CN109022527A - A kind of quinoa polypeptide and preparation method thereof with hypotensive activity - Google Patents
A kind of quinoa polypeptide and preparation method thereof with hypotensive activity Download PDFInfo
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- CN109022527A CN109022527A CN201810984240.8A CN201810984240A CN109022527A CN 109022527 A CN109022527 A CN 109022527A CN 201810984240 A CN201810984240 A CN 201810984240A CN 109022527 A CN109022527 A CN 109022527A
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- 240000006162 Chenopodium quinoa Species 0.000 title claims abstract description 99
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 70
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 54
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 52
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 230000001077 hypotensive effect Effects 0.000 title claims abstract description 27
- 230000007062 hydrolysis Effects 0.000 claims abstract description 19
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 19
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 18
- 102000004190 Enzymes Human genes 0.000 claims abstract description 16
- 108090000790 Enzymes Proteins 0.000 claims abstract description 16
- 229940088598 enzyme Drugs 0.000 claims abstract description 16
- 230000003276 anti-hypertensive effect Effects 0.000 claims abstract description 12
- 238000005238 degreasing Methods 0.000 claims abstract description 12
- 102000057297 Pepsin A Human genes 0.000 claims abstract description 11
- 108090000284 Pepsin A Proteins 0.000 claims abstract description 11
- 229940111202 pepsin Drugs 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 7
- 238000010612 desalination reaction Methods 0.000 claims abstract description 6
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims abstract description 6
- 239000003513 alkali Substances 0.000 claims abstract description 4
- 239000002994 raw material Substances 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 239000000287 crude extract Substances 0.000 claims description 24
- 239000012528 membrane Substances 0.000 claims description 22
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 108090000623 proteins and genes Proteins 0.000 claims description 19
- 239000006228 supernatant Substances 0.000 claims description 17
- 230000001376 precipitating effect Effects 0.000 claims description 14
- 238000005119 centrifugation Methods 0.000 claims description 12
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 241000871189 Chenopodiaceae Species 0.000 claims description 6
- 241000209140 Triticum Species 0.000 claims description 6
- 235000021307 Triticum Nutrition 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 235000013305 food Nutrition 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 4
- 102000035195 Peptidases Human genes 0.000 claims description 2
- 108091005804 Peptidases Proteins 0.000 claims description 2
- 239000004365 Protease Substances 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims description 2
- 238000005374 membrane filtration Methods 0.000 claims description 2
- 239000012466 permeate Substances 0.000 claims description 2
- 238000000751 protein extraction Methods 0.000 claims description 2
- 230000017854 proteolysis Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 3
- 108010072661 Angiotensin Amide Proteins 0.000 abstract description 2
- CZGUSIXMZVURDU-JZXHSEFVSA-N Ile(5)-angiotensin II Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=[NH2+])NC(=O)[C@@H]([NH3+])CC([O-])=O)C(C)C)C1=CC=C(O)C=C1 CZGUSIXMZVURDU-JZXHSEFVSA-N 0.000 abstract description 2
- 238000003916 acid precipitation Methods 0.000 abstract description 2
- 230000004531 blood pressure lowering effect Effects 0.000 abstract description 2
- 235000018102 proteins Nutrition 0.000 description 17
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 10
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 8
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 8
- 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 7
- 238000013019 agitation Methods 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000011033 desalting Methods 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 239000006260 foam Substances 0.000 description 5
- 238000010298 pulverizing process Methods 0.000 description 5
- 210000002784 stomach Anatomy 0.000 description 5
- 210000002318 cardia Anatomy 0.000 description 3
- 239000008236 heating water Substances 0.000 description 3
- 206010020772 Hypertension Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 101000984728 Chiropsoides quadrigatus Angiotensin-converting enzyme inhibitory peptide Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010028690 Fish Proteins Proteins 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 206010047139 Vasoconstriction Diseases 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229940044094 angiotensin-converting-enzyme inhibitor Drugs 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000025033 vasoconstriction Effects 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- 235000020795 whole food diet Nutrition 0.000 description 1
Classifications
-
- 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
- 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
- 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
- A23L33/185—Vegetable proteins
-
- 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
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- 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
- C07K1/30—Extraction; Separation; Purification by precipitation
-
- 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
-
- 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
Abstract
The invention discloses a kind of quinoa polypeptide and preparation method thereof with hypotensive activity, the preparation method comprises the following steps: quinoa albumen is extracted from degreasing quinoa powder using alkali extraction and acid precipitation, through complex enzyme hydrolysis, ultrafiltration, desalination, finally it is concentrated in vacuo, freeze-drying obtains quinoa active antihypertensive peptide.Method of the invention passes through using quinoa as raw material, active antihypertensive peptide, simple production process, edible safety height are prepared using food-grade pepsin and trypsase complex enzyme hydrolysis quinoa albumen, and the quinoa active peptide can effectively inhibit hypertensin conversion enzyme activity, blood pressure lowering effect is significant.
Description
Technical field
The present invention relates to technical field of plant extraction, in particular to a kind of quinoa polypeptide and its system with hypotensive activity
Preparation Method.
Background technique
Quinoa is a kind of cereal crops for originating in South America, because of the environmental suitability and high nutritive value of its wide spectrum
By common concern all over the world.In recent years, China Shanxi, Jilin, Qinghai, Gansu, Hebei, Tibet, Heilungkiang, Inner Mongol
The area such as Gu, Sichuan, Shandong, Jiangsu, Anhui, Guizhou has successively carried out the extensive introducing and planting of quinoa.Egg in quinoa seed
White matter average content is 12%~23%, it is necessary to which amino acid content is higher than common cereal, can be used as the good protein in food
Source.Protein becomes small-molecular peptides after the metabolism digestion of the enzymes such as pepsin, trypsase in human body domestic demand, can be by human body
It is absorbed and utilized.The active function for studying quinoa polypeptide makes full use of quinoa albumen to provide the biological functional activity of clear quinoa
Source, exploitation quinoa functional product are of great significance.
Hypertension is a kind of common cardiovascular disease, and angiotensin converting enzyme (ACE) inhibitor can prevent to have strong
The generation of the angiotensinⅡ of strong vasoconstriction effect is the main active drug for treating hypertension now.Wholefood comes
The ace inhibitory peptide in source is a big hot spot of biologically active peptide research field, prepares ACE using different proteasome degradation protein
The features such as peptide for inhibiting has production cost relatively low, and reaction condition is mild, and Product Safety is higher is current preparation ACE suppression
The most common production method of peptide processed.
It has isolated from the food resources such as milk protein, fish protein, soybean protein with ACE inhibitory activity at present
Polypeptide, but the research in relation to quinoa polypeptide hypotensive activity is rarely reported.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of quinoa polypeptide and its system with hypotensive activity
Preparation Method, this method can fast and efficiently prepare the quinoa polypeptide with hypotensive activity, and simple production process is eaten
It is highly-safe.
The technical problem to be solved by the present invention is to what is be achieved through the following technical solutions:
A kind of preparation method of the quinoa polypeptide with hypotensive activity, comprising the following steps: using alkali extraction and acid precipitation from
Quinoa albumen is extracted in degreasing quinoa powder, through complex enzyme hydrolysis, ultrafiltration, desalination, is finally concentrated in vacuo, freeze-drying obtains quinoa drop
Blood pressure active peptide.
Preferably, in above-mentioned technical proposal, the preparation method comprises the following steps:
1) protein extraction: raw material degreasing quinoa powder is mixed with suitable water, adjusts pH to 9~10, and constant temperature oscillation extracts
1-2h, centrifugation, takes supernatant；Supernatant pH to 4~5 is adjusted, stands, centrifugation, takes precipitating；It is adjusted after precipitating plus the dissolution of a small amount of water
PH is freeze-dried to neutrality, obtains quinoa protein crude extract administration；
2) proteolysis: taking the quinoa protein crude extract administration of step 1), be dissolved in water, and adjusts pH to 2, and pepsin is added,
30~60min is vibrated in 30 DEG C~50 DEG C water-baths, obtains quinoa albumen pepsin hydrolysis liquid；Adjust quinoa albumen pepsin
Trypsase (trypsase play active optimal pH) is added in the pH to 8 of hydrolyzate, 30 DEG C~50 DEG C water-baths oscillations 30~
60min, 95 DEG C of heating 10min inactivated proteases, centrifuging and taking supernatant obtain quinoa polypeptide crude extract；
3) ultrafiltration desalination: the quinoa polypeptide crude extract that step 2) obtains is passed sequentially through into filter membrane and ultrafiltration membrane, permeate is de-
It after salt, is concentrated in vacuo, is freeze-dried to obtain quinoa polypeptide.It is big to remove possible remaining protein, polysaccharide etc. by filter membrane
Molecular substance, then by ultrafiltration membrane, obtain molecular weight less than 5kDa polypeptide solution (physiological activity of macromolecule polypeptide compared with
Difference).
Preferably, in above-mentioned technical proposal, in the step 1), the mass ratio of the degreasing quinoa powder and the water is 1:
(10~15).The heavy pH value of the alkali carries acid is optimized by the yield of quinoa protein crude extract administration to be determined.
Preferably, in above-mentioned technical proposal, pepsin described in the step 2) and trypsase are food grade enzyme.
Preferably, in above-mentioned technical proposal, the enzymolysis time of the step 2) is selected true by the ACE inhibitory activity of enzymolysis liquid
It is fixed.
Preferably, in above-mentioned technical proposal, filter membrane described in the step 3) is 0.45 μm of filter membrane.
Preferably, in above-mentioned technical proposal, the ultrafiltration retaining molecular weight of the step 3) by filtrate ACE inhibitory activity
Selection determines.
Preferably, in above-mentioned technical proposal, the trapped molecular weight 5kDa of the ultrafiltration membrane of the step 3).
A kind of quinoa polypeptide that the preparation method of the quinoa polypeptide with hypotensive activity is prepared
A kind of application for the quinoa polypeptide that the preparation method of the quinoa polypeptide with hypotensive activity is prepared, is applied to
Prepare the drug or food with antihypertensive function.
Above-mentioned technical proposal of the present invention, has the following beneficial effects:
The present invention provides the methods that one kind can quickly and efficiently prepare quinoa active antihypertensive peptide, are to be with quinoa
Raw material prepares active antihypertensive peptide, production technology letter using food-grade pepsin and trypsase complex enzyme hydrolysis quinoa albumen
Single, edible safety is high, and the quinoa active peptide can effectively inhibit hypertensin conversion enzyme activity, and blood pressure lowering effect is aobvious
It writes.
Specific embodiment
Specific embodiments of the present invention are described in detail below, in order to further understand the present invention.
All experimental methods used are conventional method unless otherwise specified in following embodiment.In following embodiment
Material, reagent used etc. can be obtained through commercial channels unless otherwise specified.
Embodiment 1
A kind of quinoa polypeptide with hypotensive activity, preparation method are as follows:
(1) quinoa seed clear water is rinsed to no foam, dries pulverizing, and n-hexane degreasing takes 100g to be placed in conical flask,
1200mL distilled water is added, adjusts pH to 9.86, magnetic agitation is extracted twice at room temperature, each 30min；Merge twice
Filtrate adjusts pH to 4.91, and in 4 DEG C of standing 30min, centrifugation takes precipitating；Add a small amount of water to dissolve precipitating, adjust pH to 7.02,
Freeze-drying obtains quinoa protein crude extract administration.
(2) quinoa protein crude extract administration is taken, is dissolved in water, pH to 2.04 is adjusted, food-grade stomach egg is added with the ratio of 1:20
White enzyme, 40.3 DEG C of hydrolysis 30min, then pH to 8.10 is adjusted, food-grade trypsase, 40.1 DEG C of hydrolysis 30min, 95 DEG C of water are added
Bath heating 5min, is centrifuged rapidly after cooling, takes supernatant.
(3) after quinoa polypeptide crude extract being crossed 0.45um filter membrane, the Chenopodiaceae of 5kDa is less than by ultrafiltration membrane trapped molecular weight
Wheat polypeptide, dialysis desalting are concentrated in vacuo, are freeze-dried up to quinoa active antihypertensive peptide.
Embodiment 2
A kind of quinoa polypeptide with hypotensive activity, preparation method are as follows:
(1) quinoa seed clear water is rinsed to no foam, dries pulverizing, and n-hexane degreasing takes 200g to be placed in conical flask,
2000mL distilled water is added, adjusts pH to 9.53, magnetic agitation is extracted twice at room temperature, each 30min；Merge twice
Filtrate adjusts pH to 4.26, and in 4 DEG C of standing 30min, centrifugation takes precipitating；Add a small amount of water to dissolve precipitating, adjust pH to 6.97,
Freeze-drying obtains quinoa protein crude extract administration.
(2) quinoa protein crude extract administration is taken, is dissolved in water, pH to 2.11 is adjusted, food-grade stomach egg is added with the ratio of 1:20
White enzyme, 36.7 DEG C of hydrolysis 40min, then pH to 8.07 is adjusted, food-grade trypsase, 37.1 DEG C of hydrolysis 40min, 95 DEG C of water are added
Bath heating 5min, is centrifuged rapidly after cooling, takes supernatant.
(3) after quinoa polypeptide crude extract being crossed 0.45um filter membrane, the Chenopodiaceae of 5kDa is less than by ultrafiltration membrane trapped molecular weight
Wheat polypeptide, dialysis desalting are concentrated in vacuo, are freeze-dried up to quinoa active antihypertensive peptide.
Embodiment 3
A kind of quinoa polypeptide with hypotensive activity, preparation method are as follows:
(1) quinoa seed clear water is rinsed to no foam, dries pulverizing, and n-hexane degreasing takes 200g to be placed in conical flask,
2500mL distilled water is added, adjusts pH to 9.5, magnetic agitation extracts 60min at room temperature；Centrifugation, takes supernatant, adjusts
Supernatant pH to 4.5, in 4 DEG C of standing 30min, centrifugation takes precipitating；Add a small amount of water to dissolve precipitating, adjust pH to 7, freezing is dry
It is dry to obtain quinoa protein crude extract administration.
(2) quinoa protein crude extract administration is taken, is dissolved in water, pH to 2 is adjusted, food-grade stomach cardia is added with the ratio of 1:15
Enzyme, 40 DEG C of hydrolysis 45min, then pH to 8 is adjusted, food-grade trypsase, 40 DEG C of hydrolysis 45min, 95 DEG C of heating water baths are added
10min is centrifuged rapidly after cooling, takes supernatant.
(3) after quinoa polypeptide crude extract being crossed 0.45um filter membrane, the Chenopodiaceae of 5kDa is less than by ultrafiltration membrane trapped molecular weight
Wheat polypeptide, dialysis desalting are concentrated in vacuo, are freeze-dried up to quinoa active antihypertensive peptide.
Embodiment 4
A kind of quinoa polypeptide with hypotensive activity, preparation method are as follows:
(1) quinoa seed clear water is rinsed to no foam, dries pulverizing, and n-hexane degreasing takes 200g to be placed in conical flask,
2000mL distilled water is added, adjusts pH to 9, magnetic agitation extracts 1h at room temperature；Centrifugation, takes supernatant；Adjust supernatant
Liquid pH to 4, in 4 DEG C of standing 30min, centrifugation takes precipitating；Add a small amount of water to dissolve precipitating, adjusts pH to 6.97, be freeze-dried
To quinoa protein crude extract administration.
(2) quinoa protein crude extract administration is taken, is dissolved in water, pH to 2 is adjusted, food-grade stomach cardia is added with the ratio of 1:20
Enzyme, 30 DEG C of hydrolysis 60min, then pH to 8 is adjusted, food-grade trypsase, 50 DEG C of hydrolysis 30min, 95 DEG C of heating water baths are added
10min is centrifuged rapidly after cooling, takes supernatant.
(3) after quinoa polypeptide crude extract being crossed 0.45um filter membrane, the Chenopodiaceae of 5kDa is less than by ultrafiltration membrane trapped molecular weight
Wheat polypeptide, dialysis desalting are concentrated in vacuo, are freeze-dried up to quinoa active antihypertensive peptide.
Embodiment 5
A kind of quinoa polypeptide with hypotensive activity, preparation method are as follows:
(1) quinoa seed clear water is rinsed to no foam, dries pulverizing, and n-hexane degreasing takes 200g to be placed in conical flask,
3000mL distilled water is added, adjusts pH to 10, magnetic agitation extracts 2h at room temperature, and centrifugation takes supernatant；Adjust supernatant
Liquid pH to 5, in 4 DEG C of standing 30min, centrifugation takes precipitating；Add a small amount of water to dissolve precipitating, adjusts pH to 7, freeze-drying obtains
Quinoa protein crude extract administration.
(2) quinoa protein crude extract administration is taken, is dissolved in water, pH to 2 is adjusted, food-grade stomach cardia is added with the ratio of 1:20
Enzyme, 50 DEG C of hydrolysis 30min, then pH to 8 is adjusted, food-grade trypsase, 50 DEG C of hydrolysis 30min, 95 DEG C of heating water baths are added
10min is centrifuged rapidly after cooling, takes supernatant.
(3) after quinoa polypeptide crude extract being crossed 0.45um filter membrane, the Chenopodiaceae of 5kDa is less than by ultrafiltration membrane trapped molecular weight
Wheat polypeptide, dialysis desalting are concentrated in vacuo, are freeze-dried up to quinoa active antihypertensive peptide.
The hypotensive activity (ACE enzyme inhibition activity) of above-mentioned prepared quinoa polypeptide is detected below by experiment in vitro,
To illustrate beneficial effects of the present invention；
The ACE inhibitory activity of 1 quinoa polypeptide sample of table
| Sample | IC50(mg/mL) |
| Embodiment 1 | 0.187 |
| Embodiment 2 | 0.203 |
| Embodiment 3 | 0.201 |
| Embodiment 4 | 0.195 |
| Embodiment 5 | 0.199 |
The results show (as shown in table 1), the small-molecular peptides that quinoa albumen obtains after complex enzyme hydrolysis, ultrafiltration can be effective
Inhibit ACE activity.
Therefore, the quinoa polypeptide that method produced according to the present invention is prepared, can be used as effective active composition and further opens
Send out into functional food or drug with hypotensive activity.
Although the present invention is disclosed as above with embodiment, so it is not intended to limit the present invention, any those skilled in the art
Member, without departing from the spirit and scope of the present invention, can make a variety of different selections and modification, therefore protection model of the invention
It encloses and is limited by claims and its equivalents.
Claims (10)
1. a kind of preparation method of the quinoa polypeptide with hypotensive activity, which comprises the following steps: use alkali carries
The heavy method of acid extracts quinoa albumen from degreasing quinoa powder and is finally concentrated in vacuo, is freeze-dried through complex enzyme hydrolysis, ultrafiltration, desalination
To quinoa active antihypertensive peptide.
2. the preparation method of the quinoa polypeptide according to claim 1 with hypotensive activity, which is characterized in that the system
Preparation Method the following steps are included:
1) protein extraction: raw material degreasing quinoa powder is mixed with suitable water, adjusts pH to 9~10, and constant temperature oscillation extracts 1-2h,
Centrifugation, takes supernatant；Supernatant pH to 4~5 is adjusted, stands, centrifugation, takes precipitating；Adjusted after precipitating plus the dissolution of a small amount of water pH to
Neutrality, freeze-drying, obtains quinoa protein crude extract administration；
2) proteolysis: taking the quinoa protein crude extract administration of step 1), be dissolved in water, and adjusts pH to 2, pepsin is added, 30
DEG C~50 DEG C of water-baths 30~60min of oscillation, obtain quinoa albumen pepsin hydrolysis liquid；Adjust quinoa albumen pepsin hydrolysis
Trypsase is added in the pH to 8 of liquid, vibrates 30~60min in 30 DEG C~50 DEG C water-baths, 95 DEG C of heating 10min inactivated proteases,
Centrifuging and taking supernatant obtains quinoa polypeptide crude extract；
3) ultrafiltration desalination: passing sequentially through filter membrane and ultrafiltration membrane for the quinoa polypeptide crude extract that step 2) obtains, after permeate desalination,
It is concentrated in vacuo, is freeze-dried to obtain quinoa polypeptide.
3. the preparation method of the quinoa polypeptide according to claim 2 with hypotensive activity, which is characterized in that the step
It is rapid 1) in, the mass ratio of the degreasing quinoa powder and the water is 1:(10~15).
4. the preparation method of the quinoa polypeptide according to claim 2 with hypotensive activity, which is characterized in that the step
It is rapid 2) described in pepsin and trypsase be food grade enzyme.
5. the preparation method of the quinoa polypeptide according to claim 2 with hypotensive activity, which is characterized in that the step
Rapid enzymolysis time 2) is selected to determine by the ACE inhibitory activity of enzymolysis liquid.
6. the preparation method of the quinoa polypeptide according to claim 2 with hypotensive activity, which is characterized in that the step
It is rapid 3) described in filter membrane be 0.45 μm of filter membrane.
7. the preparation method of the quinoa polypeptide according to claim 2 with hypotensive activity, which is characterized in that the step
Rapid ultrafiltration retaining molecular weight 3) is selected to determine by the ACE inhibitory activity of filtrate.
8. the preparation method of the quinoa polypeptide according to claim 7 with hypotensive activity, which is characterized in that the step
The trapped molecular weight 5kDa of rapid ultrafiltration membrane 3).
9. the Chenopodiaceae that the preparation method of -8 any quinoa polypeptides with hypotensive activity is prepared according to claim 1
Wheat polypeptide.
What 10. the preparation method of the quinoa polypeptide according to claim 1 to 4 with hypotensive activity was prepared
The application of quinoa polypeptide has the drug or food of antihypertensive function applied to preparation.
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