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Method for extracting chymocotrypsin

A technology of chymotrypsin and pancreas, which is applied in the field of extracting chymotrypsin, can solve the problems of high cost, not easy to be widely used, and high technical requirements, and achieve the effect of saving cost, shortening time, and simple and feasible extraction and separation methods

Inactive Publication Date: 2011-09-07
马忠仁 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

They are first prepared into crude products of chymotrypsin, then purified by CM-cellulose column chromatography and affinity chromatography, and freeze-dried to obtain chymotrypsin with high purity. However, due to its complicated preparation process, technical requirements High, expensive, not easy to be widely used

Method used

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  • Method for extracting chymocotrypsin
  • Method for extracting chymocotrypsin
  • Method for extracting chymocotrypsin

Examples

Experimental program
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Effect test

Embodiment 1

[0015] The present invention has the following sequential steps:

[0016] a. Take fresh bovine pancreas as the raw material, remove the fat and connective tissue, wash it with clean water, and grind it with a meat grinder.

[0017] b. Take 1 kg of minced beef pancreas and move it into a stainless steel bucket, add 4000 ml of tap water, and use 15% H 2 SO 4 The solution was adjusted to a pH value of 3.0, stirred continuously at room temperature for 24 hours, and then centrifuged at a speed of 4000 rpm for 30 minutes to obtain extract I and pancreatic residue precipitate I.

[0018] c. Pour the obtained 4000 ml of extraction solution I into the extraction solution bucket for later use, stir and wash the pancreatic residue sediment I with 2000 ml of tap water with a pH value of 3.0 for 20 minutes, and then use the speed of 4000 rpm Centrifuge for 30 minutes to obtain extract II and pancreatic residue precipitate II.

[0019] d. Pour the extraction solution II into the extracti...

Embodiment 2

[0025] The present invention has the following sequential steps:

[0026] a. Take the frozen bovine pancreas as raw material, remove the fat and connective tissue, wash it with clean water, and mince it with a meat grinder.

[0027] b. Move 2 kg of minced beef pancreas into a stainless steel bucket, add 8000 ml of tap water, and use 15% H 2 SO 4 The solution was adjusted to a pH of 3.0, stirred continuously at room temperature for 24 hours, and then centrifuged at a speed of 4000 rpm for 30 minutes to obtain the extract I and the bovine pancreas residue precipitate I.

[0028]c. Pour the obtained 8000 ml of leaching solution I into the leaching solution bucket for later use. Stir and wash the bovine pancreas residue precipitate I with 4000 ml of tap water with a pH value of 3.0 for 20 minutes, and then use 4000 rpm Centrifuge at high speed for 30 minutes to obtain extract II and bovine pancreas residue precipitate II.

[0029] d. Pour the leaching solution II into the leach...

Embodiment 3

[0035] The present invention has the following sequential steps:

[0036] a. Take fresh bovine pancreas as the raw material, remove the fat and connective tissue, wash it with clean water, and grind it with a meat grinder.

[0037] b. Move 5 kg of minced beef pancreas into a stainless steel bucket, add 20,000 ml of tap water, and use 15% H 2 SO 4 solution, which was adjusted to a pH value of 3.0, stirred continuously at room temperature for 24 hours, and then centrifuged at a speed of 4000 rpm for 30 minutes to obtain the extract I and the pancreatic residue precipitate I.

[0038] c. Pour the obtained 20,000 ml of extraction solution Ⅰ into the extraction solution bucket for later use, and stir and wash the pancreatic residue sediment Ⅰ with 10,000 ml of tap water with a pH value of 3.0 for 20 minutes, and then use a speed of 4,000 rpm Centrifuge for 30 minutes to obtain extract II and pancreatic residue precipitate II.

[0039] d. Pour the extraction solution II into the ...

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Abstract

The invention relates to the field of bioengineering, in particular relating to a method for extracting chymocotrypsin. In the method, CaCl2 is used for activating chymocotrypsin zymogen into chymocotrypsin while saturated (NH4)2SO4 is added, thus CaSO4 is generated to adsorb the chymocotrypsin. The complicated steps that a salting-out multiple crystallization method combined with dialysis and an organic solvent method using ethanol, acetone and the like are adopted to prepare crude chymocotrypsin and then CM-cellulose column chromatography and affinity chromatography are adopted, are not adopted for purification and preparation; and the steps of elution, salting-out, ultrafiltration and freeze-drying are directly performed to obtain white freeze-dried powder. The extraction method is simple and feasible and is suitable for industrial production; and by adopting the method, the cost can be saved and the time can be shortened from about 7-8 days to 3-4 days. 4.0-4.5g of freeze-dried chymocotrypsin powder can be prepared from one kilogram of pancreas, wherein the specific activity of the chymotrypsin is 360U. / mg protein (nitrogen) and the specific activity of trypsin is 1380U. / mg protein (nitrogen).

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for extracting chymotrypsin. Background technique [0002] Both chymotrypsin and trypsin are typical serine proteases. Their properties are very similar and difficult to separate, but they have different specificities for proteolysis. Trypsin acts only on the C-terminal peptide bond formed by the carboxyl group of the basic amino acid L-arginine or lysine, while chymotrypsin selectively hydrolyzes aromatic or aliphatic amino acids (such as phenylalanine , tyrosine) peptide bond. Therefore, the two of them have the effect of synergistically hydrolyzing protein peptide bonds, which is better than the effect of using one enzyme alone, and has a wider application range. At present, a lot of research has been done on them at home and abroad, and they have been used clinically as a kind of anti-inflammatory and detumescence drugs. Other uses such as: leather industry to remove...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/76
Inventor 马忠仁印明善冯玉萍李明生蔡翠萍乔自林冯若飞阮小华
Owner 马忠仁
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