CN106561970A - Separation and purification method for glycinin - Google Patents
Separation and purification method for glycinin Download PDFInfo
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- CN106561970A CN106561970A CN201610789900.8A CN201610789900A CN106561970A CN 106561970 A CN106561970 A CN 106561970A CN 201610789900 A CN201610789900 A CN 201610789900A CN 106561970 A CN106561970 A CN 106561970A
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- Prior art keywords
- supernatant
- globulin
- soybean
- separation
- glycinin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
Abstract
The invention discloses a separation and purification method for glycinin. The separation and purification method comprises the following specific steps of: taking soybean meal, smashing the soybean meal until the particle diameter is 5.5-7.5mm, adding the smashed soybean meal into a petroleum ether solution, standing until the solution is layered, dumping a supernatant, performing centrifugation, then taking a precipitate, washing the precipitate with a normal hexane solution, drying the precipitate, smashing and sieving the washed precipitate, adding the smashed precipitate into distilled water so as to prepare a soybean protein extract solution, adding 30U/g of phytase, regulating the pH to 6.8, performing centrifugation, and taking a supernatant for later use, wherein the precipitate is soybean 11S globulin; and adding MgCl2 into the supernatant, regulating the pH to 5.0, taking a supernatant, adding distilled water to dilute the supernatant at a volume ratio of 1 to 1, regulating the pH to 4.8, and taking a precipitate, namely soybean7S globulin. According to the method, phytase is added into the soybean protein extract solution, phytic acid which connects soybean 7S globulin with soybean 11S globulin is cut off, and the whole separation process is performed under a condition of room temperature, so that the problem of overlong time of the globulin separation process due to an added reducing agent is avoided, and the separated soybean 11S globulin and soybean 7S globulin are relatively high in productivity and purity.
Description
Technical field
The invention belongs to protein extracting process field, more particularly to a kind of glycinin process for separation and purification.
Background technology
7S albumen and 11S albumen are the main components of soybean protein, and 70% of total protein or so is accounted for altogether.11S protein moleculars
Quality is 302-375kDa, by 6 acidics (A1, A2, A3, A5, A6) and 6 alkaline subunits (B1, B2, B3, B4, B5,
B6) constitute, with disulfide bond between acid, alkali subunit.A1, A2, A4 molecular mass is 34.8kDa, and A3 is 45kDa, and A5 is
10kDa.B1, B2, B3, B4 molecular mass is 21kDa.7S protein moleculars quality is 141-170kDa, by α ', tri- subunits of α, β
Associated by hydrophobic interaction.
Semen sojae atricolor 11S albumen is considered as a kind of single albumen, without glycosyl.Its space conformation is acid sub- in same aspect
Base and alkaline subunit are alternately arranged, by hydrophobic interaction together with disulfide bond.Contain 12 Asias in its dimer
Base, the hollow Elliptic Cylinder combined by head-head is defined by two identical six-membered cyclic list aggressiveness.The amino of 11S albumen
What content was most in acid composition is glutaminic acid residue and asparagicacid residue, and beta sheet, β-corner are mainly in its secondary structure
And random coil.
Different from 11S albumen, soybean 7S globulin contains 3.8% mannose and 1.2% glucosamine.Glycosyl part in 7S
Combined with aspartic acid, 1 molecule 7S protein binding there are 5 or 6 glycosyls.According to the difference of its physicochemical property, 7S albumen can divide
For β-companion's globulin, γ-companion's globulin and alkalescence 7S albumen.It is made up of four subunits, and each subunit is by a macromolecule matter
Amount polypeptide (26kDa) and a low molecular weight polypeptide (16kDa) are consisted of disulfide bond, and structure is similar with 11S albumen.
The content of the invention
It is an object of the invention to provide a kind of glycinin process for separation and purification, by soybean protein extracting solution
Add phytase, it is to avoid globulin separation process time longer problem, detached Semen sojae atricolor 11S caused by because adding reducing agent
Globulin and soybean 7 S globulin yield and purity are higher.
The present invention is achieved by the following technical solutions：
A kind of glycinin process for separation and purification, comprises the following specific steps that：
S1, take Semen sojae atricolor and be crushed to particle diameter for 5.5-7.5mm, with solid-to-liquid ratio as 1:5 add mechanical agitation in petroleum ether solution
For a period of time, stand to solution layering, topple over and take precipitation hexane solution and wash 2-3 post-drying after supernatant centrifugation,
It is put into 60-80 mesh sieves excessively after crushing in pulverizer and obtains defatted soybean flour；
S2, will obtain in S1 defatted soybean flour add distilled water in be configured to soybean protein extracting solution, solid-to-liquid ratio is 1:5,
30U/g phytases are added in soybean protein extracting solution, pH to 5-6 is adjusted, magnetic agitation 2-3 is little under 30 DEG C of water bath conditions
When；
S3, the enzymolysis solution obtained in S2 is adjusted into pH to 6.8 with 1-5mol/L NaOH solutions, centrifuging and taking supernatant is standby,
It is precipitated as Soy 11 S Globulin；
S4, MgCl will be added in the supernatant obtained in S32, pH is adjusted to 5.0, take supernatant；
S5, take and add in the supernatant obtained in S4 distilled water diluting, volume ratio is 1:1 adjusts pH to 4.8, takes precipitation i.e.
For soybean 7 S globulin.
Further, mechanical agitation speed is 300r/min in the S1, and the mechanical agitation time is 10min.
Further, centrifugation rate is 3000r/min in the S1, and centrifugation time is 30min.
Further, centrifugation rate is 6500r/min in the S3, and centrifugation time is 30min.
Further, MgCl is added described in S4 in supernatant2To MgCl2Concentration is 0.1mol/L.
Further, centrifugation rate is 5800r/min in the S5, and centrifugation time is 20min.
Further, centrifugation rate is 6500r/min in the S5, and centrifugation time is 30min.
The invention has the advantages that：
The present invention in soybean protein extracting solution by adding phytase, cut-out connection soybean 7 S globulin and Semen sojae atricolor 11S
Phytic acid between globulin, whole separation process carries out in normal temperature condition, it is to avoid globulin point caused by because adding reducing agent
From the longer problem of process time, detached Soy 11 S Globulin and soybean 7 S globulin yield and purity are higher.
Certainly, the arbitrary product for implementing the present invention it is not absolutely required to while reaching all the above advantage.
Specific embodiment
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only
The a part of embodiment of the present invention, rather than the embodiment of whole.Based on the embodiment in the present invention, those of ordinary skill in the art
The all other embodiment obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
Embodiment 1
S1, take Semen sojae atricolor and be crushed to particle diameter for 5.5mm, with solid-to-liquid ratio as 1:5 add in petroleum ether solution, and 300r/min is mechanical
Stirring 10min, stand 60min after solution layering, topple over the supernatant and in 3000r/min be centrifuged 30min, take precipitation with just oneself
Alkane solution is placed in 90 DEG C of baking ovens after washing 2 times and dries, and is put into 60 mesh sieves excessively after crushing in pulverizer and obtains defatted soybean flour；
S2, will obtain in S1 defatted soybean flour add distilled water in be configured to soybean protein extracting solution, solid-to-liquid ratio is 1:5,
30U/g phytases are added in soybean protein extracting solution, pH to 5 is adjusted with 1mol/L HCl solutions, under 30 DEG C of water bath conditions
Magnetic agitation 2 hours；
S3, by the enzymolysis solution obtained in S2 with 1mol/L NaOH solutions adjust pH to 6.8,6500r/min centrifugation 30min,
Soy 11 S Globulin is precipitated as, supernatant is taken standby；
S4, MgCl will be added in the supernatant obtained in S32Solution is to MgCl2Concentration is 0.1mol/L, uses 1mol/L HCl
Solution adjusts pH to 5.0,5800r/min centrifugation 20min, takes supernatant；
S5, take and add in the supernatant obtained in S4 distilled water diluting, volume ratio is 1:1, adjusted with 1mol/L HCl solution
Section pH to 4.8, in 6500r/min 30min is centrifuged, and is taken precipitation and is soybean 7 S globulin.
Embodiment 2
S1, take Semen sojae atricolor and be crushed to particle diameter for 7.5mm, with solid-to-liquid ratio as 1:5 add in petroleum ether solution, and 300r/min is mechanical
Stirring 10min, stand 60min after solution layering, topple over the supernatant and in 3000r/min be centrifuged 30min, take precipitation with just oneself
Alkane solution is placed in 90 DEG C of baking ovens after washing 3 times and dries, and is put into 80 mesh sieves excessively after crushing in pulverizer and obtains defatted soybean flour；
S2, will obtain in S1 defatted soybean flour add distilled water in be configured to soybean protein extracting solution, solid-to-liquid ratio is 1:5,
30U/g phytases are added in soybean protein extracting solution, pH to 6 is adjusted with 5mol/L HCl solutions, under 30 DEG C of water bath conditions
Magnetic agitation 3 hours；
S3, by the enzymolysis solution obtained in S2 with 5mol/L NaOH solutions adjust pH to 6.8,6500r/min centrifugation 30min,
Soy 11 S Globulin is precipitated as, supernatant is taken standby；
S4, MgCl will be added in the supernatant obtained in S32Solution is to MgCl2Concentration is 0.1mol/L, uses 5mol/L HCl
Solution adjusts pH to 5.0,5800r/min centrifugation 20min, takes supernatant；
S5, take and add in the supernatant obtained in S4 distilled water diluting, volume ratio is 1:1, adjusted with 5mol/L HCl solutions
Section pH to 4.8, in 6500r/min 30min is centrifuged, and is taken precipitation and is soybean 7 S globulin.
Embodiment 3
S1, take Semen sojae atricolor and be crushed to particle diameter for 6.5mm, with solid-to-liquid ratio as 1:5 add in petroleum ether solution, and 300r/min is mechanical
Stirring 10min, stand 60min after solution layering, topple over the supernatant and in 3000r/min be centrifuged 30min, take precipitation with just oneself
Alkane solution is placed in 90 DEG C of baking ovens after washing 2 times and dries, and is put into 70 mesh sieves excessively after crushing in pulverizer and obtains defatted soybean flour；
S2, will obtain in S1 defatted soybean flour add distilled water in be configured to soybean protein extracting solution, solid-to-liquid ratio is 1:5,
30U/g phytases are added in soybean protein extracting solution, pH to 5.5 is adjusted with 3mol/L HCl solutions, in 30 DEG C of water bath conditions
Lower magnetic agitation 2.5 hours；
S3, by the enzymolysis solution obtained in S2 with 3mol/L NaOH solutions adjust pH to 6.8,6500r/min centrifugation 30min,
Soy 11 S Globulin is precipitated as, supernatant is taken standby；
S4, MgCl will be added in the supernatant obtained in S32Solution is to MgCl2Concentration is 0.1mol/L, uses 3mol/L HCl
Solution adjusts pH to 5.0,5800r/min centrifugation 20min, takes supernatant；
S5, take and add in the supernatant obtained in S4 distilled water diluting, volume ratio is 1:1, adjusted with 3mol/L HCl solutions
Section pH to 4.8, in 6500r/min 30min is centrifuged, and is taken precipitation and is soybean 7 S globulin.
The present invention in soybean protein extracting solution by adding phytase, cut-out connection soybean 7 S globulin and Semen sojae atricolor 11S
Phytic acid between globulin, whole separation process carries out in normal temperature condition, it is to avoid globulin point caused by because adding reducing agent
From the longer problem of process time, detached Soy 11 S Globulin and soybean 7 S globulin yield and purity are higher.
Above content is only citing made for the present invention and explanation, and affiliated those skilled in the art are to being retouched
The specific embodiment stated is made various modifications or supplements or substituted using similar mode, without departing from invention or super
More scope defined in the claims, all should belong to protection scope of the present invention.
Claims (7)
1. a kind of glycinin process for separation and purification, it is characterised in that comprise the following specific steps that：
S1, take Semen sojae atricolor and be crushed to particle diameter for 5.5-7.5mm, with solid-to-liquid ratio as 1:5 add one section of mechanical agitation in petroleum ether solution
Time, stand to solution layering, topple over and take precipitation hexane solution and wash 2-3 post-drying after supernatant centrifugation, be put into
60-80 mesh sieves are crossed after crushing in pulverizer and obtains defatted soybean flour；
S2, will obtain in S1 defatted soybean flour add distilled water in be configured to soybean protein extracting solution, solid-to-liquid ratio is 1:5, Yu great
30U/g phytases are added in soybean protein extracting solution, pH to 5-6, the magnetic agitation 2-3 hour under 30 DEG C of water bath conditions is adjusted；
S3, the enzymolysis solution obtained in S2 is adjusted into pH to 6.8 with 1-5mol/L NaOH solutions, centrifuging and taking supernatant is standby, precipitation
For Soy 11 S Globulin；
S4, MgCl will be added in the supernatant obtained in S32, pH is adjusted to 5.0, take supernatant；
S5, take and add in the supernatant obtained in S4 distilled water diluting, volume ratio is 1:1 adjusts pH to 4.8, takes precipitation and is greatly
Bean 7S.
2. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Machinery is stirred in the S1
It is 300r/min to mix speed, and the mechanical agitation time is 10min.
3. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Speed is centrifuged in the S1
Rate is 3000r/min, and centrifugation time is 30min.
4. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Speed is centrifuged in the S3
Rate is 6500r/min, and centrifugation time is 30min.
5. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Supernatant described in S4
Middle addition MgCl2To MgCl2Concentration is 0.1mol/L.
6. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Speed is centrifuged in the S5
Rate is 5800r/min, and centrifugation time is 20min.
7. a kind of glycinin process for separation and purification according to claim 1, it is characterised in that：Speed is centrifuged in the S5
Rate is 6500r/min, and centrifugation time is 30min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610789900.8A CN106561970A (en) | 2016-08-31 | 2016-08-31 | Separation and purification method for glycinin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610789900.8A CN106561970A (en) | 2016-08-31 | 2016-08-31 | Separation and purification method for glycinin |
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| CN106561970A true CN106561970A (en) | 2017-04-19 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610789900.8A Pending CN106561970A (en) | 2016-08-31 | 2016-08-31 | Separation and purification method for glycinin |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107318996A (en) * | 2017-06-28 | 2017-11-07 | 合肥市凤落河豆制食品有限公司 | A kind of preparation method rich in soybean protein white spirit |
| CN112913961A (en) * | 2021-03-17 | 2021-06-08 | 长春医学高等专科学校 | Preparation method of gel-type soybean protein with high stability |
| CN116349769A (en) * | 2023-03-08 | 2023-06-30 | 东北农业大学 | Method for improving aggregation characteristic of soybean 11S globulin by using medium-intensity electric field |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1346409A (en) * | 1999-03-30 | 2002-04-24 | 不二制油株式会社 | Fractionation of soybean 7S globulin and 11S globulin and process for producing the same |
| CN102187934A (en) * | 2011-06-01 | 2011-09-21 | 华南理工大学 | Method for producing grading soy protein |
| CN102229644A (en) * | 2011-06-01 | 2011-11-02 | 华南理工大学 | Method for fractionation of soybean 7S globulins and 11S globulins of low phytic acid |
-
2016
- 2016-08-31 CN CN201610789900.8A patent/CN106561970A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1346409A (en) * | 1999-03-30 | 2002-04-24 | 不二制油株式会社 | Fractionation of soybean 7S globulin and 11S globulin and process for producing the same |
| CN102187934A (en) * | 2011-06-01 | 2011-09-21 | 华南理工大学 | Method for producing grading soy protein |
| CN102229644A (en) * | 2011-06-01 | 2011-11-02 | 华南理工大学 | Method for fractionation of soybean 7S globulins and 11S globulins of low phytic acid |
Non-Patent Citations (1)
| Title |
|---|
| 刘珊珊: "《大豆7S球蛋白亚基组成多样性的分子基础》", 31 October 2008, 黑龙江科学技术出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107318996A (en) * | 2017-06-28 | 2017-11-07 | 合肥市凤落河豆制食品有限公司 | A kind of preparation method rich in soybean protein white spirit |
| CN112913961A (en) * | 2021-03-17 | 2021-06-08 | 长春医学高等专科学校 | Preparation method of gel-type soybean protein with high stability |
| CN116349769A (en) * | 2023-03-08 | 2023-06-30 | 东北农业大学 | Method for improving aggregation characteristic of soybean 11S globulin by using medium-intensity electric field |
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