Preparation method of industrialized non-denaturing collagen II

A technology of collagen and collagen powder, applied in the field of bioengineering, can solve the problems of difficult large-scale production, cumbersome method steps, long process route, etc., and achieve the effect of facilitating post-processing, simple working hours, and simple process route

Inactive Publication Date: 2019-02-26
MEITEK TECH QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] The existing non-denatured type II collagen preparation technology basically adopts the process of degreasing cartilage, removing impurities, direct enzymatic hydrolysis, and then repeated salting out and dialysis to obtain non-denatured type II collagen, such as patent CN105132502A: A kind of The method for extracting pure type II collagen in China has just announced this method, but this method requires repeated salting out and dialysis to obtain non-denatured type II collagen, and polysaccharide impurities such as chondroitin sulfate and hyaluronic acid are difficult to remove. The steps are cumbersome and the process route is long. It is only suitable for laboratory preparation, it is difficult to produce on a large scale, and it produces a large amount of high-concentration wastewater, and the treatment cost is high.

Method used

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  • Preparation method of industrialized non-denaturing collagen II
  • Preparation method of industrialized non-denaturing collagen II
  • Preparation method of industrialized non-denaturing collagen II

Examples

Experimental program
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Embodiment 1

[0016] A method for preparing industrialized non-denatured II collagen, comprising the following steps: (1) cartilage pretreatment: after the cartilage is cleaned and fleshed, the cartilage is washed with H 2 o 2 Soak and disinfect for 18-24 hours, mash the treated cartilage with a tissue masher; (2) Alkaline extraction: soak the mashed cartilage in 0.1-4M lye at 12°C for 6-24 hours, Removal of soluble protein, fat and polysaccharide; (3) Separation and drying: separate the precipitate with a centrifuge, pulverize the precipitate with a pulverizer, add water to suspend it, and freeze-dry to obtain insoluble non-denatured II collagen powder. Wherein the lye is one of sodium hydroxide, calcium hydroxide or potassium hydroxide, the cartilage is chicken cartilage, and the centrifugal separation operating conditions of the centrifuge are 5000rpm, 4°C, and more than 10min.

Embodiment 2

[0018] Embodiment 2 On the basis of Embodiment 1, the following steps are also included: placing the insoluble non-denatured II collagen powder obtained by freeze-drying in a reaction tank, adding 1 to 5 times the amount of protein powder in deionized water, and adding 1 to 5 times the amount of protein powder. 0.05-5% pepsin, adjust the pH value to 1.5-4.5, enzymatically hydrolyze at 4°C for 24-72 hours, centrifuge or filter the enzymolyzed solution, and freeze-dry the supernatant to obtain soluble non-denatured II collagen powder.

Embodiment 3

[0020] Embodiment 3 On the basis of Embodiment 1, the following steps are also included: placing the insoluble non-denatured II collagen powder obtained by freeze-drying in a reaction tank, adding 1 to 5 times the amount of protein powder in deionized water, and adding 1 to 5 times the amount of protein powder. 0.05-5% pepsin, adjust the pH value to 1.5-4.5, enzymatically hydrolyze at 20°C for 24-72 hours, centrifuge or filter the enzymolyzed solution, and freeze-dry the supernatant to obtain soluble non-denatured II collagen powder.

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Abstract

The invention relates to the field of bioengineering, in particular to a preparation method of industrialized non-denaturing collagen II. The preparation method comprises the steps of cartilage pretreatment, alkali liquor extraction, separation, drying and enzymolysis. Most impurities such as soluble protein, fat, chondroitin sulfate and hyaluronic acid in cartilage are removed by low-temperaturealkali liquor extraction, subsequent separation and drying of insoluble non-denatured collagen II powder as well as extraction and refining of soluble non-denaturing collagen II in the enzymolysis step are facilitated, and salting out, dialysis and other operation in the enzymolysis step in the conventional method can be omitted; the preparation method has simple steps and short working hours, ismore beneficial to industrial production, avoids production of a large amount of waste water, and has potential market value accordingly.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for preparing industrialized non-denatured II collagen. Background technique [0002] The existing non-denatured type II collagen preparation technology basically adopts the process of degreasing cartilage, removing impurities, direct enzymolysis, and then repeatedly salting out and dialysis to obtain non-denatured type II collagen, such as patent CN105132502A: a kind of cartilage from chicken The method for extracting pure type II collagen in China has just announced this method, but this method requires repeated salting out and dialysis to obtain non-denatured type II collagen, and polysaccharide impurities such as chondroitin sulfate and hyaluronic acid are difficult to remove. The steps are cumbersome and the process route is long. It is only suitable for laboratory preparation, it is difficult to produce on a large scale, and it produces a large amount of high-concentr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/14C12P21/06
CPCC12P21/06C07K14/78
Inventor 张亮张雷于明晓
Owner MEITEK TECH QINGDAO
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