An anticoagulant soil element polypeptide and its enzymatic hydrolysis preparation method and application

An anti-coagulation and soil-element technology, which is applied to medical preparations containing active ingredients, blood diseases, pharmaceutical formulations, etc., can solve the problem of no further separation and purification, achieve a simple and easy-to-understand structure, and improve bioavailability. , the effect of efficient absorption

Active Publication Date: 2017-06-06
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention in this patent mainly uses pepsin and trypsin biomimetic enzymolysis to prepare mixed polypeptide substances for the treatment of cardiovascular and cerebrovascular diseases, tumors, bruises, etc., without further separation and purification, and the determination of anticoagulant activity Only PT single indicator is used

Method used

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  • An anticoagulant soil element polypeptide and its enzymatic hydrolysis preparation method and application
  • An anticoagulant soil element polypeptide and its enzymatic hydrolysis preparation method and application
  • An anticoagulant soil element polypeptide and its enzymatic hydrolysis preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033]Take 500g of clean Tuyuan dry powder, the ratio of dry powder to water is 10g:100mL, adjust the pH to 9.0, stir evenly at a speed of 200r / min for 3 hours, take the supernatant, and then adjust the pH to 4, and finally obtain protein precipitation. The pH was adjusted to 7.0, and dried to obtain earth element protein. Add the earth element protein to dissolve in water, the ratio of protein and water is 10g:100mL, control the pH value at 10, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to earth element protein weight is 1mL : 100g, the reaction temperature is 50°C, the stirring speed is 200r / min, and the reaction time is 3h. Boil for 10 minutes, pass the enzymatic solution through the macroporous adsorption resin DA201-C exchange column, gradually increase the concentration of ethanol starting from pure water, and carry out gradient elution with ethanol at different mass percentage concentrations of 0, 25, 50, 75, and 100...

Embodiment 2

[0035] Take 500g of clean Tuyuan dry powder, the ratio of dry powder to water is 15g: 100mL, adjust the pH to 9.5, stir evenly at a speed of 100r / min for 4 hours, take the supernatant, and then adjust the pH to 4.5, and finally obtain protein precipitation. The pH was adjusted to 7.0, and dried to obtain earth element protein. Add the earth element protein to dissolve in water, the ratio of protein and water is 15g:100mL, control the pH value at 9, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to earth element protein weight is 1.5 mL: 100g, the reaction temperature is 60°C, the stirring speed is 150r / min, and the reaction time is 3.5h. Boil for 10 minutes, pass the enzymatic solution through the macroporous adsorption resin DA201-C exchange column, gradually increase the concentration of ethanol starting from pure water, and carry out gradient elution with different concentrations of ethanol at 0, 25, 50, 75, and 100%, and th...

Embodiment 3

[0037] Take 500g of clean Tuyuan dry powder, the ratio of dry powder to water is 20g: 100mL, adjust the pH to 10, stir evenly at a speed of 300r / min for 5h, take the supernatant, and then adjust the pH to 5.0, and finally obtain protein precipitation. The pH was adjusted to 7.0, and dried to obtain earth element protein. Add the earth element protein to dissolve in water, the ratio of protein and water is 20g: 100mL, control the pH value at 8.5, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to earth element protein weight is 3mL : 100g, the reaction temperature is 65°C, the stirring speed is 250r / min, and the reaction time is 2h. Boil for 10 minutes, pass the enzymatic solution through the macroporous adsorption resin DA201-C exchange column, gradually increase the concentration of ethanol starting from pure water, and carry out gradient elution with different concentrations of ethanol at 0, 25, 50, 75, and 100%, and the flow ...

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Abstract

The invention discloses a ground beetle polypeptide having an anticoagulant effect, and an enzymatic hydrolysis preparation method and an application thereof. The preparation method comprises the steps: cleaning and drying ground beetle dried bodies, and crushing into a powder; adding water, stirring evenly, carrying out uniform-velocity stirring for 0.5 h-4 h, centrifuging to obtain a protein precipitate, and drying; adding water to dissolve the protein with the ratio of 4-25 g:100 mL, adjusting the pH to 7.0-11, adding an alkaline protease with the ratio of the enzyme volume to the protein weight of 0.1-5 mL:100 g, and stirring at the temperature of 30-70 DEG C; and allowing the enzymatic hydrolysate to pass through a macroporous adsorption resin exchange column, carrying out separation and purification, and collecting and drying to obtain a ground beetle anticoagulant peptide powder. A new source is added for anticoagulant active substances, the raw material is fully utilized, the product activity is high, the preparation process is convenient and feasible, and the environment is protected; and the ground beetle polypeptide is wide in application, can be used as a raw material of foods for thrombosis prevention, thrombus dissolving and blood consistency reduction, is supplemented with lactose, mannitol and the like, and is made into a powder, a tablet, a capsule or an oral liquid.

Description

technical field [0001] The present invention relates to a biological peptide and its preparation method and application, in particular to a polypeptide with anticoagulation and its enzymolysis preparation method and application. Background technique [0002] Tuyuan, also known as ground beetle or ground beetle, is the dried female body of Eupolyphagasinesis Walker or Steleophage plancyi (Boleny), a traditional Chinese medicine, and is widely used clinically. , which was first recorded in the book "Shen Nong's Materia Medica" in the Qin and Han Dynasties, has the functions of dispelling blood stasis, strengthening bones and tendons, reducing swelling and pain, and promoting menstrual flow. [0003] The traditional medicine of Tuyuan is to crush the original medicinal material and take it directly or take it after heating water extraction or alcohol extraction. The utilization rate is low, and the activity brought after entering the human body is not high. At present, someone...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K4/12C07K1/22A61K38/17A61P7/02A23L33/00
Inventor 吴晖任尧赖富饶李晓凤
Owner SOUTH CHINA UNIV OF TECH
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