Submerged fermentation method of Bacillus licheniformis and its application

A technology of submerged fermentation of Bacillus licheniformis, applied in the submerged fermentation of Bacillus licheniformis, the application field of wettable powder prepared by Bacillus licheniformis, can solve the problem of low spore content and sporulation rate, deterioration of fermentation broth, low yield of spores, etc. problems, to achieve the effect of increasing the amount of spore formation, prolonging the stable period, and increasing the rate of spores

Active Publication Date: 2021-02-09
湖南慧谷农业生态研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing powder preparation method has low efficiency, high cost, deterioration of fermentation liquid in the production process, serious loss of spores, and low yield of spores
[0004] Since the composition of the fermentation medium and the fermentation process have a great influence on the spore production in the Bacillus licheniformis fermentation broth, but the current fermentation production of Bacillus licheniformis has problems such as low spore content and sporulation rate in the fermentation broth, and high cost.
How to increase the spore content, spore yield and practical application effect as much as possible under the condition of guaranteed cost has become a bottleneck problem in the actual industrial production of wettable powder for biocontrol Bacillus licheniformis

Method used

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  • Submerged fermentation method of Bacillus licheniformis and its application
  • Submerged fermentation method of Bacillus licheniformis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation of Bacillus licheniformis wettable powder:

[0038] (1) Strain activation: Streak inoculation of Bacillus licheniformis on a nutrient agar medium plate for culture, culture at 36°C for 24 hours, then pick a single colony and transfer it to a nutrient agar slant, culture at 36°C for 24 hours Get activated bacteria.

[0039] (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 loaded into two 2000 mL Erlenmeyer flasks respectively. 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.

[0040] (3) Preparation of the secondary seed solution in the seed tank: the seed solution obtained in step (2) is microscopically checked for no bacteria contamination, and then inserted into a 2% inoculum containin...

Embodiment 2

[0048] Preparation of Bacillus licheniformis wettable powder:

[0049] (1) Strain activation is the same as in Example 1.

[0050] (2) The preparation of the primary seed solution of the shake flask is the same as in Example 1.

[0051] (3) The preparation of the secondary seed liquid in the seed tank is the same as in Example 1.

[0052] (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: bran 2%, corn flour 1%, yeast extract 0.2%, sodium chloride 0.2%, CaCO30.4%, KH2PO40.06%, magnesium sulfate 0.1%, MnCl20.03%, defoamer 0.015%. Culture conditions: fermentation temperature control: 33°C; air flow adjustment: 0h-4h control at 0.4vvm, 4h-8h control at 0.6vvm, 8h-11h control at 0.8vvm, 11h-36h control at 1.0vvm; stirring adjustment: 0h ~7h is con...

Embodiment 3

[0059] Preparation of Bacillus licheniformis wettable powder:

[0060] (1) Strain activation is the same as in Example 1.

[0061] (2) The preparation of the primary seed solution of the shake flask is the same as in Example 1.

[0062] (3) The preparation of the secondary seed liquid in the seed tank is the same as in Example 1.

[0063] (4) fermentor high-density fermentation culture: the seed liquid gained in the step (3) is inserted in the 1000L fermenter by 5% inoculum size after microscopic examination without assorted bacteria pollution, and the fermentation medium composition in the tank: bran 4%, corn flour 3%, yeast extract 0.4%, sodium chloride 0.5%, CaCO 3 0.8%, KH2PO4 0.12%, magnesium sulfate 0.2%, MnCl 2 0.05%, defoamer 0.025%. Culture conditions: fermentation temperature control: 36°C; air flow adjustment: 0h-4h control at 0.6vvm, 4h-8h control at 0.8vvm, 8h-11h control at 0.9vvm, 11h-36h control at 1.5vvm; stirring adjustment: 0h ~7h is controlled at 80r...

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Abstract

The invention provides a submerged fermentation method of Bacillus licheniformis and its application. The submerged fermentation method comprises that the seed liquid of Bacillus licheniformis is inserted into a fermentation medium for fermentation, until more than 90% of the bacteria in the obtained fermentation broth have the form of spores. The Bacillus licheniformis fermentation liquid is centrifuged to obtain a concentrated bacterial liquid, and then diatomaceous earth, cornstarch, maltodextrin and the above-mentioned concentrated bacterial liquid are mixed uniformly and then spray-dried to obtain a dry powder microbial agent. Sodium, sodium carboxymethylcellulose, β-cyclodextrin, ascorbic acid, γ-polyglutamic acid and the above-mentioned dry powder bacterial agent are mixed uniformly, pulverized, and sieved to obtain a wettable powder. The method has simple process, and the prepared bio-control bacillus licheniformis is safe and efficient, has no residue, no toxic and side effects, and can reduce the use of chemical pesticides.

Description

technical field [0001] The invention relates to the technical field of biological fermentation, in particular to a submerged fermentation method of bacillus licheniformis and its application, in particular to a submerged fermentation method of bacillus licheniformis and the application of the bacillus licheniformis obtained in the preparation of wettable powder. Background technique [0002] my country uses a large amount of chemical pesticide preparations every year, but the use of a large amount of chemical pesticides can easily lead to excessive pesticide residues and bio-enrichment, 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. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N3/00A01N25/14A01N63/22A01P3/00C12R1/10
CPCC12N1/20C12N3/00A01N25/14A01N63/10
Inventor 郑双凤谭石勇罗志威谭武贵文亚雄
Owner 湖南慧谷农业生态研究院有限公司
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