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A kind of high-density fermentation of clostridium butyricum and preparation method of microecological agent

A technology of high-density fermentation and microbial inoculum, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low activity of Clostridium butyricum strains, low de-spore rate, etc. Fermentation raw material and time cost, reducing fermentation contamination, and promoting the effect of transformation

Active Publication Date: 2021-05-25
天津市圣世莱科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] In order to solve the above problems, the present invention provides a high-density fermentation of Clostridium butyricum and a preparation method of its microecological bacterial agent, which can not only solve the problems of low activity of Clostridium butyricum species and low despore rate in the fermentation process, Moreover, the fermentation process is simple, which can effectively reduce the fermentation bacteria and reduce the cost of fermentation raw materials and time, and at the same time significantly improve the stability of the bacterial agent

Method used

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  • A kind of high-density fermentation of clostridium butyricum and preparation method of microecological agent
  • A kind of high-density fermentation of clostridium butyricum and preparation method of microecological agent
  • A kind of high-density fermentation of clostridium butyricum and preparation method of microecological agent

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Embodiment 1

[0053] The fermentation culture of embodiment 1 clostridium butyricum and the preparation of bacterial agent

[0054] 1. Primary seed liquid culture of Clostridium butyricum:

[0055] Take a frozen glycerol tube seed solution stored at -80°C, and after it melts quickly, inoculate 2% of the inoculum into the seed medium (60ml / 100ml round-bottomed flask), and culture at 37°C for 45 hours. Obtain the primary seed liquid;

[0056] 2. Secondary seed liquid culture of Clostridium butyricum:

[0057] After the primary seed solution cultivated in step 1 was bathed in 80°C for 10 minutes, inoculate it into the secondary seed medium according to the inoculum size of 5%. In the later stage of growth, when the k value of the growth curve of the strain reaches 0.7, the static culture is carried out for 20 hours, and the number of viable bacteria is 6.2×10 8 ; The bacterial growth curve of its seed liquid refers to the attached figure 2 ;

[0058] Both the primary seed medium and the ...

Embodiment 2

[0067] The fermentation culture of embodiment 2 clostridium butyricum and the preparation of bacterial agent

[0068] Concrete fermentation method comprises the following steps:

[0069] 1. Primary seed liquid culture of Clostridium butyricum:

[0070] Take the bacterial strains stored in a glycerol tube at -80°C, melt them quickly, and inoculate them in the seed medium according to the inoculum volume of 3% (filling volume 60ml / 100ml round bottom flask), and culture at 37°C for 48 hours;

[0071] 2. Secondary seed liquid culture of Clostridium butyricum:

[0072] After the primary seed liquid of Clostridium butyricum cultivated in step 1 was bathed in water at 80°C for 10 minutes, inoculate it into the secondary culture liquid according to the inoculum size of 5%, (filling volume 400 / 500ml round-bottom flask), and cultivate it statically at 37°C for 20 minutes. Hour;

[0073] Described primary and secondary seed medium composition is the same as embodiment 1

[0074] 3. T...

Embodiment 3

[0087] The fermentation culture of embodiment 3 clostridium butyricum and the preparation of bacterial agent

[0088] Concrete fermentation method comprises the following steps:

[0089] 1. Primary seed liquid culture of Clostridium butyricum:

[0090] Take the bacterial strains stored in a glycerol tube at -80°C, melt them quickly, and inoculate them in the seed medium according to the inoculation volume of 4% (filling volume 60ml / 100ml round bottom flask), and culture at 35°C for 45 hours;

[0091] 2. Secondary seed liquid culture of Clostridium butyricum:

[0092] After the primary seed liquid of Clostridium butyricum cultivated in step 1 was bathed in 85°C water bath for 5 minutes, inoculated into the secondary culture liquid according to the inoculum size 4%, (filling volume 400 / 500ml round bottom flask), and cultured at 36°C for 22 Hour;

[0093] Described primary and secondary seed medium composition is the same as embodiment 1

[0094] 3. Tertiary seed liquid cultu...

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Abstract

The invention provides a high-density fermentation of clostridium butyricum and a preparation method of a microecological bacterial agent, belonging to the field of microbial fermentation production. The present invention effectively solves the problem by controlling the seed fermentation stage of the strain to the late logarithmic growth stage, and controlling the tank pressure, ventilation rate, fermentation pH and fermentation temperature of the fermentation at the initial stage of fermentation and during the 4-5h and 12-15h fermentation processes respectively. Clostridium butyricum has low activity and low despore rate. The spore rate can reach more than 98%, and the complete spore shedding rate can reach more than 95%. Moreover, the fermentation process of the present invention is simple, which can effectively reduce fermentation contamination and reduce fermentation raw materials and Time costs. The micro-ecological bacterial agent prepared by the invention can significantly improve the stability of the bacterial agent, and the survival rate of the bacterial agent is as high as 97%, and can realize the industrialization standard of strain production.

Description

technical field [0001] The invention belongs to the field of microbial fermentation production, and in particular relates to a high-density fermentation of Clostridium butyricum and a preparation method of a microecological agent. [0002] technical background [0003] Clostridium butyricum (Clostridium butyricum) belongs to the Bacillus family, Clostridium genus, Gram-positive, with spores, spores are oval, eccentric or sub-terminal, and its diameter is (0.6-1.2)×(3.0-7.0) μm , can resist adverse environment. [0004] Clostridium butyricum is used as a feed additive in feed, and it can embody six biological characteristics in the intestinal tract of animals: First, it promotes the proliferation and development of beneficial bacteria in the intestinal tract of animals (Bifidobacteria, Lactobacillus), and inhibits harmful bacteria in the intestinal tract. The growth and reproduction of bacteria and spoilage bacteria can correct the disorder of intestinal flora and reduce the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12R1/145
CPCC12N1/04C12N1/20
Inventor 王连民刘珂飞孟铁健
Owner 天津市圣世莱科技有限公司