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Method for preparing biotoxin

A technology for biological toxins and products, applied in the field of preparation of biological toxins, can solve the problems of ion leakage, damage to intestinal wall epithelial cells, and unsatisfactory insecticidal effect, and achieve the effect of good insecticidal effect and good application prospect.

Inactive Publication Date: 2012-09-19
福建省农业科学院农业生物资源研究所
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Problems solved by technology

It is generally believed that after insects feed on crystal proteins, they digest them into active toxin proteins through their own midgut proteases, and the toxin proteins bind to receptors on the brush border membranes (BBMVs) of midgut epithelial cells, and further Inserted into the membrane, forming holes or ion channels, causing ion leakage, destroying the epithelial cells of the intestinal wall, causing dissolved substances in the intestinal tract to penetrate into the blood cavity, causing sepsis; although Bacillus thuringiensis is widely used, its insecticidal effect is not ideal

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  • Method for preparing biotoxin

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Embodiment Construction

[0023] The preparation method of a kind of biotoxin of the present invention, extracts the insecticidal protein of Bacillus thuringiensis by activating Bacillus thuringiensis bacterial classification, fermenting and cultivating and cracking, centrifugal purification; A spacer arm containing a carboxyl group is constructed; then under the joint action of the cross-linking agent EDC and N-hydroxysuccinimide (NHS), the carboxyl spacer arm at C4"-OH is constructed. Abamectin and Bacillus thuringiensis The insecticidal protein is coupled, and the product synthesized by the coupling is purified by ultrafiltration and centrifugation to obtain the biotoxin.

[0024] In the present invention: the structure of Abamectin does not have an active group that can directly combine with the Bacillus thuringiensis insecticidal protein under mild conditions, such as -COOH or NH 2 ; But there are 3 hydroxyl groups available for derivatization in Abamectin: C 4 "-OH,C 5 -OH,C 7 -OH, therefore, ...

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Abstract

The invention provides a method for preparing biotoxin. The method comprises the following steps of: activating bacillus thuringiensis strains, performing fermentation culture, cracking, and centrifuging and purifying insecticidal protein of the bacillus thuringiensis; constructing a spacer arm containing carboxyl at abamectin C4''-OH by adopting a succinic anhydride method; and coupling the abamectin and the insecticidal protein of the bacillus thuringiensis under coaction of ethylene dichloride (EDC) and n-hydroxysuccinimide (NHS) which serve as cross linkers, performing ultra-filtration and centrifugal purification on the coupled product, and thus obtaining the biotoxin. The method has the advantages that the biotoxin with a good pest killing effect is obtained and has a good application prospect in prevention and control of agricultural pests.

Description

technical field [0001] The invention relates to a preparation method of a substance in the biological field, in particular to a preparation method of a biological toxin. Background technique [0002] The growth process of crops is harmed by pests, and the control of pesticides is an important measure for a good harvest. Whether it is chemical pesticides or biological pesticides, pests can develop resistance to them. The rapid development speed requires research to provide methods for structural modification and transformation of pesticides, especially biopesticides, to deal with the development of drug resistance, so as to expand the insecticidal spectrum and improve the insecticidal effect. The resistance of pests to pesticides is to overcome the role of pesticide sites and make them ineffective and passivated. In practical applications, the rotation application of pesticides, the mixed application of pesticides, the increase of the contact area of ​​pesticides to pests, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/107C12P21/00C12R1/07
Inventor 刘波朱育菁潘志针阮传清刘芸唐建阳
Owner 福建省农业科学院农业生物资源研究所
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