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Method for extracting castor crude toxalbumin

An extraction method and a technology of toxic proteins, which are applied to peptide preparation methods, chemical instruments and methods, plant peptides, etc., can solve the problems of low extraction efficiency, high preparation cost, and long time required to achieve simple operation and maintain biological Activity, non-toxic effects of reagents

Inactive Publication Date: 2010-06-30
ZHONGKAI UNIV OF AGRI & ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the steps of the above-mentioned methods are relatively cumbersome, the required time is long, the extraction efficiency is low, the preparation cost is high, and it is difficult to industrialize

Method used

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  • Method for extracting castor crude toxalbumin
  • Method for extracting castor crude toxalbumin
  • Method for extracting castor crude toxalbumin

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Select fresh castor beans, put the castor seeds obtained after shelling or the castor cakes obtained after oil expression into a mortar or colloid mill or pounder; The mass ratio is 1: 3, adding a 20% NaCl aqueous solution as a solvent for grinding; subsequently, centrifuging (20000rmp, 4min) at -20°C to remove the grease layer and miscellaneous protein precipitation, and collect the supernatant to obtain Ricin crude protein solution. Adopt isoelectric point precipitation method, by the acetic acid-sodium acetate buffer solution that pH value is 4.6, the pH value of ricin oil solution is adjusted to the isoelectric point (being PI6.7 and PI6.1) of ricin, Then centrifuge (20000rmp, 4min) at -20°C to precipitate the ricin, discard the supernatant, and take the obtained precipitate as ricin.

Embodiment 2

[0021] Select fresh castor beans, put the castor seeds obtained after shelling or the castor cakes obtained after oil expression into a mortar or colloid mill or pounder; The mass ratio is 1: 30, adding mass percentage is 15% NaCl aqueous solution and grinds as solvent; Subsequently, carry out centrifugation (20000rmp, 4min) at 0 ℃, remove fat layer and miscellaneous protein precipitation, collect supernatant liquid, obtain castor Hemp crude toxin solution. Adopt isoelectric point precipitation method, by the acetic acid-sodium acetate buffer solution that pH value is 6.0, the pH value of castor crude toxin solution is adjusted to the isoelectric point (being PI6.7 and PI6.1) of ricin, Then centrifuge (20000rmp, 4min) at 0°C to precipitate the ricin, discard the supernatant, and take the obtained precipitate as ricin.

Embodiment 3

[0023] Select fresh castor beans, put the castor seeds obtained after shelling or the castor cakes obtained after oil expression into a mortar or colloid mill or pounder; The mass ratio is 1: 3, adding mass percentage is 20% NaCl aqueous solution and grinds as solvent; Subsequently, carry out centrifugation (20000rmp, 4min) at 20 ℃, remove fat layer and miscellaneous protein precipitation, collect supernatant liquid, obtain castor Hemp crude toxin solution. Adopt isoelectric point precipitation method, by the sodium dihydrogen phosphate buffer solution that pH value is 5.0, the pH value of ricin oil solution is adjusted to the isoelectric point (being PI6.7 and PI6.1) of ricin, Then centrifuge (20000rmp, 4min) at 20°C to precipitate the ricin, discard the supernatant, and take the obtained precipitate as ricin.

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PUM

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Abstract

The invention discloses a method for simply and rapidly extracting castor crude toxalbumin. The method comprises the following steps of: 1) shelling castor seeds or expelling oil, and backing up the castor kernel after shelling or the castor cake after expelling oil, 2) taking the castor kernel or the castor cake and adding solvent so as to grind, 3) centrifuging, removing the oil layer and the foreign protein sediment, and collecting the clear liquid so as to acquire castor crude toxalbumin solution, and 4) separating and purifying the acquired castor crude toxalbumin solution by using the isoelectric point sedimentation method so as to acquire the castor crude toxalbumin. The extracting method has simple operation, the required agent is non-toxic, environment-friendly and low-cost, and the method is fit for commercial process. The inorganic salt solution such as NaCl water solution is used as solvent to extract the castor crude toxalbumin so the bioactivity of the castor crude toxalbumin is maintained. Besides, the extracted castor crude toxalbumin is dissolved in the NaCl water solution in order to store and apply the castor crude toxalbumin.

Description

technical field [0001] The present invention relates to a kind of extraction method of protein, more specifically, the present invention relates to a kind of extraction method of ricin bean crude toxic protein. Background technique [0002] Ricin is a ribosome inactivating protein. It was first isolated from castor beans by Dixson in 1887. Its molecular weight is about 64,000-80,000. The crystal forms that have been discovered so far include BI type, TL type Type, CI type and D type, among which D type is the most toxic. Ricin with a molecular weight of 64,000 is covalently connected by two peptide chains A and B with disulfide bonds. With the assistance of chain B (RTB), its A chain (RTA) can easily pass through the cell membrane and destroy the ribosome 60S subunit. unit, inhibits protein synthesis, leading to cell death. The primary structure of Ricin shows that the A chain is composed of 263 amino acid residues and has a molecular weight of 32,000. It is an active chai...

Claims

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Application Information

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IPC IPC(8): C07K1/30C07K1/14C07K14/415
Inventor 宋光泉胡良成孙媚华陈迁梁关生张越华
Owner ZHONGKAI UNIV OF AGRI & ENG
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