CN101787387A - Acid-resistant no-bitterness soybean oligopeptide and production method and application thereof - Google Patents
Acid-resistant no-bitterness soybean oligopeptide and production method and application thereof Download PDFInfo
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- CN101787387A CN101787387A CN200910194287A CN200910194287A CN101787387A CN 101787387 A CN101787387 A CN 101787387A CN 200910194287 A CN200910194287 A CN 200910194287A CN 200910194287 A CN200910194287 A CN 200910194287A CN 101787387 A CN101787387 A CN 101787387A
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Abstract
The invention discloses acid-resistant no-bitterness soybean oligopeptide and a production method and the application thereof. The invention adopts a soy protein isoelectric point enzyme method, carries out enzymolysis simultaneously with exoproteinase and endo protease or with protease with the functions of incision enzyme and excision enzyme, so that the degree of hydrolysis of soy protein is up to 42 percent, above 85 percent of soybean oligopeptide only contains less than 8 amino acids, wherein most of the soybean oligopeptide contains 3 to 5 amino acids, the average molecular weight is below 1,000Da, and in addition, 10 to 20 percent of FAA is also contained. The acid-resistant no-bitterness soybean oligopeptide prepared by the invention has very good solubility, even at the isoelectric points of soy protein, at least 95 percent of oligopeptide is soluble; and therefore, the invention is widely applied to acid food systems, can be directly added into sports drinks, nutrition supplementary drinks and other products, and does not produce precipitation or flocculation and other unstable phenomena that protein drinks are easy to produce.
Description
Technical field
The present invention relates to a kind of intensive processing technology of soybean protein, particularly a kind of soybean oligopeptide and production method and application of acidproof and no bitter taste.
Background technology
The compound that is formed by 2～10 amino acid molecular dehydrating condensations is called oligopeptides (small-molecular peptides); The compound that 10～50 molecule dehydrating condensations form is called polypeptide; The compound that is formed by the molecule dehydrating condensation more than 50 just is called protein.Soybean oligopeptide is with after the soybean protein hydrolysis effect, again through the oligo peptide that process obtains such as separating, refining.
Discover that the oligopeptides molecular weight is little, has good solubility; Be heated or under acid-base condition, good anticoagulation arranged, good stability.The maximum characteristics of soybean oligopeptide are and can be absorbed by the body rapidly that its absorption rate is all faster than amino acid.Therefore, but directly replenish amino acid whose absorption process in the oligopeptides acceleration bodies, improve the utilization ratio of body, people's bulk-growth is provided, grows needed nutritive substance nitrogen.
Simultaneously, some physiologically active instrumentality is similar in the aminoacid sequence of some oligopeptides and the body, participates in the organism physiology active directly and regulates, and perhaps has the secretion that stimulates intestinal receptor hormone or enzyme and human body is brought into play many very important physiological actions.As blood pressure regulation, opposing free radical, improve immunizing power, alleviating fatigue, delay senility and prevent and treat hyperlipidemia etc., this is that raw material food protein or other amino acid are not available.
Oligopeptides has significant promoter action to multiple microbial growths such as milk-acid bacteria, bifidus bacillus and yeast, thereby helps digestion, the absorption of food nutrition.In addition, the low antigenicity of oligopeptides has been avoided the anaphylaxis after the food, and the osmotic pressure of little peptide is lower than amino acid, untoward reactions such as the no abdominal distension in food back, diarrhoea.
Soybean oligopeptide has broad application prospects, and it or all has its unique distinction no matter on functional performance on the trophology, be one of soybean deep processed product of foodstuffs industry focus development.The problem that present soybean peptides series products on the market mainly exists is: molecular weight is bigger, and is most 2,000～5, functional relatively poor between 000Da, and forms precipitation in the acidic drinks system easily; Local flavor is not good enough, and the hydrolysis flavor is more outstanding, and bitter taste is more obvious, has limited it in Application in Food Industry.
In disclosed patent, most patents about the soybean polypeptide preparation are not further purified and separate, the target product that obtains is the mixture of albumen and polypeptide, the a few patents such as Chinese invention patent application that for example application number is 200910071291.2, name is called " enzyme process is produced the soybean sport polypeptide method " propose to remove not by the soybean protein of enzymolysis with centrifugation, further separate with the PS film, but do not clearly define the molecular weight of product scope.
At soybean polypeptide bitter taste problem, application number is 02115912.2, name is called " production method of no bitter taste soyabean polypeptide powder " and has proposed to remove to adsorb " bitter taste polypeptides " in the soybean polypeptide with stronger hypersobers such as kaolin powder and gacs, to dispel the bitter taste of soybean polypeptide.Adopt the bitter taste that improves soybean polypeptide that this adsorption method can be bigger, but can not dispel bitter taste fully, and this kaolin powder and activated carbon are non-selective adsorbents, and the soybean polypeptide of a large amount of non-bitter tastes also is adsorbed, and cause the yield rate of soybean polypeptide to reduce.The Chinese invention patent that application number is 02139560.8, name is called " a kind of soybean polypeptide and production method thereof of not having bitter taste " has proposed to act on soybean protein with the mucor protease enzyme of preparation voluntarily, can obtain not having the soybean polypeptide of bitter taste.This mucor protease of preparation voluntarily extremely has the proteolytic enzyme of business potential, but at the further research and discovery that awaits aspect the feasibility of food safety, production and the commercial cost.
Summary of the invention
Primary and foremost purpose of the present invention is to overcome the shortcoming of prior art with not enough, and a kind of production method of soybean oligopeptide of acidproof and no bitter taste is provided.Soybean oligopeptide by this production method does not have bitter taste, acidproof, and molecular-weight average is 1, below the 000Da.
Another object of the present invention is to provide the soybean oligopeptide of the acidproof and no bitter taste that obtains by described production method.
A further object of the present invention is to provide the application of the soybean oligopeptide of described acidproof and no bitter taste.
Purpose of the present invention is achieved through the following technical solutions: a kind of production method of soybean oligopeptide of acidproof and no bitter taste comprises following several steps:
(1) soybean protein is fully disperseed with deionized water soybean protein is mixed with 5～15% protein solution of (w/w mass percents, down with), and with 1000～5000 rev/mins velocity of shear;
(2) soy bean proteinous soln after will disperseing is heated to 35～55 ℃; Be 5～8mol.L then with concentration
-1Hydrochloric acid join lentamente in the soy bean proteinous soln, reduce to soy protein isoelectric point up to pH, measure the pH value after 15～30 minutes once more, show then that as no change pH regulator finishes, if the pH value raises then needs further to add hydrochloric acid;
(3) soy bean proteinous soln that the pH value is in soy protein isoelectric point is heated to 70～100 ℃, keeps 10～30 minutes, and purpose is to make the abundant thermally denature of soybean protein, and molecular structure launches, so as further enzymolysis, cooling;
(4) get the proteolytic enzyme of soybean protein quality 0.2～2%, add step (3) after fully dissolving with deionized water to through heat treated soy bean proteinous soln, beginning iso-electric point enzymolysis, hydrolysis temperature is the optimum temperuture of proteolytic enzyme; Protein solution pH value descends to some extent in enzymolysis process, and adding basic solution is 4.3～4.7 to keep protein solution pH value, and enzymolysis time is 4～12 hours; Then in 90 ℃～100 ℃, kept 10～20 minutes, purpose is the passivation protease activities;
(5) soy bean proteinous soln after step (4) processing is centrifugal, get supernatant liquor, by the PS membrane ultrafiltration device ultrafiltration of 1000Da, then neutralization, concentrated, dry obtains acidproof and soybean oligopeptide no bitter taste.
Described basic solution be in sodium hydroxide solution or the potassium hydroxide solution one or both;
The concentration of described sodium hydroxide solution is preferably 5～10mol.L
-1
The concentration of described potassium hydroxide solution is preferably 5～10mol.L
-1
Described proteolytic enzyme is preferably the proteolytic enzyme that has inscribe effect and circumscribed effect simultaneously;
Described proteolytic enzyme is Protex 51F proteolytic enzyme (company of outstanding person's energy section) more preferably;
Centrifugal condition described in the step (5) is for being 3500～6500 rev/mins;
A kind of soybean oligopeptide and production method thereof of acidproof and no bitter taste prepare by aforementioned production method.
Described acidproof and soybean oligopeptide no bitter taste is applied in the field of food;
Described acidproof and soybean oligopeptide no bitter taste is particularly suitable for directly adding in the products such as sport-drink, nutritional supplementation type beverage, can not produce to precipitate or wild effect that protein drinkss such as flocculation are prone to.
The present invention has following advantage and effect with respect to prior art:
(1) iso-electric point enzymolysis: the present invention is different with the enzymolysis under neutrality or alkaline condition of prior art, the present invention is the method that adopts at the soy protein isoelectric point enzymolysis, thereby the degree of hydrolysis of soybean protein is up to 42%, obtain soybean oligopeptide more than 85% and contain amino acid below 8, the overwhelming majority is 3～5 amino acid, molecular-weight average is 1, below the 000Da.The soybean oligopeptide of the acidproof and no bitter taste for preparing has very good solubility, even in the iso-electric point of soybean protein, at least 95% oligopeptides is solvable, and stability is fine, has good acid resistance.Therefore be widely applied in the tart food system, as acidic drinks,
(2) the present invention is by circumscribed proteolytic enzyme and endo-protease enzymolysis or have the restriction endonuclease effect simultaneously and the proteolytic enzyme of excision enzyme effect carries out enzymolysis simultaneously, and the soybean oligopeptide that obtains does not have bitter taste.And owing to adopted excision enzyme, so finished product also contains 10～20% total free aminoacids.The soybean oligopeptide of the acidproof and no bitter taste that the present invention obtains can directly add in the products such as sport-drink, nutritional supplementation type beverage, can not produce the wild effect that protein drinkss such as precipitation or flocculation are prone to.
Embodiment
The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited thereto.
Embodiment 1
(1) with deionized water soybean protein isolate (Shandong YuWang Industry Co.,Ltd) is mixed with the protein solution of 10% (w/w mass percent, together following), and adopts 3000 rev/mins of high speed shear, soybean protein is fully disperseed;
(2) soy bean proteinous soln after will disperseing constantly is being heated to 52 ℃ under the stirring condition; Be 6mol.L then with concentration
-1Hydrochloric acid join lentamente in the protein solution, with pH regulator to 4.5, protein aggregation becomes bigger particle, needs this moment constantly to stir, and prevents that albumen precipitation is in the retort bottom.Measure the pH value after 15 minutes once more, the pH value does not change;
(3) then soy bean proteinous soln is heated to 85 ℃, kept 25 minutes, make the abundant sex change of soybean protein, then soy bean proteinous soln is cooled to 52 ℃;
(4) water fully dissolves Protex 51F proteolytic enzyme (company of outstanding person's energy section food grade commercially available protein enzyme in advance, be to originate from the polypeptide excision enzyme of aspergillus oryzae strain and the mixed enzyme of restriction endonuclease) after add again in the soy bean proteinous soln after the denaturing treatment, the proteolytic enzyme quality of using is 0.5% of soybean protein quality; The temperature of retort remains on 52 ℃, and the pH value of protein solution remains on 4.5, enzymolysis 5 hours; Then, kept the passivation protease activities 10 minutes in 90 ℃;
(5) soy bean proteinous soln after step (4) is handled is got supernatant liquor with 4000 rev/mins of centrifugal 3min of speed, by the PS membrane ultrafiltration device ultrafiltration of 1000Da, then concentrates, dry, obtains acidproof and soybean oligopeptide no bitter taste.The molecular-weight average that adopts gel permeation chromatograph to measure target product is 521Da, and 93% peptide molecular weight is less than 1, and 000Da, total free aminoacids are 11%.
Bitterness is represented with the concentration of Quinine Sulphate Di HC solution.Ten sensory evaluation personnel that pass through to train are by tasting the immediate Quinine Sulphate Di HC strength of solution of more unknown sample bitterness intensity, the oligopeptide solution bitter taste of 5% concentration that obtains is suitable with the Quinine Sulphate Di HC solution of 0.0002% concentration, a little more than the threshold value (the Quinine Sulphate Di HC solution of 0.0001% concentration) of human body to the bitter taste impression.
Embodiment 2
Present embodiment is to be raw material with the low temperature soybean meal, at first uses method extracting soybean albumen from the low temperature soybean meal of " alkali extraction and acid precipitation ", and is high-temperature denatured then, adopts the iso-electric point enzymolysis.Concrete enforcement is as follows:
With low temperature soybean meal (Shandong YuWang Industry Co.,Ltd) is raw material, add 15 times of 50～55 ℃ of hot water to the dregs of beans quality, regulate pH value 8.0～8.5, stir extraction 15 minutes, then through 4000 rev/mins of speed centrifugations, obtain soy bean proteinous soln and bean dregs, bean dregs heavily extract once; The soybean solution of extraction has good dispersiveness, and directly water is adjusted to 5% with soy bean proteinous soln concentration; Soy bean proteinous soln hydrochloric acid adjust pH 4.4～4.5, and constantly stir.After regulating pH and finishing, the temperature of protein solution further is heated to 70 ℃, and keeps 30min, make the soybean protein sex change.Then, the albumen temperature with sex change is reduced to 52 ℃.Water fully dissolves Protex51F proteolytic enzyme in advance, adds to then in the protein solution, and quality is 0.2% of a soybean protein.The temperature of retort remains on 52 ℃, and the pH value of protein solution remains on 4.5, enzymolysis 4 hours.Protein solution is heated to 95 ℃ then, the passivation protease activities, enzymolysis finishes.Enzymolysis solution through 4000 rev/mins of speed centrifugations, is discarded sedimentary protein, get supernatant liquor by 1, the PS membrane ultrafiltration device ultrafiltration of 000Da.At last, centrifuged supernatant promptly obtains target product by further concentrated, spraying drying.The molecular-weight average that adopts gel permeation chromatograph to measure target product is 890Da, and 88% peptide molecular weight is less than 1, and 000Da, total free aminoacids are 18%.
Bitterness is represented with the concentration of Quinine Sulphate Di HC solution.Ten sensory evaluation personnel that pass through to train are by tasting the immediate Quinine Sulphate Di HC strength of solution of more unknown sample bitterness intensity, the oligopeptide solution bitter taste of 5% concentration is suitable with the Quinine Sulphate Di HC solution of 0.0002% concentration, a little more than the threshold value (the Quinine Sulphate Di HC solution of 0.0001% concentration) of human body to the bitter taste impression.
Embodiment 3
With raw material, as different from Example 1, adopt different protease hydrolyzeds with soybean protein isolate.With papoid with inscribe effect and the flavor protease (Novozymes Company's food grade commercially available protein enzyme) with circumscribed effect, its quality is respectively 0.5% and 0.2% of soybean protein, and its enzyme digestion reaction temperature is 55 ℃.Other conditions are identical with embodiment 1 with working method.The molecular-weight average that adopts gel permeation chromatograph to measure target product is 960Da, and 86% peptide molecular weight is less than 1, and 000Da, total free aminoacids are 17%.
Bitterness is represented with the concentration of Quinine Sulphate Di HC solution.Ten sensory evaluation personnel that pass through to train are by tasting the immediate Quinine Sulphate Di HC strength of solution of more unknown sample bitterness intensity, the oligopeptide solution bitter taste of 5% concentration is suitable with the Quinine Sulphate Di HC solution of 0.0002% concentration, a little more than the threshold value (the Quinine Sulphate Di HC solution of 0.0001% concentration) of human body to the bitter taste impression.
Embodiment 4
(1) with deionized water soybean protein isolate (Shandong YuWang Industry Co.,Ltd) is mixed with the protein solution of 15% (w/w mass percent, together following), and adopts 5000 rev/mins of high speed shear, soybean protein is fully disperseed;
(2) soy bean proteinous soln after will disperseing constantly is being heated to 52 ℃ under the stirring condition; Be 8mol.L then with concentration
-1Hydrochloric acid join lentamente in the protein solution, with pH regulator to 4.5, protein aggregation becomes bigger particle, needs this moment constantly to stir, and prevents that albumen precipitation is in the retort bottom.Measure the pH value after 30 minutes once more, the pH value does not change;
(3) then soy bean proteinous soln is heated to 100 ℃, kept 10 minutes, make the abundant sex change of soybean protein, then soy bean proteinous soln is cooled to 52 ℃;
(4) water adds after fully dissolving has the papoid of inscribe effect and has the flavor protease (Novozymes Company's food grade commercially available protein enzyme) of circumscribed effect in advance in the soy bean proteinous soln after the denaturing treatment again, and the proteolytic enzyme quality of use is respectively 1% and 1% of soybean protein quality; The temperature of retort remains on 52 ℃, and the pH value of protein solution remains on 4.5, enzymolysis 12 hours; Then, kept the passivation protease activities 10 minutes in 99 ℃;
(5) soy bean proteinous soln after step (4) is handled is got supernatant liquor with 65000 rev/mins of centrifugal 3min of speed, by the PS membrane ultrafiltration device ultrafiltration of 1000Da, then concentrates, dry, obtains acidproof and soybean oligopeptide no bitter taste.The molecular-weight average that adopts gel permeation chromatograph to measure target product is 635Da, and 93% peptide molecular weight is less than 1, and 000Da, total free aminoacids are 18%.
Bitterness is represented with the concentration of Quinine Sulphate Di HC solution.Ten sensory evaluation personnel that pass through to train are by tasting the immediate Quinine Sulphate Di HC strength of solution of more unknown sample bitterness intensity, the oligopeptide solution bitter taste of 5% concentration that obtains is suitable with the Quinine Sulphate Di HC solution of 0.0002% concentration, a little more than the threshold value (the Quinine Sulphate Di HC solution of 0.0001% concentration) of human body to the bitter taste impression.
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (9)
1. the production method of the soybean oligopeptide of an acidproof and no bitter taste is characterized in that: comprise the steps:
(1) with deionized water soybean protein is mixed with the protein solution that concentration is mass percent 5～15%, and soybean protein is fully disperseed with 1000～5000 rev/mins velocity of shear;
(2) soy bean proteinous soln after will disperseing is heated to 35～55 ℃; Be 5～8mol.L then with concentration
-1Hydrochloric acid join lentamente in the soy bean proteinous soln, reduce to soy protein isoelectric point up to pH, measure the pH value after 15～30 minutes once more, show then that as no change pH regulator finishes, if the pH value raises then needs further to add hydrochloric acid;
(3) soy bean proteinous soln that the pH value is in soy protein isoelectric point is heated to 70～100 ℃, keeps 10～30 minutes, afterwards then cooling;
(4) get the proteolytic enzyme of soybean protein quality 0.2～2%, add step (3) after fully dissolving with deionized water to through heat treated soy bean proteinous soln, beginning iso-electric point enzymolysis, hydrolysis temperature is the optimum temperuture of proteolytic enzyme; Protein solution pH value descends to some extent in enzymolysis process, and adding basic solution is 4.3～4.7 to keep protein solution pH value, and enzymolysis time is 4～12 hours; Then, kept the passivation protease activities 10～20 minutes in 90 ℃～100 ℃;
(5) soy bean proteinous soln after step (4) processing is centrifugal, get supernatant liquor, by the PS membrane ultrafiltration device ultrafiltration of 1000Da, then neutralization, concentrated, dry obtains acidproof and soybean oligopeptide no bitter taste.
2. according to the production method of the soybean oligopeptide of the described acidproof and no bitter taste of claim 1, it is characterized in that: described basic solution be in sodium hydroxide solution or the potassium hydroxide solution one or both.
3. according to the production method of the soybean oligopeptide of the described acidproof and no bitter taste of claim 1, it is characterized in that: the concentration of described sodium hydroxide solution is 5～10mol.L
-1The concentration of described potassium hydroxide solution is 5～10mol.L
-1
4. according to the production method of the soybean oligopeptide of described acidproof and the no bitter taste of claim 1, it is characterized in that: described proteolytic enzyme is the proteolytic enzyme that has inscribe effect and circumscribed effect simultaneously.
5. according to the production method of the soybean oligopeptide of the described acidproof and no bitter taste of claim 4, it is characterized in that: described proteolytic enzyme is Protex 51F proteolytic enzyme.
6. according to the production method of the soybean oligopeptide of the described acidproof and no bitter taste of claim 1, it is characterized in that: the centrifugal condition described in the step (5) is 3500～6500 rev/mins.
7. the soybean oligopeptide of an acidproof and no bitter taste prepares by each described production method of claim 1～6.
8. the application of the soybean oligopeptide of described acidproof and the no bitter taste of claim 7 is characterized in that: describedly acidproofly be applied to field of food with soybean oligopeptide no bitter taste.
9. application according to claim 8 is characterized in that: described acidproof directly making an addition to soybean oligopeptide no bitter taste added in sport-drink or the nutritional supplementation type beverage.
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| CN102181409A (en) * | 2011-02-28 | 2011-09-14 | 湖北濠源生物科技有限公司 | Complex enzyme composition and application thereof and method for producing soybean oligopeptides |
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| JP2017528149A (en) * | 2015-04-30 | 2017-09-28 | チャイナ ナショナル リサーチ インスティテュート オブ フード アンド ファーメンテーション インダストリーズ | Hypoallergenic soy oligopeptide with reduced bitterness, preparation method thereof, and use thereof |
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| CN112322682A (en) * | 2020-11-05 | 2021-02-05 | 肽子生物科技(武汉)有限公司 | Method for preparing bitter-free plant oligopeptide |
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| JP2017528149A (en) * | 2015-04-30 | 2017-09-28 | チャイナ ナショナル リサーチ インスティテュート オブ フード アンド ファーメンテーション インダストリーズ | Hypoallergenic soy oligopeptide with reduced bitterness, preparation method thereof, and use thereof |
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| CN106974108A (en) * | 2017-04-21 | 2017-07-25 | 安徽省领航动物保健品有限责任公司 | A kind of defatted soybean meal prepares the optimize technique of small molecular protein peptide pig feed |
| CN110122656A (en) * | 2019-06-06 | 2019-08-16 | 南通光合生物技术有限公司 | Soybean protein enzyme hydrolysis process |
| CN110836936A (en) * | 2019-11-27 | 2020-02-25 | 华熙生物科技股份有限公司 | Method for measuring protein hydrolysis degree |
| CN112126668A (en) * | 2020-09-09 | 2020-12-25 | 宁波希诺亚海洋生物科技有限公司 | Method for preparing bitter-free soybean polypeptide by using specific endo-complex enzyme |
| CN112126668B (en) * | 2020-09-09 | 2024-03-01 | 宁波希诺亚海洋生物科技有限公司 | Method for preparing bitter-free soybean polypeptide by specific endo-complex enzyme |
| CN112322682A (en) * | 2020-11-05 | 2021-02-05 | 肽子生物科技(武汉)有限公司 | Method for preparing bitter-free plant oligopeptide |
| CN115211509A (en) * | 2022-06-14 | 2022-10-21 | 武汉新华扬生物股份有限公司 | Compound enzyme preparation for preparing liquid vegetable protein base material and application thereof |
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| C06 | Publication | ||
| PB01 | Publication | ||
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