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Brevibacillus brevis liquid-submerged fermentation method and wettable powder preparation method

A technology of Bacillus brevis, liquid submerged fermentation, applied in the methods of using spores, fungicides, botanical equipment and methods, etc., can solve problems such as troubles, achieve strong resistance to stress, improve the level of comprehensive utilization, and increase economy. effect of benefit

Inactive Publication Date: 2019-03-01
湖南泰谷生态工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How to increase the spore yield, recovery rate and practical application effect as much as possible under the condition of guaranteed cost has become a bottleneck problem that plagues the actual industrial production of Bacillus brevis wettable powder

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of Bacillus brevis wettable powder:

[0031] (1) Strain activation: Streak inoculation of Bacillus brevis on a nutrient agar medium plate for culture, culture at 30°C for 24h, then pick a single colony and transfer it to the slant of nutrient agar, culture at 30°C for 24h Afterwards, the activated bacteria were obtained.

[0032] (2) Preparation of primary seed liquid in shake flasks: 500 mL of seed culture medium containing 5 g of yeast extract, 1 g of peptone, 1 g of glucose, and pH 7.0 ± 0.1 were respectively loaded into two 2000 mL conical flasks, and the The activated strains were inoculated into the shake flask seed medium with an inoculation loop, and the culture conditions were as follows: temperature 36°C, shaker speed 160r / min, culture time 12h.

[0033] (3) Preparation of secondary seed liquid in seed tank: After the seed liquid obtained in step (2) is microscopically inspected without contamination by bacteria, insert 500 g of yeast extract, 100...

Embodiment 2

[0041] Preparation of Bacillus brevis wettable powder:

[0042] (1) Strain activation is the same as in Example 1.

[0043] (2) The preparation of the primary seed solution of the shake flask is the same as in Example 1.

[0044] (3) The preparation of the secondary seed liquid in the seed tank is the same as in Example 1.

[0045] (4) Fermentation tank high-density fermentation culture: the seed liquid gained in the step (3) is inserted in the 1000L fermentation tank by the inoculum size of 1% after microscopic examination without assorted bacteria pollution, and the fermentation medium composition in the tank: sucrose molasses 10g / L, soybean flour 5g / L, peptone 2g / L, MgSO 4 0.5g / L, CaCO 3 4g / L, liquid volume in fermentation tank 500L, culture conditions: temperature 30±2°C, stirring rate 160r / min, ventilation volume 15m 3 / h, tank pressure 0.05Mpa, when more than 90% thalline form in the fermented liquid is spore, stop fermentation, get the fermented liquid for subsequent ...

Embodiment 3

[0052] Preparation of Bacillus brevis wettable powder:

[0053] (1) Strain activation: same as in Example 1.

[0054] (2) Preparation of primary seed solution in shake flask: same as in Example 1.

[0055] (3) Preparation of the secondary seed liquid in the seed tank: same as in Example 1.

[0056] (4) Fermentation tank high-density fermentation culture: the seed liquid gained in the step (3) is inserted in the 1000L fermentation tank by the inoculum size of 5% after microscopic examination without assorted bacteria pollution, and the fermentation medium composition in the tank: sucrose molasses 30g / L, soybean flour 15g / L, peptone 6g / L, MgSO 4 3.5g / L, CaCO 3 8g / L, liquid volume in fermentation tank 700L, culture conditions: temperature 30±2°C, stirring rate 200r / min, ventilation volume 25m 3 / h, tank pressure 0.06Mpa, when more than 90% thalline form in the fermented liquid is spore, stop fermentation, get the fermented liquid for subsequent use, and the fermentation cycle...

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Abstract

The invention provides a brevibacillus brevis liquid-submerged fermentation method, and further provides a preparation method of a brevibacillus brevis wettable powder. The brevibacillus brevis liquid-submerged fermentation method comprises the steps: brevibacillus brevis seed liquid is inoculated into a fermentation culture medium with the volume ratio of the inoculation amount being 1-5% to be fermented till bacteria of 90% or above in obtained fermentation liquid are germs in form; the preparation method comprises the steps: the fermentation liquid is centrifuged to obtain concentrated bacterial liquid, then talcum powder, sucrose, maltodextrin and the concentrated bacterial liquid are evenly mixed and then spray-dried, a dry bacterium powder is obtained, sodium dodecyl benzene sulfonate, organosilicone, ascorbic acid and the dry bacterium powder are evenly mixed, then smashed under 30-50 DEG C, and pass through a 400-mesh sieve, and then the wettable powder is obtained. The methodis simple in process, the prepared biocontrol brevibacillus brevis wettable powder has a good control effect on tobacco black shank, safety and high efficiency are achieved, no residue and toxic and side effect exists, and using of chemical pesticides can be reduced.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a liquid submerged fermentation method of Bacillus brevis and a preparation method of wettable powder. Background technique [0002] Extensive use of chemical pesticides can easily lead to excessive pesticide residues and bio-accumulation, which not only pollutes the ecological environment, but also affects human health. Microbial pesticides refer to the use of living microorganisms and their metabolites to prevent and control agricultural diseases and insect pests. They have strong selectivity, small side effects, are harmless to humans, livestock, crops and the natural environment, and are not easy to produce drug resistance. Therefore, they are a kind of A green pesticide with broad prospects. At present, there are relatively mature microbial pesticide products on the market, such as Bacillus thuringiensis, Bacillus licheniformis, and Bacillus subtilis. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N3/00A01N25/14A01N63/00A01P3/00C12R1/01
CPCA01N25/14A01N63/00C12N1/20C12N3/00
Inventor 郑双凤郭帅谭石勇罗志威
Owner 湖南泰谷生态工程有限公司
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