under the centrifugal force of 3000g, collects 75%
Ethanol component supernatant, freeze-drying, obtains soy peptide powder II.
(4) soy peptide powder II is further purified using activated carbon adsorption chromatography, soy peptide powder II is dissolved in water, is made
Its final concentration of 5mg/ml, by substrate protein: quality of activated carbon ratio is that active carbon is added in 2:1 mass ratio, and regulation system pH value is extremely
2.0 filter after concussion absorption 1h at room temperature, collect precipitating, and rinsed to neutrality with deionized water, isometric 75% second is added
Alcoholic solution, at room temperature after concussion parsing 1h, collected by suction supernatant, one kind is obtained after freeze-drying, and there is high ACE to inhibit to live
The soybean peptide of property.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of soybean peptide, it is characterised in that include the following steps:
(1) add water to mix soybean protein isolate, alkali protease is added, be 45~55 DEG C in temperature, pH value is 7.0~8.0
Under the conditions of digest 6~10h, enzyme deactivation is collected by centrifugation supernatant, obtains soybean protein isolate enzymolysis product；
(2) by the ultrafiltration membrane of 3kDa after being diluted with water soybean protein isolate enzymolysis product, < 3kDa component is collected, freezing is dry
It is dry, obtain soy peptide powder I；
(3) soy peptide powder I is dissolved in 61~80% ethanol solutions, is stood after stirring and evenly mixing, is centrifuged, collect supernatant, freezing
It is dry, obtain soy peptide powder II；
(4) plus water soy peptide powder II is dissolved, be added activated carbon adsorption, then filter, collect precipitating, with deionized water rinse to
61~80% ethanol solutions are added in neutrality, and concussion parsing, collected by suction supernatant obtain soybean after freeze-drying at room temperature
Peptide.
2. preparation method according to claim 1, it is characterised in that: in step (1), alkali protease is
2.4L。
3. preparation method according to claim 1, it is characterised in that: in step (1), the additive amount of alkali protease is big
The 0.5~1.0% of beans protein isolate quality.
4. preparation method according to claim 1, it is characterised in that: its separation parameter of ultrafiltration described in step (2)
Are as follows: applied sample amount 5L, 4~8 DEG C of temperature, pressure 0.5MPa.
5. preparation method according to claim 1, it is characterised in that: in step (3), soy peptide powder I is dissolved in 61~
In 80% ethanol solution, make its final concentration of 5~10mg/ml.
6. preparation method according to claim 1, it is characterised in that: the additional amount of step (4) described active carbon is soybean
1~3 times of II mass of Gly-His-Lys.
7. preparation method according to claim 1, it is characterised in that: absorption described in step (4), system pH 2.0
~4.0,1~3h is adsorbed in concussion at room temperature.
8. preparation method according to claim 1, it is characterised in that: the parsing of concussion described in step (4) parses 1~3h.
9. a kind of soybean peptide, it is characterised in that: be made by the described in any item methods of claim 1-8.
10. soybean peptide as claimed in claim 9 is preparing the application in blood-pressure drug, health care product and food.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112080539A (en) * | 2020-08-25 | 2020-12-15 | 华中农业大学 | Kidney bean bioactive peptide and application thereof in preparation of uric acid reducing medicines |
| CN113215212A (en) * | 2021-04-16 | 2021-08-06 | 国肽生物工程（常德）有限公司 | Soybean protein peptide with antioxidant and ACE (angiotensin converting enzyme) inhibiting functions and preparation method thereof |
-
2019
- 2019-08-30 CN CN201910813768.3A patent/CN110499351A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112080539A (en) * | 2020-08-25 | 2020-12-15 | 华中农业大学 | Kidney bean bioactive peptide and application thereof in preparation of uric acid reducing medicines |
| CN112080539B (en) * | 2020-08-25 | 2021-12-03 | 华中农业大学 | Kidney bean bioactive peptide and application thereof in preparation of uric acid reducing medicines |
| CN113215212A (en) * | 2021-04-16 | 2021-08-06 | 国肽生物工程（常德）有限公司 | Soybean protein peptide with antioxidant and ACE (angiotensin converting enzyme) inhibiting functions and preparation method thereof |
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Application publication date: 20191126 |