Culture method and application of bacillus subtilis

A technology of Bacillus subtilis and its cultivation method, which is applied in the field of cultivation of Bacillus subtilis, can solve the problems that Bacillus subtilis is easy to produce foam and the culture conditions are stable, and achieve the effects of significant living control effect, high activity, and reduced foam generation

Active Publication Date: 2021-08-27
GUANGDONG HINAPHARM PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Based on this, in order to solve the problems of easily producing foam and stable culture conditions in the cultivation process of bacillus subtilis in the prior art, the invention provides a kind of cultivation method and application thereof of bacillus subtilis, and the specific technical scheme is as follows:

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  • Culture method and application of bacillus subtilis
  • Culture method and application of bacillus subtilis
  • Culture method and application of bacillus subtilis

Examples

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Effect test

Embodiment 1

[0068] A method for cultivating Bacillus subtilis, comprising the following steps:

[0069] Streak inoculation of Bacillus subtilis on LB solid medium plate, and activate culture at 35°C for 18 hours to obtain the activated strain;

[0070] The activated strain was inoculated in LB liquid medium, shaken and cultured at 35°C and 180r / min for 18h to obtain a seed solution;

[0071] According to the components of the liquid seed medium: corn steeping liquid 10g / L, bean cake powder 1g / L, bran 4g / L, sodium citrate 0.1g / L, calcium chloride 0.01g / L, dipotassium hydrogen phosphate 0.01g / L, Tween 0.01g / L, prepare liquid seed culture medium, and inoculate the seed liquid that is 5% inoculum in described liquid seed culture medium, control liquid seed culture medium pH7.0-7.2; Through culture system The conditions for controlling the first fermentation are as follows: the temperature is 35°C, the time is 14h, and the dissolved oxygen is 40%; after the first fermentation is completed, t...

Embodiment 2

[0075] A method for cultivating Bacillus subtilis, comprising the following steps:

[0076] Streak inoculation of Bacillus subtilis on LB solid medium plate, and activate culture at 39°C for 28 hours to obtain the activated strain;

[0077] The activated strain was inoculated in LB liquid medium, shaken and cultivated at 39°C and 250r / min for 22h to obtain a seed solution;

[0078] According to the components of the liquid seed medium: corn steeping liquid 15g / L, bean cake powder 3g / L, bran 4g / L, sodium citrate 0.4g / L, calcium chloride 0.02g / L, dipotassium hydrogen phosphate 0.30g / L, Tween 0.03g / L, prepare liquid seed culture medium, and inoculate the seed liquid that is 8% inoculum in described liquid seed culture medium, control liquid seed culture medium pH7.0-7.2; Through culture system The conditions for controlling the first fermentation are as follows: the temperature is 38°C, the time is 20h, and the dissolved oxygen is 50%; after the first fermentation is completed,...

Embodiment 3

[0082] A method for cultivating Bacillus subtilis, comprising the following steps:

[0083] Streak inoculation of Bacillus subtilis on LB solid medium plate, and activate culture at 36°C for 20 hours to obtain the activated strain;

[0084] The activated strain was inoculated in LB liquid medium, and shaken and cultured for 20 hours under the conditions of 36°C and 200r / min to obtain the seed solution;

[0085] According to the components of the liquid seed medium: corn steeping liquid 12g / L, bean cake powder 2g / L, bran 3g / L, sodium citrate 0.3g / L, calcium chloride 0.02g / L, dipotassium hydrogen phosphate 0.05g / L, Tween 0.02g / L, prepare liquid seed culture medium, and inoculate the seed liquid that inoculum is 6% in described liquid seed culture medium, control liquid seed culture medium pH7.0-7.2; Through culture system The conditions for controlling the first fermentation are as follows: the temperature is 36°C, the time is 18h, and the dissolved oxygen is 45%; after the fi...

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Abstract

The invention provides a culture method and application of bacillus subtilis. The bacillus subtilis is a strain preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No.61565. A culture system is utilized to more accurately control the culture conditions of the bacillus subtilis, so that the generation of foam in the culture process is effectively reduced, the culture temperature and the dissolved oxygen content of culture are effectively controlled, and culture fermentation liquor with fewer impurities and higher activity is obtained; besides, after the bacillus subtilis is subjected to three-stage fermentation, metabolites of the bacillus subtilis have remarkable in-vitro and in-vivo prevention and treatment effects on the clostridium perfringens, the average diameter of an inhibition zone of the bacillus subtilis on the clostridium perfringens can reach 26.78 mm, and the prevention and treatment effect on the clostridium perfringens caused by the clostridium perfringens reaches 98.71%.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a method for cultivating Bacillus subtilis and its application. Background technique [0002] Bacillus subtilis is a rod-shaped bacterium that can produce reverse-resistant endospores. It is a Gram-positive bacterium distributed in nature and can be used to produce amylase, protease, lipase and other enzymes to improve Animal intestinal flora, improve the activity of digestive enzymes, harmless to humans and animals, and do not pollute the environment. It can be widely used in the enzyme preparation industry and animal husbandry feed industry. Bacillus subtilis can produce useful metabolites during the growth process, and most of them are secreted in the culture medium. In the prior art, during the cultivation of Bacillus subtilis, the starch in the raw material absorbs water and swells and hydrolyzes to cause the viscosity of the medium system to increase significantly i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12M1/38C12M1/36C12M1/34C12M1/04C12M1/02C12M1/00A61P31/04C12R1/125
CPCC12N1/20C12M27/02C12M29/06C12M41/12C12M41/22C12M41/32C12M41/42A61P31/04
Inventor 代珍青张志清黎卓莹邓远钊朱健新
Owner GUANGDONG HINAPHARM PHARMA CO LTD
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