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Method for purifying lectin contained in peanut seeds by adopting hydroformylation hemocyte

A blood cell and lectin technology, applied in the field of aldehydized blood cell purification of lectin in peanut seeds, can solve the problems of low content, difficult purification, and obstacles to in-depth research, and achieve the effect of mild conditions and high recovery rate

Active Publication Date: 2015-03-18
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because lectin itself is very unstable, it is easily affected by changes in conditions such as pH, temperature, and ions, and the content in the organism is very low, making its purification very difficult and hindering its in-depth research.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for purifying lectin in peanut seeds by formaldehyde blood cells, characterized in that: the prepared peanut powder is degreased with petroleum ether, mixed with formaldehyde blood cells, coupled, eluted and freeze-dried to obtain lectin ; The specific steps are as follows:

[0021] Preparation of lectin crude extract: Take peanut seeds without mildew and rot and insects, wash them with running water, dry them in a blast drying oven at 30°C, remove the peanut coats from the dried raw materials, crush them with a plant grinder, and pass the crushed materials through Use a 50-mesh sieve to collect the sieved material as peanut powder for extracting lectin; add 20 times (the mass ratio of peanut powder to petroleum ether) petroleum ether to peanut powder, shake it in a constant temperature water bath at 30°C for 5 hours, filter with suction, discard the filtrate, and keep the filter residue , after drying the filter residue in a fume hood, add 5 times the volume o...

Embodiment 2

[0025] Take fresh chicken blood and add anticoagulant for later use; resuspend the blood with anticoagulant added in twice the volume of sterilized phosphate buffered saline (PBS), centrifuge at 2000 rpm for 5 minutes, resuspend with PBS for 3 times, and finally remove the blood cells The specific volume was diluted to 0.05 for use; 10 times the volume of 0.05 blood cells was mixed with 1 times the volume of neuraminidase, washed 3 times with sterilized PBS, and resuspended to the specific volume of 0.2 enzyme-treated blood cells; 4 times the volume of 0.2 specific volume After the enzyme-treated blood cells reacted with 1 volume of aldylation agent for a certain period of time, they were washed with sterilized PBS for 3-4 times, and the blood cells were diluted with PBS to have a specific volume of 0.6. 4°C refrigerator for later use;

[0026] Take the peanut seeds without mildew and rot, wash them with running water, dry them in a blast drying oven at 30°C, remove the peanut ...

Embodiment 3

[0028] Take fresh duck blood and add anticoagulant for later use; then add twice the volume of sterilized phosphate buffer solution (PBS) to resuspend, centrifuge at 2000 rpm for 5 minutes, resuspend twice with PBS, and finally dilute the hematocrit to 0.05 Standby; 10 times the volume of 0.05 blood cells mixed with 1 times the volume of neuraminidase, washed 3 times with sterilized PBS, and resuspended to a specific volume of 0.2 enzyme-treated blood cells; 4 times the volume of 0.3 specific volume of enzyme-treated blood cells mixed with 1 times After reacting for a certain period of time with a certain volume of aldehylation agent, wash with sterilized PBS for 3 times, dilute the blood cells with PBS to the formaldehyde blood cells with a specific volume of 0.5, 4°C refrigerator for later use;

[0029] Take the peanut seeds without mildew and rot and moths, wash them with running water, dry them in a blast drying oven at 30°C, remove the peanut coats from the dried raw mater...

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PUM

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Abstract

The invention discloses a method for purifying lectins contained in peanut seeds by adopting hydroformylation hemocytes. The method is characterized by comprising the following steps of: carrying out ungrease treatment on prepared peanut powder through petroleum ether; then mixing and coupling with the hydroformylation hemocytes; and eluting, and freeze-drying to obtain the lectins. The method disclosed by the invention can be used for carrying out specific adsorption on the lectins contained in the peanut seeds by utilizing the hydroformylation hemocytes and then eluting by utilizing the specific binding capacity of lactose on the lectins, thereby achieving the purpose of lectins purification; and the method disclosed by the invention has the advantages of economy, easiness, convenience, moderate condition and high recovery rate and meets the requirement for high-flux large-scale purification.

Description

technical field [0001] The invention relates to a method for purifying lectin in peanut seeds by formaldehyde blood cells, which belongs to the field of nutrition and food processing. Background technique [0002] Lectin is a kind of protein or glycoprotein that selectively binds or non-covalently binds to sugar groups widely present in animals, plants and microorganisms. The main property of these proteins is that they can specifically interact with carbohydrates and combine with carbohydrate complexes on the cell surface. Lectins play an extremely important role in the life activities of organisms. They are closely related to physiological processes such as cell adhesion, bacterial infection of the host, the body's natural immune defense, and host phagocytes clearing invading bacteria. [0003] Plant lectin was first discovered in the plant seed castor bean by Russia's Stillmark in 1888, and it was named ricin at that time. After decades of in-depth exploration, great pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/42C07K1/22
Inventor 孙杰朱凤杨庆利于丽娜毕洁张初署
Owner SHANDONG PEANUT RES INST