CN112266943A - Extraction method and application of wild goose palm clausena lansium polypeptide - Google Patents
Extraction method and application of wild goose palm clausena lansium polypeptide Download PDFInfo
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- CN112266943A CN112266943A CN202011167186.1A CN202011167186A CN112266943A CN 112266943 A CN112266943 A CN 112266943A CN 202011167186 A CN202011167186 A CN 202011167186A CN 112266943 A CN112266943 A CN 112266943A
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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
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
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Abstract
An extraction method and application of wild goose palm pellago polypeptide belong to the technical field of animal polypeptide extraction and development, and particularly relate to wild goose palm pellago polypeptide extraction and application in preparation of anti-freezing skin cream. The polypeptide extraction process comprises the following steps: degreasing by petroleum ether, hydrolyzing to obtain anser indicus flavedo protein, and carrying out balanced dialysis and purification to obtain the anser indicus flavedo polypeptide extract. The invention has the advantages of simple preparation process and low cost, and the experiment for examining the cold injury resisting activity of the in vitro cold injury cell model proves that the wild goose palm flavedo polypeptide has the advantage of certain cold injury resisting activity. The cold injury resisting skin cream is applied to cold injury resisting skin cream, and can reduce the incidence rate of cold injury. The abandoned wild goose palm peltexes are utilized, so that the waste of biological resources and the environmental pollution are reduced.
Description
Technical Field
The invention belongs to the technical field of extraction and development of animal polypeptide, and particularly relates to an extraction method of wild goose palm clausena lansium polypeptide and application thereof.
Background
Wild geese (Anser cygnoides) are anserians of the family avenae of the order avenae, which are unique species in Jilin province, live throughout the year in the natural protection area of the sea in the city of Ulmus communis of the Jilin province, and were identified as new species by the American national center for Biotechnology information 2015 (Genbank ID: 1856198). The wild geese under artificial domestication have unique characteristic that migratory birds do not migrate along with seasons, and can resist severe cold of-40 ℃ in winter.
The yellow skin of goose foot is yellow skin on the foot palm and foot web of wild goose, which is recorded in traditional ancient books of China, Ben Cao gang mu, Qian jin Shi Zhi, etc. Li Shizhen recorded it in Ben Cao gang mu as: burn and grind, and apply on the toe seam to make the toe wet and rotten. Baking, grinding, mixing with oil, and applying for treating chilblain. "
At present, research reports on wild geese at home and abroad are very rare, and the feathers, meat, blood, bones and fat of the wild geese have certain research values, but most of the wild geese are exported as raw materials, and the yellow skins of the wild geese are discarded as wastes. The number of killed wild geese to sea is about 30 thousands per year. The discarding of wild goose palm yellow skin causes great biological resource waste and environmental pollution.
Therefore, there is a need in the art for a new solution to solve this problem.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the extraction method and the application of the wild goose palm peltry polypeptide are used for solving the technical problems of great biological resource waste and environmental pollution caused by discarding the wild goose palm peltry.
A method for extracting wild goose palm flavedo polypeptide comprises the following steps,
step one, selecting the most suitable protease;
step two, hydrolyzing the degreased and dried wild goose palm pellicles for 5 hours by using selected optimal protease according to a set feed-liquid ratio, and taking hydrolysate to obtain wild goose palm pellicles protein hydrolysate;
step three, putting the wild goose palm yellow skin protein hydrolysate obtained in the step two into a dialysis bag for carrying out balance dialysis for 48 hours, wherein the obtained liquid outside the dialysis bag is the wild goose palm yellow skin protein hydrolysate;
and step four, determining the wild goose palm yellow skin protein hydrolysate obtained in the step three to be the wild goose palm yellow skin polypeptide extract through a molecular weight by utilizing gel exclusion chromatography, and freeze-drying the wild goose palm yellow skin polypeptide extract obtained in the step three for later use.
The specific method for selecting the optimal protease comprises the steps of weighing the same mass of acidic protease, neutral protease and alkaline protease, hydrolyzing the equivalent amount of anser indicus palm peltry under the same condition, and measuring the hydrolysis degree, wherein the value of the neutral protease is the highest, and the neutral protease is the optimal protease.
The acidic protease includes pepsin.
The specific method for degreasing and drying the yellow skin of the wild goose palm in the second step comprises the following steps: completely immersing the yellow skin of the wild goose palm in petroleum ether with the boiling range of 60-90 ℃ for degreasing for 48h, washing the yellow skin to be neutral by purified water, and drying the yellow skin at 60 ℃ to constant weight to obtain the degreased and dried wild goose palm yellow skin.
And the material-liquid ratio set in the second step is 1: 15.
Application of wild goose palm pelteobagrus polypeptide in cold injury resisting skin cream is provided.
Through the design scheme, the invention can bring the following beneficial effects:
the extraction method disclosed by the invention obtains the yellow skin polypeptide of the anser indicus palm by utilizing the anser indicus palm yellow skin, and has the advantages of simple preparation process and low cost. The wild goose palm peltry polypeptide has certain activity in the aspect of cold injury resistance application, and the wild goose palm peltry polypeptide extract prepared by the method disclosed by the invention is applied to cold injury resistance skin cream, so that the incidence rate of cold injury can be reduced. The abandoned wild goose palm peltexes are utilized, so that the waste of biological resources and the environmental pollution are reduced.
Drawings
The invention is further described with reference to the following figures and detailed description:
fig. 1 is a diagram of the influence of wild goose palm flavedo polypeptide on LDH in the second embodiment of the method for extracting wild goose palm flavedo polypeptide and the application thereof.
Fig. 2 is a diagram showing the influence of the anser indicus flavedo polypeptide on the NO content in an embodiment II of the method for extracting the anser indicus flavedo polypeptide and the application thereof.
Detailed Description
The present invention will be described in detail with reference to specific examples.
The first embodiment is as follows: and (3) extracting wild goose palm clausena lansium polypeptide.
Step 1, weighing different proteases with the same mass, hydrolyzing yellow skins of wild goose palms under the same conditions, and screening out the most suitable protease;
the specific method for screening the optimal protease comprises the steps of weighing the same mass of pepsin, neutral protease and alkaline protease, hydrolyzing the same amount of anser indicus palm peltry under the same condition, and measuring the hydrolysis degree, wherein the value of the neutral protease is the highest, and the neutral protease is the optimal protease.
Step 2, according to the step 1:15, taking the neutral protease screened in the step 1 to degrease petroleum ether (the boiling range is 60-90 ℃) for 48 hours, drying to constant weight wild goose palm pellicles, hydrolyzing the degreased and dried wild goose palm pellicles, and collecting hydrolysate.
And 3, carrying out equilibrium dialysis 48 on the hydrolysate obtained in the step 2 in a dialysis bag, and then taking the dialyzed external liquid.
And 4, performing gel exclusion chromatography on the dialyzed and purified external liquid in the step 3, determining the extract in the step 3 through molecular weight, namely the wild goose palm flavedo polypeptide extract, and freeze-drying for later use.
Example two: and (3) testing the cold injury resisting activity of the wild goose palm wampee polypeptide.
Cold injury resistance activity tests are respectively carried out by adopting wild goose palm yellow skin protein and wild goose palm yellow skin polypeptide.
The rat vascular endothelial cells in logarithmic phase are cooled to 0 ℃, 5 ℃, 10 ℃, 15 ℃ and 20 ℃ respectively by taking 4 ℃ as the initial temperature, kept for 5min and then cultured again at 37 ℃ for 10 min. Using the non-cooled group as a control group, 20. mu.L of MTT solution cell culture chamber was added to each well of the cold injury model group at 0 ℃, -5 ℃, -10 ℃, -15 ℃, -20 ℃ and continued culturing for 4 hours, and then the supernatant was aspirated. To each well was added 150. mu.L of dimethyl sulfoxide (DMSO), shaken at low speed for 300s, and allowed to stand for a while. And calculating the cell survival rate. The culture supernatants of each group of cells were collected to determine the serum lactate dehydrogenase LDH activity.
Respectively preparing wild goose palm flavedo polypeptide solutions with concentration of 50 μ g/mL, 100 μ g/mL, 200 μ g/mL and 400 μ g/mL, adding the above solutions into the established rat vascular endothelial cell cold injury model, and treating with 5% CO at 37 deg.C2After the culture is continued for 24 hours in the incubator, cell culture supernatant is taken to determine LDH activity and nitric oxide NO content.
And (3) detecting an activity standard: the LDH activity is reduced, and the NO content is increased, which shows that the wild goose palm flavedo polypeptide has cold injury resisting activity.
The cold injury resistance test results of the wild goose palm pellagra polypeptide are shown in fig. 1 and fig. 2. The experimental results shown in fig. 1 and fig. 2 show that the wild goose palm flavedo polypeptide has certain cold injury resistance activity, and can reduce LDH activity and increase NO content in a dose-dependent manner. Therefore, the wild goose palm yellow skin polypeptide can be applied to cold injury resistant skin cream.
The above-mentioned embodiments only represent some embodiments of the present invention, and the description thereof is more detailed and specific, but not to be construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.
Claims (6)
1. A method for extracting wild goose palm clausena lansium polypeptide is characterized by comprising the following steps: comprises the following steps which are sequentially carried out,
step one, selecting the most suitable protease;
step two, hydrolyzing the degreased and dried wild goose palm pellicles for 5 hours by using selected optimal protease according to a set feed-liquid ratio, and taking hydrolysate to obtain wild goose palm pellicles protein hydrolysate;
step three, putting the wild goose palm yellow skin protein hydrolysate obtained in the step two into a dialysis bag for carrying out balance dialysis for 48 hours, wherein the obtained liquid outside the dialysis bag is the wild goose palm yellow skin protein hydrolysate;
and step four, determining the wild goose palm yellow skin protein hydrolysate obtained in the step three to be the wild goose palm yellow skin polypeptide extract through a molecular weight by utilizing gel exclusion chromatography, and freeze-drying the wild goose palm yellow skin polypeptide extract obtained in the step three for later use.
2. The extraction method of the anser indicus palm flavedo polypeptide according to claim 1, which is characterized in that: the specific method for selecting the optimal protease comprises the steps of weighing the same mass of acidic protease, neutral protease and alkaline protease, hydrolyzing the equivalent amount of anser indicus palm peltry under the same condition, and measuring the hydrolysis degree, wherein the value of the neutral protease is the highest, and the neutral protease is the optimal protease.
3. The extraction method of the anser indicus palm flavedo polypeptide according to claim 2, which is characterized in that: the acidic protease includes pepsin.
4. The extraction method of the anser indicus palm flavedo polypeptide according to claim 1, which is characterized in that: the specific method for degreasing and drying the yellow skin of the wild goose palm in the second step comprises the following steps: completely immersing the yellow skin of the wild goose palm in petroleum ether with the boiling range of 60-90 ℃ for degreasing for 48h, washing the yellow skin to be neutral by purified water, and drying the yellow skin at 60 ℃ to constant weight to obtain the degreased and dried wild goose palm yellow skin.
5. The extraction method of the anser indicus palm flavedo polypeptide according to claim 1, which is characterized in that: and the material-liquid ratio set in the second step is 1: 15.
6. An application of wild goose palm pelteobagrus polypeptide is characterized in that: is applied to the skin cream for resisting cold injury.
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| CN202011167186.1A CN112266943B (en) | 2020-10-28 | 2020-10-28 | Extraction method and application of polypeptide from wild goose palmate yellow skin |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114235932A (en) * | 2021-12-20 | 2022-03-25 | 长春中医药大学 | Method for extracting and researching wild goose blood polypeptide by enzymolysis method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114235932A (en) * | 2021-12-20 | 2022-03-25 | 长春中医药大学 | Method for extracting and researching wild goose blood polypeptide by enzymolysis method |
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