CN106148462A - Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof - Google Patents
Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof, particularly relate to Semen sojae atricolor glycopeptide with antioxidant activity and bacteriostatic activity and preparation method thereof, first, preparation glycosylation soybean protein, utilizes glycosylation soybean protein to be respectively prepared the Semen sojae atricolor glycopeptide with antioxidant activity and the Semen sojae atricolor glycopeptide with bacteriostatic activity；Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, add Alcalase alkaline protease, after stirring, vibrate 2 hours；After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Having the glycopeptide of bacteriostatic activity: be dispersed in water by glycosylation soybean protein precipitate, obtain protein dispersion, add pepsin, after stirring, hydrolysate of soybean protein reaction adjusts the pH to 7.0 of solution after terminating.Concentrate: above-mentioned filtrate is concentrated into solid content 25 55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, and pan feeding flow velocity is 15%, and rate of air sucked in required is 80%.
Description
Technical field
The present invention relates to a kind of Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof, particularly relate to have anti-
Semen sojae atricolor glycopeptide of oxidation activity and bacteriostatic activity and preparation method thereof.
Background technology
Soybean protein is the protein resource of a kind of high-quality, and wide material sources are cheap, is the traditional food in Asia, especially
It is ensureing that China's dietary protein supply respect plays an important role.The glycosylation modified of protein is by hydrophilic
Glucide is connected among protein molecule in the way of covalent bond connection so that it is both had the macromole characteristic of protein,
There is again the water-wet behavior of glucide.The glycosylation of Maillard reaction approach, can improve the functional character of protein, but deposit
Reaction time length, product brown stain, the defect such as nutritive loss and potential safety hazard.
Use the T-5398 can not only the-NH of lysine residue of catalytic proteins molecule2Residual with glutamine
Crosslink reaction between base, prepare cross-linked proteins, moreover it is possible to-the NH of catalysis primary amine material2With protein molecule glutamine
Reaction is crosslinked between residue.(ultimate unit is D-glucosamine, primary amine by oligochitosan to utilize T-5398
Matter) it is connected with soybean protein molecule, it is achieved that soybean protein glycosylation modified, i.e. can prepare soy glycoproteins.Compare it
Under, it is strong, without side reaction, advantage that foodsafety is high that this reaction has specificity.
At present, the research utilizing this enzyme to prepare glycoprotein is limited only to several seminar in research.Most for improving albumen
The functional character of matter, improves its feasibility as functional ingredient, and the seminar and professor Betti that lead such as professor Zhao carry
The seminar of neck utilizes the catalytic action of this enzyme, and glucosamine imports to prepare in multiple proteins glycoprotein, if any dynamic
Thing albumen (casein, actomyosin), vegetable protein (soybean protein isolate)；Some are also intended to strengthen the biological of protein and live
Property, the seminar that professor Betti leads is respectively by isinglass protolysate and wheat gluten protein hydrolysate and aminoglucose
Sugar crosslinks, and strengthens gelatin peptide and the antioxidation of wheat peptide and bacteriostatic activity.It can be seen that except food-borne glycopeptide, utilize
Enzyme process beam system, for glycopeptide, can be the diet structure improving the mankind, adjusts the rationale foundation providing new, having by them
The Health and Living happiness of the mankind is safeguarded in benefit effect.
T-5398 is a kind of microbial protease, by molecule insertion, cross-linking reaction, deamination, makes albumen
Matter molecular structure changes.Alcalase alkaline protease is to be produced by the Bacillus licheniformis selected and obtain, and is a kind of
Efficient bacterialprotease, belongs to endo protease, mainly acts on the peptide bond containing hydrophobicity carboxyl.Protamex compound protein
Enzyme is subtilisin, belongs to endo protease.These three enzyme industrialized production the most, low price, it is suitable for soy sugars
The requirement of the industrialized production of peptide.
Summary of the invention
It is an object of the invention to provide a kind of Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof, the method profit
The Semen sojae atricolor glycopeptide with antioxidant activity and the Semen sojae atricolor glycopeptide with bacteriostatic activity is made with glycosylation soybean protein.
The technical solution adopted in the present invention is:
First, preparation glycosylation soybean protein:
The first step: take soybean protein isolate powder 1kg, is that 0.4 ~ 0.6:10 is dispersed in water by solid-to-liquid ratio, obtains protein
Dispersion liquid；In described solid-to-liquid ratio, solid consumption is kilogram, and volume unit is for rising；
Second step: gained protein dispersion, 90 DEG C of heating in water bath 10 minutes, is cooled to 40 DEG C afterwards；Big to heat treated
Soybean protein dispersion liquid adds isopyknic pH 7.5 oligochitosan solution, and wherein oligochitosan and soybean protein are with mass ratio 1:0.50；
3rd step: after stirring, adds T-5398, and per kilogram protein adds 10g T-5398, enzyme activity
For 1000U/g, after stirring, after 37 DEG C of vibrations 3 hours, 85 DEG C of heat treated 5 minutes, cooling.
4th step: regulation pH value of solution is to 4.5, and 4000 r/min are centrifuged 10 min, collects precipitation, washes twice.
Utilize glycosylation soybean protein to be respectively prepared there is the Semen sojae atricolor glycopeptide of antioxidant activity and there is the big of bacteriostatic activity
Bean glycopeptide:
Preparation has a glycopeptide of antioxidant activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 8.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, add Alcalase alkaline protease, add 1g Alcalase alkaline protease by per kilogram protein, stir
After, vibrate 2 hours under the conditions of 60 DEG C；After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust solution
PH to 7.0.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 7.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add
Protamex compound protease, per kilogram protein adds 10g Protamex compound protease, after stirring, 55 DEG C of vibrations
2 hours.After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
Through the glycopeptide of Alcalase hydrolysis by novo gained, degree of hydrolysis is 4%, the antioxidant activity of 2mg/mL: DPPH
Free radical scavenging activity is 59.5%；Hydroxyl radical free radical clearance rate is 38.4%；
Antioxidant activity through the glycopeptide of Protamax composite protease hydrolysis gained: DPPH free radical scavenging activity is more than 50%；Hydroxyl
Base free radical scavenging activity is more than 35%.
Preparation has a glycopeptide of bacteriostatic activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 2.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, adding pepsin, per kilogram protein adds 2g pepsin, and after stirring, 37 DEG C vibrate 2 hours；Semen sojae atricolor egg
After white enzyme digestion reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 8.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add pancreas egg
White enzyme, per kilogram protein adds 10g trypsin, and after stirring, 37 DEG C vibrate 2 hours.Hydrolysate of soybean protein reaction knot
Shu Hou, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
The invention has the beneficial effects as follows: use a species specificity strong, without the high enzyme (transglutamin-ase 9 of side reaction, safety
Enzyme) hydrophilic oligochitosan molecule covalent is linked in soybean protein molecule by the method that is catalyzed, prepares glycoprotein.And by food
Grade protease (basic protein protease enzyme/compound protease).These reaction be all conducive to provide more electron donor and
Hydrogen donor, and strengthen the oxidizing property of soybean protein.After pepsin/trypsin degradation glycosylation soybean protein, gained sugar
Peptide can play bacteriostasis, especially more efficient to gram positive bacteria, concentration be 2 mg/mL(degree of hydrolysis be 4%) time, press down
Make colibacillary ability and reach more than 90%.This, by the source of abundant food-borne glycopeptide, has promotion to exploitation food-borne glycopeptide
Effect.
Detailed description of the invention
It is an object of the invention to provide a kind of Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof, this has life
Semen sojae atricolor glycopeptide of thing active function and preparation method thereof utilizes glycosylation soybean protein to make the soy sugars with antioxidant activity
Peptide and the Semen sojae atricolor glycopeptide with bacteriostatic activity.
The technical solution adopted in the present invention is:
First, preparation glycosylation soybean protein:
The first step: take soybean protein isolate powder 1kg, is that 0.6:10 is dispersed in water by solid-to-liquid ratio, obtains protein dispersibility
Liquid；In described solid-to-liquid ratio, solid consumption is kilogram, and volume unit is for rising；
Second step: gained protein dispersion, 90 DEG C of heating in water bath 10 minutes, is cooled to 40 DEG C afterwards；Big to heat treated
Soybean protein dispersion liquid adds isopyknic pH 7.5 oligochitosan solution, and wherein oligochitosan and soybean protein are with mass ratio 1:0.50；
3rd step: after stirring, adds T-5398, and per kilogram protein adds 10g T-5398, enzyme activity
For 1000U/g, after stirring, after 37 DEG C of vibrations 3 hours, 85 DEG C of heat treated 5 minutes, cooling.
4th step: regulation pH value of solution is to 4.5, and 4000 r/min are centrifuged 10 min, collects precipitation, washes twice.
Utilize glycosylation soybean protein to be respectively prepared there is the Semen sojae atricolor glycopeptide of antioxidant activity and there is the big of bacteriostatic activity
Bean glycopeptide:
Embodiment 1
Preparation has a glycopeptide of antioxidant activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 8.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, add Alcalase alkaline protease, add 1g Alcalase alkaline protease by per kilogram protein, stir
After, vibrate 2 hours under the conditions of 60 DEG C；After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust solution
PH to 7.0.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 7.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add
Protamex compound protease, per kilogram protein adds 10g Protamex compound protease, after stirring, 55 DEG C of vibrations
2 hours.After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
Through the glycopeptide of Alcalase hydrolysis by novo gained, degree of hydrolysis is 4%, the antioxidant activity of 2mg/mL: DPPH
Free radical scavenging activity is 59.5%；Hydroxyl radical free radical clearance rate is 38.4%；
Antioxidant activity through the glycopeptide of Protamax composite protease hydrolysis gained: DPPH free radical scavenging activity is 50.4%；Hydroxyl
Base free radical scavenging activity is 37.7%.
Embodiment 2
Preparation has a glycopeptide of bacteriostatic activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 2.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, adding pepsin, per kilogram protein adds 2g pepsin, and after stirring, 37 DEG C vibrate 2 hours；Semen sojae atricolor egg
After white enzyme digestion reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 8.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add pancreas egg
White enzyme, per kilogram protein adds 10g trypsin, and after stirring, 37 DEG C vibrate 2 hours.Hydrolysate of soybean protein reaction knot
Shu Hou, 85 DEG C of enzyme denaturing 5 min, cooling.Adjust the pH to 7.0 of solution.
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer.It is spray-dried: inlet temperature is 110 DEG C, enters
Stream speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
Claims (3)
1. Semen sojae atricolor glycopeptide with bioactive functions and preparation method thereof, by soybean protein glycosylation, utilizes glycosylation
Soybean protein is respectively prepared the Semen sojae atricolor glycopeptide with antioxidant activity and the Semen sojae atricolor glycopeptide with bacteriostatic activity,
It is characterized in that:
First, preparation glycosylation soybean protein:
The first step: take soybean protein isolate powder 1kg, is that 0.4 ~ 0.6:10 is dispersed in water by solid-to-liquid ratio, obtains protein
Dispersion liquid；In described solid-to-liquid ratio, solid consumption is kilogram, and volume unit is for rising；
Second step: gained protein dispersion, 90 DEG C of heating in water bath 10 minutes, is cooled to 40 DEG C afterwards；Big to heat treated
Soybean protein dispersion liquid adds isopyknic pH 7.5 oligochitosan solution, and wherein oligochitosan and soybean protein are with mass ratio 1:0.50；
3rd step: after stirring, adds T-5398, and per kilogram protein adds 10g T-5398, enzyme activity
For 1000U/g, after stirring, after 37 DEG C of vibrations 3 hours, 85 DEG C of heat treated 5 minutes, cooling；
4th step: regulation pH value of solution is to 4.5, and 4000 r/min are centrifuged 10 min, collects precipitation, washes twice.
A kind of Semen sojae atricolor glycopeptide with bioactive functions the most according to claim 1 and preparation method thereof, is characterized in that:
Preparation has a glycopeptide of antioxidant activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 8.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, add Alcalase alkaline protease, add 1g Alcalase alkaline protease by per kilogram protein, stir
After, vibrate 2 hours under the conditions of 60 DEG C；After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling；Adjust solution
PH to 7.0；
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer；It is spray-dried: inlet temperature is 110 DEG C, enters stream
Speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide；
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 7.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add
Protamex compound protease, per kilogram protein adds 10g Protamex compound protease, after stirring, 55 DEG C of vibrations
2 hours；After hydrolysate of soybean protein reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling；Adjust the pH to 7.0 of solution；
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer；It is spray-dried: inlet temperature is 110 DEG C, enters stream
Speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
A kind of Semen sojae atricolor glycopeptide with bioactive functions the most according to claim 1 and preparation method thereof, is characterized in that:
Preparation has a glycopeptide of bacteriostatic activity:
The first:
Glycosylation soybean protein precipitate is dispersed in water, obtains protein dispersion, regulate dispersion liquid pH to 2.0, solid-liquid
Ratio is 0.4:10；In described solid-to-liquid ratio, solid consumption is soybean protein weight, kilogram, volume unit is for rising；Stirring is all
After even, adding pepsin, per kilogram protein adds 2g pepsin, and after stirring, 37 DEG C vibrate 2 hours；Semen sojae atricolor egg
After white enzyme digestion reaction terminates, 85 DEG C of enzyme denaturing 5 min, cooling；Adjust the pH to 7.0 of solution；
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer；It is spray-dried: inlet temperature is 110 DEG C, enters stream
Speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide；
The second:
Being dispersed in water by precipitate, obtain protein dispersion, regulate dispersion liquid pH to 8.0, solid-to-liquid ratio is 0.4:10；Institute
In the solid-to-liquid ratio stated, solid consumption is soybean protein weight, kilogram, volume unit is for rising；After stirring, add pancreas egg
White enzyme, per kilogram protein adds 10g trypsin, and after stirring, 37 DEG C vibrate 2 hours；Hydrolysate of soybean protein reaction knot
Shu Hou, 85 DEG C of enzyme denaturing 5 min, cooling；Adjust the pH to 7.0 of solution；
Concentrate: above-mentioned filtrate is concentrated into solid content 25-55% with vaporizer；It is spray-dried: inlet temperature is 110 DEG C, enters stream
Speed is 15%, and rate of air sucked in required is 80%, with this understanding, and the response rate 50% of soybean peptide.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106929557A (en) * | 2017-04-28 | 2017-07-07 | 安徽生物肽产业研究院有限公司 | A kind of bionic enzymatic prepares the production method of soy peptide powder |
| CN110089753A (en) * | 2019-04-16 | 2019-08-06 | 东北农业大学 | A kind of preparation method for the nanoparticle improving curcumin bioavailability |
| CN110590902A (en) * | 2019-08-26 | 2019-12-20 | 内蒙古科然生物高新技术有限责任公司 | Method for producing soybean glycopeptide by using soybean molasses and soybean protein isolate powder |
| CN113832209A (en) * | 2021-09-30 | 2021-12-24 | 吉林农业大学 | Method for preparing high-antioxidant-activity hazelnut peptide |
| CN114262371A (en) * | 2021-12-21 | 2022-04-01 | 东北农业大学 | Preparation method of casein-chitosan oligosaccharide compound with antioxidant function |
| CN114668069A (en) * | 2022-04-25 | 2022-06-28 | 齐齐哈尔大学 | Preparation method of soybean protein glycosylation modified product with different sugar contents |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106929557A (en) * | 2017-04-28 | 2017-07-07 | 安徽生物肽产业研究院有限公司 | A kind of bionic enzymatic prepares the production method of soy peptide powder |
| CN110089753A (en) * | 2019-04-16 | 2019-08-06 | 东北农业大学 | A kind of preparation method for the nanoparticle improving curcumin bioavailability |
| CN110590902A (en) * | 2019-08-26 | 2019-12-20 | 内蒙古科然生物高新技术有限责任公司 | Method for producing soybean glycopeptide by using soybean molasses and soybean protein isolate powder |
| CN113832209A (en) * | 2021-09-30 | 2021-12-24 | 吉林农业大学 | Method for preparing high-antioxidant-activity hazelnut peptide |
| CN113832209B (en) * | 2021-09-30 | 2024-05-31 | 吉林农业大学 | Method for preparing hazelnut peptide with high antioxidant activity |
| CN114262371A (en) * | 2021-12-21 | 2022-04-01 | 东北农业大学 | Preparation method of casein-chitosan oligosaccharide compound with antioxidant function |
| CN114668069A (en) * | 2022-04-25 | 2022-06-28 | 齐齐哈尔大学 | Preparation method of soybean protein glycosylation modified product with different sugar contents |
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