Method for extracting collagens from bovine achilles tendon
An extraction method and technology of bovine Achilles tendon, applied in the field of biological materials, can solve the problems of cumbersome separation process and low collagen purity, and achieve the effects of low random degradation rate, high purity and avoiding thermal degradation.
Active Publication Date: 2015-07-15
HEBEI COLLAGEN BIOTECH
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Problems solved by technology
[0005] Aiming at the problems of low collagen purity and cumbersome separation process in the traditional collagen preparation process, the purpose of the present invention is to provide a method for extracting collagen from bovine Achilles tendon. The prepared bovine collagen has high purity and maintains the natural structure of triple helix
Method used
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Experimental program
Comparison scheme
Effect test
Embodiment 1
[0025] (1) Weighing: take 50g of fresh bovine Achilles tendon, wash to remove the residue.
[0026] (2) Freezing: Store at -40°C for 12 hours.
[0027] (3) Cut into pieces: cut the frozen Achilles tendon into 0.5cm×0.5cm pieces.
[0028] (4) Pulverization: the bovine Achilles tendon cut into chunks was frozen at -40°C for 8 hours, and then placed in a pulverizer to pulverize into a slurry.
[0029] (5) Extraction: Place the pulverized bovine Achilles tendon in a temperature-controllable chromatography column (5 cm in diameter, 20 cm in height), and set a 300-mesh nylon mesh at the bottom of the extraction container. Continuously inject deionized water at a temperature of 42°C into the warm chromatography, and control the temperature of circulating water in the jacket of the chromatography column to be 42°C; collect the solution flowing out from the outlet of the chromatography column.
[0030] (6) Enzymatic hydrolysis: add pepsin to the crude collagen extract obtained in ste...
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Abstract
The invention discloses a method for extracting collagens from bovine achilles tendon. The method comprises the following steps: refrigerating the bovine achilles tendon, cutting into blocks, refrigerating, crushing, filling crushed bovine achilles tendon into a temperature-controllable chromatography column, continuously injecting an extract liquid into the temperature-controllable chromatography column, controlling the temperature of circulating water in a jacket of the chromatography column consistent to the temperature of the extract liquid, and collecting the extract liquid from an outlet of the chromatography column; carrying out enzymatic hydrolysis, carrying out chromatographic separation in an affinity chromatography mode, wherein a ligand of an affinity chromatography media is a polypeptide; and leaching out uncombined proteins, and eluting to obtain proteins which can be combined with the chromatography media, that is, highly pure collagens. The method is simple to operate, and the prepared collagens have the advantages of high purity, low random degradation and the like.
Description
technical field [0001] The invention belongs to the field of biomaterials, in particular, the invention relates to a method for extracting collagen from bovine Achilles tendon. technical background [0002] As the main component of the extracellular matrix, collagen accounts for about 33% of the total protein in mammals, exists in almost all tissues and has an important impact on the morphology, structure and function of cells and tissues. Collagen molecules are slender rods with a length of about 300nm, a diameter of about 1.5nm, and a molecular weight of about 300kDa. Collagen molecules are composed of 3 polypeptide chains, each of which has about 1000 amino acid residues. Collagen molecules have a triple helical structure. Each polypeptide chain that makes up collagen exists in the form of a left-handed rotation, and the three polypeptide chains are wound in the form of a right-handed helix. Collagen is formed. According to the difference in the amino acid sequence of t...
Claims
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IPC IPC(8): C12P21/06C07K1/22
Inventor 王云松
Owner HEBEI COLLAGEN BIOTECH