CN103484519B - A kind of wheat protein peptide and preparation method thereof and its application - Google Patents
A kind of wheat protein peptide and preparation method thereof and its application Download PDFInfo
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- CN103484519B CN103484519B CN201310412885.1A CN201310412885A CN103484519B CN 103484519 B CN103484519 B CN 103484519B CN 201310412885 A CN201310412885 A CN 201310412885A CN 103484519 B CN103484519 B CN 103484519B
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Abstract
The invention discloses a kind of wheat protein peptide and preparation method thereof and application.This wheat protein peptide prepares according to the method comprised the steps: adopt papoid to carry out enzyme digestion reaction to wheat protein concentrate, obtain described wheat protein peptide.Its relative molecular mass, mainly between 0 ~ 1000Da, forms by 2 ~ 9 amino acid.Wheat protein peptide provided by the present invention can be used for preparing the product with functions such as hypotensive, analgesia, opioid peptides sample are active, as medicine, healthcare products, food, skin care product, hair shampoo article or articles for washing etc.The preparation technology that the present invention prepares wheat protein peptide is simple and easy, less investment, instant effect, and return rate is high, input reciprocation cycle is short, is suitable for suitability for industrialized production.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of wheat protein peptide and preparation method thereof and its application.
Background technology
Peptide is the hydrolysate of protein, i.e. the structure fragment of protein.Two amino acid connect into dipeptides by peptide bond, and three amino acid connect into tripeptides by peptide bond, and multiple amino acid connects into polypeptide by peptide bond.Generally the peptide of 2-10 amino-acid residue composition, be referred to as oligopeptides.Because the molecule of the molecule opposing proteins of peptide is much smaller, but biological activity is very high, a small amount of peptide just can play an important role, so be called as bioactive peptide.
The infinite glamour of bioactive peptide is, it than protein or total free aminoacids digestion faster, absorb more, biological value and nutritive value higher.In recent years, many bioactive peptide products with different biological function are developed, develop, and obtain unusual effect for practice.Amino acid sequence different in bioactive peptide, determining them has different functions, and they are by multiple metabolism and physiological regulation, can play the effects such as Promote immunity, Green Tea Extract, hormone regulation, antibacterial, antiviral, hypotensive, reducing blood-fat
Wheat is grass, be the widest food crop that distribute in the world, sown area is the hat of food crop, is to prepare the abundant and cheap raw material of bioactive peptide, meanwhile, the exploitation of wheat bioactive peptide and the checking of biological effect also greatly improve utilization ratio and the economic worth of wheat.
Summary of the invention
An object of the present invention is to provide a kind of wheat protein peptide and preparation method thereof and its application.
A kind of method preparing wheat protein peptide provided by the invention, comprises the steps: to adopt papoid to carry out enzyme digestion reaction to wheat protein concentrate, obtains described wheat protein peptide.
In described method, the preparation process of described wheat protein concentrate comprises every 1g wheat gluten and adds the aqueous sodium hydroxide solution that 10ml concentration is 0.1mol/L, fully stirs after 1 hour and carries out centrifugation, collecting precipitation; Supernatant liquor acetic acid is adjusted to pH4, after being heated to 90 DEG C, places 10 minutes, recentrifuge is separated, collecting precipitation, namely obtains wheat protein concentrate after the precipitation of twice collection being merged.
In described method, the temperature of reaction of described enzyme digestion reaction is 65 DEG C, and the reaction times is 2 hours, and pH value in reaction is 5.5-5.9.
In described method, in the reaction system of described enzyme digestion reaction, the ratio of quality and the number of copies of wheat protein concentrate and water is 1:15; Papoid 8000U needed for enzymolysis every gram wheat protein concentrate.
Described method also comprises the steps: after enzyme digestion reaction terminates, and carries out refrigeration settlement treatment to enzymolysis solution, filters, and collects filtrate; The temperature of described refrigeration sedimentation is-4 DEG C, and the time is 12 hours.
Described method also comprises the step of described filtrate being carried out enzyme-deactivating process; The condition of described enzyme-deactivating is: temperature 100 DEG C, 20 minutes time.
The wheat protein peptide that described method prepares also belongs to the scope of protection of the invention.
The purposes of the wheat protein peptide that described method prepares in following at least one:
(1) application in hypotensive product is prepared;
(2) application in the product of analgesia and/or opioid peptides sample activity is prepared.
In described application, described product can be medicine, healthcare products, food, skin care product, hair shampoo article or articles for washing.
Another object of the present invention is to provide a kind of product with following arbitrary purposes, and its activeconstituents is the wheat protein peptide that described method prepares:
(1) hypotensive;
(2) analgesia and/or opioid peptides sample activity.
Wheat protein peptide provided by the present invention can be used for preparing the product with functions such as hypotensive, analgesia, opioid peptides sample are active, as medicine, healthcare products, food, skin care product, hair shampoo article or articles for washing etc.The preparation technology that the present invention prepares wheat protein peptide is simple and easy, less investment, instant effect, and return rate is high, input reciprocation cycle is short, is suitable for suitability for industrialized production.
Take wheat protein peptide as hypotensive, the analgesia of active fraction preparation, the medicine of opioid peptides sample activity or healthcare products, when needs, one or more pharmaceutically acceptable carriers can also be added in said medicine or healthcare products.Described carrier comprises the thinner, vehicle, weighting agent, tackiness agent, wetting agent, disintegrating agent, absorption enhancer, tensio-active agent, absorption carrier, lubricant etc. of pharmaceutical field routine.
Described hypotensive, analgesia, opioid peptides sample activity medicine can make the various ways such as oral liquid, tablet, electuary, capsule (comprising soft, hard capsule), spray, paste and coated pill.The medicine of above-mentioned various formulation all can be prepared according to the ordinary method of pharmaceutical field.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
The preparation of embodiment 1, wheat protein peptide
Wheat protein concentrate take wheat gluten as raw material, and every 1g wheat gluten adds the aqueous sodium hydroxide solution that 10ml concentration is 0.1mol/L, fully stirs after 1 hour and carry out centrifugation, collecting precipitation; Supernatant liquor acetic acid is adjusted to pH4, after being heated to 90 DEG C, places 10 minutes, recentrifuge is separated, collecting precipitation, namely obtains wheat protein concentrate after the precipitation of twice collection being merged.
The preparation of wheat protein peptide
The wheat protein concentrate of above-mentioned preparation is dropped in enzymatic vessel, in enzymatic vessel, adds the water of 15 times being equivalent to described wheat protein concentrate quality, need the amount of papoid 8000U to add papoid by enzymolysis every gram wheat protein concentrate; Stirring and evenly mixing post-heating to 65 DEG C carries out enzyme digestion reaction 2 hours, and enzymolysis process uses Glacial acetic acid adjust ph, keeps pH value at 5.5-5.9; After enzymolysis terminates, also need to carry out enzyme-deactivating process to enzymolysis solution, the condition of described enzyme-deactivating is temperature 100 DEG C, and the time is 20 minutes.Under-4 DEG C of conditions, refrigerate settlement treatment 12 hours afterwards, get supernatant liquor, more namely obtain wheat protein peptide concentrated solution with after Plate Filtration; Wheat protein peptide concentrated solution is carried out lyophilize or spray drying treatment, obtains wheat protein Gly-His-Lys.
The molecular weight distribution of embodiment 2, wheat protein peptide detects
The measuring method of the molecular weight distribution of wheat protein peptide adopts high performance gel filtration chromatography, test result shows that the relative molecular mass of wheat protein peptide provided by the invention mainly concentrates on small-molecular peptides and the aminoacid mixture of 0-1000Da, and little peptide content is more than 70%.
The preparation of comparative example 1-3, wheat protein peptide
According to the enzyme reaction condition described in table 1, by method described in embodiment 1, carry out the preparation of wheat protein peptide.
Table 1
| Enzyme digestion reaction pH value | Enzyme digestion reaction temperature | The enzyme digestion reaction time | |
| Comparative example 1 | 6.0-6.5 | 60℃ | 3 hours |
| Comparative example 2 | 5.0-5.5 | 55℃ | 2.5 hour |
| Comparative example 3 | 6.5-7.0 | 70℃ | 1 hour |
Calculate the content mainly concentrating on the small-molecular peptides of 0-1000Da by the wheat protein peptide Middle molecule quality prepared by the reaction conditions of each comparative example, result shows, according to comparative example 1-3 reaction conditions prepared by the little peptide content of wheat protein peptide be followed successively by 44%, 42%, 36%.The preparation method of the wheat protein peptide that these data show described in embodiment 1 is better than comparative example 1-3.
The hypotensive pharmacodynamics test of embodiment 3, wheat protein peptide
The hypotensive pharmacodynamics merit of wheat protein peptide is verified by the test experience of Zinc metallopeptidase Zace1 (ACE) inhibit activities.
(1) preparation of reagent and testing sample solution
The preparation of ACE solution: 1UACE be dissolved in 2ml0.1mol/L borate buffer (pH8.3, containing 0.4mol/LNaCl) and get final product.
The preparation of HHL solution: get HHL appropriate, dissolves with 0.1mol/L borate buffer (pH8.3, containing 0.4mol/LNaCl) and is made into 5mmol/LHHL solution.
The preparation of urobenzoic acid reference liquid: get urobenzoic acid standard substance appropriate, be made into the urobenzoic acid reference liquid of different concns with 0.1mol/L borate buffer (pH8.3, containing 0.4mol/LNaCl).
The preparation of wheat protein peptide sample: wheat protein peptide prepared by the wheat protein peptide of Example 1 preparation, comparative example 1-3 is appropriate, dissolves mixing, be made into the sample solution of different concns with 0.1mol/L borate buffer (pH8.3, containing 0.4mol/LNaCl).
(2) enzyme reaction
Get the wheat protein peptide sample solution 15 μ L of different concns, add the ACE solution of 15 μ L, add 50 μ LHHL solution after 37 DEG C of water bath heat preservation 3min and start reaction, react 30min under 37 DEG C of conditions after, add 100 μ L1.0mol/LHCl solution termination reactions; Use 15 μ L0.1mol/L borate buffers (pH8.3, containing 0.4mol/LNaCl) to replace wheat protein peptide sample solution, as blank group simultaneously.
(3) HPLC analyzing and testing result
Above-mentioned enzyme reaction solution is carried out HPLC analysis after 0.45 μm of membrane filtration.Determined wavelength: 228nm; Mobile phase A is water (containing 0.05% trifluoroacetic acid), and Mobile phase B is acetonitrile (containing 0.05% trifluoroacetic acid); Flow velocity 0.4ml/min; Column temperature 30 DEG C.
(4) interpretation
The calculating of ACE inhibiting rate:
ACE inhibiting rate (%)=(a-b)/a × 100%
In formula, a is the peak area of blank group urobenzoic acid; B is the peak area of sample sets urobenzoic acid.
ACE inhibiting rate experiment and HPLC analytical results display, wheat protein peptide sample solution concentration and ACE inhibiting rate linear, increasing namely along with wheat protein peptide concentration, its ACE inhibiting rate also raises; The IC of the ACE suppression of wheat protein peptide prepared by embodiment 1
50for 5.24mg/ml; The IC of the ACE suppression of wheat protein peptide prepared by comparative example 1-3
50be followed successively by 2.84mg/ml, 2.45mg/ml, 2.01mg/ml.
The analgesia pharmacodynamics test of embodiment 4, wheat protein peptide
The wheat protein peptide adopting embodiment 1 to prepare, tests the impact of the mouse threshold of pain, investigates the analgesic activity of medicine of the present invention.Specific experiment process is as follows:
Get female mice, body weight 18 ~ 22g.Its normal pain threshold surveyed by the hot plate before experiment mouse being placed in 55 ± 0.2 DEG C of pain threshold detectors, and lick with mouse the index that metapedes required time (second) occurs for the threshold of pain, all threshold of pain are all given it up more than 30 seconds or lower than 5 seconds or leaper.Get the mouse of threshold of pain between 5 ~ 30 seconds 60, body weight 20.0 ± 0.9g, often only survey its normal pain threshold secondary, get its average as the normal pain threshold before this mouse administration, animal is divided at random 4 experimental group, often organize 10, comprise blank group, the dosage using the wheat protein peptide of embodiment 1 is respectively 100mg/kg, 50mg/kg, three groups of 25mg/kg, each group of equal gastric infusion, control group gives equal-volume distilled water, once a day, continuous 5 days, after last administration 30 minutes, the threshold of pain of every treated animal is respectively surveyed once when 90 minutes, and compare with control group, carry out statistical procedures, the results are shown in Table 2.
Table 2(χ ± SD)
Compare with control group in table 2, * P < 0.05, * * P < 0.01)
Table 2 result shows, wheat protein Toplink of the present invention obviously increases the threshold of pain of mouse, has obvious analgesic activity.
Claims (5)
1. prepare a method for wheat protein peptide, comprise the steps: to adopt papoid to carry out enzyme digestion reaction to wheat protein concentrate, obtain described wheat protein peptide; The preparation process of described wheat protein concentrate comprises every 1g wheat gluten and adds the aqueous sodium hydroxide solution that 10ml concentration is 0.1mol/L, fully stirs after 1 hour and carries out centrifugation, collecting precipitation; Supernatant liquor acetic acid is adjusted to pH4, after being heated to 90 DEG C, places 10 minutes, recentrifuge is separated, collecting precipitation, namely obtains wheat protein concentrate after the precipitation of twice collection being merged;
The temperature of reaction of described enzyme digestion reaction is 65 DEG C, and the reaction times is 2 hours, and pH value in reaction is 5.5-5.9;
In the reaction system of described enzyme digestion reaction, the ratio of quality and the number of copies of wheat protein concentrate and water is 1:15; Papoid 8000U needed for enzymolysis every gram wheat protein concentrate;
Described method also comprises the steps: after enzyme digestion reaction terminates, and carries out refrigeration settlement treatment to enzymolysis solution, filters, and collects filtrate; The temperature of described refrigeration sedimentation is-4 DEG C, and the time is 12 hours;
Described method also comprises the step of described filtrate being carried out enzyme-deactivating process; The condition of described enzyme-deactivating is: temperature 100 DEG C, 20 minutes time.
2. the wheat protein peptide for preparing of method described in claim 1.
3. the purposes of wheat protein peptide according to claim 2 in following at least one:
(1) application in hypotensive product is prepared;
(2) application in the product of analgesia and/or opioid peptides sample activity is prepared.
4. application according to claim 3, is characterized in that: described product is medicine, healthcare products, food, skin care product, hair shampoo article or articles for washing.
5. have a product for following arbitrary purposes, its activeconstituents is wheat protein peptide according to claim 2:
(1) hypotensive;
(2) analgesia and/or opioid peptides sample activity.
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| CN105961819A (en) * | 2016-05-05 | 2016-09-28 | 巢湖市鑫宇良种农民专业合作社 | Method for extracting wheat oligopeptide from wheat |
| CN107712065A (en) * | 2016-08-11 | 2018-02-23 | 宏乐集团有限公司 | A kind of person in middle and old age are formulated peptide milk powder and preparation method thereof |
| CN107319099A (en) * | 2017-07-13 | 2017-11-07 | 安徽生物肽产业研究院有限公司 | It is a kind of that there is rich potassium wheat small peptide of reduction hypertension effect and preparation method thereof |
Citations (2)
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| CN101156645A (en) * | 2007-11-06 | 2008-04-09 | 江苏大学 | A wheat gluten proteolysis matter and method for making same as well as its usage |
| CN103173511A (en) * | 2011-12-23 | 2013-06-26 | 中国食品发酵工业研究院 | Method for industrially producing wheat glutamine peptide |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101156645A (en) * | 2007-11-06 | 2008-04-09 | 江苏大学 | A wheat gluten proteolysis matter and method for making same as well as its usage |
| CN103173511A (en) * | 2011-12-23 | 2013-06-26 | 中国食品发酵工业研究院 | Method for industrially producing wheat glutamine peptide |
Non-Patent Citations (3)
| Title |
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| 不同方法分离小麦麸皮中蛋白质的研究，;陈凤莲;《粮食加工》;20090430;第34卷(第4期);28-30 * |
| 酶法水解小麦面筋蛋白研究;孔祥珍;《粮食与油脂》;20060430(第4期);21-23 * |
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Effective date of registration: 20190704 Address after: 430071 Building A3 of Guanggu No. 7 Biomedical Park, Donghu New Technology Development Zone, Wuhan City, Hubei Province Patentee after: Th Wuhan biological Polytron Technologies Inc Address before: Room 2102, Huayin Building, 786 Democracy Road, Wuchang District, Wuhan City, Hubei Province Patentee before: Zou Yuandong |
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