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Method for preparing carboxypeptidase B and composition of carboxypeptidase B

A technology of carboxypeptidase and balance, which is applied in the field of preparation of carboxypeptidase B and its composition, can solve problems such as the instability of carboxypeptidase B preparations, and achieve the effects of shortening the production cycle, being easy to control, and accelerating release

Inactive Publication Date: 2010-12-22
JIANGSU WANBANG BIOPHARMLS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 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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  • Method for preparing carboxypeptidase B and composition of carboxypeptidase B
  • Method for preparing carboxypeptidase B and composition of carboxypeptidase B
  • Method for preparing carboxypeptidase B and composition of carboxypeptidase B

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The preparation of embodiment one acetone powder

[0028]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

[0029] The preparation of embodiment diacetone powder

[0030] 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

[0031] The preparation of embodiment triacetone powder

[0032] 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

The invention discloses a method for preparing carboxypeptidase B and a composition of the carboxypeptidase B, and relates to the technical fields of enzymology, compositions containing enzymes, and separation or purification methods. The invention aims to solve the technical problem of providing the method for preparing the carboxypeptidase B and the composition of the carboxypeptidase B to overcome the defects of low yield and unstable carboxypeptidase B in the conventional method for preparing the carboxypeptidase B. The invention discloses the method for preparing the carboxypeptidase B, which comprises the following steps of: salting out, hydrophobic chromatography and anion exchange chromatography, and is characterized by treating 2 to 3 times by using 1 to 5 times of acetone to prepare acetone powder; and the invention discloses a carboxypeptidase B composition which contains 3 to 5 weight parts of mannitol. The carboxypeptidase B has the yield of more than 60 U / g of pancreas, which is improved by over 20 percent, the specific activity of more than 200 U / mg, and the stability of 2 years.

Description

[0001] This application is a divisional application of 200510030397.X (application date: October 11, 2005). technical field [0002] 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 [0003] 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...

Claims

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Application Information

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IPC IPC(8): C12N9/48
Inventor 李显林杨炜王伟刚
Owner JIANGSU WANBANG BIOPHARMLS
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