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Method for preparing collagen from deer sinew

A technology of deer tendon collagen and deer tendon, applied in the field of collagen purification, can solve the undiscovered problems of collagen extraction process research and reporting, and achieve the effects of stable experimental results, short enzymatic hydrolysis time and mild enzymatic hydrolysis conditions

Inactive Publication Date: 2012-03-14
赵雨
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on the extraction process of collagen, but no relevant reports on the extraction process of collagen in deer tendon have been found.

Method used

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  • Method for preparing collagen from deer sinew
  • Method for preparing collagen from deer sinew

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1) After removing the fat, skin and fascia from fresh deer tendons, cut them into 0.5~1cm 3 Use 0.9% normal saline to repeatedly clean the small pieces of serum and fat, then rinse with distilled water, and store in a refrigerator at -20°C. The moisture content is about 65%;

[0034] 2) Take 30g of fresh deer tendon processed above, add 300ml, 0.1% volume concentration of hydrochloric acid solution according to the material-to-liquid ratio of 1g: 10ml, and put it in the refrigerator at 4℃ for 12 hours to make deer tendon absorb acid water and acid swell, 12 hours Afterwards, take out the deer tendons and place them in boiling water at 100°C until the deer tendons are completely dissolved. The time required is 75min. Then filter with gauze with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and save the supernatant. ;

[0035] 3) Add 1M NaOH solution to the supernatant to adjust the pH to 7.0, centrifuge (12000rpm / min, 30min), discar...

Embodiment 2

[0041] 1) After removing the fat, skin and fascia from fresh deer tendons, cut them into 0.5~1cm 3 Use 0.9% normal saline to repeatedly clean the small pieces of serum and fat, then rinse with distilled water, and store in a refrigerator at -20°C. The moisture content is about 65%;

[0042] 2) Take 30g of the fresh deer tendon processed above, add 0.2% hydrochloric acid solution at a material-to-liquid ratio of 1g: 10ml and place it in a refrigerator at 4°C for 36 hours to make the deer tendon absorb acid water and acid swell. Take out the deer tendon after 36 hours Place it in boiling water at 100°C until the deer tendon is completely dissolved. The time required is 55 minutes, then filter with gauze with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and save the supernatant;

[0043] 3) Add 1M NaOH solution to the supernatant of step 2) to adjust the pH to 7.0, centrifuge (12000rpm / min, 30min), discard the precipitate, and retain the sup...

Embodiment 3

[0049] 1) After removing the fat, skin and fascia from fresh deer tendons, cut them into 0.5-1cm 3 Use 0.9% normal saline to repeatedly clean the small pieces of serum and fat, then rinse with distilled water, and store in a refrigerator at -20°C. The moisture content is about 65%;

[0050] 2) Take 30g of the fresh deer tendon processed above, add 0.8% hydrochloric acid solution at a material-to-liquid ratio of 1g:15ml and place it in a refrigerator at 4°C for 60 hours to make the deer tendon absorb acid water and acid swell. Take out the deer tendon after 60 hours Place it in boiling water at 100°C until the deer tendon is completely dissolved, the time is 32 minutes, then filter with gauze with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and save the supernatant;

[0051] 3) Add 1M KOH solution to the gelatin solution in step 2) to adjust the pH to 7.0, centrifuge (12000rpm / min, 30min), discard the precipitate and retain the supernatan...

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Abstract

The invention belongs to the technical field of collagen extraction and particularly relates to a method for preparing collagen from deer sinew by using the processes of extraction and purification through hydrochloric acid hydrolysis and trypsin enzymolysis. The method comprises the following steps: removing fat, skin and sinew membrane from the deer sinew; cutting the deer sinew into small pieces with the size being 0.5cm<3> to 1cm<3>; then, repeatedly washing the cut deer sinew with normal saline to remove sera and fat; further rinsing the washed deer sinew with distilled water; adding hydrochloric acid with the volume concentration thereof being 0.05% to 1.0% to the rinsed deer sinew by the solid-liquid ratio being 1g:5ml to 1g:15ml, and soaking the deer sinew in the hydrochloric acid for 12h to 60h; then, feeding the deer sinew in boiling water (100 DEG C) until the deer sinew is completely dissolved; centrifuging; neutralizing the supernatant with alkaline solution; adding trypsin for conducting the enzymolysis process with the ratio between the trypsin and the zymolyte being 1g:5,000g to 1g:50,000g; concentrating the supernatant and filtering with a 0.2mum-0.45mum filter membrane; and freeze-drying the filtrate in a vacuum to obtain the collagen powder. The method of the invention has the advantages of simple and easy-to-operate process and stable experimental result. Therefore, the method is suitable for large-scale industrialized production.

Description

Technical field [0001] The invention belongs to the technical field of collagen purification, and specifically relates to a method for preparing deer tendon collagen with good water solubility by adopting an extraction and purification process of hydrochloric acid hydrolysis and trypsin hydrolysis. Background technique [0002] The English word for collagen is collagen, which is derived from the Greek, meaning "the product of the formation of glue". It is an important type of protein. In 1940, Orekhovich et al. used citric acid buffer (pH 3.0~4.5) to dissolve water-insoluble protein from rat skin, which contained keratin and elastin. In addition, there was also a precursor of collagen. The collagen extracted from the acidic salt solution is named "procollagen". Later, through the efforts of many researchers, it was found that collagen can also be extracted from neutral salt and alkaline salt solutions. In 1953, Gross named the protein monomer that builds collagen "tropocollagen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K1/12
Inventor 赵雨张鹤徐云凤惠歌
Owner 赵雨
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