CN103242423A - Method for extracting silybum marianum protein - Google Patents
Method for extracting silybum marianum protein Download PDFInfo
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- CN103242423A CN103242423A CN2013101732958A CN201310173295A CN103242423A CN 103242423 A CN103242423 A CN 103242423A CN 2013101732958 A CN2013101732958 A CN 2013101732958A CN 201310173295 A CN201310173295 A CN 201310173295A CN 103242423 A CN103242423 A CN 103242423A
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Abstract
The invention discloses a method for extracting silybum marianum protein and belongs to the technical field of application of biotechnology in foods. The method comprises the following steps: dissolving silybum marianum seed kenel spent meal in distilled water, adding mixed enzyme of cellulose and amylase according to a mass ratio of the enzyme to the substrate of 4.5-12 percent so as to carry out enzymolysis for 0.5-3 hours under the conditions of the pH of 4-7 and the temperature of 40-70 DEG C, heating and inactivating the enzymes, regulating the pH value of the solution to be 10 at the water bath temperature of 50 DEG C, leaching for 60 minutes, centrifuging, regulating the pH value of the liquid supernatant to be 5.5 by using hydrochloric acid, centrifuging and collecting the precipitate, dissolving the precipitate by using water, regulating the pH to be neutral, and performing vacuum freeze drying to obtain the silybum marianum protein powder. The silybum marianum seed kenel spent meal is treated by employing cellulose and amylase, and the cellulose and amylase can hydrolyze cellulose, starch and other polysaccharide components, so that the constraint of the polysaccharide on the protein can be relieved, the dissolution of the protein is promoted, and the protein extracting rate is improved.
Description
Technical field
The present invention relates to a kind of extracting method of Silymarin albumen, particularly carry out the pre-treatment method of alkali extraction and acid precipitation method extraction Silymarin albumen again with cellulase and amylase, belong to biotechnology in the Application in Food technical field.
Background technology
Silymarin is catananche's medicinal material, and Silymarin mainly utilizes its seed as a kind of traditional medicinal plant.Silybum marianum seed is mainly used to its main component silymarin of extraction separation, and then purifying is silibinin.Silibinin mainly is present in the seed shell, and Silymarin oil and albumen then are present in the seed benevolence.Studies have shown that protein content is up to about 47% in the silybum marianum seeds spent meal, amino acid A wide selection of colours and designs in the Silymarin albumen be a kind of quality protein, and Silymarin albumen can prepare the polypeptide that has than the strong anti-oxidation activity through enzymolysis.Therefore, be that raw material extracts albumen with the silybum marianum seeds spent meal, will be to improving the comprehensive utilization of silybum marianum seeds rate and reducing the wasting of resources and play positive pushing effect.
At present the extracting method of albumen mainly is the alkali extraction and acid precipitation method, but because of the influence of plant cell wall, the extraction of albumen is still insufficient.In recent years, protease preparation also is applied to the extraction research of vegetable-protein, and it is by becoming proteolysis the polypeptide than small molecules to improve extraction rate of protein, but the processing characteristics of this method products obtained therefrom (as emulsifying property, foaminess) can be affected.Cellulase, amylase and Viscozyme enzyme etc. also have application in the extraction of vegetable-protein, thereby it mainly is by removing polyose compositions such as Mierocrystalline cellulose improve albumen to the constraint of protein extraction yield, and generally do not influence the structure of protein, the processing characteristics that has kept protein has unique effect in the extraction of vegetable-protein.The present invention proposes with the silybum marianum seeds spent meal is raw material, with after cellulase and the amylase pre-treatment again alkali extraction and acid precipitation extract the new operational path of Silymarin albumen.
Summary of the invention
The present invention is to provide a kind of extracting method of Silymarin albumen, be raw material with the silybum marianum seeds spent meal, by adding cellulase and amylase the silybum marianum seeds spent meal is carried out pre-treatment, obtain Silymarin albumen by the alkali extraction and acid precipitation method again, this method can significantly improve the extraction yield of Silymarin albumen.
Above-mentioned purpose of the present invention realizes by following technique means.
A kind of extracting method of Silymarin albumen, carry out according to following step:
(1) silybum marianum seeds is pulverized the back according to (ZL201010152017.0) method in is carried out degreasing for a kind of comprehensive utilization of silybum marianum seeds method, China, and the gained spent meal is crossed 60 mesh sieves;
(2) silybum marianum seeds spent meal and distilled water are pressed quality and 1: 16 mixing of volume ratio (w/v), add cellulase and diastatic mixed enzyme according to enzyme-to-substrate mass ratio 4.5%～12% then, wherein cellulase and amylase mass ratio are 1:2, be 4～7 at pH, temperature is under 40 ℃～70 ℃ the condition, carried out the enzymolysis pre-treatment 0.5～3 hour, heat the enzyme that goes out, regulate pH value to 10, bath temperature is 50 ℃, and lixiviate 60 min are centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing precipitates adjust pH to neutral again, and vacuum lyophilization is the Silymarin protein powder.
Preferably, Silymarin protein preparation method of the present invention is:
Silybum marianum seeds spent meal and distilled water are by quality and 1: 16 mixing of volume ratio (w/v), concentration according to enzyme-to-substrate mass ratio 9% adds cellulase and diastatic mixed enzyme then, wherein cellulase and amylase mass ratio are 1:2, the enzyme pretreatment temperature is 50 ℃, pH is 6.0, the enzyme pretreatment time is 150 min, heat the enzyme that goes out, regulate pH value to 10, alkali water lift bath temperature is 50 ℃, extraction time is 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.
Compare with traditional alkaline extraction, adopt cellulase and amylase that the silybum marianum seeds spent meal is carried out pre-treatment, can remove polysaccharose substances such as Mierocrystalline cellulose and starch to the constraint of protein, be beneficial to the stripping of albumen, thereby extraction yield, economic benefit and the obvious social benefit of Silymarin albumen have been improved.
Embodiment
Be further elaborated below in conjunction with the present invention of embodiment:
Cellulase is available from Amresco company, and enzyme activity is 3.5 ten thousand U/g; Amylase is available from Shanghai chemical reagents corporation, and enzyme activity is 0.2 ten thousand U/g.
Embodiment 1
Silybum marianum seeds is pulverized the back according to (ZL201010152017.0) method is carried out degreasing for a kind of comprehensive utilization of silybum marianum seeds method, China, and the gained spent meal is crossed 60 mesh sieves;
Silybum marianum seeds spent meal and distilled water are by quality and 1: 16 (g/mL) mixing of volume ratio (w/v), concentration according to enzyme-to-substrate mass ratio 7.5% adds cellulase and diastatic mixed enzyme then, wherein cellulase and amylase mass ratio are 1:2, at pH value of solution 5.5, under the condition that the enzyme pretreatment temperature is 50 ℃, behind enzymolysis 60 min, heat the enzyme that goes out, regulate pH value to 10, alkali water lift bath temperature is 50 ℃, lixiviate 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.With this understanding, the extraction yield of Silymarin albumen is 50.73%.
Embodiment 2
Silybum marianum seeds is pulverized the back according to (method ZL201010152017.0) is carried out degreasing for a kind of comprehensive utilization of silybum marianum seeds method, China, and the gained spent meal is crossed 60 mesh sieves;
Silybum marianum seeds spent meal and distilled water are by quality and 1: 16 (g/mL) mixing of volume ratio (w/v), concentration according to enzyme-to-substrate mass ratio 9% adds cellulase and diastatic mixed enzyme then, wherein cellulase and amylase mass ratio are 1:2, at pH value of solution 6.5, under the condition that the enzyme pretreatment temperature is 50 ℃, behind enzymolysis 60 min, heat the enzyme that goes out, regulate pH value to 10, alkali water lift bath temperature is 50 ℃, lixiviate 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.With this understanding, the extraction yield of Silymarin albumen is 53.57%.
Embodiment 3
Silybum marianum seeds is pulverized the back according to (ZL201010152017.0) method in is carried out degreasing for a kind of comprehensive utilization of silybum marianum seeds method, China, and the gained spent meal is crossed 60 mesh sieves;
Silybum marianum seeds spent meal and distilled water are by quality and 1: 16 (g/mL) mixing of volume ratio (w/v), concentration according to enzyme-to-substrate mass ratio 9% adds cellulase and diastatic mixed enzyme then, wherein cellulase and amylase mass ratio are 1:2, at pH value of solution 6.0, under the condition that the enzyme pretreatment temperature is 50 ℃, behind enzymolysis 150 min, heat the enzyme that goes out, regulate pH value to 10, alkali water lift bath temperature is 50 ℃, lixiviate 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.With this understanding, the extraction yield of Silymarin albumen is 66.75%.
Comparative Examples (without the enzyme pre-treatment)
Silybum marianum seeds is pulverized the back according to (ZL201010152017.0) method in is carried out degreasing for a kind of comprehensive utilization of silybum marianum seeds method, China, and the gained spent meal is crossed 60 mesh sieves;
Silybum marianum seeds spent meal and distilled water are by quality and 1: 16 (g/mL) mixing of volume ratio (w/v), transfer pH value of solution to 6.0, be under 50 ℃ the condition behind water-bath 150 min in temperature, regulate pH value to 10, in temperature be lixiviate 60 min under 50 ℃ the condition, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing precipitates adjust pH to neutral again, and vacuum lyophilization is the Silymarin protein powder.With this understanding, the extraction yield of Silymarin albumen is 53.62%.
Claims (3)
1. the extracting method of a Silymarin albumen is characterized in that carrying out according to following step:
(1) will carry out degreasing according to a conventional method after the silybum marianum seeds pulverizing, the gained spent meal is crossed 60 mesh sieves;
(2) silybum marianum seeds spent meal and distilled water are pressed quality and 1: 16 mixing of volume ratio (w/v), add cellulase and diastatic mixed enzyme according to enzyme-to-substrate mass ratio 4.5%～12% then, be 4～7 at pH, temperature is under 40 ℃～70 ℃ the condition, carried out the enzymolysis pre-treatment 0.5～3 hour, heat the enzyme that goes out, regulate pH value to 10, bath temperature is 50 ℃, lixiviate 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.
2. the extracting method of a kind of Silymarin albumen according to claim 1, it is characterized in that carrying out according to following step: silybum marianum seeds spent meal and distilled water are by quality and 1: 16 mixing of volume ratio (w/v), concentration according to enzyme-to-substrate mass ratio 9% adds cellulase and diastatic mixed enzyme then, the enzyme pretreatment temperature is 50 ℃, pH is 6.0, the enzyme pretreatment time is 150 min, heat the enzyme that goes out, regulate pH value to 10, alkali water lift bath temperature is 50 ℃, extraction time is 60 min, centrifugal, supernatant liquor salt acid for adjusting pH value to 5.5, centrifugal and collecting precipitation thing again, the precipitation adjust pH is to neutral, and vacuum lyophilization is the Silymarin protein powder.
3. the extracting method of a kind of Silymarin albumen according to claim 1 and 2 is characterized in that wherein cellulase and amylase mass ratio are 1:2.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104621339A (en) * | 2015-01-23 | 2015-05-20 | 沈阳化工大学 | Method for extracting silybum marianum protein isolate |
| CN105285314A (en) * | 2015-09-14 | 2016-02-03 | 哈尔滨工业大学 | Extraction method of 13KDa rice alcohol soluble protein |
| CN107141342A (en) * | 2017-06-21 | 2017-09-08 | 江苏中兴药业有限公司 | A kind of method that soybean dietary fiber method extracts silybum marianum seeds albumen |
| CN107802820A (en) * | 2017-12-12 | 2018-03-16 | 江西省三抗保健品有限公司 | A kind of Traditional Chinese medicine compound composition for alleviating mental fatigue |
| CN109266432A (en) * | 2018-09-26 | 2019-01-25 | 江苏大学 | Method that is a kind of while extracting Silybum Marianum Gaertn Seed Oil and protein hydrolysate |
| CN111171110A (en) * | 2020-02-26 | 2020-05-19 | 大连理工大学 | Method for extracting silybum marianum protein from silybum marianum pharmaceutical slag |
| CN111269285A (en) * | 2020-02-26 | 2020-06-12 | 大连理工大学 | Method for extracting vegetable protein from silybum marianum seed kernels |
| CN112359085A (en) * | 2020-11-06 | 2021-02-12 | 江苏大学 | Method for improving antioxidant activity of silybum marianum protein zymolyte |
| CN116391817A (en) * | 2023-02-23 | 2023-07-07 | 大连理工大学 | Beverage rich in silybin and silybum marianum protein and preparation method thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104621339A (en) * | 2015-01-23 | 2015-05-20 | 沈阳化工大学 | Method for extracting silybum marianum protein isolate |
| CN105285314A (en) * | 2015-09-14 | 2016-02-03 | 哈尔滨工业大学 | Extraction method of 13KDa rice alcohol soluble protein |
| CN107141342A (en) * | 2017-06-21 | 2017-09-08 | 江苏中兴药业有限公司 | A kind of method that soybean dietary fiber method extracts silybum marianum seeds albumen |
| CN107141342B (en) * | 2017-06-21 | 2020-06-23 | 江苏中兴药业有限公司 | Method for extracting silybum marianum seed kernel protein by wet superfine grinding method |
| CN107802820A (en) * | 2017-12-12 | 2018-03-16 | 江西省三抗保健品有限公司 | A kind of Traditional Chinese medicine compound composition for alleviating mental fatigue |
| CN109266432A (en) * | 2018-09-26 | 2019-01-25 | 江苏大学 | Method that is a kind of while extracting Silybum Marianum Gaertn Seed Oil and protein hydrolysate |
| CN111171110A (en) * | 2020-02-26 | 2020-05-19 | 大连理工大学 | Method for extracting silybum marianum protein from silybum marianum pharmaceutical slag |
| CN111269285A (en) * | 2020-02-26 | 2020-06-12 | 大连理工大学 | Method for extracting vegetable protein from silybum marianum seed kernels |
| CN111171110B (en) * | 2020-02-26 | 2022-09-16 | 大连理工大学 | Method for extracting silybum marianum protein from silybum marianum pharmaceutical slag |
| CN112359085A (en) * | 2020-11-06 | 2021-02-12 | 江苏大学 | Method for improving antioxidant activity of silybum marianum protein zymolyte |
| CN116391817A (en) * | 2023-02-23 | 2023-07-07 | 大连理工大学 | Beverage rich in silybin and silybum marianum protein and preparation method thereof |
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Application publication date: 20130814 |