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Method for promoting clostridium butyricum to form spores and microecological preparation and application thereof

A technology of Clostridium butyricum and inactivation of Clostridium butyricum, which is applied in the direction of using spores, methods based on microorganisms, applications, etc., can solve the problems of low total yield, short shelf life, late Clostridium butyricum, etc., and achieve Increase the amount of viable bacteria and spore rate, increase feed intake, and reduce diarrhea

Pending Publication Date: 2022-03-22
茂名傲新生物科技有限公司 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] my country's research on Clostridium butyricum started relatively late. At present, domestic Clostridium butyricum preparations have low output, high price, and short shelf life, and it is difficult to meet the market demand for Clostridium butyricum.
These problems are mainly due to the complex culture conditions and immature technology of Clostridium butyricum.
In addition, since the stress resistance of Clostridium butyricum mainly comes from its sporulation ability, how to increase the spore rate of Clostridium butyricum is also the main factor restricting the production of Clostridium butyricum, but because Clostridium butyricum is an obligate anaerobic Bacteria, culture conditions, high requirements on equipment, it is difficult to produce products with high bacterial count and high spore rate under normal equipment conditions, resulting in the current inability to popularize and low total output

Method used

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  • Method for promoting clostridium butyricum to form spores and microecological preparation and application thereof
  • Method for promoting clostridium butyricum to form spores and microecological preparation and application thereof
  • Method for promoting clostridium butyricum to form spores and microecological preparation and application thereof

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preparation example Construction

[0047]Specifically, the preparation method of the inactivated Clostridium butyricum sludge comprises: inoculating the Clostridium butyricum into the induction medium at a volume ratio of 1-2%, and statically culturing at 36-38° C. for 16-18 hours; In the boiling water bath for 20-40min, the solid-liquid separation is carried out after detecting no live bacteria. There are many ways of solid-liquid separation, including but not limited to centrifugation or filtration; in this application, it is preferred to use centrifugation for solid-liquid separation, centrifuge at 8000-12000rpm for 8-12min, remove the supernatant to obtain inactivated butyric acid shuttle Bacteria slime.

[0048] When the bacillus group forms spores under the harsh environment in nature, only part of the bacteria will form spores, and the other part of the bacteria will disintegrate and die. Studies have shown that this is mainly because during the spore formation process of bacillus, It will release subst...

Embodiment 1

[0056] The present embodiment provides a method for promoting the formation of spores by Clostridium butyricum, comprising the steps of:

[0057] S1. Prepare seed solution.

[0058] Clostridium butyricum was inoculated into a 1L conventional shake flask with a liquid content of 800ml according to a volume ratio of 1%, and cultured statically at 36-38° C. for 30 hours to obtain a Clostridium butyricum seed liquid. The seed medium in the shake flask was RCM liquid medium.

[0059] S2, expansion of fermenter.

[0060] The seed solution is inoculated into the fermentation medium at a volume ratio of 0.3%. When the seed solution is inoculated, the temperature is controlled at 36-38° C., the rotational speed is controlled at 50 rpm, and there is no ventilation. After seed liquid inoculation is completed, adjust the pH of the fermentation medium to ≥6.5 with 10% sodium hydroxide solution, and continuously control the pH of the fermentation medium to ≥6.5 during the entire expansion...

Embodiment 2

[0066] This embodiment is the same as other step parameters of Embodiment 1, the difference is only in:

[0067] The fermentation medium in the fermenter in step S2 consists of: glucose 8g / L, yeast powder 3g / L, peptone 8g / L, corn flour 8g / L, beef extract powder 8g / L, sodium acetate 3g / L, hydrogen phosphate Dipotassium 1.8g / L, ferrous sulfate 1.8g / L, sodium alginate 4g / L, agar 0.6g / L.

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Abstract

The invention discloses a method for promoting clostridium butyricum to form spores and a microecological preparation and application thereof, and relates to the technical field of microbial preparations. The method for promoting the clostridium butyricum to form the spores comprises the steps that a spore production inducer is added in the fermentation culture process, the spore production inducer comprises inactivated clostridium butyricum bacterial sludge, and the inactivated clostridium butyricum bacterial sludge is obtained by inoculating clostridium butyricum into an induction culture medium to be cultured and inactivated and then conducting solid-liquid separation. According to the application, the situation that part of thalli which form spores firstly release substances such as spore production delay protein and the like to inhibit other thalli from forming spores due to an overlong spore forming process can be effectively prevented, so that the viable bacterium amount and the spore rate of the clostridium butyricum are effectively improved, the problem that high bacterium amount and high spore rate of the clostridium butyricum are difficult to coexist is solved, and the yield of the clostridium butyricum is improved. The preservation, the transportation and the use of the clostridium butyricum are greatly promoted; in addition, the microecological preparation prepared by the invention has a remarkable effect when being applied to piglet feeding, and can effectively improve the feed intake, promote digestion and reduce the occurrence of diarrhea.

Description

technical field [0001] The invention relates to the technical field of microbial preparations, in particular to a method for promoting the formation of spores by Clostridium butyricum and its microecological preparation and application. Background technique [0002] As one of antibiotic replacement products, probiotics can regulate intestinal health, improve immunity, improve feed digestion and utilization, and reduce waste discharge. The commonly used strains of microecological preparations include Bacillus subtilis, Bacillus licheniformis, Enterococcus faecalis, Enterococcus faecium, Lactobacillus acidophilus, Lactobacillus plantarum, Saccharomyces cerevisiae, etc. There are few reports on the use of Clostridium butyricum to prepare microecological preparations. [0003] Clostridium butyricum, also known as Clostridium butyricum, is an obligate anaerobic Gram-positive bacillus. The bacterium is straight or slightly curved and can produce spores. Mesophytic endophytic spore...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12N1/20A23K10/18A23K50/30A23K50/60C12R1/145
CPCC12N3/00C12N1/20A23K10/18A23K50/30A23K50/60
Inventor 张志榕覃智斌郑甜力方勇军严二存李本松吴有林
Owner 茂名傲新生物科技有限公司
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