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Method for increasing hirudin extraction yield

A technology of hirudin and leech, which is applied to the preparation method of peptides, leech inhibitors, chemical instruments and methods, etc., can solve the problems of unsuccessful realization, excessive dependence on leech resource supply, unstable content of hirudin, etc. To achieve the effect of increasing the concentration of active ingredients

Inactive Publication Date: 2015-09-09
广西复鑫益生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Most of the methods for industrial production of hirudin are to obtain hirudin by destroying the living leech, which affects the purity of the obtained hirudin. At the same time, it is too dependent on the supply of leech resources, and it is especially difficult to avoid the seasonal problem of hirudin production.
Blood-sucking leeches have a high content of anticoagulant active substances, but during the feeding process of blood-sucking leeches, the content of hirudin in the blood-sucking leeches is unstable due to the feeding and fishing time of the same batch of leeches
[0004] Utilize the stimulating effect of electric current on the glands of organisms, directly use DC weak current to stimulate living leeches to extract high-purity hirudin in batches and maintain the living traits of leeches in order to simplify the production process of hirudin, improve product purity, and maintain wild leeches The method of reusing resources, reducing production costs, and overcoming seasonal obstacles in production has not yet been successfully realized.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The method for improving hirudin extraction yield of the present invention comprises the following steps:

[0029] A. Supplementary feeding: supplementary feed at least once 12 hours before fishing; or fishing after 12 hours of normal feeding;

[0030] B. Fishing and electrical stimulation: extract the live leech from the breeding tank to the storage tank at room temperature, rinse with deionized water to remove impurities on the surface, filter out the water, then immerse in normal saline for rinsing, add an electrolyte layer to cover, and pass through Inject direct current to stimulate, collect the liquid secreted by leeches, and put the live leeches back into the breeding tank from the filter tank;

[0031] C. Subsequent treatment: The collected secretions are subjected to subsequent treatment with 800 μm nanofiltration membrane for 30 minutes to remove 80% of the water, add 8% stabilizer, decompress to 0.9 atmospheric pressure, and spray dry to obtain 800AT-U / g of...

Embodiment 2

[0039] The method for improving hirudin extraction yield of the present invention comprises the following steps:

[0040] A. Supplementary feeding: supplementary feeding at least once 24 hours before fishing; or fishing after 24 hours of normal feeding;

[0041] B. Fishing and electrical stimulation: extract the live leech from the breeding tank to the storage tank at room temperature, rinse with deionized water to remove impurities on the surface, filter out the water, then immerse in normal saline for rinsing, add an electrolyte layer to cover, and pass through Inject direct current for stimulation while ensuring appropriate current intensity to avoid the death of leeches, collect the liquid secreted by leeches, and then put the live leeches back into the breeding tank from the filter tank;

[0042] C. Subsequent treatment: The collected secretions were treated with 800 μm nanofiltration membrane for 100 minutes to remove 85% of the water, add 10% stabilizer, depressurize to...

Embodiment 3

[0050] The method for improving hirudin extraction yield of the present invention comprises the following steps:

[0051] A. Supplementary feeding: 36 hours before fishing, supplementary feeding; or fishing after 36 hours of normal feeding;

[0052] B. Fishing and electrical stimulation: extract the live leech from the breeding tank to the storage tank at room temperature, rinse with deionized water to remove impurities on the surface, filter out the water, then immerse in normal saline for rinsing, add an electrolyte layer to cover, and pass through Inject direct current for stimulation while ensuring appropriate current intensity to avoid the death of leeches, collect the liquid secreted by leeches, and then put the live leeches back into the breeding tank from the filter tank;

[0053] C. Subsequent treatment: The collected secretions were treated with 800 μm nanofiltration membrane for 150 minutes to remove 90% of the water, add 12% stabilizer, depressurize to 0.9 atmosphe...

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Abstract

The invention discloses a method for increasing hirudin extraction yield. The method includes: supplementary feeding 12-54 hours before fishing; or fishing 12-54 hours after normal feeding; extracting live leeches to a storage tank under room temperature, rinsing with deionized water, filtering water, immersing into normal saline for rising, adding electrolyte for covering, applying direct currents for stimulation, collecting liquid secreted by the leeches, and returning the live leeches to a cultivation tank from a filtering tank; performing 800-micrometer nano-filtration film subsequent processing on the collected secretion to remove part of moisture, adding stabilizer, and performing spraying drying to obtain antithrombase-active hirudin dry powder. The method has the advantages that leech feeding time is changed to improve the hirudin contents in the leeches, electricity is used to stimulate the live leeches so as to extract the hirudin, the production process of the hirudin is simplified, the live leeches can be kept, and high-purity hirudin can be extracted for many times.

Description

technical field [0001] The invention relates to the technical field of hirudin extraction, in particular to a method for improving the yield of hirudin extraction. Background technique [0002] Hirudin is named after it was first isolated from leeches. Natural hirudin consists of 65 amino acid residues with a molecular weight of 7000 Daltons. Hirudin can irreversibly combine with the catalytic center of thrombin and the binding center of fibrinogen to inactivate thrombin. It is the most powerful thrombin inhibitor found so far. In addition, it has a strong antithrombotic effect, and has inhibitory effects on various experimental thrombus models. Hirudin has preventive and therapeutic effects on various thrombotic diseases such as venous thrombosis, disseminated intravascular coagulation, cerebral coagulation, thrombophlebitis and coronary artery thrombosis, especially the treatment of venous thrombosis and disseminated intravascular coagulation; it can also be used for Pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/815C07K1/34A61N1/18A01K67/033
CPCC07K14/815A01K67/033A61N1/18
Inventor 谢海林
Owner 广西复鑫益生物科技有限公司
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