CN106434810A - Preparation method of cordyceps militaris polypeptide dried powder - Google Patents
Preparation method of cordyceps militaris polypeptide dried powder Download PDFInfo
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- CN106434810A CN106434810A CN201611015400.5A CN201611015400A CN106434810A CN 106434810 A CN106434810 A CN 106434810A CN 201611015400 A CN201611015400 A CN 201611015400A CN 106434810 A CN106434810 A CN 106434810A
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- Prior art keywords
- cordyceps militaris
- link
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- polypeptide
- dry powder
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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
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
Abstract
The invention relates to a preparation method of cordyceps militaris polypeptide dried powder. The preparation method comprises the following steps: crushing dried cordyceps militaris into powder; adding water with a certain volume and immersing for 1h to enable the dried cordyceps militaris to be sufficiently swollen; adding two types of proteinase according to a certain ratio in sequence and carrying out enzymolysis; after sufficiently carrying out the enzymolysis, raising the temperature to inactivate the proteinase; centrifuging and taking supernatant; concentrating in vacuum; carrying out ultrafiltration on a concentrated solution; performing spray-drying to obtain the cordyceps militaris polypeptide dried powder. By utilizing two types of biological enzymes to carry out the enzymolysis, the efficiency is high, the process is simple and the time is saved when the preparation method is compared with outdated water extract and ethanol precipitation methods, acid soaking extracting methods and the like. Meanwhile, the index of extracted cordyceps militaris polypeptide is high and can reach 90 percent or more; raw materials can be provided for the fields of cosmetics, food industries and medicines, and an additional value of the cordyceps militaris can also be improved; the cordyceps militaris polypeptide dried powder can be produced in an amplified large scale and the practicability is great.
Description
Technical field
Present invention relates particularly to a kind of preparation method of Cordyceps militaris (L.) Link. polypeptide dry powder.
Background technology
Cordyceps militaris (L.) Link., also known as Cordyceps militaris (L.) Link., Cordyceps militaris, belong to xenogenesis together with Cordyceps.Containing rich in protein, fat
Fat, cordycepic acid, cordycepin, Cordyceps polysaccharide, SOD enzyme and some trace element.Have and effectively the becoming of Cordyceps fairly similar
Divide and medicinal efficacy, such as beneficial vital essence, tonifying deficiency, antibacterial, blood pressure lowering, calmness, cooling, immunization etc..
Polypeptide is that body completes a kind of requisite material of various physiologically actives, is the rudimentary form of protein, is giving birth to
Also play a part of in the metabolic activity of thing and disease treatment not replacing.It is the important chemistry that links up between cell, organ
Courier, the life process such as scalable growth, development, breeding metabolism.Biologically active polypeptide it need not digest, directly absorb, and
And absorption is fast and complete, belongs to active, preferential absorption, does not expend body energy during absorption.
Cordyceps militaris (L.) Link. contains abundant amino acid and protein, and protein content accounts for 30% or so, is the strong work of Cordycepses tonification
With with one of the material base for strengthening immunologic function.High efficiency extraction Cordyceps militaris (L.) Link. protein polypeptide, can be cosmetics, food service industry and
Field of medicaments provides raw material, can lift the added value of Cordyceps militaris (L.) Link. again, but Hot water extraction of the prior art, and acidleach lifting manipulation etc. is carried
Take purity relatively low, relatively costly.
Content of the invention
The present invention is directed to above-mentioned problems of the prior art, it is therefore an objective to provide a kind of Cordyceps militaris (L.) Link. polypeptide biological dual life
Thing enzymatic extraction method and the preparation method of dry powder, make full use of Cordyceps militaris (L.) Link. resource, improve the added value of Cordyceps militaris (L.) Link., simple easy
OK, reduces cost, it is easy to the marketization.Cordyceps militaris (L.) Link. polypeptide extractive technique is using 2 kinds of biological enzyme resolving tech, efficiency high, simple, province
When, using many polypeptide index height of the Cordyceps militaris (L.) Link. of compound bio enzymolysis technology extraction, up to more than 90%, human body cell is had larger
It is worth.
The present invention is that the technical scheme for reaching above-mentioned purpose comprises the steps：
Fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked out, put 40-70 DEG C and dry to constant weight, crushed with pulverizer, mistake
Add water to soak 1h after 80 mesh sieves, successively 2 kinds of protease hydrolyzeds are added, heating up after enzymolysis inactivates enzyme, centrifuging and taking supernatant, very
Empty concentration, concentrated solution ultrafiltration, it is spray-dried and obtains Cordyceps militaris (L.) Link. polypeptide dry powder.
Advantages of the present invention effect is as follows：
The present invention extracts polypeptide in Cordyceps militaris (L.) Link.sporophore using the method for dual biological enzyme hydrolysis, and method is simple, convenient, high
Effect, pollution-free, production raw material sources enrich, low cost, large-scale production can be amplified, Practical significance is big.
Specific embodiment
Embodiment 1
(1) fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked, put 50 DEG C and dry to constant weight, crushed with pulverizer, cross 80
Mesh sieve, soaks 1h, Cordyceps militaris (L.) Link. powder and water (W:V) ratio is 1:10, make Cordyceps militaris (L.) Link. powder fully swelling.
(2) it is 2.0 to adjust PH, adds 2.0% pepsin of Cordyceps militaris (L.) Link. grain weight amount, and 37 DEG C of temperature is kept for 3 hours not
Disconnected stirring.
(3) PH to 8.0 is adjusted, 2.0% trypsin of Cordyceps militaris (L.) Link. grain weight amount is added, temperature 45 C is kept for 2 hours not
Disconnected stirring.
(4) enzymolysis feed temperature to 100 DEG C is improved, keeps making enzyme lose activity in 10 minutes.
(5) enzymolysis feed liquid is after enzyme-deactivating after 8000r/min centrifugation 10min, and supernatant is through concentrated in vacuo, and polypeptide is molten
Liquid solid content reaches 38%-40%.
(6) 300,000 ultrafiltration post ultrafiltration of molecular weight is used after enzymolysis solution is concentrated, and filtrate is filtered again through 100,000 ultrafiltration post ultrafiltration of molecular weight
Go out after liquid is spray-dried and obtain Cordyceps militaris (L.) Link. polypeptide dry powder.
Embodiment 2
(1) fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked, put 50 DEG C and dry to constant weight, crushed with pulverizer, cross 80
Mesh sieve, adds water to soak 1h, makes Cordyceps militaris (L.) Link. powder and water (W:V) ratio is 1:15, make Cordyceps militaris (L.) Link. powder fully swelling.
(2) it is 2.0 to adjust PH, adds 2.5% pepsin of Cordyceps militaris (L.) Link. grain weight amount, and 37 DEG C of temperature is kept for 3 hours not
Disconnected stirring.
(3) PH to 8.0 is adjusted, 2.5% trypsin of Cordyceps militaris (L.) Link. grain weight amount is added, temperature 45 C is kept for 2 hours not
Disconnected stirring.
(4) enzymolysis feed temperature to 100 DEG C is improved, keeps making enzyme lose activity in 10 minutes.
(5) enzymolysis feed liquid is after enzyme-deactivating after 8000r/min centrifugation 10min, and supernatant is through concentrated in vacuo, and polypeptide is molten
Liquid solid content reaches 38%-40%.
(6) 300,000 ultrafiltration post ultrafiltration of molecular weight is used after enzymolysis solution is concentrated, and filtrate is filtered again through 100,000 ultrafiltration post ultrafiltration of molecular weight
Go out after liquid is spray-dried and obtain Cordyceps militaris (L.) Link. polypeptide dry powder.
Embodiment 3
(1) fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked, put 50 DEG C and dry to constant weight, crushed with pulverizer, cross 80
Mesh sieve, soaks 1h, makes Cordyceps militaris (L.) Link. powder and water (W:V) ratio is 1:12, make Cordyceps militaris (L.) Link. powder fully swelling.
(2) it is 2.0 to adjust PH, adds 2.5% pepsin of Cordyceps militaris (L.) Link. grain weight amount, and 37 DEG C of temperature is kept for 2 hours not
Disconnected stirring.
(3) extracting solution adjusts PH to 8.0, adds 2.0%% trypsin of Cordyceps militaris (L.) Link. grain weight amount, and temperature 45 C keeps 2
Hour is simultaneously stirred continuously.
(4) enzymolysis feed temperature to 100 DEG C is improved, keeps making enzyme lose activity in 10 minutes.
(5) enzymolysis feed liquid is after enzyme-deactivating after 8000r/min centrifugation 10min, and supernatant is through concentrated in vacuo, and polypeptide is molten
Liquid solid content reaches 38%-40%.
(6) 300,000 ultrafiltration post ultrafiltration of molecular weight is used after enzymolysis solution is concentrated, and filtrate is filtered again through 100,000 ultrafiltration post ultrafiltration of molecular weight
Go out after liquid is spray-dried and obtain Cordyceps militaris (L.) Link. polypeptide dry powder.
Embodiment 4
(1) fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked, put 40 DEG C and dry to constant weight, crushed with pulverizer, cross 80
Mesh sieve, adds water to soak 1h, makes Cordyceps militaris (L.) Link. powder and water (W:V) ratio is 1:12, make Cordyceps militaris (L.) Link. powder fully swelling.
(2) it is 2.0 to adjust PH, adds 1.5% pepsin of Cordyceps militaris (L.) Link. grain weight amount, and 37 DEG C of temperature is kept for 3 hours not
Disconnected stirring.
(3) PH to 8.0 is adjusted, 1.5% trypsin of Cordyceps militaris (L.) Link. grain weight amount is added, temperature 45 C is kept for 2 hours not
Disconnected stirring.
(4) enzymolysis feed temperature to 100 DEG C is improved, keeps making enzyme lose activity in 10 minutes.
(5) enzymolysis feed liquid is after enzyme-deactivating after 8000r/min centrifugation 10min, and supernatant is through concentrated in vacuo, and polypeptide is molten
Liquid solid content reaches 38%-40%.
(6) 300,000 ultrafiltration post ultrafiltration of molecular weight is used after enzymolysis solution is concentrated, and filtrate is filtered again through 100,000 ultrafiltration post ultrafiltration of molecular weight
Go out after liquid is spray-dried and obtain Cordyceps militaris (L.) Link. polypeptide dry powder..
Embodiment 5
(1) fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked, put 70 DEG C and dry to constant weight, crushed with pulverizer, cross 80
Mesh sieve, adds water to soak 1h, makes Cordyceps militaris (L.) Link. powder and water ratio for 1:12, make Cordyceps militaris (L.) Link. powder fully swelling.
(2) it is 2.0 to adjust PH, adds 2.5% pepsin of Cordyceps militaris (L.) Link. grain weight amount, and 37 DEG C of temperature is kept for 3 hours not
Disconnected stirring.
(3) PH to 8.0 is adjusted, 2.5% trypsin of Cordyceps militaris (L.) Link. grain weight amount is added, temperature 45 C is kept for 2 hours not
Disconnected stirring.
(4) enzymolysis feed temperature to 100 DEG C is improved, keeps making enzyme lose activity in 10 minutes.
(5) enzymolysis feed liquid is after enzyme-deactivating after 8000r/min centrifugation 10min, and supernatant is through concentrated in vacuo, and polypeptide is molten
Liquid solid content reaches 38%-40%.
(6) 300,000 ultrafiltration post ultrafiltration of molecular weight is used after enzymolysis solution is concentrated, and filtrate is filtered again through 100,000 ultrafiltration post ultrafiltration of molecular weight
Go out after liquid is spray-dried and obtain Cordyceps militaris (L.) Link. polypeptide dry powder..
Subordinate list：Different enzymes digest Cordyceps militaris (L.) Link.sporophore effectiveness comparison
Claims (9)
1. a kind of preparation method of Cordyceps militaris (L.) Link. polypeptide dry powder, it is characterised in that comprise the following steps：
Fresh Cordyceps militaris (L.) Link. is taken, macroscopic impurity is picked out, put 40-70 DEG C and dry to constant weight, crushed with pulverizer, mistake
Add water to soak 1h after 80 mesh sieves, successively 2 kinds of protease hydrolyzeds are added, heating up after enzymolysis inactivates enzyme, centrifuging and taking supernatant
Liquid, concentrated in vacuo, concentrated solution ultrafiltration, it is spray-dried, obtains dry powder.
2. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 1, it is characterised in that：Cordyceps militaris (L.) Link. and addition
The quality volume of soaking water（W:V）Proportionate relationship is, Cordyceps militaris (L.) Link.：Water=1:10-1:15.
3. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 1, it is characterised in that：2 kinds of protease divide
Wei not pepsin and trypsin.
4. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 3, it is characterised in that：Pepsic most
Good enzymatic hydrolysis condition is 37 DEG C of temperature, pH value 2.0, enzyme concentration 1.5-2.5wt%, concentration of substrate 8wt %, 3 h of enzymolysis time.
5. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 3, it is characterised in that：Tryptic
Optimum enzymolysis condition is temperature 45 C, pH value 8.0, enzyme concentration 1.5-2.5wt %, concentration of substrate 8wt %, enzymolysis time
2h.
6. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 1, it is characterised in that：Improve enzymolysis liquid temperature
Spend to 100 DEG C, keep making enzyme-deactivating in 10 minutes.
7. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 1, it is characterised in that：Enzymolysis solution is passed through
After enzyme-deactivating after 8000r/min centrifugation 10min, supernatant is through concentrated in vacuo, and polypeptide solution solid content reaches 38wt %-
40 wt %, can be spray-dried after ultrafiltration.
8. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 7, it is characterised in that：With molecular weight 30
Ten thousand ultrafiltration post ultrafiltration, filtrate is with 100,000 ultrafiltration post ultrafiltration of molecular weight.
9. the preparation method of a kind of Cordyceps militaris (L.) Link. polypeptide dry powder according to claim 7, it is characterised in that：It is spray-dried bar
Part inlet temperature 125-130 DEG C, temperature 75-78 DEG C in tower, exhaust outlet temperature is 80-85 DEG C, obtains Cordyceps militaris (L.) Link. polypeptide and does
Powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611015400.5A CN106434810A (en) | 2016-11-18 | 2016-11-18 | Preparation method of cordyceps militaris polypeptide dried powder |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611015400.5A CN106434810A (en) | 2016-11-18 | 2016-11-18 | Preparation method of cordyceps militaris polypeptide dried powder |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107602660A (en) * | 2017-10-31 | 2018-01-19 | 湖北省农业科学院农产品加工与核农技术研究所 | A kind of Cordyceps militaris polypeptide product and preparation method and application |
| CN109329900A (en) * | 2018-12-10 | 2019-02-15 | 丽江程海湖天然螺旋藻生产基地有限公司 | A kind of Cordyceps militaris spirulina powder preparation method and applications |
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| CN103194516A (en) * | 2013-03-17 | 2013-07-10 | 赤峰鼎基生物工程科技有限公司 | Preparation method of cordyceps militaris polypeptide |
| CN105063141A (en) * | 2015-07-30 | 2015-11-18 | 北京天肽生物科技有限公司 | Small-molecule Cordyceps militaris active peptide and preparing method thereof |
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| CN103194516A (en) * | 2013-03-17 | 2013-07-10 | 赤峰鼎基生物工程科技有限公司 | Preparation method of cordyceps militaris polypeptide |
| CN105734099A (en) * | 2014-12-10 | 2016-07-06 | 黑龙江众生生物工程有限公司 | Preparation method of cordyceps militaris polypeptide |
| CN105063141A (en) * | 2015-07-30 | 2015-11-18 | 北京天肽生物科技有限公司 | Small-molecule Cordyceps militaris active peptide and preparing method thereof |
| CN105349606A (en) * | 2015-12-16 | 2016-02-24 | 黑龙江众生生物工程有限公司 | Method for extracting cordyceps militaris polypeptides by co-enzyme method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107602660A (en) * | 2017-10-31 | 2018-01-19 | 湖北省农业科学院农产品加工与核农技术研究所 | A kind of Cordyceps militaris polypeptide product and preparation method and application |
| CN109329900A (en) * | 2018-12-10 | 2019-02-15 | 丽江程海湖天然螺旋藻生产基地有限公司 | A kind of Cordyceps militaris spirulina powder preparation method and applications |
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Application publication date: 20170222 |