Preparation method of bacillus thuringiensis insecticidal protein and spore endospore adsorption composite dosage form

A technology of Bacillus thuringiensis and insecticidal protein, which is applied in insecticides, botanical equipment and methods, biocides, etc., can solve the problems affecting the unit mass titer of adsorbent preparations, low adsorption capacity, etc., to improve insecticidal activity, The effect of large adsorption capacity and improved insecticidal activity

Inactive Publication Date: 2011-05-04
TIANJIN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The adsorption amount of protoxin protein on the mineral carrier is low, which will inevitably affect the unit mass titer of the adsorption preparation

Method used

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  • Preparation method of bacillus thuringiensis insecticidal protein and spore endospore adsorption composite dosage form

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Bacillus thuringiensis insecticidal protein and spore montmorillonite adsorption composite formulation

[0027] Take 1000 mL of Bacillus thuringiensis fermentation broth (solid content 6.5%, w / v), and detect the insecticidal protein 0.51% by SDS-PAGE. Weigh 30 g of montmorillonite (particle size < 2 microns), add 500 mL of deionized water to make a 6% (w / v) suspension. The montmorillonite suspension was added into the fermentation broth of Bacillus thuringiensis under stirring. Heat the fermentation broth in a water bath to raise the temperature to 37°C, slowly add 1mol / L sodium hydroxide solution dropwise under stirring to adjust the pH to 11-12, cool down to 25°C after 5 hours, adjust the suspension to pH 6-7 with 1mol / L HCl, Stirring was continued for 3 h.

[0028] Put the Bacillus thuringiensis adsorption suspension into a centrifuge tube, centrifuge at 15,000 r / min for 10 min, and pour off the supernatant to obtain the Bacillus thuringiensis insecticid...

Embodiment 2

[0029] Example 2: Bacillus thuringiensis insecticidal protein and spore kaolin adsorption compound formulation

[0030]Take 1000 mL of Bacillus thuringiensis fermentation broth (solid content 6.5%, w / v), and detect the insecticidal protein 0.51‰ by SDS-PAGE. Weigh 80 g of kaolin (particle size < 2 microns) and add 1000 mL of deionized water to make an 8% (w / v) suspension. The kaolin suspension was added into the fermentation broth of Bacillus thuringiensis under stirring. Heat the fermentation broth in a water bath to raise the temperature to 60°C, slowly add 1mol / L sodium hydroxide solution dropwise under stirring to adjust the pH to 11-12, and cool down to 20°C after 4 hours, adjust the suspension to pH 7-8 with 1mol / L HCl, Stirring was continued for 2 h. Put the adsorbed suspension into a centrifuge tube, centrifuge at 15,000 r / min for 10 min, and pour off the supernatant to obtain the Bacillus thuringiensis insecticidal protein and spore kaolin adsorption composite formu...

Embodiment 3

[0031] Example 3: Bacillus thuringiensis insecticidal protein and spore sepiolite adsorption composite formulation

[0032] Take 1500 mL of Bacillus thuringiensis fermentation liquid (solid content 6.8%, w / v), and detect insecticidal protein 0.56% by SDS-PAGE. Weigh 117 g of sepiolite (particle size < 2 microns), add 1200 mL of deionized water to make a 9.75% (w / v) suspension. Add the sepiolite suspension into the Bacillus thuringiensis fermentation broth under stirring. Heat the fermentation broth in a water bath to raise the temperature to 50°C, slowly add 1mol / L sodium hydroxide solution dropwise under stirring to adjust the pH to 11-12, cool down to 20°C after 5 hours, adjust the suspension to pH 7-8 with 1mol / L HCl, Stirring was continued for 6 h. Put the adsorbed suspension into a centrifuge tube, centrifuge at 15,000 r / min for 10 min, and pour off the supernatant to obtain a composite formulation of Bacillus thuringiensis insecticidal protein and spore sepiolite a...

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Abstract

The invention discloses a preparation method of a bacillus thuringiensis insecticidal protein and spore endospore adsorption composite dosage form. The method comprises: adding a micron-level mineral adsorbent in bacillus thuringiensis fermentation liquor in advance; then, directly adding alkali-dissolving parasporal crystals; meanwhile, adsorbing the generated active insecticidal protein by a mineral carrier; and realizing fixed load to the active insecticidal protein. The bacillus thuringiensis insecticidal protein and spore endospore adsorption composite dosage form prepared with the method has simple method and low cost, omits the process that the bacillus thuringiensis cell crystal mixture is washed by a NaCl solution and sterile deionized water, and realizes synchronous preparation and adsorption of the active insecticidal protein. The obtained adsorption composite dosage form has large adsorption quantity and high insecticidal activity. In addition, because of the synergy of the spore endospore, the insecticidal activity of the active insecticidal protein adsorption preparation is further improved as compared with the original toxin adsorption dosage form.

Description

【Technical field】 [0001] The invention relates to the technical field of biological control of harmful organisms, especially by omitting the extraction of insecticidal proteins and the separation of spores, directly adding minerals to the fermentation broth, and the active insecticidal proteins generated by alkaline hydrolysis of parasporal crystals are adsorbed by mineral carriers. The Bacillus thuringiensis insecticidal protein adsorption composite dosage form containing spores is obtained. 【Background technique】 [0002] Bacillus thuringiensis ( Bacillus thuringiensis , referred to as Bt) is a bacterium that is closely related to Bacillus cereus, produces spores and parasporal crystals, and can parasitize insects and cause insect disease. In 1938, the first Bacillus thuringiensis preparation appeared in France, and it was realized in the middle of the 20th century. Industrialized production, its output accounts for 97% of the total amount of biopesticides, and it has bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/02A01N25/08A01P7/04
Inventor 周学永董庆洁董建萍马锦明
Owner TIANJIN UNIVERSITY OF TECHNOLOGY
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