Cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption

A technology of mucosal absorption and velvet antler polypeptide, which is applied in the preparation method of peptide, chemical instrument and method, peptide and other directions, can solve the problems of difficult dissolution, waste of resources, difficult dissolution, etc., and achieves a simple preparation process, improved utilization, and improved The effect of extraction rate

Inactive Publication Date: 2014-10-08
JILIN UNIV
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Problems solved by technology

Chinese patent CN 100339387C discloses the technology of extracting peptides with 40-60% ethanol after acid hydrolysis at pH 3.5, and obtains an extract below 500 000 Da with a 4% extraction rate, but in practical applications, it is found that the extract is in Easy to dissolve in pH3.5 acidic solution, but not easy to dissolve in neutral or alkaline solution
In 2011, Guangzhou Huangsha Cosmetics Co., Ltd. (application number: 102225967 A) proposed a strategy of 50-75% ethanol extraction after rough extraction with pH 7.2 buffer. The molecular weight range and extraction rate of the prepared polypeptide were not mentioned in the article And solubility, after experimental testing, it was found that the polypeptide obtained by ethanol precipitation still has the disadvantages of being difficult to dissolve in neutral solvents stably and having a wide molecular weight distribution, which limit its application to the skin
Chinese patent CN 103169942 A discloses an enzymatic hydrolysis method for preparing velvet collagen. Under the conditions of pH 6-8, 20-35 ℃, a clarified golden yellow velvet hydrolyzate is obtained by hydrolysis with a composite biocatalyst, but no ultrafiltration of the product is carried out. Refined, there are still insufficiently hydrolyzed macromolecular substances in the product, which seriously affects the effective utilization of its transdermal absorption
[0004]The methods used in the above patented technologies are all one-time extractions, which fail to make full use of the raw materials, and the residues of the remaining raw materials still contain a large number of polypeptide growth factors, resulting in an extreme shortage of resources. waste

Method used

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  • Cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption
  • Cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption
  • Cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Purchase chilled velvet velvet, slice the velvet velvet, crush it and weigh it, a total of 1089g, add 20 times the pre-cooled acetic acid-sodium acetate buffer solution with pH 4.2 to make a homogenate, place it at 4 ℃ and stir it at a speed increase Stir and extract at a constant speed for 4 h, filter with a 500-mesh filter cloth (collect the filter residue in this step for subsequent enzymatic hydrolysis extraction), and refine the filtrate by graded ultrafiltration: 1 μm filter membrane coarse filtration (collect the concentrated solution in this step for subsequent enzymatic hydrolysis and extraction) Enzymatic hydrolysis extraction), the filtrate was ultrafiltered through a 10 000 Da filter membrane (collect the concentrate at this step for subsequent enzymatic hydrolysis extraction), and the filtrate was concentrated to 1 000 mL by 300 Da nanofiltration. The nanofiltration concentrate was added with 4% lyoprotectant (40 g) and freeze-dried (pre-freeze at -45 °C for...

Embodiment 2

[0037] Purchase chilled velvet antler, slice the velvet antler, crush it and weigh it, a total of 1013g, add 20 times the pre-cooled pH4.6 acetic acid-sodium acetate buffer solution to make a homogenate, place it at 4 ℃ and stir it at a speed increase Stir and extract at a constant speed for 6 hours, filter with a 100-mesh filter cloth (collect the filter residue in this step for subsequent enzymatic hydrolysis and extraction), and refine the filtrate by graded ultrafiltration: 1 μm filter membrane coarse filtration (collect the concentrated solution in this step for subsequent enzymatic hydrolysis and extraction) hydrolysis and extraction), the filtrate was ultrafiltered through a 10 000 Da filter membrane (collect the concentrate at this step for subsequent enzymatic extraction), and the filtrate was concentrated to 1 000 mL by 300 Da nanofiltration. The nanofiltration concentrate was added with 4% lyoprotectant (40 g) and freeze-dried (pre-freeze at -45 °C for 1 h, and freez...

Embodiment 3

[0040] Purchase fresh velvet antler, slice the velvet antler, crush it and weigh it, totaling 998 g, add 30 times the pre-cooled acetic acid-sodium acetate buffer solution with pH 4.8 to make a homogenate, and place it at 4 °C and stir it at an increased speed Stir and extract at a constant speed for 8 hours, filter with a 1000-mesh filter cloth (collect the filter residue in this step for subsequent enzymatic hydrolysis extraction), coarse filter the filtrate with a filter cloth, and refine by graded ultrafiltration: coarse filter with a 1 μm filter membrane (collect this step to concentrate liquid for subsequent enzymatic extraction), the filtrate was ultrafiltered through a 10 000 Da filter membrane (collecting the concentrate at this step for subsequent enzymatic extraction), and the permeate was concentrated to 1000 mL by 300 Da nanofiltration. The nanofiltration concentrate was added with 4% lyoprotectant (40 g) and freeze-dried (pre-freeze at -45 °C for 1 h, and freeze-d...

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Abstract

The invention provides a cornua cervi pantotrichum polypeptide serial extraction method beneficial to skin or mucosa absorption. A weak acid solution is adopted for extracting cornua cervi pantotrichum natural polypeptide, residual byproducts are subjected to enzymolysis by adopting neutral protease, ultrafiltration and refining are combined, so that the cornua cervi pantotrichum polypeptide is extracted, and higher solubility is obtained. The extracted cornua cervi pantotrichum polypeptide is close to a pH tolerance value of human skin, higher solubility can be easily obtained in preparation of a product, and stimulation of pH value of the polypeptide, extracted by adopting the method provided by the invention, to skin is small. An acidolysis-enzymolysis two-step method is adopted for extracting the cornua cervi pantotrichum polypeptide, extraction ratio of the cornua cervi pantotrichum polypeptide is greatly increased, utilization ratio of raw materials is higher, extraction rate of polypeptide below 1000Da is increased to 2.64% from 0.44%, and polypeptide prepared through enzymolysis by virtue of the byproducts of the cornua cervi pantotrichum polypeptide has biological activity; hierarchical ultrafiltration refining is adopted, molecular weight of 90% of the prepared polypeptide is lower than 1000Da, and the prepared polypeptide is beneficial to absorption of skin and mucosa of human; wide-range application of the cornua cervi pantotrichum polypeptide in cosmetic or external skin preparation is provided.

Description

technical field [0001] The invention provides a method for extracting velvet polypeptide series which is beneficial for skin or mucous membrane absorption, and belongs to the technical field of biotechnology extraction. Background technique [0002] Antler grows rapidly (1-2 cm / d) and has the characteristics of regeneration. It is a natural cell growth factor library. With the in-depth study of velvet, people have begun to pay attention to the development and utilization of biologically active polypeptides and proteins in velvet, that is, growth factors. Chinese patent CN 100339387C discloses the technology of extracting peptides with 40-60% ethanol after acid hydrolysis at pH 3.5, and obtains an extract below 500 000 Da with a 4% extraction rate, but in practical applications, it is found that the extract is in It is easy to dissolve in acidic solution with pH3.5, but not easy to dissolve in neutral or alkaline solution. The pH range that the skin can withstand is 5.5-7.5...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34
Inventor 郝林琳李思明刘松财陆超焦安龙张鑫丁克祥
Owner JILIN UNIV
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