Earthworm protein EP-2 and its separation method and uses

A technology of EP-2 and protein, which is applied in the field of earthworm protein EP-2 and its separation, can solve the problems of no separation of the active ingredient protein and the mechanism of action

Inactive Publication Date: 2007-05-23
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no research report on the separation and mechanism of the active ingredient protein that has medicinal effects on earthworms.

Method used

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  • Earthworm protein EP-2 and its separation method and uses

Examples

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Comparison scheme
Effect test

Embodiment 1

[0043] (1) Preparation of crude protein solution of Eisenia chinensis

[0044] Take fresh earthworms and wash them repeatedly in water, drain their digestive tract dirt, wash them, weigh them, add 2 times the amount of 20mM phosphate buffer with a pH value of 6.8, and mash them in a tissue grinder to make Homogenize, extract at room temperature for 4-6 hours, centrifuge at 4°C for 20 minutes at high speed, take the supernatant, slowly add the ground ammonium sulfate solid at 4°C, and stir while adding, so that the saturation of ammonium sulfate reaches 80 %, place overnight at 4°C, centrifuge at 4°C for 10 minutes at high speed, take the precipitate, suspend the precipitate with 20mM phosphate buffer saline with a pH value of 6.8, and put it in a dialysis bag with a molecular weight of 3500 for 20mM phosphate buffer with a pH value of 6.8 Dialyze for 48 hours, change the medium 4-5 times in the middle, centrifuge the obtained protein solution at a high speed for 20 minutes at ...

Embodiment 2

[0048] (1) Preparation of crude protein solution of Eisenia chinensis

[0049] Take fresh earthworms and wash them repeatedly in water to drain their digestive tract dirt, wash them, weigh them, add 3 times the amount of water, mash them in a tissue grinder, make a homogenate, and extract them at room temperature for 4-6 hours , centrifuge at 4°C for 20 minutes at high speed, take the supernatant, slowly add an equal amount of acetone at 4°C, stir evenly, leave it at 4°C overnight, centrifuge at 4°C for 10 minutes at high speed, take the precipitate, suspend it with water, and high-speed at 4°C Centrifuge for 20 minutes, and the supernatant is earthworm crude protein solution.

[0050] (2) Purification of crude protein from Eisenia chinensis

[0051] Take the crude protein solution of Eisenia chinensis, centrifuge at 4°C for 20 minutes at high speed, put it on a DEAE SepharoseCL-6B weak anion exchange resin column, elute with 300mM NaCl, collect the eluate, dialyze to remove ...

Embodiment 3

[0053] (1) Preparation of crude protein solution of Eisenia chinensis

[0054] Take fresh earthworms and wash them repeatedly in water to drain the dirt from their digestive tract, wash them, weigh them, add 1 times the amount of sodium chloride solution, mash them in a tissue blender to make a homogenate, and extract them at room temperature for 4 -6 hours, high-speed centrifugation at 4°C for 20 minutes, take the supernatant, slowly add an equivalent amount of 95% ethanol at 4°C, stir well, leave it overnight at 4°C, centrifuge at 4°C for 10 minutes, take the precipitate, and dissolve the precipitate in water Suspend and centrifuge at a high speed at 4°C for 20 minutes, and the supernatant is earthworm crude protein solution.

[0055] (2) Purification of crude protein from Eisenia chinensis

[0056] Take the crude protein solution of Ulceria chinensis, centrifuge at 4°C for 20 minutes at high speed, put it on a DEAE Sepharose A-50 anion exchange resin column, elute with 200...

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Abstract

The invention relates to protein earthworms EP-2, as well as its methods and applications. The disclosed earthworms protein EP-2 is based on fresh Eiseniafoetida as raw materials. The said method includes extracting by water-buffer homogenate, precipitating with precipitant, purifying with weak anion exchange resin to obtain a mixture of proteins, including protein F-I-0 and F-I-1 and fibrinolytic enzyme A component of 30%-40%. The earthworms protein EP-2 has a significant role in anti-asthma, and is a kind of active proteins with high medicinal value.

Description

technical field [0001] The invention relates to the technical field of active ingredients of traditional Chinese medicine and its extraction and separation, in particular to earthworm protein EP-2 with antiasthma effect, its separation method and application. Background technique [0002] The name of the earthworm medicine is Dilong, and its medicinal use is first recorded in "The Book of Songs", and "Shen Nong's Herbal Classic" is listed as a lower grade. The traditional Chinese medicine Dilong has the effects of clearing away heat and calming convulsions, dredging collaterals, relieving asthma and diuresis, and is a commonly used animal medicinal material. The terrestrial dragons recorded in the 2005 edition of the Chinese Pharmacopoeia are Pheretima aspergillum (E.Perrier), Pheretima vulagris Chen, Pheretima guillelm (Michaelsen) or Pheretima pectinifera Michaelsen. dry body. In addition to the species recorded in the Pharmacopoeia: P.hupeiensis Hubei, P.tschiliensis Zh...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K14/435A61K38/17A61P11/06A61P11/00
Inventor 冯怡徐德生
Owner SHANGHAI UNIV OF T C M
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