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Malania oleifera lectin and method for preparing same

A technology of lectin and nuts, which is applied to the preparation method of peptides, chemical instruments and methods, and pharmaceutical formulations, can solve the problems of research and application of lectins from garlic nuts, and achieve comprehensive and efficient utilization, high resistance HIV activity, simple and rapid effect of extraction and purification process

Inactive Publication Date: 2011-09-07
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, until now, there is no report on the research and application of garlic nut lectin

Method used

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  • Malania oleifera lectin and method for preparing same
  • Malania oleifera lectin and method for preparing same
  • Malania oleifera lectin and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A. Crush the dried garlic nuts to 100 mesh, weigh 100g of powder, dissolve in 800g of water at 4°C, stir at 4°C for 24 hours, then filter the mixture to obtain a yellow transparent filtrate;

[0024] B. Add ammonium sulfate powder to the filtrate, stir and dissolve until the saturation of ammonium sulfate reaches 40%, stop adding, let stand at 4°C for 2 hours, then centrifuge at 0°C at 8000rpm, take the supernatant to continue Add ammonium sulfate powder until the saturation reaches 70%, and after standing for 2 hours, perform centrifugation at 4°C and 4000 rpm, remove the supernatant, and take the precipitate, which is the crude lectin;

[0025] C. The crude lectin obtained in step B is dissolved in distilled water, and then dialyzed to remove salt, and the membrane permeation molecular weight is ≤ 10kd to obtain a concentrated solution;

[0026] D. Purify the concentrated solution obtained in step C through an ion exchange column, hang the column with 0.02mol / L Tr...

Embodiment 2

[0029] A. Crush the dried garlic nuts to 200 mesh, weigh 100g of powder, dissolve in 300g of water at 0°C, stir for 48 hours at 0°C, and then filter the mixture to obtain a yellow transparent filtrate;

[0030] B. Add ammonium sulfate powder to the filtrate, stir and dissolve until the saturation of ammonium sulfate reaches 25%, stop adding, let it stand at 0°C for 1 hour, centrifuge at 4°C at 1000rpm, and take the supernatant Continue to add ammonium sulfate powder until the saturation reaches 75%. After standing still for 1 hour, perform centrifugation at 0°C and 1000 rpm, remove the supernatant, and take the precipitate, which is the crude lectin;

[0031] C. The crude lectin obtained in step B is dissolved in distilled water, and then dialyzed to remove salt, and the membrane permeation molecular weight is ≤ 10kd to obtain a concentrated solution;

[0032] D. Purify the concentrated solution obtained in step C through an ion exchange column, hang the column with 0.02mo...

Embodiment 3

[0035] A. Crush the dried garlic nuts to 40 mesh, weigh 100g of powder, dissolve in 500g of water at a temperature of 10°C, stir at 10°C for 10h, then filter the mixture to obtain a yellow transparent filtrate;

[0036]B. Add ammonium sulfate powder to the filtrate, stir and dissolve until the saturation of ammonium sulfate reaches 30%, stop adding, let stand at 10°C for 1.5 hours, then centrifuge at 10°C at 4000rpm, and take the supernatant Continue to add ammonium sulfate powder until the saturation reaches 60%. After standing still for 1.5 hours, perform centrifugation at 10°C and 8000 rpm, remove the supernatant, and take the precipitate, which is the crude lectin;

[0037] C. The crude lectin obtained in step B is dissolved in distilled water, and then subjected to ultrafiltration to remove salt, and the membrane permeation molecular weight is ≤10kd to obtain a concentrated solution;

[0038] D. Purify the concentrated solution obtained in step C through an ion exchan...

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Abstract

The invention provides malania oleifera lectin and a method for preparing the same. The method comprises the following steps: grinding malania oleifera, stirring and socking the ground malania oleifera into cold water, grading and precipitating by using ammonium sulfate, ultrafiltering to remove salt, concentrating, carrying out ion exchange chromatography, purifying, ultrafiltering to remove salt, concentrating, freezing and drying to obtain the refined malania oleifera lectin. The prepared malania oleifera lectin has the molecular weight of about 60kd, the electrophoresis purity of above 90% and the potency of 1:10<5> or more, namely the HIV infected cell (inhibition concentration 50) IC50 of 1:10<5>, can agglutinate the red blood cells of a rabbit, a pig, a chicken, a duck and a mouse and can be used for preventing HIV infection.

Description

Technical field [0001] The present invention relates to a kind of Agaricaceae garlic nut lectin and its preparation method, and belongs to the technical field of plant lectin extraction or preparation technology. Background technique [0002] Lectins are a type of non-enzymatic and non-immune proteins or multivalent sugar-binding proteins secreted by animal, plant and microbial cells. They are called because they can generally agglutinate certain cells (such as red blood cells). Their family members are constantly increasing and their functions are complex. Various, such as mitogenesis, virus virulence determination, agglutination of malignant cells, sperm, bone marrow cells, etc. Due to their high sugar-binding specificity, lectins have become very useful tools for studying biology and clinical medicine. They are used to isolate, purify and determine glycocomplexes, cell division, identify microorganisms, tumor diagnosis, treatment and prevention of bone marrow transplantat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/42A61K38/16A61P31/18C07K1/36C07K1/34C07K1/30C07K1/18
Inventor 陈朝银赵声兰徐进葛锋刘迪秋韩本勇熊向峰
Owner KUNMING UNIV OF SCI & TECH
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