Mixed cooperative fermentation process of bacillus subtilis

A technology of Bacillus subtilis and fermentation process, which is applied in the field of mixed bacteria synergistic fermentation process of Bacillus subtilis, can solve problems such as instability, poor control effect, and few biological control products and varieties, so as to enhance adaptability, improve inhibition Bacterial effect, enhanced effect of antibacterial potency

Inactive Publication Date: 2010-06-02
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005]At present, the number and variety of biological control products in my country are relatively small, and the control effect is poor or unstable

Method used

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  • Mixed cooperative fermentation process of bacillus subtilis
  • Mixed cooperative fermentation process of bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Step 1. Seed activation

[0026] The three fermentation strains that will be stored in the seed tube: Bacillus subtilis B9601-Y2, Bacillus subtilis NCD-2 and Bacillus subtilis BAB-1 were respectively streaked on the slant of the seed activation medium test tube and cultivated in a 28°C incubator 24h, obtain the inclined-plane bacterial classification for production;

[0027] Seed activation medium: beef extract 3g / L, peptone 5g / L, sodium chloride 5g / L, glucose 10g / L, agar 15g, pH 7.0-7.2, medium sterilization conditions: 0.15MPa steam sterilization for 30min, Sterilization temperature 121°C;

[0028] Step 2, preparation of primary seed solution

[0029] Use the inoculation loop to pick the single colonies of the slant strains of each strain of Bacillus subtilis for production respectively, and inoculate them in three 250ml Erlenmeyer flasks with 50ml primary seed culture medium respectively, at 28~30°C, 180~ Shake culture at 200r / min for 18h to obtain the first-grade...

Embodiment 2

[0041] Steps one, two and three are the same as steps one, two and three in Example 1;

[0042] Step 4. Seed tank cultivation

[0043] Add 20L seed tank culture medium in the 25L stirring type fermentation tank as seed tank, cool to 30 ℃ after the culture medium is sterilized, pH adjustment is 7.0, the secondary seed liquid of each strain Bacillus subtilis single strain, Inoculate into the seed tank respectively, the inoculation ratio is Bacillus subtilis B9601-Y2: Bacillus subtilis NCD-2: Bacillus subtilis BAB-1 equals 2: 1: 3, and the total inoculum amount is 8% (v / v) of the medium of the seed tank ; The control conditions for the mixed bacteria fermentation in the seed tank are 30°C, the ventilation rate is 10L / min, the tank pressure is 80kPa, the stirring speed is 200r / min, the aerobic mixed bacteria fermentation culture is cultivated for 20 hours, and the third-grade seed liquid is obtained;

[0044] Seed tank medium is: bean cake powder 10g / L, beef extract 5g / L, yeast e...

Embodiment 3

[0049] Steps one, two and three are the same as steps one, two and three in Example 1;

[0050] Step 4. Seed tank cultivation

[0051] Add 20L seed tank culture medium in the 25L stirring type fermentation tank as seed tank, cool to 30 ℃ after the culture medium is sterilized, pH adjustment is 8.0, the secondary seed liquid of each strain Bacillus subtilis single strain, Inoculate into the seed tank respectively, the inoculation ratio is Bacillus subtilis B9601-Y2: Bacillus subtilis NCD-2: Bacillus subtilis BAB-1 is equal to 3: 1: 4, and the total inoculum amount is 10% (v / v) of the medium of the seed tank ;The control conditions of the mixed bacteria fermentation in the seed tank are 30°C, the ventilation rate is 8L / min, the tank pressure is 80kPa, the stirring speed is 200r / min, the aerobic mixed bacteria fermentation culture is cultivated for 18 hours, and the third-grade seed liquid is obtained;

[0052] Seed tank culture medium: bean cake powder 10g / L, beef extract 4g / L,...

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Abstract

The invention relates to the mixed cooperative fermentation method of bacillus subtilis, and belongs to the field of microbe engineering. The scheme comprises the following steps of: selecting three different kinds of bacillus subtilis having strong inhibiting effects on different pathogenic bacteria; performing slant cultivation and shaking cultivation on the three kinds of bacillus subtilis respectively by using the mixed fermentation method; inoculating the three kinds of bacillus subtilis in certain proportion; and performing seeding tank cultivation and fermentation tank cultivation on the mixed bacteria. Methods such as culture medium selection, pH value, temperature, ventilatory capacity and inoculum concentration proportion of the three strains of bacteria are optimized during fermentation. Compared with the conventional single bacterial strain fermentation method, the mixed cooperative fermentation method is simplified, improves efficiency, reduces production cost and has effective bacteria quantity in a fermentation liquor of more than or equal to 1,012 cfu / ml.

Description

technical field [0001] The technical proposal relates to a mixed-bacteria synergistic fermentation process of Bacillus subtilis, which belongs to the field of microbial engineering. Background technique [0002] The important development trend of biological control and product development at home and abroad is from single bacterial agent fermentation to mixed bacterial fermentation, using the functional mechanism of different microorganisms, through the synergistic effect of multiple strains, improving the biomass and functional characteristics of the strains, simplifying production process, reducing fermentation batches, improving fermentation efficiency and shortening the fermentation cycle provide broad prospects for the industrial production and application of biological pesticides and biological bacterial fertilizers. [0003] For the co-cultivation and synergistic fermentation research of multiple strains, as early as 1925, when Sack investigated bacterial metabolism, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N1/20C12R1/125
Inventor 李春武占省赵雅峰陶晶李艳菊
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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