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Preparation technology of high-activity trypsin

A technology of trypsin and preparation process, which is applied in the field of trypsin preparation process, which can solve the problems of difficult control, low trypsin activity, and limited use of trypsin, so as to achieve the effect of improving activity and improving work efficiency

Inactive Publication Date: 2017-10-24
天津科仪嘉欣科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the preparation process is relatively simple, the trypsin activity thus prepared is relatively low and difficult to control, thus limiting the use of trypsin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A preparation technique of highly active trypsin is carried out successively according to the following steps:

[0020] Step 1: dissolving the crude trypsin product to obtain the crude trypsin product dissolving solution;

[0021] Step 2: Place the crude trypsin solution at 60-70°C for 0.5-3 hours;

[0022] Step 3: Buffer and dissolve with phosphate buffer to reach equilibrium; the concentration of phosphate buffer is 1 mmol / L.

[0023] Step 4: after balancing, add sodium alginate and calcium chloride solution to the trypsin crude product dissolving solution, stir evenly; the volume ratio of the sodium alginate and calcium chloride solution is 1:5.

[0024] Step 5: adding glutaraldehyde to the well-stirred solution for reaction, and removing excess glutaraldehyde with water;

[0025] Step 6: put the solution into a dialysis bag, immerse it in the external permeate, filter to obtain crystals, dissolve in water, and adjust the pH value to 8; dialyze the solution to remo...

Embodiment 2

[0028] A preparation technique of highly active trypsin is carried out successively according to the following steps:

[0029] Step 1: dissolving the crude trypsin product to obtain the crude trypsin product dissolving solution;

[0030] Step 2: Place the crude trypsin solution at 60-70°C for 0.5-3 hours;

[0031] Step 3: Buffer and dissolve with phosphate buffer to reach equilibrium; the concentration of phosphate buffer is 3mmol / L.

[0032] Step 4: after equilibration, add sodium alginate and calcium chloride solution to the trypsin crude product dissolving solution, and stir evenly; the volume ratio of the sodium alginate and calcium chloride solution is 1:6.

[0033] Step 5: adding glutaraldehyde to the well-stirred solution for reaction, and removing excess glutaraldehyde with water;

[0034] Step 6: put the solution into a dialysis bag, immerse it in the external permeable liquid, filter to obtain crystals, dissolve in water, and adjust the pH value to 8.3; dialyze the...

Embodiment 3

[0037] A preparation technique of highly active trypsin is carried out successively according to the following steps:

[0038] Step 1: dissolving the crude trypsin product to obtain the crude trypsin product dissolving solution;

[0039] Step 2: Place the crude trypsin solution at 60-70°C for 0.5-3 hours;

[0040] Step 3: Buffer and dissolve with phosphate buffer to reach equilibrium; the concentration of phosphate buffer is 5mmol / L.

[0041] Step 4: after equilibration, add sodium alginate and calcium chloride solution to the trypsin crude product dissolving solution, and stir evenly; the volume ratio of the sodium alginate and calcium chloride solution is 1:7.

[0042] Step 5: adding glutaraldehyde to the well-stirred solution for reaction, and removing excess glutaraldehyde with water;

[0043] Step 6: put the solution into a dialysis bag, immerse it in the external permeate, filter to obtain crystals, dissolve in water, and adjust the pH value to 8.5; dialyze the solutio...

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Abstract

The invention belongs to the technical field of biochemical medicine and pharmacy, and in particular relates to a preparation process of highly active trypsin. The crude trypsin solution is obtained by dissolving the crude trypsin; place the crude trypsin solution at 60-70°C for 0.5-3 hours; buffer and dissolve with phosphate buffer to reach equilibrium; Add sodium alginate and calcium chloride solution to the crude product solution of protease, and stir well; add glutaraldehyde to the well-stirred solution for reaction, and remove excess glutaraldehyde with water; put the solution into a dialysis bag, and soak it in the Liquid, filter to obtain crystals, add water to dissolve, adjust the pH value; dialyze the solution to remove impurities; take out the dialysate, filter with suction to obtain the filtrate, adjust the pH value, and obtain the finished product of crystalline trypsin after freeze-drying. It can improve the activity of trypsin and improve work efficiency. The activity of the obtained trypsin reaches above 4000U / mg.

Description

technical field [0001] The invention belongs to the technical field of biochemical medicine and pharmacy, and particularly relates to a preparation process of highly active trypsin. Background technique [0002] Trypsin (Trypsin, EC 3.4.21.4) is synthesized in the pancreas as an enzyme precursor trypsinogen, secreted as a component of pancreatic juice, and is enzymatically hydrolyzed by enterokinase or activated by trypsin itself. Active trypsin is an endopeptidase that cleaves the carboxyl side of lysine and arginine residues in polypeptide chains. It not only acts as a digestive enzyme, but also restricts the decomposition of precursors of other enzymes such as chymotrypsinogen, carboxypeptidase, and phospholipase, and plays an activating role. It is the most specific protease and is important in protein sequencing. tool enzyme. It can hydrolyze proteins into polypeptides or amino acids, and has a selective hydrolysis effect on the peptide chains of arginine and lysine, ...

Claims

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Application Information

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IPC IPC(8): C12N9/76
CPCC12N9/6427C12Y304/21004
Inventor 黄金科
Owner 天津科仪嘉欣科技有限公司
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