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Method for extracting trypsin from animal pancreas

A technique for trypsin and pancreas, applied in the field of separation of trypsin

Active Publication Date: 2013-04-24
青岛华迈士药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The products are all mixtures of chymotrypsin, there is no simple extraction of trypsin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The zymogen contained in the pancreatic cells is extracted by using dilute acid solution, and then according to the principle of isoelectric point precipitation, the pH is adjusted to precipitate and remove a large amount of acidic miscellaneous protein and non-protein impurities, and then graded salting out with ammonium sulfate Precipitation of trypsinogen etc., including the following steps:

[0019] 1. Extraction process:

[0020] 1.1 For fresh pancreas, use tweezers and scissors to remove fatty connective tissue, etc., put it in a grinder and grind it, and weigh 15Kg;

[0021] 1.2 Put the minced material in a plastic bucket, soak it with 30L of 0.125mol / L sulfuric acid solution for 24 hours, and stir once an hour during the soaking period;

[0022] 1.3 Filter the impregnation with a filter bag, then soak the filter residue with 15L of 0.125mol / L sulfuric acid solution for 1 hour, and discard the filter residue after drying;

[0023] 2. Salting and salting out pro...

Embodiment 2

[0035] 1. Extraction process:

[0036] 1.1 For fresh pancreas, use tweezers and scissors to remove fatty connective tissue, etc., put it in a grinder and grind it, and weigh 10Kg;

[0037] 1.2 Put the minced material in a plastic bucket, soak it with 25L of 0.150mol / L sulfuric acid solution for 24 hours, and stir it every hour during the soaking period;

[0038] 1.3 Filter the impregnation with a filter bag, then soak the filter residue with 12.5L of 0.150mol / L sulfuric acid solution for 1 hour, and discard the filter residue after drying;

[0039] 2. Salting and salting out process:

[0040] 2.1 Combine the two impregnating solutions with a volume of 37.4L, put them in a plastic bucket, add 225g of solid ammonium sulfate to every 1000ml of solution to reach 35% saturation, and add 8.415Kg of solid ammonium sulfate in total, and let it stand for 12 hours;

[0041] 2.2 Filter, keep the filtrate, the volume of the filtrate is 36.8L, discard the solid;

[0042] 2.3 Add 186g of...

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PUM

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Abstract

The invention discloses a method for extracting trypsin from animal pancreas. According to the technical scheme of the invention, the zymogen in pancreas cells is extracted by using a sulfuric acid solution; subsequently according to the principle of isoelectric precipitation, the pH is adjusted, and the trypsinogen is deposited and precipitated in a classified salting-in and salting out mode by using ammonium sulfate. By utilizing the method, 6g trypsin can be extracted from 1kg of pancreas.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating trypsin from animal pancreas. Background technique [0002] Trypsin (Parenzyme) is a kind of protease, EC3.4.21.4. In vertebrates, acts as a digestive enzyme. Synthesized in the pancreas as the precursor of the enzyme trypsinogen. Secreted as a component of pancreatic juice, it is decomposed into activated trypsin by the restriction of enterokinase or trypsin, which is an endopeptidase that can cut off the carboxyl side of lysine and arginine residues in the polypeptide chain. It not only acts as a digestive enzyme, but also restricts and decomposes the precursors of other enzymes such as chymotrypsinogen, carboxypeptidase, and phospholipase, and activates them. It is the most specific protease, and it becomes an indispensable tool in determining the amino acid arrangement of proteins. [0003] Chymotrypsin and elastase are also closely related in terms of ...

Claims

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Application Information

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IPC IPC(8): C12N9/76
Inventor 王议锋刘翠珍葛翠凤宋超龙
Owner 青岛华迈士药业有限公司
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