Method of extracting and purifying trypsin in pancrease slag

A technology of trypsin and pancreatic residue, applied in the field of enzymology, and its separation or purification, can solve problems such as pollution and complicated activation procedures of trypsin, reduce environmental pollution, reduce the risk of clogging and scaling, and be simple to operate Effect

Active Publication Date: 2009-05-27
JIANGSU WANBANG BIOPHARMLS +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Therefore, the technical problem to be solved by the present invention is to provide a method for extracting and purifying trypsin from pancreatic residue, so as to solve the problems in the prior art that the activation procedure of trypsin is complicated, the trypsin affinity chromatography column is easy to scale and the The organic solvent in the purification process pollutes the technical defect of the environment, finally the method of the present invention reaches the purpose of improving the yield and activity of trypsin, and reducing cost and reducing pollution

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  • Method of extracting and purifying trypsin in pancrease slag
  • Method of extracting and purifying trypsin in pancrease slag
  • Method of extracting and purifying trypsin in pancrease slag

Examples

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Embodiment 1

[0030] Example 1 Extraction and activation of trypsin

[0031] Mix 250 g of pancreatic residue from which insulin has been extracted with 1000 mL of water, adjust the pH to 1.5 with 6 mol / L HCl solution, and stir for 2.5 hours; filter and collect the supernatant. Adjust the pH of the extracted supernatant to 6.0 with 10mol / L sodium hydroxide solution, and add calcium chloride to make Ca 2+ The final concentration is 10mmol / L, stir for 2.5 hours; filter, collect the supernatant, and operate at a temperature of 4-25°C.

Embodiment 2

[0032] Example 2 Extraction and activation of trypsin

[0033] Directly mix 1000mL of pH 2.0 hydrochloric acid and 250g of pancreatic residue, stir for 2.5hr, extract trypsin, filter, and collect the supernatant. Adjust the pH of the extracted supernatant to 7.0 with 10mol / L sodium hydroxide solution, and add calcium chloride to make Ca 2+ The final concentration was 10mmol / L, stirred for 2.5 hours; filtered, and the supernatant was collected. The operating temperature is 4-25°C.

Embodiment 3

[0034] Example 3 Extraction and activation of trypsin

[0035] Mix 250 g of pancreatic residue from which insulin has been extracted with 1000 mL of water, adjust the pH to 2.5 with 6 mol / L HCl solution, and stir for 2 hours; filter and collect the supernatant. Adjust the pH of the extracted supernatant to 7.0 with 10mol / L sodium hydroxide solution, and add calcium chloride to make Ca 2+ The final concentration was 20mmol / L, stirred for 3 hours; filtered, and the supernatant was collected. The operating temperature is 4-25°C.

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Abstract

The invention relates to the preparation of extracting and purifying tryptic from slag, to solve the problem of the complicated pancreatin activation procedures, technical defects of organic solvent pollution of the environment in purification process. Steps include use inorganic acid to extract trypsin from pancreas residue; Adjust pH of extract to 6.0 to 8.0, with Ca2+ activated; adjust the pH of supernatant to 3.0-4.0.By the way of cation exchange chromatography to collect activity elution containing trypsin; Using affinity chromatography purification that is a further purification of trypsin.

Description

technical field [0001] The invention relates to the technical field of enzymology and its separation or purification method, in particular to a method for extracting and purifying trypsin from pancreatic residue. Background technique [0002] Trypsin is a commonly used proteolytic enzyme that catalyzes the hydrolysis of peptide bonds composed of carboxy-terminal lysine or arginine residues, regardless of the length of the peptide chain or its amino acid sequence. However, trypsin itself is easy to degrade spontaneously. The original β-trypsin is converted into α-trypsin, and then further degraded into pseudotrypsin and even fragments, and its activity gradually decreases until it is completely lost. [0003] The extraction and purification of trypsin from pancreas is of great industrial value, because trypsin as a proteolytic enzyme has been widely used in medical fields, beer fog removal and meat tenderization. Trypsin is now widely used in the field of protein chemistry, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/76
Inventor 吴世斌王伟刚
Owner JIANGSU WANBANG BIOPHARMLS
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