CN1285731C - Method for enzyme hydrolyzing zein through two steps method - Google Patents
Method for enzyme hydrolyzing zein through two steps method Download PDFInfo
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- CN1285731C CN1285731C CNB2004100775181A CN200410077518A CN1285731C CN 1285731 C CN1285731 C CN 1285731C CN B2004100775181 A CNB2004100775181 A CN B2004100775181A CN 200410077518 A CN200410077518 A CN 200410077518A CN 1285731 C CN1285731 C CN 1285731C
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Abstract
The present invention relates to a method for the enzymatic hydrolysis of corn alcohol soluble protein in two steps, which comprises the steps: adding proteinase according to the dosage of 2000 to 10000U/ substrate into 150 ml of corn alcohol soluble protein solution for hydrolysis; evaporating 1/5 to 1/2 of the organic solvent in the mixture, filling the mixture with deionized water to 150 ml to form water phase, adjusting the pH value of the mixture to 8.0, and adding proteinase liquid according to the dosage of 2000 to 10000U/ substrate into the mixture for secondary hydrolysis; and decompressing the hydrolysis liquid obtained by concentration, and freezing and drying the hydrolysis liquid to obtain hydrolyzate, wherein the organic solvent for dissolving the corn alcohol soluble protein is etanol or isopropanol with the concentration of 60 to 80%(v/v), and the corn alcohol soluble protein hydrolyzate has the hydrolysis degree of 20%. The method can effectively hydrolyze corn alcohol soluble protein, and has the advantage of high hydrolysis degree. The obtained hydrolyzate which can be completely dissolved in water contains depressor peptide components with high activity, and can be applied to functional health-care food.
Description
Technical field
The present invention relates to the Technology field of zein hydrolysis, specifically be meant the production technique of a kind of zein in organic phase and aqueous phase two step enzyme method hydrolysis.
Background technology
Zein is a kind of high value-added product that the by product Zein powder from maize raw material or W-Gum production process extracts.Because zein is water insoluble, does not have edibleness.If can be with its hydrolysis, just can increase that it is water-soluble, and can improve its functional performance, just might in foodstuff production, be used widely.
Find according to some recent research reports, can obtain a kind of little peptide after the zein hydrolysis, this material is at present in the step-down peptide product that makes with the bioprotein material, to ACE (angiotensin-converting enzyme) inhibition the strongest active a kind of inhibitor.This discovery makes the value of zein hydrolyzate double.
But because zein only is dissolved in 60%～90% alcoholic solution, the enzymic hydrolysis of zein just must be carried out in organic phase, and this has just increased the difficulty of hydrolysis greatly.Because most enzymes lost efficacy with this understanding, can not have an effect, therefore require to select the hydrolase of some anti-alcohol for use, and adopt special hydrolysis process could really realize the hydrolysis of zein.Yet the screening of this kind of enzyme and discovery work are quite difficult.
The main method of domestic production zein step-down peptide is to be raw material with the Zein powder at present, adopts proteolytic enzyme to be hydrolyzed at aqueous phase.The major defect of this method is: the prolamine component in the zein can not be dissolved in the water, and proteolytic enzyme can not be hydrolyzed to prolamine, and the main ingredient of corn step-down peptide all stems from zein.Therefore this method is not the method for effectively producing corn step-down peptide.
Summary of the invention
The present invention is exactly that providing a kind of can be hydrolyzed zein smoothly, and makes hydrolytic process have the method for hydrolysis of the zein of controllability in order to solve above-mentioned the deficiencies in the prior art part.
The method of method for enzyme hydrolyzing zein through two steps of the present invention comprises the steps:
(1) get zein solution 150mL, press 2000-10000U/ substrate consumption and add proteolytic enzyme, be hydrolyzed, hydrolysis temperature is 45 ℃-65 ℃, and the time is 20 minutes;
(2) evaporate the wherein organic solvent of 1/5-1/2, be supplemented to 150mL with deionized water again, become water, transfer pH to 8.0, add described liquid of protease by 2000-10000U/ substrate consumption and carry out the secondary hydrolysis, hydrolysis temperature is 45 ℃-65 ℃, and hydrolysis time is 40-120 minute;
(3) hydrolyzed solution that obtains of concentrating under reduced pressure (2), lyophilize.
Used zein extracts from corn or Zein powder and obtains.
Zein solution concentration is 1%-6% (g/mL), and the pH value is 8.0, and the organic solvent of dissolving zein is that concentration is ethanol or the Virahol of 60%-80% (v/v).
Described proteolytic enzyme is Sumizyme MP or Alcalase, and its pH is 8.0, is the proteolytic enzyme that is mixed with 1% (v/v) with phosphate buffered saline buffer.
The present invention adopts freeze-drying method to reclaim drying products, can keep the biological activity of products obtained therefrom to greatest extent.
Preferred plan of the present invention is as follows:
(1) get zein solution 150mL, the consumption of pressing 6000-7200U/ gram solid-state protein adds proteolytic enzyme, is hydrolyzed, and hydrolysis temperature is 55 ℃-60 ℃, and the time is 20 minutes;
(2) evaporate the wherein organic solvent of 11/30-13/30, be supplemented to 150mL with deionized water again, become water, transfer pH to 8.0, add described liquid of protease by the consumption of 6000-7200U/ gram solid-state protein and carry out the secondary hydrolysis, hydrolysis temperature is 50 ℃-55 ℃, and hydrolysis time is 70-90 minute;
(3) hydrolyzed solution that obtains of concentrating under reduced pressure (2), lyophilize;
Zein solution concentration is 3%-5% (g/mL), and the pH value is 8.0, and the organic solvent of dissolving zein is that concentration is ethanol or the Virahol of 70%-75% (v/v);
Described proteolytic enzyme is Sumizyme MP or Alcalase, and its pH is 8.0, is the proteolytic enzyme that is mixed with 1% (v/v) with Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution.
The degree of hydrolysis DH of gained hydrolysate is 2%-20%.
Present technique compared with prior art has following advantage and effect:
(1) be raw material directly, and technology in the past is to be raw material with the Zein powder with the zein;
(2) raw material is dissolved with organic dissolving earlier, under the effect of enzyme, divided for two steps carried out hydrolysis again, have the hydrolysis efficiency height, degree of hydrolysis reaches as high as DH20%;
(3) the zein hydrolyzate both can be used as foodstuff additive, had foaming emulsification and local flavor and function such as sheltered, and the corn peptide of gained also is a kind of high-quality hypertensin enzymeinhibitor simultaneously, has hypotensive function.
(4) zein is a kind of albumen of high value, and its production cost religion is high, if can be used for making the functional food ingredient of high value, then can effectively expand the range of application of zein.Owing to zein is that by product Zein powder from corn processed extracts, increased the application of zein, just equaled to improve the added value of agricultural byproducts, have very high social benefit and economic benefit.
Embodiment
Embodiment 1
1 takes by weighing 3 gram zein fully dissolves evenly with 150mL 65% ethanolic soln, is mixed with 2% albumen ethanolic soln, and with the NaOH solution adjust pH to 8.0 of 0.2mol/L;
2 is that Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution of 8.0 is mixed with 1% enzyme liquid (enzyme activity unit is 1800U/mL) with a kind of Sumizyme MP with the pH value;
3 enzymolysis for the first time: measure protein solution 150mL, add 1% liquid of protease 6mL, evenly mixed, enzymolysis in 48 ℃ of water-baths drips the NaOH solution of 0.1mol/L, makes the pH value of solution value maintain 8.0;
4 enzyme digestion reactions carried out underpressure distillation with reaction solution rapidly after 20 minutes under 50 ℃, stop after steaming the 35mL ethanolic soln;
5 add deionized water again to 150mL in above-mentioned concentrated solution, transferring pH with the NaOH solution of 0.1mol/L is 8.0, adds 14mL 1% liquid of protease hydrolysis again, and hydrolysis temperature is 48 ℃;
6 hydrolysis for the second time are after 60 minutes, and recording degree of hydrolysis DH is 17.6%, termination reaction, and reduction vaporization concentrates, and reclaims zein hydrolyzate product 2.6 grams through lyophilize.
Embodiment 2
1 takes by weighing 7.5 gram zein fully dissolves evenly with 150mL 75% ethanolic soln, is mixed with 5% albumen ethanolic soln, and with the KOH solution adjust pH to 8.0 of 0.25mol/L;
2 is that Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution of 8.0 is mixed with 1% enzyme liquid (enzyme activity unit is 1800U/mL) with a kind of Sumizyme MP with the pH value;
3 enzymolysis for the first time: measure protein solution 150mL, add 1% liquid of protease 25mL, evenly mixed, enzymolysis in 55 ℃ of water-baths drips the NaOH solution of 0.1mol/L, makes the pH value of solution value maintain 8.0;
4 enzyme digestion reactions carried out underpressure distillation with reaction solution rapidly after 20 minutes under 50 ℃, stop after steaming the 65mL ethanolic soln;
5 add deionized water again to 150mL in above-mentioned concentrated solution, transferring pH with the KOH solution of 0.25mol/L is 8.0, adds 25mL 1% liquid of protease hydrolysis again, and hydrolysis temperature is 55 ℃;
6 hydrolysis for the second time are after 90 minutes, and recording degree of hydrolysis DH is 18.4%, termination reaction, and reduction vaporization concentrates, and reclaims zein hydrolyzate product 6.8 grams through lyophilize.
Embodiment 3
1 takes by weighing 4.5 gram zein fully dissolves evenly with 150mL 70% different ethanolic soln, is mixed with 3% albumen ethanolic soln, and with the NaOH solution adjust pH to 8.0 of 0.5mol/L;
2 is that Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution of 8.0 is mixed with 1% enzyme liquid (enzyme activity unit is 1800U/mL) with a kind of Sumizyme MP with the pH value;
3 enzymolysis for the first time: measure protein solution 150mL, add 1% liquid of protease 18mL, evenly mixed, enzymolysis in 60 ℃ of water-baths drips NaOH or the KOH solution of 0.1mol/L, makes the pH value of solution value maintain 8.0;
4 enzyme digestion reactions carried out underpressure distillation with reaction solution rapidly after 20 minutes under 50 ℃, stop after steaming the 55mL ethanolic soln;
5 add deionized water again to 150mL in above-mentioned concentrated solution, transferring pH with the NaOH solution of 0.5mol/L is 8.0, adds 18mL 1% liquid of protease hydrolysis again, and hydrolysis temperature is 60 ℃;
6 hydrolysis for the second time are after 70 minutes, and recording degree of hydrolysis DH is 20.3%, termination reaction, and reduction vaporization concentrates, and reclaims zein hydrolyzate product 4.1 grams through lyophilize.
Embodiment 4
1 takes by weighing 6 gram zein fully dissolves evenly with 150mL 60% different ethanolic soln, is mixed with 3% albumen ethanolic soln, and with the KOH solution adjust pH to 8.0 of 1mol/L;
2 is that Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution of 8.0 is mixed with 1% enzyme liquid (enzyme activity unit is 1800U/mL) with a kind of Sumizyme MP with the pH value;
3 enzymolysis for the first time: measure protein solution 150mL, add 1% liquid of protease 17mL, evenly mixed, enzymolysis in 65 ℃ of water-baths drips the NaOH solution of 0.1mol/L, makes the pH value of solution value maintain 8.0;
4 enzyme digestion reactions carried out underpressure distillation with reaction solution rapidly after 20 minutes under 50 ℃, stop after steaming the 45mL ethanolic soln;
5 add deionized water again to 150mL in above-mentioned concentrated solution, transferring pH with the KOH solution of 1mol/L is 8.0, adds 17mL 1% liquid of protease hydrolysis again, and hydrolysis temperature is 65 ℃;
6 hydrolysis for the second time are after 80 minutes, and recording degree of hydrolysis DH is 15.4%, termination reaction, and reduction vaporization concentrates, and reclaims zein hydrolyzate product 5.2 grams through lyophilize.
Claims (2)
1, a kind of method of method for enzyme hydrolyzing zein through two steps is characterized in that comprising the steps:
(1) get zein solution 150mL, the consumption of pressing 2000-10000U/ gram solid-state protein adds proteolytic enzyme, is hydrolyzed, and hydrolysis temperature is 45 ℃-65 ℃, and the time is 20 minutes;
(2) evaporate the wherein organic solvent of 1/5-1/2, be supplemented to 150mL with deionized water again, become water, transfer pH to 8.0, add described liquid of protease by the consumption of 2000-10000U/ gram solid-state protein and carry out the secondary hydrolysis, hydrolysis temperature is 45 ℃-65 ℃, and hydrolysis time is 40-120 minute;
(3) hydrolyzed solution that obtains of concentrating under reduced pressure (2), lyophilize;
Zein solution concentration is 1%-6%g/mL, and the pH value is 8.0, and the organic solvent of dissolving zein is that concentration is ethanol or the Virahol of 60%-80%v/v;
Described proteolytic enzyme is Sumizyme MP or Alcalase, and its pH is 8.0, is the proteolytic enzyme that is mixed with 1%v/v with phosphate buffered saline buffer.
2, the method for method for enzyme hydrolyzing zein through two steps according to claim 1 is characterized in that comprising the steps:
(1) get zein solution 150mL, the consumption of pressing 6000-7200U/ gram solid-state protein adds proteolytic enzyme, is hydrolyzed, and hydrolysis temperature is 55 ℃-60 ℃, and the time is 20 minutes;
(2) evaporate the wherein organic solvent of 11/30-13/30, be supplemented to 150mL with deionized water again, become water, transfer pH to 8.0, add described liquid of protease by the consumption of 6000-7200U/ gram solid-state protein and carry out the secondary hydrolysis, hydrolysis temperature is 50 ℃-55 ℃, and hydrolysis time is 70-90 minute;
(3) hydrolyzed solution that obtains of concentrating under reduced pressure (2), lyophilize;
Zein solution concentration is 3%-5%g/mL, and the pH value is 8.0, and the organic solvent of dissolving zein is that concentration is ethanol or the Virahol of 70%-75%v/v;
Described proteolytic enzyme is Sumizyme MP or Alcalase, and its pH is 8.0, is the proteolytic enzyme that is mixed with 1%v/v with Sodium phosphate dibasic-phosphate sodium dihydrogen buffer solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100775181A CN1285731C (en) | 2004-12-22 | 2004-12-22 | Method for enzyme hydrolyzing zein through two steps method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100775181A CN1285731C (en) | 2004-12-22 | 2004-12-22 | Method for enzyme hydrolyzing zein through two steps method |
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| CN1661031A CN1661031A (en) | 2005-08-31 |
| CN1285731C true CN1285731C (en) | 2006-11-22 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101560252B (en) * | 2009-05-08 | 2012-01-11 | 广东省食品工业研究所 | Method for extracting zein by microwave |
| CN108004287A (en) * | 2017-11-13 | 2018-05-08 | 江苏大学 | A kind of preparation method of the macromolecular polypeptides based on gastro-intestinal digestion |
| CN108003230B (en) * | 2017-11-13 | 2020-11-03 | 江苏大学 | Method for preparing zein antihypertensive peptide |
| CN114656548A (en) * | 2022-04-13 | 2022-06-24 | 练文飞 | Antihypertensive peptide, and preparation method and application thereof |
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Assignee: Guangdong Huanxi Health Technology Inc. Assignor: South China University of Technology Contract fulfillment period: 2008.11.5 to 2013.11.4 contract change Contract record no.: 2009440000608 Denomination of invention: Method for enzyme hydrolyzing zein through two steps method Granted publication date: 20061122 License type: Exclusive license Record date: 2009.7.13 |
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Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2008.11.5 TO 2013.11.4; CHANGE OF CONTRACT Name of requester: GUANGDONG HUANXI HEALTH SCI-TECH CO., LTD. Effective date: 20090713 |
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