CN108004287A - A kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion - Google Patents
A kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion, it is related to field of polypeptide preparation.Weigh albumen (casein, zeins and soybean protein isolate etc.), add a certain amount of distilled water, carry out solubilization, specific protease is added, by factor controlling inside or outside system, makes reaction system close to the optimal catalyst system and catalyzing of protease, and maintain to reaching predetermined enzymolysis degree of hydrolysis, enzyme deactivation, it is dry, it is prepared into finished product.The technology of the present invention has the following advantages：Using solubilized technical finesse albumen so that substrate is not all the time in dissolved state (or forming big particle precipitation) in enzymolysis process, and the protein content not digested is remarkably decreased, and the homogeneity of enzymolysis product is high；In terms of macromolecular functional polypeptide is prepared based on gastro-intestinal digestion, its restriction enzyme site of protease used is complementary with pepsin, therefore its enzymolysis product is by can quickly absorbability, biological activity significantly improve after gastro-intestinal digestion.
Description
Technical field
The present invention relates to field of polypeptide preparation, particularly relates to a kind of preparation side of the macromolecular polypeptides based on gastro-intestinal digestion
Method.
Background technology
China's protein resource enriches, and has 90,000,000 tons of protide processing of farm products accessory substance every year, but it is integrated
Utilization rate only has 40%, less than the half of developed country, causes waste and the economic loss of resource.Main reason is that at present I
The processing method of state's protein is extensive, lacks the correlation technique of high integration.Therefore, the profound comprehensive process production of protein resource is carried out
The key technology research of industry upgrading synergy is very urgent, meets National modern agricultural development requirement.
Largely scientific investigations showed that, by selecting appropriate protease, the protein of hydrolysis can largely be had
The biologically active peptide of various biological functions, these active peptides not only have extremely extensive activity and diversity, but also its source
Enrich, cost is low, security is good.It is easy to operate, easy to industrialized production, therefore become research new hot spot (Pang Guangchang etc.,
The progress theoretical foundation of biologically active peptide and prospect, 2001).Its Related product has been widely used in nutraceutical, spy
Cure the fields such as food, health products, cosmetics.
Enzymatic isolation method prepares active peptide at present, mainly to prepare based on micromolecule polypeptide (molecular weight is less than 500~3000Da),
Because by the speed of intestinal wall than amino acid faster, its preparation process is all based on expectation target small molecule to body transport small peptide
Polypeptide is directly absorbed as the theory of basic norm, i.e. " small molecule prepares small molecule and absorbs " by internal alimentary canal, due to its enzymolysis
Degree is excessive (if to prepare two, tripeptides as ultimate aim, its degree of hydrolysis is up to more than 33.3%), follow-up centrifugation, UF membrane and
The links such as concentration also cause its preparation process complicated, and cost is excessive.In addition micromolecule polypeptide disappears by the protease in intestines and stomach
The problem of changing, also producing excessive degradation (Katherina Fern á ndez, Simulated digestion of
proanthocyanidins in grape skin and seed extracts and the effects of
digestion on the angiotensin I-converting enzyme(ACE)inhibitory activity,
2013).From the angle consider, using enzymatic isolation method prepare based on gastro-intestinal digestion Large molecule active peptide (molecular weight is more than 6000Da,
Degree of hydrolysis is in 2%-20%) tool has great advantage, and the program is the theory based on " macromolecular prepare small molecule absorb ", by albumen
Macromolecular polypeptides are prepared by low degree enzymolysis in vitro, are taken orally after entering stomach, using the proteolysis ability of internal stomach and intestine,
The small active peptides for forming effect at least up to similar with tradition " small molecule prepares small molecule and absorbs " are absorbed by organisms, at the same time
The effect of similar sustained release can also be played.
The raw material of polypeptide prepared by enzymatic isolation method mainly has milk protein, casein, zein, rice protein, paddy at present
Protein powder etc., wherein most albumen is water-soluble undesirable under normal circumstances, if directly carrying out low degree enzyme in short time
Solution, insoluble albumen can cause proteolysis degree inside and outside particle to differ, i.e. extra-granular albumen quilt since granularity is larger
Excessively enzymolysis, internal albumen are not digested, cause the uniformity of enzymolysis product low.The hydrolysis degree of usual albumen is to its product
Bioactivity has significant impact.Therefore, the homogeneity for improving enzymolysis product improves macromolecular polypeptides product quality with important
Meaning.
The method for improving protein solubility has very much, such as：(1) temperature is changed, as soybean protein is dissolved in higher temperature
In water (pH 8.0), not only solubility greatly improves, but also protein solution colloidization degree and viscosity can be remarkably decreased；(2)
Change pH value, be dissolved in as casein is extremely difficult in neutral or acid water, when pH is more than 8, dissolving is preferable；(3) non-pole is changed
Property solution proportion, as zeins is insoluble in water, but is soluble in 60% ethanol solution.Although these methods can be effective
The problems of dissolution of albumen is solved, but the corresponding system of these methods not necessarily commonly uses the optimal catalyst system and catalyzing of protease,
For example in certain temperature range, soybean protein solubility with temperature is raised and raised, but temperature is most preferably urged more than protease
When changing temperature, digest for a long time, proteinase activity can be caused to decline to a great extent or even inactivation can be caused.Consider substrate protein
The catalytic activity of dissolubility and enzyme, the research for preparing the Large molecule active polypeptide based on gastro-intestinal digestion of high homogeneity yet there are no report
Road.
The content of the invention
The purpose of the present invention in order to overcome the shortcomings of conventional art, there is provided a kind of high homogeneity based on gastro-intestinal digestion
Macromolecular Large molecule active polypeptide preparation method.
In order to realize foregoing invention purpose, its specific technical solution is as follows：
Albumen is weighed, carries out solubilized pretreatment, adds specific protease, by factor control inside or outside system
System, makes reaction system be carried out with enzyme digestion reaction close to the optimal catalyst system and catalyzing of (or reaching) protease, and maintains to reaching pre-
Fixed enzymolysis degree of hydrolysis, enzyme deactivation is dry after cooling, is prepared into finished product.
Wherein above-mentioned albumen, can be the albumen for being not readily dissolved in water, such as casein, zeins, and be dissolved in water
Degree of gelation and the larger albumen of viscosity afterwards, such as soybean protein isolate.
Wherein above-mentioned solubilized preprocessing means include and are not limited to adjust nonpolar solvent ratio, temperature or pH.
If the soluble preferably gelation of wherein above-mentioned albumen is not serious, such as lactalbumin, oralbumin can be with
Solubilized pretreatment is not used.
Wherein above-mentioned protease, be preferably its proteolysis sites and pepsin hydrolytic sites (selective hydrolysis by
Two hydrophobic amino acid residues form peptide bond, such as Phe-Phe) there is relatively strong complementary protease, such as neutral proteinase, pancreas
Protease, pancreatin (predominantly trypsase), papain, bromelain, ficin etc..
Wherein casein macromolecular polypeptides preparation method, carries out according to above-mentioned steps：Take a certain amount of casein, 50 DEG C of water
Bath preheating 10min, adjusts pH to 8.0-8.5, is allowed to be completely dissolved, and regulatory protein concentration is 50g/L, in being added by 5% (E/S)
Property protease, when pH drops to 6.5-7.5, constantly be added dropwise NaOH maintain pH it is constant, question response to certain degree of hydrolysis (10%-
20%), enzymolysis terminates, enzyme deactivation, dry after cooling, is prepared into finished product.
Wherein zein macromolecules polypeptide production methods, carry out according to above-mentioned steps：With the ethanol of 60%-70%
Zeins is dissolved, 50 DEG C of water-baths preheat 10min, are slowly added to wait warm water, the concentration of alcohol of solution is diluted to 15%-
25%, protein concentration is adjusted to 15g/L, and it is 8.0 to adjust pH immediately, and adding pancreatin by 5% (E/S) is digested, and is being digested
Constantly dropwise addition NaOH maintenances pH is constant in journey, and bath temperature is constant.Certain degree of hydrolysis (5%-15%) is reacted to, enzymolysis terminates,
Tune pH is 4-5, and after rotary evaporation falls ethanol, moisturizing enzyme deactivation is dry, is prepared into finished product.
Wherein soybean protein macromolecular polypeptides preparation method, carries out according to subordinate's step：Take a certain amount of soybean separation protein
In vain, it is 50g/L to add water to be made into protein concentration, and 63-68 DEG C of water-bath preheats 10min, adjust pH to 8.0-8.5, adds 5% (E/S)
Neutral proteinase is digested, and when pH declines 6.5-7.5, NaOH is constantly added dropwise and maintains pH constant, then reduces temperature to 50-55
℃.Certain degree of hydrolysis (10%-20%) is reacted to, enzymolysis terminates, enzyme deactivation, dry after cooling, is prepared into finished product.
Advantages of the present invention：
Compared with prior art, the present invention on the one hand so that the dissolubility of protein substrate and the catalytic activity of enzyme obtain
It is unified so that substrate in enzymolysis process all the time in dissolved state (or not forming big particle precipitation), product it is homogeneous
Property be improved, the protein content that is not digested in enzymatic hydrolysis system declines 9.34%-54.39% and (is prepared with direct enzymolysis protein
Micromolecule polypeptide is compared)；On the other hand, the digestive functions of human body are made full use of, make macromolecular polypeptides further in vivo
Degraded, compared with direct enzymolysis protein prepares micromolecule polypeptide, polypeptide (molecular weight 200-500Da) content that can directly absorb carries
High 4.66%-17.67%, hypotensive activity improves 19.74%-81.68% (using ACE inhibiting rates as index), and is adopted
Neutral proteinase (its restriction enzyme site is complementary with pepsin), its enzymolysis product pass through the biology work after gastro-intestinal digestion
Property is better than alkali protease.
Below by embodiment, technical scheme is described in further detail.
Embodiment
Used term in the present invention, it is unless otherwise specified, generally usual with those of ordinary skill in the art
The implication of understanding.The present invention is described in further detail with reference to specific embodiment, and with reference to data.It is to be understood that these
Embodiment is that the present invention is further described, it is impossible to is interpreted as limiting the scope of the present invention, the skill in the field
Art engineer can make the present invention some nonessential modifications and adaptations according to the content of foregoing invention.
Below in an example, the various processes and method not being described in detail are conventional methods as known in the art.
The source of agents useful for same, trade name and it is necessary to list its constituent person, indicates on the first appearance, thereafter phase used
With reagent unless otherwise specified, it is identical with the content indicated first.
The protease and casein of the present embodiment and reference examples are purchased from Sigma companies.
The present embodiment and the degree of hydrolysis of reference examples add neutral proteinase start enzymolysis up to pH reach setting pH value it
Before, the degree of hydrolysis of casein is measured using formol titration, is measured after setting pH is reached using pH-stat methods, total hydrolysis
Spend for both sums.
The present embodiment and the in-vitro simulated pipe intestinal digesting of reference examples are with the following method：Two parts of enzymolysis product is taken, is pressed
Pepsin is added according to 2% certain (E/S) enzyme concentration, when enzymolysis 2 is small, after enzymolysis, portion is used for enzyme deactivation, centrifuges, and surveys
Determine the hypotensive activity of peptic digest product.Another adds pancreatin according to 4% (E/S) enzyme concentration, and when enzymolysis 4 is small, enzymolysis terminates
Afterwards, enzyme deactivation, centrifugation, the hypotensive activity of measure gastro-intestinal digestion product.
The present embodiment and the hypotensive activity of reference examples measure are with angiotensin converting enzyme (angiotensin
Converting enzyme, ACE) inhibiting rate is index, with reference to Dingqing sesame, (prepared by impulse ultrasound assistance enzymolysis method for concrete operations
The research of maize yellow-powder ACEI active peptides, 2008) method.
The content of peptides that can directly absorb of the present embodiment and reference examples, is simulated by measuring protein zymolyte by stomach and intestine
The molecular weight distribution in 200-500Da of digestion product.Experiment uses efficient liquid phase size exclusion chromatography, chromatographic column：TSK
Gel G2000 (300mm × 7.8mm), the composition of mobile phase is acetonitrile：Water：Trifluoroacetic acid=45:55:0.1, elution flow rate：
0.5mL/min, Detection wavelength：220nm, 30 DEG C of column temperature, 10 μ L of sample size, the control and data processing of liquid phase systems use software
Breeze (Waters, MA, USA).With retention time (t) for abscissa, molecular weight is ordinate to numerical value (lg Mr), and
Standard items using following known molecular amount make standard curve：Bacillus enzyme (1450Da), aminoacetic acid-aminoacetic acid-tyrosine-essence ammonia
Sour (451Da) and aminoacetic acid-aminoacetic acid-aminoacetic acid (189Da).The chromatographic peak of each sample is integrated, obtains 200-500Da
The relative amount of polypeptide.
The protein assay method not digested in the present embodiment and reference examples is：Using trichloroacetic acid precipitation (three chloroethenes
Acid it is final concentration of 10%).
Reference examples 1
20g caseins are weighed, 400mL distilled water is added, adjusts the temperature to 50 DEG C, it is 6.5 to adjust pH with 3M NaOH, is put down
Weigh after 15min, add 1mL neutral proteinases, it is 6.5 that 1M NaOH, which are constantly added dropwise, and maintain pH, when degree of hydrolysis is 26%, enzymolysis knot
Beam, the enzyme deactivation 15min under the conditions of 100 DEG C, centrifuging and taking supernatant, and 3000Da ultrafiltration membranes are crossed, filtered solution is spray-dried, is prepared into
Finished product.
Non- enzymolysis protein is 9.64g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 31.23%, can directly be absorbed
Content of peptides be 43.17%.
Reference examples 2
20g caseins are weighed, 400mL distilled water is added, adjusts the temperature to 50 DEG C, it is 6.5 to adjust pH with 3M NaOH, is put down
Weigh after 15min, add 1mL neutral proteinases, it is 6.5 that 1M NaOH, which are constantly added dropwise, and maintain pH, when degree of hydrolysis is 20%, enzymolysis knot
Beam, the enzyme deactivation 15min under the conditions of 100 DEG C is dry after cooling, is prepared into finished product.
Non- enzymolysis protein is 11.30g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 46.23%, can directly be absorbed
Content of peptides be 44.82%.
Reference examples 3
20g caseins are weighed, 400mL distilled water is added, adjusts the temperature to 50 DEG C, it is 6.5 to adjust pH with 3M NaOH, is put down
Weigh after 15min, add 960mL alkali proteases (identical with above-mentioned added neutral proteinase enzyme activity), 1M is constantly added dropwise
It is 6.5 that NaOH, which maintains pH, and when degree of hydrolysis is 20%, enzymolysis terminates, the enzyme deactivation 15min under the conditions of 100 DEG C, dry after cooling, system
It is standby into finished product.
Non- enzymolysis protein is 10.99g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 32.26%, can directly be absorbed
Content of peptides be 35.68%.
Embodiment 1
20g caseins are weighed, add 400mL distilled water, it is 8.5 that 3M NaOH, which are constantly added dropwise, and maintain pH, in the process of dropwise addition
In be stirred continuously, treat its all dissolving after, by temperature adjustment to 50 DEG C, add 1mL neutral proteinases, after pH is down to 7.5,
It is 7.5 that 1M NaOH, which are constantly added dropwise, and maintain pH, and when degree of hydrolysis is 10%, enzymolysis terminates, and enzyme deactivation 15min, cold under the conditions of 100 DEG C
But dry afterwards, be prepared into finished product.
Non- enzymolysis protein is 8.74g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 51.37%, can directly be absorbed
Content of peptides be 45.18%.
Embodiment 2
20g caseins are weighed, add 400mL distilled water, it is 8.0 that 3M NaOH, which are constantly added dropwise, and maintain pH, in the process of dropwise addition
In be stirred continuously, treat its all dissolving after, by temperature adjustment to 50 DEG C, add 1mL neutral proteinases, after pH is down to 6.5,
It is 6.5 that 1M NaOH, which are constantly added dropwise, and maintain pH, and when degree of hydrolysis is 20%, enzymolysis terminates, and enzyme deactivation 15min, cold under the conditions of 100 DEG C
But dry afterwards, be prepared into finished product.
Non- enzymolysis protein is 4.53g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 56.74%, can directly be absorbed
Content of peptides be 47.36%.
Reference examples 4
Zeins 6.75g is weighed, adds 450mL water, 37 DEG C of water-baths preheat 10min, and it is 8.0 to adjust pH, is added
The pancreatin for entering 337.5mg is digested, and 1mol/L NaOH are constantly added dropwise in enzymolysis process and maintain pH constant, bath temperature is permanent
It is fixed.It is 28% to be reacted to degree of hydrolysis, and enzymolysis terminates, and it is 4 to adjust pH, after rotary evaporation falls ethanol, moisturizing enzyme deactivation under the conditions of 100 DEG C
15min, centrifuging and taking supernatant, and 3000Da ultrafiltration membranes are crossed, filtered solution is spray-dried, is prepared into finished product.
Non- enzymolysis protein is 4.21g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 39.62%, can directly be absorbed
Content of peptides be 37.42%.
Reference examples 5
Zeins 6.75g is weighed, adds 450mL water, 37 DEG C of water-baths preheat 10min, and it is 8.0 to adjust pH, is added
The pancreatin for entering 337.5mg is digested, and 1mol/L NaOH are constantly added dropwise in enzymolysis process and maintain pH constant, bath temperature is permanent
It is fixed.It is 15% to be reacted to degree of hydrolysis, and enzymolysis terminates, and it is 4 to adjust pH, after rotary evaporation falls ethanol, moisturizing enzyme deactivation under the conditions of 100 DEG C
15min, it is dry after cooling, it is prepared into finished product.
Non- enzymolysis protein is 4.76g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 44.35%, can directly be absorbed
Content of peptides be 39.61%.
Embodiment 3
Zeins 6.75g is weighed, 60% ethanol of 150mL is added and is completely dissolved, 37 DEG C of water-baths preheat 10min, adjust
It is 15% to save the concentration of alcohol in solution, system 450mL.It is 8.0 to adjust pH immediately, and the pancreatin for adding 337.5mg carries out enzyme
Solution, constantly dropwise addition 1mol/L NaOH maintenances pH is constant in enzymolysis process, and bath temperature is constant.It is 15% to be reacted to degree of hydrolysis,
Enzymolysis terminates, and it is 4 to adjust pH, and after rotary evaporation falls ethanol, moisturizing enzyme deactivation 15min under the conditions of 100 DEG C is dry after cooling, system
It is standby into finished product.
Non- enzymolysis protein is 1.92g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 65.17%, can directly be absorbed
Content of peptides be 43.19%.
Embodiment 4
Zeins 6.75g is weighed, 70% ethanol of 150mL is added and is completely dissolved, 37 DEG C of water-baths preheat 10min, adjust
It is 25% to save the concentration of alcohol in solution, system 450mL.It is 8.0 to adjust pH immediately, and the pancreatin for adding 337.5mg carries out enzyme
Solution, constantly dropwise addition 1mol/L NaOH maintenances pH is constant in enzymolysis process, and bath temperature is constant.It is 5% to be reacted to degree of hydrolysis,
Enzymolysis terminates, and it is 5 to adjust pH, and after rotary evaporation falls ethanol, moisturizing enzyme deactivation 15min under the conditions of 100 DEG C is dry after cooling, system
It is standby into finished product.
Non- enzymolysis protein is 3.47g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 54.68%, can directly be absorbed
Content of peptides be 41.72%.
Reference examples 6
20g soybean protein isolates are taken, add 400mL water, 50 DEG C of water-baths preheat 10min, and adjust pH to 7.5, add 1mL
Neutral proteinase is digested, and 1mol/L NaOH are constantly added dropwise and maintain pH constant, and it is 25% to be reacted to degree of hydrolysis, and enzymolysis terminates,
The enzyme deactivation 15min under the conditions of 100 DEG C, centrifuging and taking supernatant, and 3000Da ultrafiltration membranes are crossed, filtered solution is spray-dried, is prepared into
Product.
Non- enzymolysis protein is 12.63g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 29.38%, can directly be absorbed
Content of peptides be 36.27%.
Reference examples 7
20g soybean protein isolates are taken, add 400mL water, 50 DEG C of water-baths preheat 10min, and adjust pH to 7.5, add 1mL
Neutral proteinase is digested, and 1mol/L NaOH are constantly added dropwise and maintain pH constant, and it is 20% to be reacted to degree of hydrolysis, and enzymolysis terminates,
The enzyme deactivation 15min under the conditions of 100 DEG C, it is dry after cooling, it is prepared into finished product.
Non- enzymolysis protein is 14.21g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 31.47%, can directly be absorbed
Content of peptides be 37.30%.
Embodiment 5
20g soybean protein isolates are taken, add 400mL water, 63 DEG C of water-baths preheat 10min, and adjust pH to 8.5, add 1mL
Neutral proteinase is digested, and when pH declines near 7.5,1mol/L NaOH is constantly added dropwise and maintain pH constant, then reduce temperature
To 50 DEG C (cooling rate is 5 DEG C/min), it is 20% to be reacted to degree of hydrolysis, and enzymolysis terminates, the enzyme deactivation 15min under the conditions of 100 DEG C,
It is dry after cooling, it is prepared into finished product.
Non- enzymolysis protein is 6.41g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 36.97%, can directly be absorbed
Content of peptides be 42.68%.
Embodiment 6
20g soybean protein isolates are taken, add 400mL water, 68 DEG C of water-baths preheat 10min, and adjust pH to 8.0, add 1mL
Neutral proteinase is digested, and when pH declines near 6.5,1mol/L NaOH is constantly added dropwise and maintain pH constant, then reduce temperature
To 55 DEG C (cooling rate is 5 DEG C/min), it is 10% to be reacted to degree of hydrolysis, and enzymolysis terminates, the enzyme deactivation 15min under the conditions of 100 DEG C,
It is dry after cooling, it is prepared into finished product.
Non- enzymolysis protein is 7.28g, and the hypotensive activity for measuring its gastro-intestinal digestion product is 35.18%, can directly be absorbed
Content of peptides be 40.33%.
Claims (7)
- A kind of 1. preparation method of the macromolecular polypeptides based on gastro-intestinal digestion, it is characterised in that：Albumen is subjected to solubilized pre- place Reason, adds specific protease, by factor controlling inside or outside system, makes reaction system during the reaction close to albumen The optimal catalyst system and catalyzing of enzyme, and maintain to predetermined enzymolysis degree of hydrolysis is reached, enzyme deactivation is dry after cooling, is prepared into finished product.
- 2. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, including and The protein that is water-soluble poor or easily forming gelling system in water in neutral either acid water is not limited to, such as junket egg In vain, zeins and soybean protein isolate etc..
- 3. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, including and It is not limited to adjust solvent polarity, temperature or pH.
- 4. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, is preferably The hydrolytic sites of its proteolysis sites and pepsin have relatively strong complementary protease, such as neutral proteinase, tryptose Enzyme, pancreatin (predominantly trypsase), papain, bromelain, ficin etc..
- 5. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, take certain The casein of amount, 50 DEG C of water-baths preheat 10min, adjust pH and are allowed to be completely dissolved to 8.0-8.5, and neutral proteinase, treats that pH declines When near 6.5-7.5, constantly dropwise addition NaOH maintenances pH is constant, and to certain degree of hydrolysis (10-20%), enzymolysis terminates question response, goes out Enzyme, it is dry after cooling, it is prepared into finished product.
- 6. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, use The ethanol dissolving zeins of 60%-70%, 50 DEG C of water-baths preheat 10min, are slowly added to wait warm water, by the ethanol of solution Concentration dilution is 15%-25%, and it is 8.0 to adjust pH immediately, adds pancreatin to be digested, and NaOH dimensions are constantly added dropwise in enzymolysis process It is constant to hold pH, bath temperature is constant；Certain degree of hydrolysis (5-15%) is reacted to, enzymolysis terminates, and adjusts pH to fall for acidity, rotary evaporation After ethanol, moisturizing enzyme deactivation is dry, is prepared into finished product.
- 7. a kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion according to claim 1, is characterized in that, take certain The soybean protein isolate of amount, 63-68 DEG C of water-bath preheat 10min, adjust pH to 8.0-8.5, add neutral proteinase and are digested, When pH declines near 6.5-7.5, NaOH is constantly added dropwise and maintains pH constant, then reduces temperature to 50-55 DEG C；It is reacted to certain water Xie Du (10%-20%), enzymolysis terminates, enzyme deactivation, dry after cooling, is prepared into finished product.
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| CN103352064A (en) * | 2013-06-25 | 2013-10-16 | 天津大学 | Method for preparing corn protein active peptide by using composite carrier immobilized double enzymes |
| CN103719534A (en) * | 2013-12-23 | 2014-04-16 | 江南大学 | Method for carrying out enzymatic hydrolysis on isolated soybean protein |
| CN103923964A (en) * | 2014-04-17 | 2014-07-16 | 江南大学 | Method for preparing casein hydrolysate with stable functionality by using different hydrolysis conditions |
| CN104938765A (en) * | 2015-07-17 | 2015-09-30 | 东北农业大学 | Preparation meted for high-stability soybean protein emulsion |
| CN107163133A (en) * | 2017-06-26 | 2017-09-15 | 大连工业大学 | A kind of biologically active peptide and preparation method thereof |
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| WO2019091471A1 (en) * | 2017-11-13 | 2019-05-16 | 江苏大学 | Preparation of macromolecular polypeptide based on gastrointestinal digestion and method for in situ real-time monitoring process of same |
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