Preparation method of protaminase B and its composition

A technology of carboxypeptidase and balance, applied in the field of preparation method of carboxypeptidase B and its composition, can solve problems such as instability of carboxypeptidase B preparations, achieve the effects of shortening the production cycle, reducing production costs, and facilitating control

Active Publication Date: 2010-09-01
JIANGSU WANBANG BIOPHARMLS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to overcome the unstable defect of carboxypeptidase B preparation in the prior art, the inventor has carried out extensive research

Method used

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  • Preparation method of protaminase B and its composition
  • Preparation method of protaminase B and its composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation of embodiment one acetone powder

[0027]Select 1 kg of frozen pig pancreas, cut into thin slices about 2 mm thick, and autolyze at room temperature for 16-24 hours. After autolysis, add pre-cooled acetone for two times, each time add 3 times the amount of acetone (V / W), stir, centrifuge to collect the filter residue, and the filter residue is naturally air-dried to obtain acetone powder containing carboxypeptidase B.

Embodiment 2

[0028] The preparation of embodiment diacetone powder

[0029] Select 1 kg of fresh porcine pancreas, grind it with a meat grinder, add pre-cooled acetone for three times, use 5 times the amount of acetone (v / w) for each time, stir continuously, collect the filter residue by centrifugation, and the filter residue is naturally air-dried to contain Carboxypeptidase B in acetone powder.

Embodiment 3

[0030] The preparation of embodiment triacetone powder

[0031] Select 1 kg of frozen pig pancreas, cut into thin slices about 2 mm thick, and autolyze at room temperature for 16-24 hours. After autolysis, add pre-cooled acetone for two times, each time add 3 times the amount of acetone (V / W), stir, centrifuge to collect the filter residue, and then treat with 2 times the amount of acetone: ether (1:1) mixed solution, stirring continuously , centrifuge to collect the filter residue, and air-dry the filter residue naturally to obtain acetone powder containing carboxypeptidase B.

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Abstract

This invention discloses a preparation of carboxyl peptidase B and its combination relating to enzymology, combination containing enzyme and technology field of disconnecting and depurating method. The technology problem which needs to be solved by this invention is that providing a preparation of carboxyl peptidase B and its combination to solve the yield of carboxyl peptidase B preparation is low and carboxyl peptidase B is astable in present technology. Disclosing a preparation of carboxyl peptidase B, including the steps: salting out, hydrophobic chromatography, anion-exchange chromatography, its characteristic is that using 1-5 times of acetone to process 2-3 times, producing acetone powder; it also discloses a carboxyl peptidase B combination which contains 3-5%(w / v, g / 100ml) manicol. The yield of the invented carboxyl peptidase B achieves more than 60U / g pancreas, raising more than 20%, specific activity of carboxyl peptidase B exceeds 200U / mg, stability of carboxyl peptidase Bis 2 years.

Description

technical field [0001] The present invention relates to the technical field of enzymology, enzyme-containing composition and separation or purification method, in particular to a preparation method of carboxypeptidase B and its composition. Background technique [0002] Carboxypeptidase B (CPB for short, EC3.4.17.2) is a zinc-containing exopeptidase that specifically hydrolyzes basic amino acids at the C-terminus of the peptide chain: arginine, lysine or ornithine acid. Carboxypeptidase B contains 307 amino acid residues and a molecular weight of 35KDa. Carboxypeptidase B present in porcine pancreas is produced by preprocarboxypeptidase B (preprocarboxypeptidase B). The N-terminal of procarboxypeptidase B contains a fragment of 108 amino acids (of which 13 amino acids are the signal peptide sequence and 95 amino acid residues form the active peptide part) connected to the carboxypeptidase B at the C-terminal, and the procarboxypeptidase B In the process of transporting B ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/48
Inventor 李显林杨炜王伟刚
Owner JIANGSU WANBANG BIOPHARMLS
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