Preparation method of Bacillus licheniformis and raw pesticide

A technology of Bacillus licheniformis and spores, which is applied in the field of preparation of Bacillus licheniformis and original medicine, can solve the problems of being susceptible to contamination by miscellaneous bacteria, extensive management, unstable product quality, etc.

A technology of Bacillus licheniformis and spores, which is applied in the field of preparation of Bacillus licheniformis and original medicine, can solve the problems of being susceptible to contamination by miscellaneous bacteria, extensive management, unstable product quality, etc.

CN101775360AActive Publication Date: 2010-07-14WUHAN KERNEL BIO-TECH CO LTD
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Experimental program
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Embodiment Construction

[0048] A method for preparing a former drug from bacillus licheniformis, the steps are:

[0049] 1. Screening of B1 strains:

[0050] Collect samples from different soils and water bodies across the country, weigh 1.0g of the sample into a small triangle flask on a sterile ultra-clean bench, add 9mL of sterile water, shake it fully and let it stand, take the supernatant as a concentration gradient Dilution (1Γ—10 -4 , 1Γ—10 -5 ,, 1Γ—10 -6 , 1Γ—10 -7 , 1Γ—10 -8 ). Use a pipette gun to draw 100 ul of the diluent and evenly spread it on the separation medium. After culturing in a temperature-controlled incubator at 30Β°C for 3-5 days, pick out the strains with high spore synchronization rate (spore formation rate β‰₯ 90%), and then test Microscopic examination after Lang's staining, the bacterial strain with Bl morphological characteristics was selected and transferred to B 1 In the liquid shake flask fermentation medium, the filling volume of 500mL shake flask is 25mL, the shaker...

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Abstract

The invention discloses a preparation method of Bacillus licheniformis (Bacilluslicheniformis subsp. Licheniformis, BL KN-08, CCTCC: NO. M208251) raw pesticide, comprising the following steps: A. screening B1 strains, collecting samples from soil and water body, adding sterile water, and standing; B. conforming the B1 strain the 16 SrDNA of which totally measures 1514 effective basic groups, and identifying; C. for B1 culture medium, adopting the response surface experiment designed by Box-Behken, taking the amount of spores as a screening index, and using a single factor experiment to select factors from multiple carbon sources, nitrogen sources and microelement factors; D. improving ferment biomass, and using the culture medium of the response surface method for batch fermentation; E. extracting and recovering spores; and F. detecting the amount of live spores. The invention can accurately detect the amount of the effective live spore in a product, has favourable repeatability, low detection cost, simple operation and convenient popularization and can simultaneously detect 7-8 samples; the amount of the live spores in the Bacillus licheniformis raw pesticide of the invention can reach 500 billion / g.

Description

technical field [0001] The present invention relates to new fields such as purification of polluted water bodies, garbage deodorization, etc., more specifically relates to a kind of bacillus licheniformis, and also relates to a kind of method for preparing the original drug (500 billion living spores / gram) of bacillus licheniformis, which will be used in the feed industry and Widely used in aquaculture. As the functions of bacillus are still being discovered, the scope of application is also expanding, and it has strong competitiveness in the domestic and international markets. Background technique [0002] Since the 1950s, antibiotics have played a major role as disease treatments and animal growth promoters. However, due to the increasingly serious negative effects of antibiotics, the World Health Organization has banned the use of any antibacterial drugs in feed, and some developed countries have clear regulations on the use of veterinary antibiotics. Feed and aquacultu...

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Application Information

Patent Timeline
14 Jul 2010
Publication
CN101775360A
IPC
C12N1/20; A61K35/74; A61P1/00; A61P31/04; A61P37/04; A23K1/16; C12R1/10; A61K35/742
Inventors
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