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Culture medium for promoting growth of bacillus and application thereof

A technology of bacillus and culture medium, which is applied in the field of medium for promoting the growth of bacillus, can solve the problems of unclear influence on the growth rate of bacillus, microbial toxicity, lack of live bacteria as the target product research, etc., and shorten the time for spore formation , production cost reduction, good promotion value effect

Active Publication Date: 2012-09-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, excessive addition can also cause toxicity to microorganisms
[0003] At present, the research hotspots of Bacillus fermentation at home and abroad are the screening of carbon and nitrogen sources targeting metabolites and the optimization of culture conditions. There is a lack of relevant research on live bacteria as the target product, and the impact of mineral nutrients on the growth rate of Bacillus is still unclear.

Method used

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  • Culture medium for promoting growth of bacillus and application thereof
  • Culture medium for promoting growth of bacillus and application thereof
  • Culture medium for promoting growth of bacillus and application thereof

Examples

Experimental program
Comparison scheme
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preparation example Construction

[0047] The preparation of the seed liquid involved in the following examples is carried out according to the following method:

[0048] Bacillus subtilis (Bacillus subtilis) B201 CGMCC No.5786 was activated on a tryptone soybean agar (Tryptic Soy Agar, TSA) plate. After culturing overnight at 30°C, a single colony was picked from the plate and inoculated in 15 mL of TS liquid medium Cultivate overnight at 32°C and 160r / min (12h). The morphology of the bacteria is correct and free of pollution by microscope inspection. Centrifuge the cultured bacteria liquid at 6000r / min for 5min, wash it twice with sterile water, and make a bacterial suspension. liquid. Measure the OD value with a spectrophotometer at a wavelength of 600 nm, and make the OD value reach 1 to obtain the seed solution of Bacillus subtilis B201 CGMCC No.5786.

Embodiment 1

[0049] Embodiment 1, the determination of the mineral nutrient used in the substratum that promotes the growth of bacillus

[0050] 1. Determination of the basic concentration range of mineral nutrients

[0051] 1. The preparation of different concentrations of mineral nutrients and the preparation of corresponding medium

[0052] Since the quality of the mineral nutrients required for each treatment is very small and the weighing error is large, the mother liquor is prepared first. The mother liquor requires the solute to be completely dissolved, and the volume required for each treatment is converted according to the concentration of the mother liquor. The increase in this volume is very important for the culture medium The influence of the volume should be ignored; the content of the mineral nutrients in the Tryptone Broth medium itself was detected with an atomic absorption spectrophotometer (see Table 1 for the contents of each element in the Tryptone Broth medium), accor...

Embodiment 2

[0093] Embodiment 2, cultivate bacillus with the Tryptone Broth medium that contains optimal mineral nutrient ratio

[0094] 1. Culture in growth curve analyzer

[0095] The Tryptone Broth medium containing the optimal mineral nutrient ratio optimized in Example 1 was used to cultivate Bacillus subtilis (Bacillus subtilis) in the culture plate matched with the growth curve analyzer (bioscreen, Finland, Oy Growth Curves Ab Ltd) B201 CGMCC No.5786, add culture solution with a volume of 150 μL to each well, repeat 3 times for each treatment, put the added culture plate into a growth curve analyzer, culture at 32°C for 20 hours with continuous shaking, measure OD every 15 minutes 600 value. At the same time, the Tryptone Broth medium without any mineral nutrients was set as a control, and the growth curve was drawn.

[0096] The result is as image 3 As can be seen from the figure, Bacillus subtilis (Bacillus subtilis) B201 CGMCC No.5786 grows in the Tryptone Broth medium conta...

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Abstract

The invention discloses a culture medium for promoting growth of bacillus and application thereof. According to the culture medium, K+ is added into a Tryptone Broth culture medium until the final concentration is 2.2mM, and Cu2+ is added until the final concentration is 8 mu M. K2HPO4 at the final concentration of 0.3g / L and CuSO4.5H2O at the final concentration of 0.002g / L are added into the culture medium. Experiments prove that when the culture medium is used for culturing Bacillus subtilis B201 CGMCC No.5786, the bacteria content of a fermentation liquor can be improved double, and the spore forming time is shortened by 15 percent. Therefore, the production cost is reduced by 30 percent. The culture medium has high promotion value. The invention provides a reference for the optimization of a Bacillus subtilis fermentation culture medium, and provides theoretical guidance for the research and development of related biocontrol agents.

Description

technical field [0001] The invention relates to a culture medium for promoting the growth of Bacillus subtilis and its application, in particular to a medium for promoting the growth of Bacillus subtilis (Bacillus subtilis) added with mineral nutrients and its application. Background technique [0002] Bacillus sp. is an important resource for biological agents because it can produce stress-resistant spores, can tolerate a variety of adverse environments, and has the functions of promoting growth and preventing diseases. Mineral nutrients are essential nutrients for microbial growth. They are mainly involved in the composition of cell structures, and are related to energy transfer and cell permeability regulation. They are active components or activators of enzymes. However, excessive addition can also cause toxicity to microorganisms. [0003] At present, the research hotspots of Bacillus fermentation at home and abroad are the screening of carbon and nitrogen sources targ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/38C12N1/20C12R1/125
Inventor 王琦张丽霞李燕郑绍芳金洪段玲玲李超
Owner CHINA AGRI UNIV