CN101509029A - Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite - Google Patents
Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite Download PDFInfo
- Publication number
- CN101509029A CN101509029A CNA200910025324XA CN200910025324A CN101509029A CN 101509029 A CN101509029 A CN 101509029A CN A200910025324X A CNA200910025324X A CN A200910025324XA CN 200910025324 A CN200910025324 A CN 200910025324A CN 101509029 A CN101509029 A CN 101509029A
- Authority
- CN
- China
- Prior art keywords
- rapeseed
- hydrolysis
- enzyme
- peptide
- wat
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a method which uses sodium sulfite for speeding up the enzymatic hydrolysis of rapeseed meal so as to prepare a rapeseed peptide, comprising the use of protease for conducting hydrolysis, enzyme destroying, removal of insoluble matter by decentralization, and obtaining of the rapeseed peptide by ultrafiltration and drying; the method is characterized in that after the rapeseed meal is crushed and then added in Na2SO3, the rapeseed peptide is obtained by enzyme hydrolysis. The method has the advantages that Na2SO3, as a reducing agent, can break the disulfide bonds among and in the protein molecules, causes the broken rapeseed mea molecules to be more easily contacted with the protease, improves the enzyme hydrolysis rate and the hydrolysis degree, and simultaneously destroys the protease inhibitor molecules which exist in the rapeseed protein, and thus leads the protease inhibitor molecules to lose the inhibiting effect on the activity of the protease; as the reducing agent, Na2SO3 can also prevent the polyphenols in the rapeseed meal to be oxidized during the hydrolysis process so as to cause the color of the obtained rapeseed peptide to be lighter.
Description
Technical field
The present invention relates to a kind of method of producing rapeseed peptide (rsp), relate in particular to a kind of at Na
2SO
3Effect uses the protease hydrolysis rapeseed meal to produce the method for rapeseed peptide (rsp) down.
Background technology
Vegetable seed is important oil crops, and world's annual production surpasses 4,000 ten thousand tons, and China is the vegetable seed producing country of output maximum in the world.Vegetable seed is carried in the dregs of rice behind the oil and is contained 40-50% albumen, and rape seed protein amino acid Compositional balance, so rapeseed meal may become the suitable source of food protein.But there are antinutritional factor such as glucosinolate, phytic acid and Polyphenols in the rapeseed meal, have only and reduce as far as possible or remove these materials, rapeseed meal is originated as food protein.In recent years, vegetable protein hydrolyzate is considered to the source of biologically active peptides, and they are easy to be entered blood by the colorectal cell absorption, produce useful physiological action.Physiological functions such as the biologically active peptides of having reported has immunomodulatory, antibiotic, antithrombotic forms and hypotensive.The traditional method of producing polypeptide is to produce protein isolate by the alkaline extraction acid precipitation method earlier, re-uses different proteolytic enzyme separation proteolysis is produced peptide.Because the rape seed protein complicated component is insoluble in water, need to use the above highly basic of pH11 to extract usually; Rape seed protein iso-electric point scope is wide in addition, and adding acid after the extraction, to regulate the protein isolate yield that iso-electric point separates out lower.
The CN1884572A patent documentation discloses " a kind of method for preparing rapeseed peptide (rsp) with the direct enzymic hydrolysis of rapeseed meal ", this method is used sour water that rapeseed meal is soaked and is washed removing anti-nutrient substances such as phytic acid, sulphur glucoside, and directly uses production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs.This method with produce rape seed protein earlier, the traditional method of producing rapeseed peptide (rsp) with the enzymic hydrolysis rape seed protein is compared again, has simplified technological process.But sour water has also been taken away the part soluble proteins when removing anti-nutrient substances such as phytic acid, sulphur glucoside, produced more waste water.
Summary of the invention
The objective of the invention is to overcome above-mentioned prior art and have deficiency, the novel method of the production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs that a kind of hydrolysis rate is fast, loss of proteins is few, product color is good is provided.
We discover, add the Na of lower concentration when the protease hydrolysis rapeseed meal
2SO
3The aqueous solution can significantly be accelerated hydrolysis rate, improves protein hydrolysis degree.Because Na
2SO
3Can rupture between protein molecule and intramolecular disulfide linkage as a kind of reductive agent, make easier contact of post-rift rape seed protein molecule and improve enzymic hydrolysis speed and degree of hydrolysis, while Na with proteolytic enzyme
2SO
3The effect of fracture disulfide linkage has also destroyed the proteinase inhibitor molecule that exists in the rape seed protein, makes it to lose the restraining effect for protease activity.Na
2SO
3To the booster action of protease hydrolysis rapeseed meal the gel chromatography figure of hydrolysis curves and proteolysate confirm.Na
2SO
3Can also prevent polyphenol oxidation in hydrolytic process in the rapeseed meal as reductive agent, thereby make the rapeseed peptide (rsp) paler colour that obtains.In addition, the present invention adopts membrane separation technique can remove anti-nutrient substance and inorganic salt such as sulphur glucoside, phytic acid simultaneously after hydrolysis, reduces loss of proteins.
The object of the present invention is achieved like this: a kind of method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs, comprise utilize proteolytic enzyme to rapeseed meal be hydrolyzed, go out enzyme, centrifugally remove insolubles, again through ultrafiltration and the dry rapeseed peptide (rsp) that obtains, it is characterized in that: will rapeseed meal pulverize the back and add Na
2SO
3The aqueous solution obtains rapeseed peptide (rsp) through enzymic hydrolysis.
In the present invention: the process of specifically producing rapeseed peptide (rsp) is:
A) rapeseed meal was pulverized 60～100 mesh sieves, its Na2SO3 aqueous solution with 0.1%～0.6% is joined in the reaction vessel, making substrate concn is 5～10%w/v, mixes to be placed on the temperature of reaction of heating in water bath to proteolytic enzyme, and regulates the pH value according to the reaction conditions of proteolytic enzyme;
B) add proteolytic enzyme, making proteolytic enzyme and matrix ratio is 1～6%, and under agitation the hydrolysis rapeseed meal obtains the rape seed protein hydrolyzate;
C) regulate pH to 4.0～4.5 of hydrolyzate and keep the 30min enzyme that goes out, or be heated to 80 ℃～100 ℃ and keep 5～15min enzyme that goes out;
D) the centrifugal insolubles of removing of hydrolyzate is obtained supernatant liquid;
E) with supernatant liquid ultrafiltration and diafiltration, elimination anti-nutrient substance and inorganic salt;
F) solution that will be not see through filter membrane carries out spraying drying and obtains rapeseed peptide (rsp).
In the present invention: described proteolytic enzyme is Sumizyme MP, or neutral protease, or papoid, or flavor protease.
In the present invention: that uses filter membrane in the ultrafiltration holds back Fen Liang ≦ 5000Da; Described anti-nutrient substance is glucosinolate, phytic acid and polyphenols, and described inorganic salt are S-WAT and sodium-chlor.
In the present invention: when carrying out spraying drying, the inlet temperature of warm air is 180 ℃, and temperature out is 85 ℃.
The invention has the advantages that: because Na
2SO
3Can rupture between protein molecule and intramolecular disulfide linkage as a kind of reductive agent, make easier contact of post-rift rape seed protein molecule and improve enzymic hydrolysis speed and degree of hydrolysis, while Na with proteolytic enzyme
2SO
3The effect of fracture disulfide linkage has also destroyed the proteinase inhibitor molecule that exists in the rape seed protein, makes it to lose the restraining effect for protease activity, Na
2SO
3Can also prevent polyphenol oxidation in hydrolytic process in the rapeseed meal as reductive agent, thereby make the rapeseed peptide (rsp) paler colour of producing.
Description of drawings
Fig. 1 is process flow sheet of the present invention;
Fig. 2 is the hydrolysis curves of Alcalase hydrolysis rapeseed meal;
Fig. 3 is the gel chromatography figure of the rapeseed peptide (rsp) product produced of the present invention;
Fig. 4 is the gel chromatography figure of the rapeseed peptide (rsp) product produced of control group.
Embodiment
The present invention further specifies with the following example, but protection scope of the present invention is not limited to embodiment.
Embodiment 1
Rapeseed meal was pulverized 80 mesh sieves, again with 0.25% Na
2SO
3The aqueous solution joins in the reaction vessel together, places heating in water bath to 50 ℃, regulates pH to 8.0 with 4NNaOH, add Sumizyme MP Alcalase again in stirring hydrolysis down, the enzymic hydrolysis parameter is: substrate concn 5%w/v, and enzyme and matrix are than 3%, pH8.0,50 ℃ of temperature of reaction.It is constant that the NaOH that adds 4mol/L in the reaction process keeps the pH value, and with pH-stat method assaying reaction degree of hydrolysis (DH).With hydrochloric acid conditioned reaction thing pH to 4.3, and keep the 30min enzyme that goes out behind the reaction 120min in 50 ℃.Hydrolyzate is removed insolubles (dregs of rice slag) with the centrifugal 20min of 4000 * g, obtain supernatant liquid.With the supernatant liquid molecular weight cut-off is that the film of 3000Da carries out ultrafiltration.Membrane area is 0.1M
2, the temperature of ultrafiltrated is 400C, carries out diafiltration after the ultrafiltration again.The solution that will not see through filter membrane at last carries out spraying drying and obtains the rapeseed peptide (rsp) product.The inlet temperature of warm air is 180 during spraying drying
0C, temperature out is 85
0C (technical process of present embodiment such as accompanying drawing 1).
The index of the rapeseed peptide (rsp) product of producing by present embodiment is as follows:
| Outward appearance | Moisture/% | Protein content/% | Glucosinolate content/mg/g | Phytic acid content/% |
| Faint yellow powdery | 7.04 | 60.35 | 0.39 | 0.41 |
0.25% Na among the water replacement embodiment 1
2SO
3The aqueous solution is organized in contrast, carries out enzymic hydrolysis and hydrolysis aftertreatment by above-mentioned identical test conditions and step, and the rapeseed peptide (rsp) product that obtains is thing in contrast.
Add Na during hydrolysis
2SO
3(the present invention, down with) and do not add Na
2SO
3Two groups of rapeseed meal hydrolysis curves of (control group, down with) are recorded in the accompanying drawing 2 together, the hydrolysis time that shows from accompanying drawing 2 and the mutual relationship of degree of hydrolysis as seen, Na
2SO
3Sumizyme MP Alcalase hydrolysis rapeseed meal is had obvious booster action, and when hydrolysis 120min, the degree of hydrolysis of the present invention and control group is respectively 12.1% and 7.7%, and the former is 1.6 times of the latter.
Embodiment 3
The molecular weight distribution of the contrast of producing among rapeseed peptide (rsp) product that use gel chromatography mensuration embodiment 1 produces and the embodiment 2.In the present embodiment, used gel column be Superose 1210/300GL (10 * 300mm, GE Healthcare Bio-Science Co., Piscataway, NJ USA), uses UV-detector to detect in 215nm.Elutriant is the 50mM phosphate buffered saline buffer that pH equals 7 0.15MNaCl, and sample liquid is earlier through 0.45 μ M membrane filtration, and sample size is 500 μ l, and eluent flow rate is 0.8ml/min.Get bovine serum albumin (67000Da), cytopigment (12700Da), VB
12(1355Da) and Sleep-promoting factor B (612Da) be molecular weight standard.Test result is noted down respectively in accompanying drawing 3 and accompanying drawing 4.The above peak area of 10000Da only accounts for 7.8% in the accompanying drawing 3, and the above peak area of 10000Da has accounted for 33.5% in the accompanying drawing 4, and the rapeseed peptide (rsp) that the quantity of protein in the rapeseed peptide (rsp) product that the present invention produces and macromole peptide is produced less than control group is described.As seen, can obviously improve the degree of hydrolysis of rape seed protein, make the molecular weight of hydrolyzate reduce with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs.
During concrete enforcement, described proteolytic enzyme not only can adopt Sumizyme MP, can also select neutral protease, or papoid, or proteolytic enzyme such as flavor protease, when selecting different proteolytic enzyme, should be according to basic general knowledge well known in the art, working conditions according to different proteolytic enzyme, regulate the temperature and the pH value of rapeseed meal hydrolysis,, select suitable substrate concn and proteolytic enzyme and matrix ratio simultaneously within the scope of the claims to satisfy the condition that proteolytic enzyme carries out enzymic hydrolysis to rapeseed meal, give full play to the effect of proteolytic enzyme, improve the utilization ratio of proteolytic enzyme.When going out enzyme, the 30min enzyme that goes out is also kept in pH to 4.0～4.5 that can be by regulating hydrolyzate, or is heated to 80 ℃～100 ℃ and keep 5～15min enzyme that goes out.
Claims (5)
1, a kind of method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs, comprise utilize proteolytic enzyme to rapeseed meal be hydrolyzed, go out enzyme, centrifugally remove insolubles, produce rapeseed peptide (rsp) through ultrafiltration and drying again, it is characterized in that: rapeseed meal is pulverized the back add Na
2SO
3The aqueous solution obtains rapeseed peptide (rsp) through enzymic hydrolysis.
2, the method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs according to claim 1, it is characterized in that: the method for producing rapeseed peptide (rsp) is:
A) rapeseed meal was pulverized 60～100 mesh sieves, with itself and 0.1%～0.6% Na
2SO
3The aqueous solution joins in the reaction vessel together, and making substrate concn is 5～10%w/v, mixes to be placed on the temperature of reaction of heating in water bath to proteolytic enzyme, and regulates the pH value according to the reaction conditions of proteolytic enzyme;
B) add proteolytic enzyme, making proteolytic enzyme and matrix ratio is 1～6%, and under agitation the hydrolysis rapeseed meal obtains the rape seed protein hydrolyzate;
C) regulate pH to 4.0～4.5 of hydrolyzate and keep the 30min enzyme that goes out, or be heated to 80 ℃～100 ℃ and keep 5～15min enzyme that goes out;
D) the centrifugal insolubles of removing of hydrolyzate is obtained supernatant liquid;
E) with supernatant liquid ultrafiltration and diafiltration, elimination anti-nutrient substance and inorganic salt;
F) solution that will be not see through filter membrane carries out spraying drying and obtains rapeseed peptide (rsp).
3, the method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs according to claim 2, it is characterized in that: described proteolytic enzyme is Sumizyme MP, or neutral protease, or papoid, or flavor protease.
4, the method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs according to claim 2 is characterized in that: that uses filter membrane in the ultrafiltration holds back Fen Liang ≦ 5000Da; Described anti-nutrient substance is glucosinolate, phytic acid and polyphenols, and described inorganic salt are S-WAT and sodium-chlor.
5, according to the described method with the S-WAT accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs of one of claim 2～4, it is characterized in that: when carrying out spraying drying, the inlet temperature of warm air is 180 ℃, and temperature out is 85 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200910025324XA CN101509029A (en) | 2009-03-23 | 2009-03-23 | Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200910025324XA CN101509029A (en) | 2009-03-23 | 2009-03-23 | Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101509029A true CN101509029A (en) | 2009-08-19 |
Family
ID=41001561
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA200910025324XA Pending CN101509029A (en) | 2009-03-23 | 2009-03-23 | Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101509029A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676621A (en) * | 2012-04-28 | 2012-09-19 | 南京财经大学 | Antihypertensive rapeseed peptide and preparation method and application of antihypertensive rapeseed peptide |
| CN103710416A (en) * | 2013-12-20 | 2014-04-09 | 安徽省农业科学院农产品加工研究所 | Preparation method and applications of rapeseed cake-sourced metal chelating peptide and peptide metal chelate |
| CN103911420A (en) * | 2014-04-08 | 2014-07-09 | 南京财经大学 | Method for preparing rapeseed peptide through synergistic fermentation of lysozyme and rapeseed dregs |
| CN107400158A (en) * | 2017-09-28 | 2017-11-28 | 南京财经大学 | A kind of method that stepwise discretization prepares high yield anti-inflammatory rapeseed peptide (rsp) |
| CN107628876A (en) * | 2017-10-26 | 2018-01-26 | 安徽耘泰农业发展有限公司 | A kind of composite fertilizer's preparation method for improving rice yield |
| CN107673846A (en) * | 2017-10-26 | 2018-02-09 | 安徽耘泰农业发展有限公司 | A kind of high usage Rice composite fertilizer preparation method |
| CN111521666A (en) * | 2020-04-07 | 2020-08-11 | 清华大学 | Rapid hydrolysis analysis method for protein under high-temperature and high-pressure state |
-
2009
- 2009-03-23 CN CNA200910025324XA patent/CN101509029A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676621A (en) * | 2012-04-28 | 2012-09-19 | 南京财经大学 | Antihypertensive rapeseed peptide and preparation method and application of antihypertensive rapeseed peptide |
| CN102676621B (en) * | 2012-04-28 | 2014-03-19 | 南京财经大学 | Antihypertensive rapeseed peptide and preparation method and application of antihypertensive rapeseed peptide |
| CN103710416A (en) * | 2013-12-20 | 2014-04-09 | 安徽省农业科学院农产品加工研究所 | Preparation method and applications of rapeseed cake-sourced metal chelating peptide and peptide metal chelate |
| CN103710416B (en) * | 2013-12-20 | 2016-08-17 | 安徽省农业科学院农产品加工研究所 | Rapeseed cake source metal chelating peptide, the Preparation method and use of peptide metallo-chelate |
| CN103911420A (en) * | 2014-04-08 | 2014-07-09 | 南京财经大学 | Method for preparing rapeseed peptide through synergistic fermentation of lysozyme and rapeseed dregs |
| CN107400158A (en) * | 2017-09-28 | 2017-11-28 | 南京财经大学 | A kind of method that stepwise discretization prepares high yield anti-inflammatory rapeseed peptide (rsp) |
| CN107628876A (en) * | 2017-10-26 | 2018-01-26 | 安徽耘泰农业发展有限公司 | A kind of composite fertilizer's preparation method for improving rice yield |
| CN107673846A (en) * | 2017-10-26 | 2018-02-09 | 安徽耘泰农业发展有限公司 | A kind of high usage Rice composite fertilizer preparation method |
| CN111521666A (en) * | 2020-04-07 | 2020-08-11 | 清华大学 | Rapid hydrolysis analysis method for protein under high-temperature and high-pressure state |
| CN111521666B (en) * | 2020-04-07 | 2022-03-18 | 清华大学 | Rapid hydrolysis analysis method for protein under high-temperature and high-pressure state |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101509029A (en) | Method for accelerating production of rapeseed peptide with enzyme hydrolysis of rapeseed dregs by using sodium sulfite | |
| US8142832B2 (en) | Vegetable protein fractionization process and compositions | |
| NZ600548A (en) | Protein concentrates and isolates, and processes for the production thereof from toasted oilseed meal | |
| US4409248A (en) | Heat-gelling and foam-stabilizing enzymatically modified vegetable isolates | |
| CN102010460B (en) | Tea seed polypeptide and preparation method thereof | |
| EP2591683A3 (en) | Oilseed protein concentrates and isolates, and processes for the production thereof | |
| JPS5836345A (en) | Separation and isolation 7s and 11s protein from isoelectrically precipitated vegetable protein mixture | |
| CN101390564B (en) | Production method of corn separation protein | |
| CN100451022C (en) | Process for extracting vegetable seed protein and calcium phytate from vegetable seed cake | |
| CN111528332A (en) | Bird's nest peptide and preparation method and application thereof | |
| CN105918606A (en) | Extracting method of proteins in green tea dregs | |
| CN105420325A (en) | Placenta polypeptide preparation method | |
| CN104630319B (en) | The method that biological enzyme prepares medical bone gelatin | |
| WO2004104036A1 (en) | Process for producing soyeban whey protein and digested soybean whey protein | |
| CN101988080A (en) | Method for producing soybean small peptide | |
| Yust et al. | Improvement of protein extraction from sunflower meal by hydrolysis with alcalase | |
| CN107723328B (en) | Guar active peptide and preparation method thereof | |
| RU2233097C2 (en) | Method for producing of protein isolate from proteinaceous substance | |
| CN116035218A (en) | Pea oligopeptide health care product and preparation method thereof | |
| CN101487003B (en) | Acid protease | |
| CN1534093A (en) | Synthesis of batroxobin gene and purification preparation of its expresson product | |
| CN113151382B (en) | Method for producing wheat oligopeptide by precipitation method | |
| CN110241163B (en) | Method for extracting quinoa polypeptide by alkali extraction-membrane method | |
| CN103074404A (en) | Method of using compound enzyme for hydrolysis of whey protein and preparation of peptone | |
| CN101492664B (en) | Acid protease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20090819 |