CN106086132A - A kind of utilize ginkgo nut leather for the method for anti-oxidation peptide - Google Patents
A kind of utilize ginkgo nut leather for the method for anti-oxidation peptide Download PDFInfo
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- CN106086132A CN106086132A CN201610420906.8A CN201610420906A CN106086132A CN 106086132 A CN106086132 A CN 106086132A CN 201610420906 A CN201610420906 A CN 201610420906A CN 106086132 A CN106086132 A CN 106086132A
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- 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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Abstract
This application discloses and a kind of utilize ginkgo nut leather for the method for anti-oxidation peptide, specifically include following steps: (1), will put in NaOH aqueous solution through the Semen Ginkgo peel being dried, pulverize, extract Semen Ginkgo peel crude protein；(2), by Semen Ginkgo peel crude protein being formulated as mass concentration is 1 10%, and pH value is the aqueous solution of 9 10；Being subsequently adding protease, the temperature of regulation aqueous solution is 50 60 DEG C, is hydrolyzed after reacting 25 hours, then aqueous solution keeps at 95 100 DEG C 10 15min that seethe with excitement, and carries out enzyme denaturing, it is thus achieved that hydrolyzed solution；(3), by said hydrolyzed liquid centrifugation, after taking supernatant vacuum lyophilization, anti-oxidation peptide is prepared.Therefore the anti-oxidation peptide prepared by the present invention can use as antioxidant, it is higher to the clearance rate of DPPH free radical, when anti-oxidation peptide concentration in the solution reaches 0.4 more than g/L, stable more than 80% to the clearance rate of DPPH free radical, human body is had higher health-care effect.
Description
Technical field
The present invention relates to a kind of method that enzyme process prepares anti-oxidation peptide, especially one utilizes ginkgo nut leather for antioxidation
The method of peptide, belongs to biotechnology manufacture field.
Background technology
At present, the annual Semen Ginkgo total output of China is more than 1.2 ten thousand tons, and Folium Ginkgo yield reaches 1.5 ten thousand tons.And ginkgo nut
Skin yield is relatively big, reaches 1.5-2.0 ten thousand tons.Semen Ginkgo peel contains protein, fat, saccharide, tannin, trace element, aminoacid,
And flavonoid and phenolic acid, such as ginkgol, bilobol, ginkgolic acid, hydroginkgoic acid, hydroginkgolinic acid, Semen Ginkgo
Alcohol, its acidic materials are shikimic acid, D-saccharic acid, orange such as acid.
Research to Semen Ginkgo the most both at home and abroad is concentrated mainly on Folium Ginkgo, and ginkgo nut crust is main still with garbage
Form is thrown away, and is not effectively utilized, and has both wasted ample resources, has polluted again environment.Therefore Semen Ginkgo peel is comprehensive
Utilize, turn waste into wealth and just seem particularly important.At present both at home and abroad to the comprehensive utilization of Semen Ginkgo peel also in the blank stage, and
There is no the application of reality.
Containing higher protein in Semen Ginkgo peel, naturally after discharge, not only waste resource, also result in environment dirty simultaneously
Dye.The proposition of plant active peptides concept and the raising of separation detection technique, make people start gradually and pay attention to rich in proteins
The deep processing of agricultural byproducts also therefrom obtains various active peptide, such as phosphopeptide caseinate, blood pressure lowering peptide and absorption peptide easy to digest etc.,
The research and development of plant origin bioactive peptide has important function to the added value improving agricultural products in China deep processing.
Antioxidant the most at home and abroad develops quickly, and purposes is more and more wider.Antioxidant is applied not only to containing fat
The antioxidation of fat food, and the exploitation of health food and cosmetics etc. it is used for as functional factor.The kind of antioxidant is very
Many, but the antioxidant of chemosynthesis such as BHA, BHT etc., owing to itself having toxic and side effects, national governments are one after another to it
Its addition of mandatory provision.People progressively turn to the extraction Natural antioxidant from various plant and animal tissues sight.Anti-
Oxidation activity polypeptide is due to low toxicity, the feature such as efficiently, as the antioxidant of food and body it is considered to be synthetic antioxygen
The preferable replacer of agent.
Therefore, the present invention utilizes enzyme process preparation to have the polypeptide of antioxidant activity, for antioxidation with Semen Ginkgo peel for raw material
The exploitation of functional food provides theoretical foundation and technical support.
Summary of the invention
The purpose of the application is that providing a kind of utilizes ginkgo nut leather for the method for anti-oxidation peptide, specifically includes following step
Rapid:
(1), will put in NaOH aqueous solution through the Semen Ginkgo peel being dried, pulverize, extract Semen Ginkgo peel crude protein；
(2), by Semen Ginkgo peel crude protein being formulated as mass concentration is 1-10%, and pH value is the aqueous solution of 9-10；It is subsequently adding egg
White enzyme, the temperature of regulation aqueous solution is 50-60 DEG C, is hydrolyzed after reacting 2-5 hour, then is protected at 95-100 DEG C by aqueous solution
Hold boiling 10-15min, carry out enzyme denaturing, it is thus achieved that hydrolyzed solution；In this step, the aqueous solution that Semen Ginkgo peel crude protein is configured to is excellent
Choosing uses NaOH to regulate pH value；PH value after regulation most preferably 9；
(3), by said hydrolyzed liquid centrifugation, after taking supernatant vacuum lyophilization, anti-oxidation peptide is prepared.
Further, after being put in NaOH aqueous solution by Semen Ginkgo peel in step (1), the 1-5 that vibrates at 55-65 DEG C is little
Time, it is then centrifuged for precipitation, takes supernatant；By supernatant salt acid for adjusting pH value to 2-3, recentrifuge precipitates, by precipitation water
After washing, obtain Semen Ginkgo peel crude protein after drying；The concentration of described NaOH aqueous solution is 0.05-0.1mol/L；Ginkgo nut
Skin is 1:10-1:100 with the mass ratio of NaOH aqueous solution.After Semen Ginkgo peel is put in NaOH aqueous solution, during vibration
Optimum temperature is 60 DEG C.In above-mentioned steps (1), after being put in NaOH aqueous solution by Semen Ginkgo peel, temperature during vibration is preferred
It it is 60 DEG C；And after supernatant regulates with hydrochloric acid, its pH value most preferably 2.6.
Use the method to extract Semen Ginkgo peel crude protein, the albumen in Semen Ginkgo peel can be extracted to greatest extent
Come, and obtain the Semen Ginkgo peel crude protein that purity is higher.
The Main Function of NaOH is by the proteolytic in Semen Ginkgo peel out, is beneficial to extract, when the concentration of NaOH
After reaching finite concentration, the dissolution velocity for protein has increased limited, controls the concentration of NaOH at 0.05-0.1mol/
L, on the premise of ensureing that protein is dissolved out, reduces the consumption of NaOH.Suitable solid-to-liquid ratio can make in ginkgo nut skin
Protein can be extracted, and enters in NaOH aqueous solution, for avoiding the excessive concentration of protein in NaOH aqueous solution, and
Affecting the extraction ratio of protein in Semen Ginkgo peel, NaOH aqueous solution can not exceed very little certainly relative to the amount of Semen Ginkgo peel
After certain ratio, too much NaOH aqueous solution can not extract more protein, by Semen Ginkgo peel and NaOH solution
Mass ratio controls between 1:10-1:100, can ensure that the protein in Semen Ginkgo peel can be extracted as much as possible.
Further, described protease is alkaline protease.Through screening, alkaline protease is optimal for decomposition of protein
The protease of matter.
Further, the addition of protease is the 1-5% of Semen Ginkgo peel thick protein amount, more preferably 2-3%.This model
The consumption enclosing interior protease can preferably complete the decomposition of protein, it is not necessary to adds too much.
Anti-oxidation peptide prepared by the present invention can use as antioxidant, and it is to the clearance rate of DPPH free radical relatively
Height, when anti-oxidation peptide concentration in the solution reaches 0.4 more than g/L, to the clearance rate of DPPH free radical stable 80% with
On, human body is had higher health-care effect.
Detailed description of the invention
Embodiment 1
By Semen Ginkgo peel remove impurity, drying 12 hours, pulverize with pulverizer for 40 DEG C, taking 10kg, to put into 100kg NaOH water-soluble
In liquid, after vibrating 2 hours at 60 DEG C, centrifugation takes supernatant, then by supernatant salt acid for adjusting pH value to 2.6, so
Rear centrifugation, by sediment undergoes washing and prepare Semen Ginkgo peel crude protein after drying；Wherein, the concentration of NaOH aqueous solution is 0.08
mol/L。
Prepared Semen Ginkgo peel crude protein is made mass concentration be 1%, pH value be the aqueous solution of 9, be subsequently adding relatively
In Semen Ginkgo peel crude protein quality 1% alkaline protease, 50 DEG C be hydrolyzed reaction 2 hours after, aqueous solution is existed
Keep boiling 10 min at 100 DEG C, carry out enzyme denaturing, it is thus achieved that hydrolyzed solution.The aqueous solution that wherein Semen Ginkgo peel crude protein is made is used
NaOH regulates pH value.
By hydrolyzed solution centrifugation, take supernatant after vacuum lyophilization, prepare anti-oxidation peptide A.
Embodiment 2
By Semen Ginkgo peel remove impurity, dry 8 hours, pulverize with pulverizer, take 1kg and put into 100kg NaOH aqueous solution for 50 DEG C
In, vibrate after 5 hours at 50 DEG C, centrifugation takes supernatant, then by supernatant salt acid for adjusting pH value to 2, then from
The heart precipitates, by sediment undergoes washing and prepare Semen Ginkgo peel crude protein after drying；Wherein, the concentration of NaOH aqueous solution is 0.1 mol/
L。
Prepared Semen Ginkgo peel crude protein is made mass concentration be 10%, pH value be the aqueous solution of 10, be subsequently adding relatively
In the alkaline protease of the 2% of Semen Ginkgo peel thick protein amount, after being hydrolyzed react 5 hours at 60 DEG C, by aqueous solution at 95 DEG C
Lower holding 12 min, carries out enzyme denaturing, it is thus achieved that hydrolyzed solution.
By hydrolyzed solution centrifugation, take supernatant after vacuum lyophilization, prepare anti-oxidation peptide B.
Embodiment 3
By Semen Ginkgo peel remove impurity, dry 8 hours, pulverize with pulverizer, take 2kg and put into 100kg NaOH aqueous solution for 50 DEG C
In, vibrate after 3 hours at 55 DEG C, centrifugation takes supernatant, then by supernatant salt acid for adjusting pH value to 3, then from
The heart precipitates, by sediment undergoes washing and prepare Semen Ginkgo peel crude protein after drying；Wherein, the concentration of NaOH aqueous solution is 0.05
mol/L。
Prepared Semen Ginkgo peel crude protein is made mass concentration be 5%, pH value be the aqueous solution of 9.5, be subsequently adding relatively
In the alkaline protease of the 4% of Semen Ginkgo peel thick protein amount, after being hydrolyzed react 2 hours at 50 DEG C, by aqueous solution at 98 DEG C
Lower holding 11 min, carries out enzyme denaturing, it is thus achieved that hydrolyzed solution.
By hydrolyzed solution centrifugation, take supernatant after vacuum lyophilization, prepare anti-oxidation peptide C.
Embodiment 4
By Semen Ginkgo peel remove impurity, dry 12 hours, pulverize with pulverizer, take 1kg and put into 90kg NaOH aqueous solution for 40 DEG C
In, after vibrating 4 hours at 60 DEG C, centrifugation takes supernatant, then by supernatant salt acid for adjusting pH value to 2.6, then
Centrifugation, by sediment undergoes washing and prepare Semen Ginkgo peel crude protein after drying；Wherein, the concentration of NaOH aqueous solution is 0.09
mol/L。
Prepared Semen Ginkgo peel crude protein is made mass concentration be 8%, pH value be the aqueous solution of 9, be subsequently adding relative to
The alkaline protease of the 5% of Semen Ginkgo peel thick protein amount, after being hydrolyzed react 3 hours at 50 DEG C, by aqueous solution at 97 DEG C
Keep 15 min, carry out enzyme denaturing, it is thus achieved that hydrolyzed solution.
By hydrolyzed solution centrifugation, take supernatant after vacuum lyophilization, prepare anti-oxidation peptide D.
The anti-oxidation peptide obtained in embodiment is carried out Activity determination, uses the scavenging action to DPPH free radical to carry out table
Showing, concrete operations are as follows:
Anti-oxidation peptide is respectively prepared the sample of variable concentrations, takes 0.4 mL sample and isopyknic 40g/L DPPH the most respectively
Ethanol solution is sufficiently mixed, and under room temperature, lucifuge stands 30 min, measures light absorption value under wavelength 517 nm.Use deionized water
Sample is replaced to make blank and positive control respectively with reduced glutathion.The DPPH free radical scavenging vigor of sample is pressed
Row formula (1) calculates:
(1)
In formula, AiFor adding the light absorption value of the DPPH ethanol solution of sample；AjFor adding the ethanol solution of sample
Light absorption value；A0For substituting the light absorption value of the DPPH ethanol solution of sample with deionized water.
Testing result is shown in Table 1.
Table 1 anti-oxidation peptide elimination effect to DPPH free radical
| Sample concentration (g/L) | 0.1 | 0.2 | 0.4 | 0.6 | 0.8 | 1.0 |
| The anti-oxidation peptide A removing (%) to DPPH free radical | 49 | 62 | 87 | 86 | 87 | 87 |
| The anti-oxidation peptide B removing (%) to DPPH free radical | 45 | 60 | 82 | 81 | 82 | 88 |
| The anti-oxidation peptide C removing (%) to DPPH free radical | 47 | 61 | 85 | 84 | 84 | 87 |
| The anti-oxidation peptide D removing (%) to DPPH free radical | 50 | 63 | 87 | 87 | 86 | 87 |
| The reduced glutathion removing (%) to DPPH free radical | 69 | 83 | 91 | 92 | 93 | 93 |
This application discloses and a kind of utilize ginkgo nut leather for the method for anti-oxidation peptide, specifically include following steps: (1), will be through
Cross Semen Ginkgo peel dry, that pulverize and put in NaOH aqueous solution, extract Semen Ginkgo peel crude protein；(2), by thick for Semen Ginkgo peel egg
Being formulated as mass concentration in vain is 1-10%, and pH value is the aqueous solution of 9-10；Being subsequently adding protease, the temperature of regulation aqueous solution is
50-60 DEG C, it is hydrolyzed after reacting 2-5 hour, then aqueous solution is kept at 95-100 DEG C the 10-15min that seethes with excitement, go out
Enzyme, it is thus achieved that hydrolyzed solution；(3), by said hydrolyzed liquid centrifugation, after taking supernatant vacuum lyophilization, anti-oxidation peptide is prepared.Cause
Anti-oxidation peptide prepared by this present invention can use as antioxidant, and it is higher to the clearance rate of DPPH free radical, when
When anti-oxidation peptide concentration in the solution reaches 0.4 more than g/L, stable more than 80% to the clearance rate of DPPH free radical, right
Human body has higher health-care effect.
Claims (8)
1. one kind utilizes ginkgo nut leather for the method for anti-oxidation peptide, it is characterised in that comprise the steps:
(1), will put in NaOH aqueous solution through the Semen Ginkgo peel being dried, pulverize, extract Semen Ginkgo peel crude protein；
(2), by Semen Ginkgo peel crude protein being formulated as mass concentration is 1-10%, and pH value is the aqueous solution of 9-10；It is subsequently adding albumen
Enzyme, the temperature of regulation aqueous solution is 50-60 DEG C, is hydrolyzed after reacting 2-5 hour, then is kept at 95-100 DEG C by aqueous solution
Boiling 10-15min, carries out enzyme denaturing, it is thus achieved that hydrolyzed solution；
(3), by said hydrolyzed liquid centrifugation, after taking supernatant vacuum lyophilization, anti-oxidation peptide is prepared.
Method the most according to claim 1, it is characterised in that
After Semen Ginkgo peel is put in NaOH aqueous solution by step (1), vibrate 1-5 hour at 55-65 DEG C, be then centrifuged for sinking
Form sediment, take supernatant；By supernatant salt acid for adjusting pH value to 2-3, recentrifuge precipitates, after precipitation is washed with water, and drying
After obtain Semen Ginkgo peel crude protein；
The concentration of described NaOH aqueous solution is 0.05-0.1mol/L；Semen Ginkgo peel is 1:10-1 with the mass ratio of NaOH aqueous solution:
100。
Method the most according to claim 2, it is characterised in that in step (1), Semen Ginkgo peel is put into NaOH aqueous solution
After in, temperature during vibration is 60 DEG C.
Method the most according to claim 2, it is characterised in that in step (1), after supernatant regulates with hydrochloric acid, pH value is
2.6。
Method the most according to claim 1, it is characterised in that described protease is alkaline protease.
Method the most according to claim 1, it is characterised in that the addition of protease is Semen Ginkgo peel thick protein amount
1-5%。
Method the most according to claim 1, it is characterised in that in step (2), the water that Semen Ginkgo peel crude protein is configured to
Solution NaOH regulates pH value.
Method the most according to claim 1, it is characterised in that in step (2), the water that Semen Ginkgo peel crude protein is configured to
The pH value of solution is 9.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610420906.8A CN106086132A (en) | 2016-06-16 | 2016-06-16 | A kind of utilize ginkgo nut leather for the method for anti-oxidation peptide |
| PCT/CN2017/078555 WO2017215313A1 (en) | 2016-06-16 | 2017-03-29 | Method for the preparation of antioxidant peptide using gingko nut shells |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610420906.8A CN106086132A (en) | 2016-06-16 | 2016-06-16 | A kind of utilize ginkgo nut leather for the method for anti-oxidation peptide |
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| CN106086132A true CN106086132A (en) | 2016-11-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2017215313A1 (en) * | 2016-06-16 | 2017-12-21 | 如皋福大工程技术研究院有限公司 | Method for the preparation of antioxidant peptide using gingko nut shells |
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| CN111616362B (en) * | 2020-06-29 | 2022-03-08 | 山东省林业保护和发展服务中心 | Walnut male flower antioxidant and preparation method and application thereof |
| CN114774499A (en) * | 2022-04-24 | 2022-07-22 | 山东省农业科学院 | Extraction method and application of pearl oil apricot antioxidant peptide APHs-1-c |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102337318A (en) * | 2011-07-26 | 2012-02-01 | 南京林业大学 | Gingko antioxidant active peptide and preparation method thereof |
| CN106119324A (en) * | 2016-06-16 | 2016-11-16 | 如皋福大工程技术研究院有限公司 | A kind of utilize ginkgo nut leather for the method for calcium chelating peptide |
| CN106086132A (en) * | 2016-06-16 | 2016-11-09 | 如皋福大工程技术研究院有限公司 | A kind of utilize ginkgo nut leather for the method for anti-oxidation peptide |
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- 2016-06-16 CN CN201610420906.8A patent/CN106086132A/en active Pending
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017215313A1 (en) * | 2016-06-16 | 2017-12-21 | 如皋福大工程技术研究院有限公司 | Method for the preparation of antioxidant peptide using gingko nut shells |
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