Lumbricus polypeptide having anticoagulant and thrombolytic effects, and enzymatic hydrolysis preparation method and application thereof

An anticoagulant and thrombin technology, applied in the preparation method, application, food preparation and other directions of peptides, can solve the problems of easy to cause immunogenicity, low activity, reduced activity, etc., to improve bioavailability, structure Easy-to-understand, highly controllable effects

Active Publication Date: 2014-08-27
广州速科食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these functional ingredients are extracted by decoction or powdered orally directly. The utilization rate of the former is low, and the dosage of the latter is large, and the activity after entering the human body is not high.
Some people extract a single protein from Earthworm to make preparations, but the oral preparations are easily digested by gastric enzymes and pancreatin after passing through the gastrointestinal tract, and the activity is greatly reduced; and when made into injections, macromolecular proteins are also It is easy to cause immunogenicity and has potential safety hazards
The functional substances obtained by these two m

Method used

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  • Lumbricus polypeptide having anticoagulant and thrombolytic effects, and enzymatic hydrolysis preparation method and application thereof
  • Lumbricus polypeptide having anticoagulant and thrombolytic effects, and enzymatic hydrolysis preparation method and application thereof
  • Lumbricus polypeptide having anticoagulant and thrombolytic effects, and enzymatic hydrolysis preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 20g: 100mL, adjust the pH to 9.0, stir evenly at a speed of 200r / min for 3h, 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 ground dragon protein. Add groundworm protein to water to dissolve, the ratio of protein to water is 20g: 100mL, control the pH value at 10, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to the weight of groundworm protein is 0.6 mL: 100g, the reaction temperature is 50°C, the stirring speed is 200r / min, and the reaction time is 1h. Boil for 10min; then adjust the pH value to 7.5, add trypsin (Sigma company, the U.S.), the ratio of enzyme to ground dragon protein weight is 0.5g: 100g, the reaction temperature is 50 ° C, the stirring speed is 200r / min, the reaction time 1h. Boil for 10min. Pass the enzymatic solution through a DEAE ...

Embodiment 2

[0036] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 25g:100mL, adjust the pH to 9.5, stir evenly at a speed of 250r / min for 4h, 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 ground dragon protein. Add ground dragon protein to water and dissolve, the ratio of protein to water is 25g: 100mL, control the pH value at 9.5, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to ground dragon protein weight is 0.8 mL: 100g, the reaction temperature is 60°C, the stirring speed is 250r / min, and the reaction time is 1.5h. Boil for 10 minutes; then adjust the pH value to 8.0, add trypsin (Sigma company, the United States), the weight ratio of enzyme and earthworm protein is 1g:100g, the reaction temperature is 50°C, the stirring speed is 200r / min, and the reaction time is 1h . Boil for 10min. Pass the enzymatic soluti...

Embodiment 3

[0038] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 30g:100mL, adjust the pH to 10, stir evenly at a speed of 300r / min for 5h, take the supernatant, then adjust the pH to 5.0, and finally obtain protein precipitation, The pH was adjusted to 7.0 and dried to obtain ground dragon protein. Add groundworm protein to water to dissolve, the ratio of protein to water is 30g: 100mL, control the pH value at 8.5, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to the weight of groundworm protein is 1.0 mL: 100g, the reaction temperature is 60°C, the stirring speed is 300r / min, and the reaction time is 1h. Boil for 10min; then adjust the pH value to 8.5, add trypsin (Sigma company, the U.S.), the ratio of enzyme to earthworm protein weight is 1.5g: 100g, the reaction temperature is 45 ° C, the stirring speed is 300r / min, the reaction time 1h. Boil for 10min. Pass the enzymatic solution through a DEAE sepharo...

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Abstract

The invention discloses a lumbricus polypeptide having anticoagulant and thrombolytic effects, and an enzymatic hydrolysis preparation method and an application thereof. The preparation method comprises the steps: cleaning and drying lumbricus dried bodies, and crushing into a powder; adding water, stirring, and centrifuging to obtain a protein precipitate; adding water to dissolve the protein, adding an alkaline protease with the ratio of the enzyme volume to the protein weight of 0.1-3 mL:100 g, and stirring; exterminating enzyme activity, adding trypsin with the ratio of the enzyme mass to the protein weight of 0.5-5 g:100 g, and stirring at the temperature of 25-60 DEG C; allowing the enzymatic hydrolysate to pass through an anion exchange column, collecting a part of eluate; and then passing through a macroporous adsorption resin, carrying out separation and purification, and collecting and drying to obtain a lumbricus anticoagulant peptide powder. A new source is added for high-activity anticoagulant active substances, the product activity is high, the preparation process is convenient and feasible, and the environment is protected; and the lumbricus polypeptide can be used as a raw material of foods for thrombosis prevention, thrombolysis 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 an anticoagulant and thrombus-dissolving earthworm polypeptide and its enzymatic preparation method and application. Background technique [0002] Dilong, that is, earthworm, is cold in nature and slightly salty in taste. It is a traditional Chinese medicinal material in my country. According to records, more than 2,000 years ago, our ancestors used earthworm to treat qi deficiency and blood stagnation, stroke, hemiplegia and other diseases caused by vein stasis. "Compendium of Materia Medica" lists more than 40 kinds of earthworm prescriptions. Modern pharmacological research has found that it not only anticoagulant, inhibits platelet aggregation, antiarrhythmia, but also has anticancer and analgesic effects. Most of these functional ingredients are extracted by decoction or powdered orally directly. The utilization rate of the former is...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K4/12C07K1/22C07K1/18A61P7/02A23L1/29A23L33/00
Inventor 吴晖任尧赖富饶李晓凤
Owner 广州速科食品有限公司
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