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Collagen purification method

A collagen and purification method technology, applied in peptide preparation methods, chemical instruments and methods, animal/human proteins, etc., can solve the problems of collagen yield and purity not meeting the requirements, low risk, etc., to improve the yield and purity, increase the amount of precipitation, low cost

Active Publication Date: 2021-07-13
浙江崇山生物制品有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Chinese patent application 201610893949.8 provides an efficient and simple collagen ultrafiltration purification method, which mainly includes the following steps: dilute the dissolved collagen solution; turn on the delivery pump, and the collagen solution is purified through the purification system. Purification can produce purified collagen. This purification method is safe, effective, low-risk, simple, energy-saving, high production efficiency, does not pollute the environment, and is suitable for industrial production. However, the yield and purity of the obtained collagen are low. can not meet the requirements

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  • Collagen purification method

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Effect test

Embodiment 1

[0050] Embodiment 1 A kind of collagen purification method

[0051] Specifically include the following steps:

[0052] (1) Take the crude collagen extract (210L, containing about 400g of collagen), dilute it with water, add acetic acid to adjust the pH value to 2.5, add diatomaceous earth (2.1kg) and biological flocculant ( 0.7kg), stirred for 30min and adsorbed, followed by plate and frame filtration to obtain mixed solution A;

[0053] (2) Add an equal volume to the mixed solution A and the volume ratio is 5:1, the mass fraction is 15% ammonium chloride and the mass fraction is 30% ammonium sulfate mixed solution to carry out salting out for 2h to obtain solid B, and the solid B After reconstitution, the obtained reconstituted solution is first filtered with a 5-micron filter membrane, the obtained filtrate a is then filtered with a 3-micron filter membrane, and the obtained filtrate b is then filtered with a 0.45-micron filter membrane to obtain a mixture C;

[0054] (3) ...

Embodiment 2

[0056] Embodiment 2 A kind of collagen purification method

[0057] Specifically include the following steps:

[0058] (1) Take the crude collagen extract (210L, containing about 400g of collagen), dilute it with water, add acetic acid to adjust the pH value to 3.5, add diatomaceous earth (8.4kg) and biological flocculant ( 1.4kg), plate and frame filtration was carried out after adsorption, and then 0.1 μm membrane filtration was carried out to obtain the mixed solution A;

[0059] (2) Add an equal volume of ammonium chloride with a volume ratio of 10:1 and a mixed solution of ammonium sulfate with a mass fraction of 25% and 40% ammonium sulfate in the mixed solution A for salting out to obtain a solid B, and redissolve the solid B Finally, the obtained complex solution is first filtered with a 5-micron filter membrane, the obtained filtrate a is then filtered with a 3-micron filter membrane, and the obtained filtrate b is then filtered with a 0.45-micron filter membrane to ...

Embodiment 3

[0062] Embodiment 3 A kind of collagen purification method

[0063] Specifically include the following steps:

[0064] (1) Take the crude collagen extract (210L, containing about 400g of collagen), dilute it with water, add acetic acid to adjust the pH value to 3.0, add diatomaceous earth (6.3kg) and biological flocculant ( 1.26kg), after adsorption, perform plate and frame filtration, and then perform 0.1 μm membrane filtration to obtain mixed solution A;

[0065] (2) adding volume ratio to mixed solution A is that 8:1 mass fraction is 20% ammonium chloride and mass fraction is the ammonium sulfate mixed solution of 35% to obtain solid B after salting out, after solid B is redissolved, The obtained complex solution is first filtered with a 5-micron filter membrane, the obtained filtrate a is then filtered with a 3-micron filter membrane, and the obtained filtrate b is then filtered with a 0.45-micron filter membrane to obtain a mixed solution C;

[0066] (3) The mixture C w...

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Abstract

The invention provides a collagen purification method, and relates to the technical field of protein processing. According to the method, components such as insoluble substances and non-acid-soluble proteins in the solution are precipitated by adopting a manner of combining the diatomite and the biological flocculant, and then are removed by plate and frame filtration, so that the operation is simple, the subsequent ultrafiltration time can be saved, and the yield and purity of the collagen are improved; in the salting-out process, a mixture of ammonium chloride with the mass fraction of 15-25% and ammonium sulfate with the mass fraction of 30-40% is used, and the volume ratio of the ammonium chloride to the ammonium sulfate is controlled to be (5-10): 1, so that the precipitation amount of the collagen can be obviously increased, and the yield of the collagen is further increased; and finally, the mixed solution is desalted in a manner of combining an ultrafiltration membrane with a tubular ultrafiltration membrane, so that all salts in the salting-out process can be removed, the purity of the obtained collagen is ensured, and the condition that the stability is reduced due to the fact that the collagen contains the salts is avoided.

Description

technical field [0001] The invention relates to the technical field of protein processing, in particular to a collagen purification method. Background technique [0002] Collagen, a natural biopolymer produced in animal fibroblasts, accounts for one-third of the total protein in mammals. Structurally, collagen molecules exist in the extracellular matrix in a supramolecular structure. The most common structural feature of collagen is the triple helix structure, which is composed of three α-polypeptide chains, and the polypeptide chains are intertwined in a right-handed helical configuration to form a superhelical structure, which makes the collagen molecule very stable. Due to a series of characteristics in its structure and composition, collagen has good properties such as repair, biocompatibility, mechanical elasticity, and low immune prototype. Collagen and related derivative products are widely used in food, beauty, medicine, biology, etc. It has a wide range of applica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/34C07K1/30C07K1/36
CPCC07K14/78
Inventor 张智武陈新亮彭新生周艳芳
Owner 浙江崇山生物制品有限公司
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