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Method for culturing bacillus subtilis through high-density fermentation

A high-density fermentation technology of Bacillus subtilis, which is applied in the field of microbial fermentation, can solve the problems of death, reduced bacterial agent activity and quality stability, low bacterial count and spore rate of Bacillus subtilis, and achieves simple process parameters and short fermentation cycle Short and high spore rate

Inactive Publication Date: 2014-08-20
WEIHAI GOLD FEED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are generally problems of low bacterial count and spore rate in the production process of Bacillus subtilis, and the general spore content is 5.0×10 9 Below cfu / ml, bacteria without spores will die in a short time, greatly reducing the activity and quality stability of the bacterial agent

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Bacillus subtilis stored at -70°C was inoculated with a sterile streak on a sterilized solid medium (the solid medium was composed of the following components in terms of mass percentage: 0.3% beef extract, 0.6% peptone, 1.5% agar, and the rest The amount is water, PH7.0, sterilized at 121°C, 0.11Mpa for 20min.) 37°C for 4 days.

[0021] After the colony on the solid medium was washed with sterile physiological saline, it was inoculated into the sterilized medium of 1000ml shake flask (the medium was composed of the following components by mass percentage: glucose 0.4%, beef extract 0.3%, peptone 1.0%, yeast extract 0.5%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, sodium chloride 0.5%, defoamer 0.15%, the balance is water, pH7.0, 121℃, 0.11Mpa sterilization for 20min) , the loading capacity of the medium is 200ml, and the shaker culture conditions are: temperature 37°C, rotation speed 220rpm, and culture for 16 hours.

[0022] will step The obt...

Embodiment 2

[0025] Bacillus subtilis stored at -70°C was inoculated with a sterile streak on a sterilized solid medium (the solid medium was composed of the following components in terms of mass percentage: 0.3% beef extract, 0.6% peptone, 1.5% agar, and the rest The amount is water, PH7.0, sterilized at 121°C, 0.11Mpa for 20min.) 37°C for 4 days.

[0026] After the colony on the solid medium was washed with sterile physiological saline, it was inoculated into the sterilized medium of 1000ml shake flask (the medium was composed of the following components by mass percentage: glucose 0.4%, beef extract 0.3%, peptone 1.0%, yeast extract 0.5%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, sodium chloride 0.5%, defoamer 0.15%, the balance is water, pH7.0, 121℃, 0.11Mpa sterilization for 20min) , the loading capacity of the medium is 200ml, and the shaker culture conditions are: temperature 37°C, rotation speed 220rpm, culture for 18 hours.

[0027] will step The obtaine...

Embodiment 3

[0030] Bacillus subtilis stored at -70°C was inoculated with a sterile streak on a sterilized solid medium (the solid medium was composed of the following components in terms of mass percentage: 0.3% beef extract, 0.6% peptone, 1.5% agar, and the rest The amount is water, PH7.0, sterilized at 121°C, 0.11Mpa for 20min.) 37°C for 4 days.

[0031] After the colony on the solid medium was washed with sterile physiological saline, it was inoculated into the sterilized medium of 1000ml shake flask (the medium was composed of the following components by mass percentage: 0.4% glucose, 0.3% beef extract, peptone 1.0%, yeast extract 0.5%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, sodium chloride 0.5%, defoamer 0.15%, the balance is water, pH7.0, 121℃, 0.11Mpa sterilization for 20min) , the loading capacity of the medium is 200ml, and the shaker culture conditions are: temperature 37°C, rotation speed 220rpm, culture for 18 hours.

[0032] will step The obtaine...

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Abstract

The invention discloses a method for culturing bacillus subtilis through high-density fermentation, belonging to the field of microbial fermentation engineering. The method is used for increasing the thalli yield and spore rate of the bacillus subtilis. The method comprises the steps of activating the bacillus subtilis and performing two-step fermentation culture to produce a target product. The method has the advantages that the components of a culture medium are readily available and cheap, the process parameters are simple, the fermentation period is short, the cell biomass of the obtained bacillus subtilis can reach over 9*10<9>cfu / ml, and the spore rate is over 90%. The quality and stability of the product are significantly improved.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and more specifically relates to a method for high-density fermentation and cultivation of Bacillus subtilis. Background technique [0002] Bacillus subtilis ( Bacillus subtilis ) is a genus of Bacillus, Gram-positive aerobic bacteria, non-capsulated, peri-flagellated, and able to move. It is widely distributed in soil and decaying organic matter, and is easy to reproduce in withered grass juice. Subtilisin, polymyxin, nystatin, gramicidin and other active substances produced by Bacillus subtilis during the growth process, these active substances have obvious effects on pathogenic bacteria or opportunistic pathogenic bacteria of endogenous infection inhibitory effect. It can quickly consume free oxygen in the environment, cause intestinal hypoxia, promote the growth of beneficial anaerobic bacteria, and produce organic acids, reduce intestinal pH, and indirectly inhibit the growth of othe...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/125
Inventor 李贵杰王燕张群峰耿建
Owner WEIHAI GOLD FEED
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