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 problems of reported and undiscovered collagen extraction technology research, and achieve the effect of short enzymatic hydrolysis time, stable experimental results and high content

Inactive Publication Date: 2010-08-18
赵雨
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are many studies on the extraction process of collagen, but no re

Method used

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

Examples

Experimental program
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Effect test

Embodiment 1

[0034] 1) Take fresh deer tendon, remove fat, skin and fascia, and cut into 0.5-1cm 3 The small pieces were washed repeatedly with 0.9% normal saline to remove serum and fat, then rinsed with distilled water, and stored in a -20°C refrigerator with a water content of about 65%;

[0035] 2) Take 30g of fresh deer tendon that has been processed above, add 300ml of hydrochloric acid solution with a volume concentration of 0.1% according to the ratio of material to liquid 1g: 10ml, and put it in a refrigerator at 4°C for 12 hours, so that the deer tendon absorbs acid water and undergoes acid swelling for 12 hours Finally, take out the deer tendon and place it in boiling water at 100°C until the deer tendon is completely dissolved. It takes 75 minutes, then filter it with gauze cloth sandwiched with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and keep the supernatant ;

[0036] 3) Add 1M NaOH solution to the supernatant to adjust the p...

Embodiment 2

[0042] 1) Take fresh deer tendon, remove fat, skin and fascia, and cut into 0.5-1cm 3 The small pieces were washed repeatedly with 0.9% normal saline to remove serum and fat, then rinsed with distilled water, and stored in a -20°C refrigerator with a water content of about 65%;

[0043] 2) Take 30g of the fresh deer tendon that has been processed above, add 0.2% hydrochloric acid solution according to the ratio of material to liquid 1g: 10ml, put it in the refrigerator at 4°C for 36 hours, make the deer tendon absorb acid water and cause acid swelling, take out the deer tendon after 36 hours Place in boiling water at 100°C until the deer tendon is completely dissolved, which takes 55 minutes, then filter with gauze cloth sandwiched with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and keep the supernatant;

[0044] 3) Add 1M NaOH solution to the supernatant in step 2) to adjust the pH to 7.0, centrifuge (12000rpm / min, 30min), discar...

Embodiment 3

[0050] 1) Take fresh deer tendon, remove fat, skin and fascia, and cut into 0.5-1cm 3 The small pieces were washed repeatedly with 0.9% normal saline to remove serum and fat, then rinsed with distilled water, and stored in a -20°C refrigerator with a water content of about 65%;

[0051] 2) Take 30g of fresh deer tendon that has been processed above, add 0.8% hydrochloric acid solution with a volume concentration of 1g according to the ratio of material to liquid, and put it in a refrigerator at 4°C for 60 hours to make the deer tendon absorb acid water and cause acid swelling, and take out the deer tendon after 60 hours Place in boiling water at 100°C until the deer tendons are completely dissolved, which takes 32 minutes, then filter with gauze cloth sandwiched with absorbent cotton, centrifuge the filtrate (12000rpm / min, 30min), discard the precipitate, and keep the supernatant;

[0052] 3) Add 1M KOH solution to the gelatin solution in step 2) to adjust the pH to 7.0, centr...

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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 in particular relates to a method for preparing deer tendon collagen with good water solubility by adopting hydrochloric acid hydrolysis and trypsin enzymolysis extraction and purification processes. Background technique [0002] The English of collagen is collagen, which is derived from Greek and means "the product of forming glue". It is an important class of protein. In 1940, Orekhovich et al. used citric acid buffer (pH 3.0-4.5) to dissolve water-insoluble proteins from rat skin, which contained keratin and elastin, and there was also a precursor of collagen. Collagen extracted in acidic saline 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 as "tropocollagen", which is the basic struc...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/12
Inventor 赵雨张鹤徐云凤惠歌
Owner 赵雨
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