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Technique of enzyme

A process method and ph value technology, applied in biochemical equipment and methods, enzymes, peptidases, etc., can solve the problems of low yield and purity of defibrase, unstable quality, difficult to master, etc., achieve short separation time, reagent The effect of small dosage and stable quality

Inactive Publication Date: 2017-11-24
芜湖通全科技有限公司
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, due to the complex process, difficult to master, low yield, unstable quality and other problems, the yield and purity of defibrase are low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0009] A kind of enzyme processing method, comprises the following steps:

[0010] (1) DEAE-sephadex A50 balance method: Take an appropriate amount of DEAE-sephadex A50, add fresh distilled water, heat and boil for 50 minutes, let it cool to room temperature, add pH 8, 0.08mol / L Tris buffer, stir, let stand for several hours, discard the upper Clear liquid, then add pH value 8, 0.08mol / L Tris buffer solution, repeat this several times, and measure pH value = 8;

[0011] (2) Linear elution method: Add snake venom evenly to the DEAE-sephadex A50 column, the first gradient, add 1000ml of pH 7.2, 0.1mol / L NaCl Tris buffer in a stirring bottle, wash at a speed of 10-15 drops / min For the second gradient, add 1000ml of pH 7.2, 0.1mol / L NaCl Tris buffer solution to the stirring bottle, and add 2000ml of pH 7.0-7.8, 0.8mol / L NaCl Tris buffer solution to the storage bottle, at 10-15 drops / min Speed ​​elution; a total of 9 peaks were separated, and the qualified peaks were combined.

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PUM

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Abstract

The invention relates to a technique of an enzyme. The technique of the enzyme comprises the following steps: (1) carrying out a DEAE-sephadex A50 balance method; and (2) carrying out a linear elution method, wherein nine peaks are separated in total, and qualified peaks are combined. The technique provided by the invention has the advantages that a technique for extracting defibrase by virtue of snake venom is adopted, and compared with the original technology, separation time is short, usage amount of a reagent is small, process is simple, yield is high, and quality is stable.

Description

technical field [0001] The invention relates to the field of biochemical technology, in particular to an enzyme process method. Background technique [0002] Defibrase can dissolve thrombus, inhibit thrombus formation, and improve microcirculation; main treatment (1) acute cerebral infarction, including cerebral thrombosis, cerebral embolism, transient ischemic attack (TIA), and prevention of recurrence of cerebral infarction. (2) Prevention of myocardial infarction, unstable angina and recurrence of myocardial infarction. (3) Vascular diseases of the extremities, including femoral artery embolism, thromboangiitis obliterans, and Raynaud's disease. (4) The blood is in a hyperviscosity state, a hypercoagulable state, and a prethrombotic state. (5) Sudden deafness. (6) Pulmonary embolism. However, due to the complex process, difficult to master, problems such as low yield and unstable quality, the yield and purity of defibrase are relatively low. Contents of the inventio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
CPCC12N9/6418C12Y304/00
Inventor 季一全汤佳
Owner 芜湖通全科技有限公司
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