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Method for purifying collagen derived from pelodiscus sinensis

A purification method, a technology of soft-shelled turtle source collagen, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., to achieve the effects of increasing product added value, extending the industrial chain, and high extraction efficiency

Inactive Publication Date: 2015-11-11
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The connective soft tissue around the soft-shelled turtle is usually called "skirt", and the content of collagen is very rich. According to previous experiments, its dry weight content exceeds 70%. According to literature search, the research on collagen in Chinese soft-shelled turtle is limited to its isolation There are reports on extraction and structural characterization, but there are no reports on the purification process and biological properties of soft-shelled turtle collagen

Method used

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  • Method for purifying collagen derived from pelodiscus sinensis
  • Method for purifying collagen derived from pelodiscus sinensis
  • Method for purifying collagen derived from pelodiscus sinensis

Examples

Experimental program
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Effect test

Embodiment 1

[0027] A method for purifying collagen from Chinese soft-shelled turtle, comprising the following steps:

[0028] (1) Soaking: Take soft-shelled soft-shelled turtle skirt homogenate and crush it, put it in alkaline soaking salt solution at 4°C for 24 hours, wash it repeatedly with distilled water, drain it fully with gauze and set it aside;

[0029] (2) Fat removal: wash the soft-shelled soft-shelled turtle skirt soaked in step (1), add 10% isopropanol solution and soak at 4°C for 24 hours to remove fat, wash repeatedly with distilled water, fully drain with gauze and set aside;

[0030] (3) Enzymolysis: put the de-impured soft-shelled soft-shelled turtle skirt into 0.5mol / L acetic acid, the ratio of solid to liquid is 1:25, add it into pepsin enzymatic hydrolysis solution for enzymolysis treatment, extract at 4°C for 24 hours, and obtain Zhonghua Soft-shelled turtle skirt collagen crude extract;

[0031] (4) Salting out purification: take the crude extract of step (3) and di...

Embodiment 2

[0033] A method for purifying collagen from Chinese soft-shelled turtle, comprising the following steps:

[0034] (1) Soaking: Take soft-shelled soft-shelled turtle skirt homogenate and crush it, put it in alkaline soaking salt solution at 10°C for 48 hours, wash it repeatedly with distilled water, drain it fully with gauze and set it aside;

[0035] (2) Fat removal: Clean the soft-shelled soft-shelled turtle skirt after soaking in step (1), add 15% isopropanol solution and soak at 10°C for 48 hours to remove fat, wash repeatedly with distilled water, fully drain with gauze and set aside;

[0036] (3) Enzymolysis: put the de-impured soft-shelled soft-shelled turtle skirt into 0.5mol / L acetic acid, the ratio of solid to liquid is 1:25, add it into pepsin enzymolysis solution for enzymolysis treatment, extract at 10°C for 48h, and obtain Zhonghua Soft-shelled turtle skirt collagen crude extract;

[0037] (4) Salting out purification: take the crude extract of step (3) and disso...

Embodiment 3

[0039] A method for purifying collagen from Chinese soft-shelled turtle, comprising the following steps:

[0040](1) Soaking: Take soft-shelled soft-shelled turtle skirt homogenate and crush it, put it in alkaline soaking salt solution at 8°C for 30 hours, wash it repeatedly with distilled water, drain it fully with gauze and set it aside;

[0041] (2) Fat removal: Clean the soft-shelled turtle skirt soaked in step (1), add 12% isopropanol solution and soak at 8°C for 30 hours to remove fat, wash repeatedly with distilled water, fully drain with gauze and set aside;

[0042] (3) Enzymolysis: put the de-impured soft-shelled soft-shelled turtle skirt into 0.5mol / L acetic acid, the ratio of solid to liquid is 1:25, add it into pepsin enzymatic hydrolysis solution for enzymolysis treatment, extract at 8°C for 30 hours, and obtain Zhonghua Soft-shelled turtle skirt collagen crude extract;

[0043] (4) Salting out purification: take the crude extract of step (3) and dissolve it in ...

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Abstract

The invention relates to a method for purifying collagen derived from pelodiscus sinensis. The method is characterized by comprising the steps of soaking, fat removal and enzymolysis; in an enzymolysis process, carinas of the pelodiscus sinensis, of which impurities are removed, are put into acetic acid with the concentration of 0.5mol / L, wherein the ratio of the carinas to the acetic acid is 1:25, pepsase enzymatic hydrolysate is added for enzymolysis treatment, and extraction is carried out at 4-10 DEG C for 24-48 h, so that a pelodiscus sinensis carina collagen coarse extracting solution is obtained; in a salting-out purification process, the coarse extracting solution is dissolved into a Tris-HCL buffer solution containing NaCl, the mixed solution is centrifuged to obtain supernatant, NaCl is added into the supernatant, salting-out is carried out, centrifugation is carried out again, and precipitates and the supernatant are respectively collected; and precipitates are dissolved into acetic acid with the concentration of 0.5 mol / L, and freeze-drying is carried out after dialysis, so that the pelodiscus sinensis collagen freeze-dried product is obtained. Compared with the prior art, the method provided by the invention has the advantages that collagen extracted through enzymolysis is purified through a salting-out purification method, so that no environment pollution is realized, the extraction efficiency is high and the obtained product has good biological characteristics.

Description

technical field [0001] The invention relates to a method for purifying collagen from soft-shelled turtle, which belongs to the field of natural active substances. Background technique [0002] Collagen is an important structural protein in living organisms and a major component of supporting and connective tissues. The unique triple helical amino acid structure of collagen gives it many useful properties, such as high tensile strength, good biocompatibility, low antigenicity, low irritation and low cytotoxicity, and the ability to promote cell growth. All these properties make it an ideal biomedical material. Biomedical materials derived from collagen have become one of the fastest growing points in the field of medical materials, and the annual sales in the global market have reached more than 4 billion US dollars. [0003] The raw materials used in the traditional extraction of collagen mainly come from connective tissues such as the skin of animals such as pigs and catt...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/30
Inventor 李彩燕杨桠楠钱国英宋伟
Owner ZHEJIANG WANLI UNIV
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