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Extraction method for bioactive collagen

A technology of biological activity and extraction method, applied in the field of biological active collagen extraction, can solve problems such as insufficient hydrolysis, and achieve the effect of reducing risks

Active Publication Date: 2014-08-06
SHENZHEN LANDO BIOMATERIALS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantage is that the hydrolysis is not thorough enough

Method used

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  • Extraction method for bioactive collagen
  • Extraction method for bioactive collagen
  • Extraction method for bioactive collagen

Examples

Experimental program
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Effect test

Embodiment 1

[0049] (1) In the pretreatment step, the fresh beef Achilles tendon is washed with cooling pure water and cut into small pieces, and the cells in the animal tissue are removed by using the decellularization method. The specific method is to use a sodium chloride solution with a mass percentage of 10% Soak and stir for 2 hours, then use ultrasonic treatment for 0.5 hours, then soak and stir with 0.3% sodium dodecylsulfonate solution for 2 hours, then soak and stir with 5% sodium chloride solution for 2 hours, and then use ultrasonic Treat for 0.5 hours, then soak and stir for 2 hours with 0.2% sodium dodecylsulfonate solution by mass percentage, and finally wash with cooling pure water, put it into a tissue grinder and pound into tissue debris, and the tissue debris After cleaning and filtering the scraps, soak them in 1.5% sodium hydroxide solution for 2 hours, wash and filter again, and put the tissue scraps for use;

[0050] (2) Rough extraction by acid method, take 1 part b...

Embodiment 2

[0056] (1) The pretreatment step is to wash the fresh cowhide with cooling pure water and cut it into small pieces, and use the decellularization method to remove the cells in the animal tissue. The specific method is to soak and stir with 5% sodium chloride solution 2 hours, then ultrasonic treatment for 1 hour, then immerse and stir with 0.2% sodium dodecylsulfonate solution for 3 hours, then soak and stir with 1% sodium chloride solution for 2 hours, and then use ultrasonic treatment for 1 hour hours, then soak and stir with 0.05% sodium dodecylsulfonate solution by mass percentage for 2 to 3 hours, and finally wash with cooling pure water, put it into a tissue masher and mash it into tissue debris, and the tissue debris After the scraps are cleaned and filtered, they are soaked in 0.1% sodium hydroxide solution for 4 hours, washed and filtered again, and the tissue scraps are used for later use;

[0057] (2) Rough extraction by acid method, take 1 part by weight of filtere...

Embodiment 3

[0063] (1) The pretreatment step is to wash the fresh pigskin with cooling pure water and cut it into small pieces, and use the decellularization method to remove the cells in the animal tissue. The specific method is, for example, soak it in a 7% sodium chloride solution by mass Stir for 1.5 hours, then use ultrasonic treatment for 0.8 hours, then soak and stir with 0.25% sodium dodecylsulfonate solution for 2.5 hours, then soak and stir with 2.5% sodium chloride solution for 1.5 hours, and then use ultrasonic treatment 0.8 hours, then soaked and stirred for 2.5 hours with 0.1% sodium dodecylsulfonate solution by mass percentage, and finally washed with cooling pure water, put into a tissue grinder and smashed into tissue debris, the tissue debris After cleaning and filtering, soak in 0.12% sodium hydroxide solution for 3 hours, wash and filter again, and set aside tissue debris;

[0064] (2) Rough extraction by acid method, take 1 part by weight of filtered tissue debris in ...

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Abstract

The invention relates to an extraction method for bioactive collagen. The method comprises: a pretreatment step, i.e. pretreating fresh bovine tendon into fragments for standby use; an acid process crude extraction step, i.e. taking 1 part by weight of fragment, adding 4-8 parts by weight of an acetic acid or citric acid solution and conducting stirring, then carrying out coarse filtration, performing high speed centrifugation on a coarse filtrate, then taking the supernatant for standby use; a step of end group removal by an enzymatic process, i.e. adjusting the pH of the supernatant to 9.0, then adding 0.1-0.3% part by weight of Ficus proteinase to conduct enzymolysis digestion; a gradient salting-out step, i.e. using a precipitating agent to conduct multi-gradient salting-out treatment on the solution subjected to enzymolysis to obtain a collagen dissolved solution; and a dialysis step, i.e. loading the collagen dissolved solution into a dialysis bag, using an acetic acid solution and pure water as outer dialysate in order to conduct dialysis, thus obtaining a collagen stock solution with bioactivity. The method provided by the invention combines two existing collagen extraction processes, i.e. the acid process and the enzymatic process, overcomes the low yield defect of the acid process and the incomplete hydrolysis defect of the enzymatic process, and adopts non-animal derived protease, thus reducing the risk of animal origin.

Description

technical field [0001] The invention relates to the technical field of collagen extraction, in particular to a bioactive collagen extraction method. Background technique [0002] Collagen is a structural protein of the extracellular matrix with a triple helix structure. Collagen contains 18 kinds of amino acids, does not contain tryptophan, and is an incomplete protein. The special structure of collagen makes it widely used in diagnosis, treatment, rehabilitation and beauty. [0003] Collagen is the most abundant protein in mammals. It is widely distributed in cornea, skin, bones, ligaments, tendons and other parts. It is an important structural protein that constitutes connective tissue. Therefore, extracting collagen from animal tissue is an important way to obtain collagen. [0004] In the production process of extracting collagen, some by-products collagen and gelatin will be produced. Gelatin is composed of peptide chains that are no longer biologically active after ...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/36C07K1/34C07K1/30
Inventor 谭荣伟
Owner SHENZHEN LANDO BIOMATERIALS
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