Preparation method of raw powder of each gram of bacillus subtillis with five hundred billion of live germs

A technology of Bacillus subtilis and spores, which is applied in the field of feed industry and water purification agent, can solve the problems of extensive management, unstable product quality, difficult control of bacterial content, etc., and achieves the effect of low cost

Active Publication Date: 2010-01-06
WUHAN KERNEL BIO-TECH CO LTD
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Problems solved by technology

The solid fermentation method is to inoculate probiotics on solid medium for fermentation and cultivation. Its advantages are relatively extensive management, simple production process, and low investment. Disadvantages of unstable product quality

Method used

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  • Preparation method of raw powder of each gram of bacillus subtillis with five hundred billion of live germs

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Embodiment Construction

[0039] A kind of preparation method of 500 billion live spores / gram Bacillus subtilis former medicine, its steps are:

[0040] 1. Screening of Bs strains:

[0041] 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 (1×10 -4 , 1×10 -5 ,, 1×10 -6 , 1×10 -7 , 1×10 -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 30°C for 3-5 days, pick out the strains with high spore synchronization rate (spore formation rate ≥ 90%), and then test Microscopic examination after Lang's staining, the selected strains with Bs morphological characteristics were transferred to the liquid shake flask fermentation medium, the volume of the 500mL shake flask was 25mL, the...

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Abstract

The invention discloses a preparation method of raw powder of each gram of bacillus subtillis with five hundred billion of live germs, which comprises the following steps: (A) the screening of a Bs strain: collecting a sample from soil and water; weighing the sample on a sterile and ultraclean table and then loading the sample in a small triangular flask; adding sterile water into the triangular flask and standing still the sample; (B) the confirmation of the Bs strain: carrying out molecular classification according to a 16S rDNA sequence and together testing effective basic groups of the 16S rDNAs of the strain and identifying the strain; (C) a culture medium of the Bs: utilizing a single factor experiment to select a factor from numerous carbon sources, nitrogen sources and trace element factors firstly by adopting a response surface experiment designed by Box-Behken and using the germ number as a screening index; (D) using a high-density liquid feed-batch fermentation technology for improving the quantity of fermentation organisms so as to use the culture medium of a response surface method to carry out batch fermentation by using a fermentation early phase, an exponential phase, a stationary phase OD, dissolved oxygen, pH and germ numbers as indexes; (E) carrying out a germ extraction and recovery process; and (F) detecting the number of live germs. The number of the live germs in raw medicine prepared by the process of the bacillus subtillis reaches five hundred billion/gram.

Description

technical field [0001] The invention relates to a preparation method of a bacillus subtilis original drug (500 billion live spores / gram) as a feed industry and water body purifying agent, and will be widely used in the feed industry and aquaculture. As the function of bacillus is still being discovered, the scope of application is also expanding. This product will be applied to new fields such as purification of polluted water, fruit preservation, garbage deodorization, etc., and 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 aquaculture have been greatly impacte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/125
Inventor 刘华梅陈振民李青谢翔杨克华程治国陈又香缪华军余娜王巧梅黄志远
Owner WUHAN KERNEL BIO-TECH CO LTD
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