Bacillus thuringiensis protoxin microcapsule and preparation method thereof

A technology of Bacillus thuringiensis and Bacillus aureus, which is applied in the field of Bacillus thuringiensis protoxin microcapsules and its preparation, can solve the problems of high cost and narrow pH range, achieve protein protection, prolong the effective period, and have a wide range of applications Effect

Active Publication Date: 2014-08-27
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The microcapsule uses two kinds of polymers as the wall material, which realizes the preservation of the insecticidal protein of Bacillus thuringiensis in neutral conditions, releases in alkaline (pH 10) conditions, and has a certain ability to resist high temperature and anti-drying. Insecticidal protein has a good protective effect, but its cost is high, and the pH range of slow release is narrow

Method used

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  • Bacillus thuringiensis protoxin microcapsule and preparation method thereof
  • Bacillus thuringiensis protoxin microcapsule and preparation method thereof
  • Bacillus thuringiensis protoxin microcapsule and preparation method thereof

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Embodiment Construction

[0026] The present invention will be further described in detail below in conjunction with the embodiments and accompanying drawings.

[0027] 1 Materials and methods

[0028] 1.1 Materials:

[0029] CaCl 2 / PSS solution: weigh 0.29g CaCl 2 2H 2 O was mixed with 0.25g PSS and dissolved in 10mL distilled water;

[0030] Na 2 CO 3 Solution: weigh 0.21g Na 2 CO 3 The solution was dissolved in 10mL distilled water;

[0031] Tris-HCL buffer, PBS buffer, Na 2 CO 3buffer

[0032] 1mg / mL PAH (pH6.5): 20mg PAH was dissolved in 100mM Tris, and the pH was adjusted to 6.5 with concentrated hydrochloric acid;

[0033] 1mg / mL Cry protein (pH6.5): 20mg Cry protein dissolved in 50mM Na 2 CO 3 In, adjust the pH to 6.5 with Tris-HCL buffer;

[0034] 0.15M EDTA solution, NaOH solution to adjust pH to 7.0

[0035] 1.2 Preparation method of microcapsules

[0036] 1) Preparation of CaCO 3 core

[0037] Equivalent amount of CaCl that has been configured 2 / PSS solution (0.1M) and...

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Abstract

The invention relates to a bacillus thuringiensis protoxin microcapsule which is prepared by self-assembly. Characteristics of the bacillus thuringiensis protoxin microcapsule are that: bacillus thuringiensis protoxin protein is encapsulated in the microcapsule, and the wall of the microcapsule is formed by successive self-assembly of poly(allylamine) (PAH) and bacillus thuringiensis insecticidal protein (Cry). When the pH value of the assembled microcapsule is 6.0, stable encapsulation of the microcapsule for bacillus thuringiensis protoxin protein can be realized in a long term, bacillus thuringiensis protoxin protein is protected, and the effective duration of bacillus thuringiensis protoxin protein is prolonged; and the controlled releasing effect is realized when the microcapsule is in environment with a pH value of more than 6.5. The difference of the intestinal pH value of bacillus thuringiensis insecticidal protein target larvae is great, for example, the intestinal pH value of lepidopterous larvae is 8.0 to 10.5, the intestinal pH value of coleoptera larvae is 6.5 to 7.5, and the intestinal pH value of diptera larvae is 7.0 to 8.0, so that application range of the controlled releasing effect of the microcapsule is wide. It is shown, by various characteristics, that the microcapsule possesses significant importance in further practical application of insecticidal protein Cry.

Description

technical field [0001] The invention relates to the field of biological pharmaceutics, in particular to a bacillus thuringiensis protoxin microcapsule and a preparation method thereof. Background technique [0002] Bacillus thuringiensis (Bt) can produce a crystal protein with insecticidal activity, called Cry protein, while forming spores. Such crystals are dissolved into soluble protoxins under alkaline conditions, and then activated by proteases in the midgut environment of insects to become active toxin proteins. Because it has specific insecticidal activity against most agricultural pests, is safe for humans and animals, and does not pollute the environment, it has become the most productive and widely used microbial insecticide in the world. However, the short duration of action has become an important factor restricting the application of Cry protein preparations. Various uncertain factors in the external environment will lead to the inactivation of crystal proteins....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N25/28A01N63/02A01P7/04B01J13/02
Inventor 郭淑元李峰杨文慧
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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