CN105420321A - Preparation method of weakly-alkaline soybean peptide - Google Patents
Preparation method of weakly-alkaline soybean peptide Download PDFInfo
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- CN105420321A CN105420321A CN201510861343.1A CN201510861343A CN105420321A CN 105420321 A CN105420321 A CN 105420321A CN 201510861343 A CN201510861343 A CN 201510861343A CN 105420321 A CN105420321 A CN 105420321A
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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
Abstract
The invention provides a preparation method of weakly-alkaline soybean peptide. The preparation method includes the following steps of 1, performing enzymolysis, wherein soybean protein isolate is dissolved, the protein concentration is controlled to be 1-12 weight%, protease is added, enzymolysis treatment is performed for 1-6 h under the conditions that the temperature ranges from 35 DEG C to 65 DEG C and the pH ranges from 5.5 to 8.0, and a soybean peptide solution is prepared; 2, performing chromatographic separation, wherein the soybean peptide solution obtained in the step 1 is introduced into a cation-exchange chromatography column, the introduction quantity of the soybean peptide solution is 2-12 times the volume of cation-exchange resin, and after the chromatography column is leached through clean water, a sodium chloride solution with the volume 1-6 times that of the cation-exchange resin is adopted as an eluant; 3, performing desalination, wherein the eluant obtained in the step 2 is introduced into a macroporous adsorption resin chromatography column, an organic solvent aqueous solution with the volume being 1-3 times that of macroporous adsorption resin and the volume concentration being 10-90% is adopted as an eluant, and the macroporous adsorption resin chromatography column is eluted; 4, performing concentration, wherein all the eluants obtained in the step 3 are concentrated under the pressure reduction condition, and the concentration of peptide is controlled to be 20-30 weight% after concentration; 5, performing drying.
Description
Technical field
The invention belongs to field of food science, particularly, the present invention relates to a kind of preparation method of weakly alkaline soybean polypeptide.
Technical background
Polypeptide is that the amino acid that quantity does not wait is formed by peptide bond condensation, and amino acid quantity is generally within 20, and an important sources of polypeptide is exactly that protein bio method or chemical method are degraded.Much research shows that protein mainly absorbs with the form of peptide after digestive tube enzymically hydrolyse, and absorbs more easily, sooner by body than complete total free aminoacids and utilize.Many polypeptide can not only provide growth in humans, grow required for nutritive substance, and there is unique biological function can prevent and treat thrombus, hyperlipidemia and hypertension, delay senility, resisting fatigue, raising immunity of organisms.Polypeptide has extremely strong activity and diversity, and some polypeptide has former food protein or the unexistent important physiological function of its composition amino acid.Biologically active polypeptides has caused in current scientific and technological circle, medical circle, nutrition educational circles and field of food science and has paid attention to widely.Along with deepening continuously of research, peptide art worldwide receives publicity day by day.After numerous protein converts polypeptide to, its nutritive value will significantly promote.
In soybean, protein content accounts for 35-40%, it is more than 2 times of cereal crop protein content, and soybean protein amino acid ratio also comparatively balances, close to the desired proportions of needed by human body, especially lysine content is higher, therefore, the nutritive value of soybean protein can match in excellence or beauty with animal proteinum, is the protein resource of human foods middle ideal.In theory, soybean protein to be degraded into after polypeptide its functional and nutritive effect higher than soybean protein.The soybean polypeptide prepared fully can not meet the food of the special conditions personnel of proteinaceous nutrient demand as those usual diet, for the nutritional fortification of food and the base-material of functional foodstuff, polypeptide specific absorption is high, so can be used as intestinal absorption nutrition.The elderly of the reason poor appetite of some diseases or aging and be operating patient and directly take in this nutrition and more promptly can return to normal nutrition state from mouth, stomach than through intravenous amino acid, in addition, soybean polypeptide can also be applied to body building and weight reducing food, protective foods etc.
From human consumption's structure composition and functional perspective analysis thereof, proteins and peptides is degraded into polypeptide and the lower polypeptide of molecular weight respectively by stomach en-under stomach sour environment, and the characteristic of this process degraded is not strong.Under the Chymotrypsin of pancreas secretion and trypsin acting, form oligopeptides and amino acid after polypeptide enters enteron aisle, this process completes in the basic conditions.Polypeptide containing arginine and Methionin in aminoacid sequence could by trypsin degradation, in peptide sequence containing arginine and Methionin then this polypeptide be weakly alkaline, therefore, weakly alkaline polypeptide should have higher specific absorption and utilization ratio, therefore its nutritive value and functional higher than common not containing the polypeptide of the basic aminoacids such as arginine and Methionin.
Summary of the invention
The present invention aims to provide a kind of preparation method of weakly alkaline soybean polypeptide, according to the feature that human body protein and polypeptide absorb, take soybean protein isolate as raw material, utilize based on the peptide separation method of chromatography, prepare and be rich in Methionin and arginic weakly alkaline soybean polypeptide and be prepared into solid beverage.
The preparation method of a kind of weakly alkaline soybean polypeptide of the present invention, it is characterized in that, this preparation method comprises the following steps:
(1) enzymolysis: dissolved by soybean protein isolate, protein concentration controls, 1 ~ 12 % by weight, to add proteolytic enzyme, 35 ~ 65 DEG C, enzymolysis processing 1 ~ 6 hour under the condition of pH5.5 ~ 8.0, prepares soybean polypeptide solution;
(2) chromatographic separation: soybean polypeptide solution step (1) obtained imports cation-exchange chromatography post, the import volume of soybean polypeptide solution is 2 ~ 12 times of Zeo-karb volume, the sodium chloride solution of 1-6 times of Zeo-karb volume is adopted to be elutriant after adopting clear water drip washing chromatography column, elution chromatography post, collect whole elutriant, containing weakly alkaline soybean polypeptide in this elutriant;
(3) desalination: elutriant step (2) obtained imports macroporous adsorption resin chromatography post, adopt 1-3 times of macroporous adsorbent resin, the concentration of volume is 10 ~ 90% aqueous solutions of organic solvent is elutriant, and wash-out macroporous adsorption resin chromatography post, collects whole elutriant;
(4) concentrated: to be concentrated at reduced pressure conditions by whole elutriants that step (3) obtains, after concentrated, the concentration of polypeptide controls 20 ~ 30 % by weight;
(5) dry: the elutriant that step (4) obtains is carried out spraying dry, size controlling is at 20-160 order, and water content control, 1 ~ 6 % by weight, obtains weakly alkaline soybean polypeptide, at least containing a Methionin or Histidine in each polypeptide.
Preferably, in the enzymolysis process of described step (1), the proteolytic enzyme added and the ratio of soybean protein isolate are 5:1 ~ 200:1 ~ 50:1.
Preferably, in the chromatographic separation process of described step (2), the concentration of sodium chloride solution can control at 0.1 ~ 1.5mol/L.
Preferably, in described step (3) demineralising process, organic solvent can be ethanol, also can be methyl alcohol or acetone.
The preparation method of a kind of weakly alkaline soybean polypeptide provided by the invention, the polypeptide prepared can as beverage, foodstuff additive or emulsifying agent etc., its advantage is that preparation process is simple, in its aminoacid sequence of the polypeptide of preparation Methionin or arginine content high, the absorption rate of polypeptide is high.
Figure of description
Fig. 1 is preparation method's schema of a kind of weakly alkaline soybean polypeptide of the present invention
Specific embodiment
Below by embodiment, the present invention is specifically described; what be worth proposition is; the explanation of embodiment just to the preparation method of a kind of weakly alkaline soybean polypeptide of the present invention; the whole of patent of the present invention can not be represented; more limiting the scope of the invention can not be interpreted as; under the precondition of design not departing from essence of the present invention, some adjustment or improvement can also be made, all belong to protection scope of the present invention.
Embodiment 1
(1) enzymolysis: be dissolved into by 1kg soybean protein isolate in 19kg water, soybean protein isolate concentration is 5 % by weight, adds subtilisin 10g, enzymolysis processing 2 hours under 50 DEG C and pH6.5 condition, obtains soybean polypeptide solution 20kg.
(2) chromatographic separation: soybean polypeptide solution step (1) obtained imports the cation-exchange chromatography post that resin volume is 2L, adopt 6L deionized water drip washing chromatography column, employing 5L concentration is that the sodium chloride solution of 1.5mol/L carries out wash-out, collect whole elutriant 5L, containing weakly alkaline soybean polypeptide in this elutriant.
(3) desalination: 5L elutriant step (2) obtained imports the macroporous adsorption resin chromatography post that resin volume is 1L, employing 3L volumetric concentration is the aqueous ethanolic solution wash-out macroporous adsorption resin chromatography post of 60%, collects whole elutriant 3L.
(4) concentrated: to be concentrated at reduced pressure conditions by whole elutriants that step (3) obtains, after concentrated, the concentration of polypeptide is 26 % by weight.
(5) dry: the elutriant that step (4) obtains is carried out spraying dry, size controlling is at 60 orders, water content is 4 % by weight, obtains weakly alkaline soybean polypeptide, at least containing a Methionin or Histidine in each polypeptide of mass spectrometry results surface.
Embodiment 2
(1) enzymolysis: be dissolved into by 2kg soybean protein isolate in 9kg water, soybean protein isolate concentration is 10 % by weight, adds subtilisin 20g, enzymolysis processing 1 hour under 55 DEG C and pH7.0 condition, obtains soybean polypeptide solution 10kg.
(2) chromatographic separation: soybean polypeptide solution step (1) obtained imports the cation-exchange chromatography post that resin volume is 1L, adopt 2L deionized water drip washing chromatography column, employing 2L concentration is that the sodium chloride solution of 1.0mol/L carries out wash-out, collect whole elutriant 2L, containing weakly alkaline soybean polypeptide in this elutriant.
(3) desalination: 2L elutriant step (2) obtained imports the macroporous adsorption resin chromatography post that resin volume is 500ml, employing 1L volumetric concentration is the aqueous ethanolic solution wash-out macroporous adsorption resin chromatography post of 90%, collects whole elutriant 1L.
(4) concentrated: to be concentrated at reduced pressure conditions by whole elutriants that step (3) obtains, after concentrated, the concentration of polypeptide is 20 % by weight.
(5) dry: the elutriant that step (4) obtains is carried out spraying dry, size controlling is at 80 orders, water content is 3 % by weight, obtains weakly alkaline soybean polypeptide, at least containing a Methionin or Histidine in each polypeptide of mass spectrometry results surface.
Claims (2)
1. a preparation method for weakly alkaline soybean polypeptide, is characterized in that, this preparation method comprises the following steps:
(1) enzymolysis: dissolved by soybean protein isolate, protein concentration controls, 1 ~ 12 % by weight, to add proteolytic enzyme, 35 ~ 65 DEG C, enzymolysis processing 1 ~ 6 hour under the condition of pH5.5 ~ 8.0, prepares soybean polypeptide solution;
(2) chromatographic separation: soybean polypeptide solution step (1) obtained imports cation-exchange chromatography post, the import volume of soybean polypeptide solution is 2 ~ 12 times of Zeo-karb volume, the sodium chloride solution of 1-6 times of Zeo-karb volume is adopted to be elutriant after adopting clear water drip washing chromatography column, elution chromatography post, collect whole elutriant, containing weakly alkaline soybean polypeptide in this elutriant;
(3) desalination: elutriant step (2) obtained imports macroporous adsorption resin chromatography post, adopt 1-3 times of macroporous adsorbent resin, the concentration of volume is 10 ~ 90% aqueous solutions of organic solvent is elutriant, and wash-out macroporous adsorption resin chromatography post, collects whole elutriant;
(4) concentrated: to be concentrated at reduced pressure conditions by whole elutriants that step (3) obtains, after concentrated, the concentration of polypeptide controls 20 ~ 30 % by weight;
(5) dry: the elutriant that step (4) obtains is carried out spraying dry, size controlling is at 20-160 order, and water content control, 1 ~ 6 % by weight, obtains weakly alkaline soybean polypeptide, at least containing a Methionin or Histidine in each polypeptide.
2. the preparation method of a kind of weakly alkaline soybean polypeptide as claimed in claim 1, is characterized in that at least containing a Methionin or Histidine in each polypeptide in this weakly alkaline polypeptide.
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| CN201510861343.1A CN105420321A (en) | 2015-12-01 | 2015-12-01 | Preparation method of weakly-alkaline soybean peptide |
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| CN201510861343.1A CN105420321A (en) | 2015-12-01 | 2015-12-01 | Preparation method of weakly-alkaline soybean peptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106072666A (en) * | 2016-06-03 | 2016-11-09 | 李继富 | A kind of low ash, the production method of soybean peptide without bitter powder and production system |
| 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 |
|---|---|---|---|---|
| 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 |
| CN104593317A (en) * | 2013-10-31 | 2015-05-06 | 中国食品发酵工业研究院 | Soybean bioactive peptide additive used for cell medium |
-
2015
- 2015-12-01 CN CN201510861343.1A patent/CN105420321A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN104593317A (en) * | 2013-10-31 | 2015-05-06 | 中国食品发酵工业研究院 | Soybean bioactive peptide additive used for cell medium |
Non-Patent Citations (3)
| Title |
|---|
| 张晓梅: "降血压和降胆固醇大豆肽的分离纯化", 《中国优秀博硕士学位论文全文数据库（硕士） 工程科技I辑》 * |
| 徐曼等: "大豆多肽分离纯化技术研究进展", 《中国酿造》 * |
| 郭红莲: "大豆蛋白生物降解及大豆肽的研究进展", 《粮油加工与食品机械》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106072666A (en) * | 2016-06-03 | 2016-11-09 | 李继富 | A kind of low ash, the production method of soybean peptide without bitter powder and production system |
| 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: 20160323 |