CN103627764A - Method for preparing antioxidant peptide by stepwise enzymolysis of corn germ meal - Google Patents
Method for preparing antioxidant peptide by stepwise enzymolysis of corn germ meal Download PDFInfo
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- CN103627764A CN103627764A CN201310610305.XA CN201310610305A CN103627764A CN 103627764 A CN103627764 A CN 103627764A CN 201310610305 A CN201310610305 A CN 201310610305A CN 103627764 A CN103627764 A CN 103627764A
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- corn germ
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Abstract
The invention relates to a technology for preparing an antioxidant peptide by stepwise enzymolysis of corn germ meal. The method comprises the following steps: (1) crushing the corn germ meal; (2) carrying out stepwise enzymolysis on multienzyme; (3) carrying out centrifugal separation to obtain supernatant; (4) carrying out ultrafiltration and nanofiltration on the supernatant; and (5) drying to obtain a product. According to the method provided by the invention, corn germ oil byproducts are fully used to bring the corn germ protein function into full play. Byproducts in a corn deep processing process are effectively appreciated, and the social benefit and economic benefit are very significant.
Description
?
technical field:
The invention belongs to biological technical field, be specifically related to take corn germ cake as raw material, by the method for substep enzymolysis, prepare the technology of maize germ antioxidation biology bioactive peptide.
background technology:
Maize germ accounts for 11%～14% of corn kernel quality, contains in corn kernel 22% protein, 83% mineral substance and 84% fat.When producing corn starch sugar and corn industrial spirit, by wet method and dry method, put forward embryo technology, from corn kernel, rejected.The crude fat content of maize germ approximately 35%～56%, protein content 13-18%, is mainly as a kind of oil resource, for producing Fructus Maydis oil.Corn germ protein is high-quality protein, and essential amino acid rich content can compare favourably with soybean protein.
Corn germ cake is that maize germ is carried the remainder after oil, wherein contains the protein of 20% left and right and the carbohydrate of 40% left and right.Owing to carrying in embryo oil expression process, receive pyroprocessing, the sex change of most of protein, declines to a great extent corn germ protein function, and corn germ cake can only be as feed applications.China produces maize germ per year 800,000 tons, nearly 2,000,000 tons of the corn germ cake of generation, but almost completely for feed, thereby wasted a large amount of high-quality proteins.
Corn germ cake protein content is higher, though be denatured protein, its hydrolyzate can be described as biological oligopeptide, and the more former albumen of the albumen after sex change is easier to hydrolysis.Maize germ biologically active peptides is the polypeptide mixture of the certain molecular weight scope that forms after protease hydrolysis of corn germ protein, there is good solubility, mobility, thermostability, and its viscosity is low, concentration is high, absorb in body fast, utilization ratio is high, have oxidation-resistance, Ginseng Extract, reduction blood sugar concentration, hypotensive, promote unique physiologically actives such as alcohol metabolism (sobering up).
To sum up, the present invention will be take corn germ cake as raw material, by multienzyme substep enzymolysis, improve as far as possible productive rate and purity, the preparation of hydrolysis plumule cake protein is compared with high antioxidant biologically active peptides, thereby realized the further exploitation of maize germ resource, plumule nutritive value is more fully brought into play.
summary of the invention:
The object of this invention is to provide a kind of corn germ cake antioxidation biology bioactive peptide extracts and technology of preparing, the product of gained has higher purity and yield, and there is oxidation-resistance, Ginseng Extract, the physiological function such as hypotensive, the present invention by the following technical solutions:
Multienzyme substep enzymolysis corn germ cake is prepared a method for antioxidation biology bioactive peptide, comprises the following steps: (1) corn germ cake is pulverized substep enzymolysis (3) centrifugation supernatant liquor (4) the supernatant liquor ultrafiltration of (2) multienzyme and nanofiltration (5) is dried to obtain product.
Grinding particle size in described step (1) is: 80-120 order, this size range is to be both suitable for fully contacting of the plumule dregs of rice and water, makes again to pulverize the suitableeest scope of Energy Intensity Reduction.
Proteolytic enzyme in described step (2) is: papoid, neutral protease and Sumizyme MP, substep enzymolysis is sequentially neutral protease → Sumizyme MP → papoid or Sumizyme MP → papoid → neutral protease, and these two kinds of enzyme-added orders obtain peptides extraction rate higher than other enzyme-added order.
In described step (2), various protease hydrolyzed conditions are: papain enzymolysis temperature 55-65 ℃, and pH6-7, time 1-3h, enzyme concentration 0.05-0.5%(w/w, enzyme-added quality is with respect to raw materials quality per-cent), enzyme 500000u/g alive; 50～60 ℃ of Sumizyme MP hydrolysis temperatures, pH8-9, time 1-2h, enzyme concentration 0.1-1%(w/w), enzyme 200000u/g alive; Neutral protease enzymolysis temperature 40-50 ℃, pH7-7.8, time 2-3h, enzyme concentration 0.5-1%(w/w), enzyme 60000u/g alive; Material-water ratio is 1:12-1:8(w/v).
The middle centrifugation of described step (3) need make to precipitate moisture and be down to below 80%.
In described step (4), ultrafiltration needs 100,000 u and 5000u two times of ultrafiltration to remove the larger impurity of molecular weight; Nanofiltration needs 200u desalination.
In affiliated step (5), drying mode is dry for spraying, and inlet temperature is no more than 180 ℃, makes final moisture content below 10%.
Embodiment
Embodiment 1:
A corn germ cake antioxidation biology bioactive peptide technology of preparing, comprises the following steps: (1) carries corn germ cake grinding particle size after oil to granularity 120 orders through squeezing and solvent extraction.(2) press neutral protease → Sumizyme MP → papoid order enzymolysis.Papain enzymolysis temperature 60 C, pH6.5, time 2h, enzyme concentration 0.08%(w/w), enzyme 500000u/g alive; 55 ℃ of Sumizyme MP hydrolysis temperatures, pH9, time 2h, enzyme concentration 0.02%(w/w), enzyme 200000u/g alive; Neutral protease enzymolysis temperature 45 C, pH7.5, time 2h, enzyme concentration 0.6%(w/w), enzyme 60000u/g alive; Material-water ratio 1:8(w/v) (3) enzymolysis is complete, feed liquid is carried out centrifugal, makes to precipitate moisture and reaches 80%.(4) centrifuged supernatant is respectively through 100,000 u and 5000u ultrafiltration and the spray-dried product that obtains of the 200u nanofiltration removal of impurity (5) nanofiltration concentrated solution.
Embodiment 2:
A corn germ cake antioxidation biology bioactive peptide technology of preparing, comprises the following steps: (1) carries corn germ cake grinding particle size after oil to granularity 100 orders through squeezing and solvent extraction.(2) press Sumizyme MP → papoid → neutral protease order enzymolysis.Papain enzymolysis temperature 60 C, pH6.5, time 2h, enzyme concentration 0.1%(w/w), enzyme 500000u/g alive; 60 ℃ of Sumizyme MP hydrolysis temperatures, pH8.5, time 1h, enzyme concentration 0.5%(w/w), enzyme 200000u/g alive; Neutral protease enzymolysis temperature 45 C, pH7, time 2h, enzyme concentration 1%(w/w), enzyme 60000u/g alive; Material-water ratio 1:10(w/v) (3) enzymolysis is complete, it is carried out centrifugal, makes to precipitate moisture and reaches 70%.(4) centrifuged supernatant is respectively through 100,000 u and 5000u ultrafiltration and the spray-dried product that obtains of the 200u nanofiltration removal of impurity (5) nanofiltration concentrated solution.
The composition measurement of raw material and product:
Corn germ cake peptide content and extraction yield are measured and are pressed GB 5009.5-2010 mensuration bioactive peptide purity
Bioactive peptide yield is calculated and is pressed formula: P=m/M
Wherein: P-bioactive peptide extraction yield
Bioactive peptide quality/g in m-product
Protein mass/g in M-raw material
GB 5009.3-2010 is pressed in moisture determination
GB 5009.4-2010 is pressed in determination of ash
Carbohydrate content is measured: anthrone colorimetry: carbohydrate can be dewatered by vitriol oil effect and generate after furfural or hydroxymethylfurfural, with anthrone (C under comparatively high temps
14h
10o) dehydrating condensation, the derivative of formation furfural, is blue-greenish colour.This material has maximum absorption at 620 nm places, and within the scope of 150 μ g/ml, the depth of its color is directly proportional to soluble sugar content.
Little peptide molecular weight is measured: adopt SDS-PAGE gel electrophoresis, and acrylamide concentration 15%, albumen Marker molecular weight is respectively 66,45,35,27,20,14.4,9.5,6.5,4.1ku.
Oxidation-resistance measuring method is: take dehydrated alcohol as solvent, configuration concentration is 0.1mmol/L DPPH solution.Preparation massfraction is 1% corn germ cake peptide solution, gets 2ml solution, with distilled water, is settled to 100ml.Get the solution after 2ml constant volume, add the DPPH ethanolic soln preparing, mix, dark place's lucifuge reaction 30min, the sample of take adds the mixed solution of 2ml ethanol as blank, adjusts photometer zero point, in 517nm place reader absorbancy, is At, meanwhile, surveying the mixed absorbancy of 2mlDPPH+2ml distilled water is Ao.Calculate as follows clearance rate: clearance rate (%)=[1 one At/Ao] * 100%.
Table 1 raw material and product composition content
| ? | Albumen and little peptide | Moisture | Ash content | Carbohydrate | Yield |
| Raw material | 18.4% | 8.7% | 15.4% | 52.5% | ? |
| Embodiment 1 | 75% | 8.2% | 6.5% | 10% | 82% |
| Embodiment 2 | 72% | 8% | 7.7% | 9.8% | 85% |
From table 1, after enzymolysis purifying, little peptide content significantly improves, and little peptide purity and extraction yield all reach higher level.By electrophorogram, make Marker molecular weight logarithm and relative mobility relational expression, thus the molecular weight that calculates plumule dregs of rice peptide between 4.5-5.6 ku, and major part concentrates on 5.2 ku.By measuring, DPPH clearance rate is respectively 48.5% and 49.1%, illustrates that corn germ cake peptide has higher oxidation-resistance.
Claims (7)
1. substep enzymolysis corn germ cake is prepared a method for antioxidation biology bioactive peptide, comprises the following steps: (1) corn germ cake is pulverized substep enzymolysis (3) centrifugation supernatant liquor (4) the supernatant liquor ultrafiltration of (2) multienzyme and nanofiltration (5) is dried to obtain product.
2. as claimed in claim 1, multienzyme substep enzymolysis corn germ cake is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: described step (1) corn germ cake grinding particle size is 80-200 order.
3. as claimed in claim 1, multienzyme substep enzymolysis corn germ cake is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: it is papoid, neutral protease and Sumizyme MP that described step (2) is used proteolytic enzyme.
4. as claimed in claim 1, multienzyme substep enzymolysis corn germ cake is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: (2) three kinds of enzyme-added orders of enzyme of described step: neutral protease → Sumizyme MP → papoid or Sumizyme MP → papoid → neutral protease.
5. multienzyme substep enzymolysis corn germ cake as claimed in claim 1 is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: three kinds of protease hydrolyzed conditions in described step (2): papain enzymolysis temperature 55-65 ℃, pH6-7, time 1-3h, enzyme concentration 0.05-0.5%(w/w, be that enzyme-added quality is with respect to raw materials quality per-cent), enzyme 500000u/g alive; 50～60 ℃ of Sumizyme MP hydrolysis temperatures, pH8-9, time 1-2h, enzyme concentration 0.1-1%(w/w), enzyme 200000u/g alive; Neutral protease enzymolysis temperature 40-50 ℃, pH7-7.8, time 2-3h, enzyme concentration 0.5-1%(w/w), enzyme 60000u/g alive; Material-water ratio is 1:12-1:8(w/v).
6. multienzyme substep enzymolysis corn germ cake as claimed in claim 1 is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: described step (4) ultra-filtration membrane molecular weight is 100,000 u and 5000u; Nanofiltration membrane molecular weight is 200u.
7. multienzyme substep enzymolysis corn germ cake as claimed in claim 1 is prepared the method for antioxidation biology bioactive peptide, it is characterized in that: described step (5) drying means is dry for spraying, and dry temperature in is no more than 180 ℃.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073540A (en) * | 2014-07-05 | 2014-10-01 | 广东润科生物工程有限公司 | Polypeptide production method for enhancing utilization ratio of protein |
| CN104544080A (en) * | 2015-01-09 | 2015-04-29 | 孙欣 | Production process of composite germ powder |
| CN110387395A (en) * | 2019-08-23 | 2019-10-29 | 武汉轻工大学 | The preparation method and selenium-rich solid beverage of mushroom with abundant selenium protein peptide powder |
Citations (3)
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|---|---|---|---|---|
| CN1396175A (en) * | 2002-07-15 | 2003-02-12 | 徐昌洪 | Process for extracting both active embryonic peptide and natural corn oil from embryo bud of corn |
| CN101914604A (en) * | 2010-08-02 | 2010-12-15 | 江南大学 | Antioxidant peptide active protection in protein enzymolysis process and preparation method thereof |
| CN102018092A (en) * | 2010-12-21 | 2011-04-20 | 山东省鲁洲食品集团有限公司 | Method for extracting protein from maize germ dregs |
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- 2013-11-26 CN CN201310610305.XA patent/CN103627764A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1396175A (en) * | 2002-07-15 | 2003-02-12 | 徐昌洪 | Process for extracting both active embryonic peptide and natural corn oil from embryo bud of corn |
| CN101914604A (en) * | 2010-08-02 | 2010-12-15 | 江南大学 | Antioxidant peptide active protection in protein enzymolysis process and preparation method thereof |
| CN102018092A (en) * | 2010-12-21 | 2011-04-20 | 山东省鲁洲食品集团有限公司 | Method for extracting protein from maize germ dregs |
Non-Patent Citations (2)
| Title |
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| 王晓杰 等: "双酶复合水解对玉米肽抗氧化活性的影响", 《食品与发酵工业》 * |
| 王霞 等: "玉米胚芽蛋白水解物的抗氧化活性研究", 《中国酿造》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073540A (en) * | 2014-07-05 | 2014-10-01 | 广东润科生物工程有限公司 | Polypeptide production method for enhancing utilization ratio of protein |
| CN104544080A (en) * | 2015-01-09 | 2015-04-29 | 孙欣 | Production process of composite germ powder |
| CN110387395A (en) * | 2019-08-23 | 2019-10-29 | 武汉轻工大学 | The preparation method and selenium-rich solid beverage of mushroom with abundant selenium protein peptide powder |
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Application publication date: 20140312 |