CN109097424A - A kind of preparation method of anti-oxidant soy peptide powder - Google Patents
A kind of preparation method of anti-oxidant soy peptide powder Download PDFInfo
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- CN109097424A CN109097424A CN201810926514.8A CN201810926514A CN109097424A CN 109097424 A CN109097424 A CN 109097424A CN 201810926514 A CN201810926514 A CN 201810926514A CN 109097424 A CN109097424 A CN 109097424A
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- oxidant
- feed liquid
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- soy peptide
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
Abstract
The invention discloses a kind of preparation methods of anti-oxidant soy peptide powder, using Soybean Meal and/or soybean protein isolate as raw material, it is digested using compound protease system, and is main indicator to influencing the enzymolysis time of enzymolysis efficiency, enzymatic hydrolysis PH, enzyme concentration, hydrolysis temperature, concentration of substrate etc. because optimizing for element using inoxidizability.Soy peptide powder produced by the present invention has very strong inoxidizability, can be used for making various cosmetics, healthy food, special medicine formula food and health care product.
Description
Technical field
The invention belongs to soybean further process and field of comprehensive utilization, and in particular to a kind of preparation method of soy peptide powder.
Background technique
Protein is mainly absorbed in the form of peptide and free amino acid in human body as nutriment, and the suction of peptide
Receipts mechanism is different from amino acid again, has the advantages that amino acid and protein is incomparable.Peptide is two or more ammonia
The compound that base acid is connected with peptide bond, is the intermediate material between amino acid and protein, is the function fragment and knot of protein
Tile section is the active gene part of protein, is the nutriment and basic substance of life.Soybean peptide is high-quality protein,
The ratio of each amino acid is in admirable proportion, so a kind of product of the soybean peptide as high nutrition surcharge, in recent years by
Social extensive concern, and production technology is constantly updating, and functional and safety is stepped up.
Soybean peptide has the characteristics such as good, the strong, hypoallergenic of water suction moisture holding capacity of highly dissoluble, low viscosity, stability, makes it
It is upper very flexibly and extensive in application；It has been investigated that soybean peptide relieve fatigue, enhance muscular endurance, promote fat metabolism,
Aided blood pressure-lowering, reducing blood lipid, blood pressure lowering etc. have good effect, and future can become main in special medicine food
Protein (amino acid) source.So the soybean peptide for how producing higher quality is the important research and development of current Health Products Enterprises
One of project.
On the other hand, human body is because of the continuous contact with the external world, including breathing (oxidation reaction), outside contamination, radioactive ray
The factors such as irradiation constantly generate free radicals in body.When interior free yl is more than a certain amount, free radical will be gone
Attack cells film reacts with serum antiprotease, or even goes to rob electronics with gene, to cause to hurt to our body
Evil, generate various difficult and complicated cases, scientific investigations showed that, cancer, aging or Other diseases mostly with excessive free radicals
It generates relevant.Study it is anti-oxidant can effectively overcome harm brought by its, so anti-oxidant looked forward to by health food, cosmetics
Industry is classified as one of most important functional demand of one of main R&D direction and market.
Summary of the invention
The present inventor for the demand to soybean peptide and antioxidant food in the market, have developed it is a kind of with Soybean Meal and/or
Soybean protein isolate is raw material, the method for preparing anti-oxidant soy peptide powder with enzymatic isolation method, and the soy peptide powder produced has very
Strong inoxidizability improves the functionality and nutritive value of product, can be in anti-oxidant healthy food, cosmetics, special medicine
It is widely applied with fields such as formula foods.
The preparation method for realizing a kind of anti-oxidant soy peptide powder that the object of the invention provides, includes the following steps:
Step 1, feed liquid is configured:
Using Jing Guo pulverization process Soybean Meal and/or soybean protein isolate as raw material, according to concentration of substrate be 3%~15% be added go
Ionized water stirs evenly, and obtains feed liquid；
Step 2, feed liquid is handled:
Feed liquid prepared by step 1 is heated to 90 DEG C, keeps 10min, is cooled to room temperature；
Step 3, composite protease hydrolysis is handled:
The feed liquid PH that step 2 obtains is adjusted to 6.5~9.5,0.02~0.08 gram of compound egg then is added by every 100 grams of raw materials
Feed liquid is heated to 55 DEG C~75 DEG C, 80min~160min is hydrolyzed by white enzyme；
Step 4, purifying filtering:
The above-mentioned feed liquid handled by protease hydrolytic is heated to 90 DEG C of holding 10min, carries out destroy the enzyme treatment, then with centrifugation
Machine separation, obtains supernatant；
Supernatant is heated to 50 DEG C~70 DEG C again, clarification hydrolyzate is obtained by filtration by membrane filter system, finally uses gel color
Column refined filtration is composed, obtains molecular weight in 3000Da oligopeptide solution below；
Step 5: it is concentrated and dried:
By above-mentioned oligopeptide solution, after low temperature is concentrated and is spray-dried, obtained dry matter is anti-oxidant soybean peptide
Powder.
In the above-mentioned methods, the membrane filter system is composed of 0.45 μM of filtration membrane and plate filter.
In the above-mentioned methods, the compound protease is made of alkali protease, plant rennet, flavor protease,
The weight ratio of said components is 2:1:1~4:3:1.
The oligopeptide solution of the method for the present invention step 4 preparation, the clearance rate to DPPH free radical are 11.3%~17.08%,
Antioxidant activity is high.
The method of the present invention it is profitable:
A kind of preparation method for anti-oxidant soy peptide powder that the present invention researches and develops, using Soybean Meal and/or soybean protein isolate as raw material,
It does not need to add any free radical scavenger (antioxidant) during the preparation process, the soy peptide powder being prepared being prepared
With very strong inoxidizability, can be answered extensively in fields such as anti-oxidant healthy food, cosmetics, special medicine formula foods
With.The present invention also improves the recycle value of Soybean Meal and the nutritive value of soybean protein isolate simultaneously.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
Embodiment 1
A kind of preparation method of anti-oxidant soy peptide powder provided by the invention, includes the following steps:
Step 1, feed liquid is configured:
Using the Soybean Meal Jing Guo pulverization process as raw material, it is 15% addition deionized water according to concentration of substrate, stirs evenly, obtain
Feed liquid；
Step 2, feed liquid is handled:
Feed liquid prepared by step 1 is put into 90 DEG C of water-baths, is kept 10min, is cooled to room temperature；
Step 3, composite protease hydrolysis is handled:
The feed liquid PH that step 2 obtains is adjusted to 6.5~9.5,0.02~0.08 gram of compound egg then is added by every 100 grams of raw materials
White enzyme is heated to 65 DEG C in thermostat water bath, 160min is hydrolyzed；
Step 4, purifying filtering:
It will be placed in water-bath by the feed liquid of protease hydrolytic processing, be heated to 90 DEG C of holding 10min and carry out destroy the enzyme treatment, so
After be put in centrifuge, be centrifuged 20min under the revolving speed of centrifuge 4000r/ min, abandon precipitating, obtain supernatant；
Supernatant is heated to 55 DEG C again, is filtered with 0.45 μM of membrane filter system, clarification hydrolyzate is obtained；Gel chromatographic columns are used again
Refined filtration obtains molecular weight in 3000Da oligopeptide solution below；
Step 5, it is concentrated and dried:
The oligopeptide solution that step 4 is obtained is concentrated and is spray-dried by low temperature, obtained dry matter, as anti-oxidant big
Beans Gly-His-Lys.
In the preparation step of the anti-oxidant soy peptide powder of the present invention, the oligopeptide solution that can be prepared to step 4 is hydrolyzed
The measurement of the clearance rate of the measurement and DPPH free radical of degree, to ensure to obtain the anti-oxidant soy peptide powder of high-quality.Specifically
Measuring method is as follows:
(1) to the measurement of the oligopeptide solution degree of being hydrolyzed prepared in step 4
By oligopeptide solution obtained in step 4, its degree of hydrolysis is measured using PH-stat method,
The calculation formula of degree of hydrolysis is as follows:
N: concentration of lye (1M NaOH)
B: lye dosage, ml
1/: correction coefficient
M: hydrolyzed albumen quality, g(soybean meal amyloid proteins content 50%)
Htot: the quantity (soy proteinaceous Htot, value 7.75) of peptide bond in unit mass albumen
The degree of dissociation of α amino is calculated as follows:
P K is the dissociation constant of α amino in formula, is generally calculated with 7.0 for average value.
In order to measure the content of polypeptide in soy hydrolysate peptide, understands the ability of such method enzymatic hydrolysis dregs of beans, can be used to lower section
Method measures content of peptides in hydrolyzate:
(1) Coomassie brilliant blue reagent is added in supernatant, 595nm measures light absorption value, then measures standard protein data, is made
Standard curve searches the content of sample polypeptide on curve；
(2) the specific mensuration program of content of peptides is as follows, sample solution 0.05ml, and 0.95ml deionized water is added, and 5ml is added and examines
Mas bright blue reagent mixes, and 595nm surveys light absorption value；
(3) prepared by standard curve: taking 14 test tubes, sequentially adds standard BSA solution 0,0.1,0.2,0.4,0.6,0.8,1ml.
1ml is complemented to deionized water respectively, 5ml Coomassie brilliant blue reagent is added in each pipe, mixes, and 595nm surveys light absorption value.Contained with albumen
Amount is ordinate, and light absorption value is abscissa, makes standard curve, obtains regression equation Y=0.1244X-0.0739. according to formula
It calculates, obtained peptide concentration is 4.749mg/ml, percent hydrolysis 32%.
(2) measurement of the clearance rate of DPPH free radical is carried out to the oligopeptide solution prepared in step 4
The methanol or ethanol solution of DPPH free radical are in royal purple, and have maximum absorption peak in 519nm range, when to DPPH
When free radical scavenger (antioxidant) being added in free-atom aqueous solution, Cheng Dan is paired, and the DPPH free radical of darkviolet is reduced
Yellowly DPPH-H molecule, the absorption value at 519nm wavelength reduce, and reduction degree is with the electron amount that is received at quantitative pass
System, thus quantitative analysis can be carried out by the variation of absorbance.
(1) it weighs a certain amount of DPPH to be dissolved in dehydrated alcohol, mix well, be made into 0.1mmol/l solution, be kept in dark place；
(2) concentration of oligopeptide solution is adjusted to 0.5mg/ml, and 2.8mlDPPH solution, room temperature is added in 0.1ml oligopeptide solution
It is protected from light reflection 30min, light absorption value Ai is measured at 517nm.
Blank group: oligopeptide 0.1ml+2.8ml ethyl alcohol, blank group light absorption value Aj；
Control group: water 0.1ml+2.8mlDPPH solution, control group light absorption value Ao；
Blank zeroing group: 1:1 water+ethyl alcohol
Clearance rate=(1-(Ai-Aj)/Ao) * 100%
The clearance rate that such method measures is 11.36%.
The present embodiment passes through said determination using Soybean Meal as raw material, it can be deduced that the soy peptide powder of the method for the present invention preparation,
With very strong anti-oxidant.
Embodiment 2
Step 1, sample liquid configures:
Using soybean protein isolate as raw material, which is added deionized water according to concentration of substrate 3%, stirs evenly, is expected
Liquid；
Step 2, feed liquid is handled:
The feed liquid that step 1 is obtained is put into 90 DEG C of water-baths, and heating water bath 10min is cooled to room temperature；
Step 3, protease hydrolytic is handled:
The material liquid PH value that step 2 is obtained is adjusted to 8.5, and 65 DEG C are heated in thermostat water bath, is added according to every 100 grams of raw materials
Enter 0.06 gram of compound protease to be hydrolyzed, stops reaction when hydrolyzing 120min；
Step 4, purifying filtering:
It will be placed in water-bath by the feed liquid of protease hydrolytic processing, be heated to 90 DEG C of holding 10min and carry out destroy the enzyme treatment, so
After be put in centrifuge, be centrifuged 20min under the revolving speed of centrifuge 4000r/ min, abandon precipitating, obtain supernatant；
Supernatant is heated to 55 DEG C again, is filtered with 0.45 μM of membrane filter system, clarification hydrolyzate is obtained；Gel chromatographic columns are used again
Refined filtration obtains molecular weight in 3000Da oligopeptide solution below；
Step 5: it is concentrated and dried:
By above-mentioned oligopeptide solution, after low temperature is concentrated and is spray-dried, obtained dry matter, as inoxidizability soybean
Gly-His-Lys.
In the present embodiment, the clearance rate of the oligopeptide solution degree of being hydrolyzed and DPPH free radicals that prepared to step 4
Measuring method is using the measuring method provided in embodiment 1.
In the present embodiment, the degree of hydrolysis of oligopeptide solution prepared by step 4, according to the measuring method provided in embodiment 1,
Obtained peptide concentration is 0.874mg/ml, percent hydrolysis 29%.
In the present embodiment, the clearance rate of the DPPH free radical of oligopeptide solution prepared by step 4, according to being mentioned in embodiment 1
The measuring method of confession measures its Scavenging activity to DPPH free radical: obtained clearance rate is 17.08%.
The present embodiment passes through said determination using soybean protein isolate as raw material, it can be deduced that the method for the present invention is prepared big
Beans Gly-His-Lys, have it is very strong anti-oxidant, can be wide in fields such as anti-oxidant healthy food, cosmetics, special medicine formula foods
General application.
Above-described embodiment is only used as the explanation to the method for the present invention, technical solution of the present invention can there are many embodiment,
No further details to be given herein, and all transformation using equivalent technologies mean technical solution obtained belongs to the claims in the present invention
The range of protection.
Claims (4)
1. a kind of preparation method of anti-oxidant soy peptide powder, it is characterized in that including the following steps:
Step 1, configure feed liquid: using Jing Guo pulverization process Soybean Meal and/or soybean protein isolate as raw material, according to concentration of substrate
Deionized water is added for 3%~15%, stirs evenly, obtains feed liquid；
Step 2, feed liquid is handled: feed liquid prepared by step 1 is heated to 90 DEG C, keeps 10min, is cooled to room temperature；
Step 3, composite protease hydrolysis is handled: the feed liquid PH that step 2 obtains being adjusted to 6.5~9.5, then presses every 100 grams
0.02~0.08 gram of compound protease is added in raw material, and feed liquid is heated to 55 DEG C~75 DEG C, 80min~160min is hydrolyzed；
Step 4, purifying filtering: the above-mentioned feed liquid handled by protease hydrolytic is heated to 90 DEG C of holding 10min, carries out enzyme deactivation
Processing, is then separated with centrifuge, obtains supernatant；
Supernatant is heated to 50 DEG C~70 DEG C again, clarification hydrolyzate is obtained by filtration by membrane filter system, finally uses gel color
Column refined filtration is composed, obtains molecular weight in 3000Da oligopeptide solution below；
Step 5, it is concentrated and dried: above-mentioned oligopeptide solution is obtained into anti-oxidant soybean after low temperature is concentrated and is spray-dried
Gly-His-Lys.
2. the preparation method of a kind of anti-oxidant soy peptide powder according to claim 1, it is characterized in that: the compound protein
Enzyme is made of alkali protease, plant rennet, flavor protease, and the weight ratio of said components is 2:1:1~4:3:1.
3. the preparation method of a kind of anti-oxidant soy peptide powder according to claim 1, it is characterized in that: the film filtering is set
It is standby, it is composed of 0.45 μM of filtration membrane and plate filter.
4. a kind of preparation method of anti-oxidant soy peptide powder according to claim 1, it is characterized in that: prepared in step 4
Antioxygen oligopeptide solution, the clearance rate to DPPH free radical are 11.3%~17.08%.
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| CN201810926514.8A CN109097424A (en) | 2018-08-15 | 2018-08-15 | A kind of preparation method of anti-oxidant soy peptide powder |
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| CN201810926514.8A CN109097424A (en) | 2018-08-15 | 2018-08-15 | A kind of preparation method of anti-oxidant soy peptide powder |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020173112A1 (en) * | 2019-02-28 | 2020-09-03 | 江南大学 | Active peptide having anti-oil oxidation function, preparation method therefor and use thereof |
| CN114717285A (en) * | 2022-04-12 | 2022-07-08 | 重庆大学 | Method for extracting soybean antioxidant peptide by enzymolysis method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1593646A (en) * | 2004-07-16 | 2005-03-16 | 吉林大学 | Soybean peptide, its preparation and application |
| CN1932027A (en) * | 2006-09-20 | 2007-03-21 | 吉林农业大学 | Double enzyme hydrolysis process for preparing soybean peptide without bitter |
-
2018
- 2018-08-15 CN CN201810926514.8A patent/CN109097424A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1593646A (en) * | 2004-07-16 | 2005-03-16 | 吉林大学 | Soybean peptide, its preparation and application |
| CN1932027A (en) * | 2006-09-20 | 2007-03-21 | 吉林农业大学 | Double enzyme hydrolysis process for preparing soybean peptide without bitter |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020173112A1 (en) * | 2019-02-28 | 2020-09-03 | 江南大学 | Active peptide having anti-oil oxidation function, preparation method therefor and use thereof |
| US11319569B2 (en) | 2019-02-28 | 2022-05-03 | Jiangnan University | Preparation method and application thereof of peptides with anti-lipid-oxidation capability |
| CN114717285A (en) * | 2022-04-12 | 2022-07-08 | 重庆大学 | Method for extracting soybean antioxidant peptide by enzymolysis method |
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Effective date of registration: 20191203 Address after: 409, floor 4, building 30, Ruiqi Jiayuan, Huilongguan town, Changping District, Beijing 102200 Applicant after: Beijing Tengyue High Tech Co.,Ltd. Address before: 466600, Henan, Zhoukou province Xihua AI Gang county, Pan South executive Village Pan Village Applicant before: Yan Yunxia |
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Application publication date: 20181228 |