Large-scale production method for clostridium butyricum

Clostridium butyricum, a production method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of many butyric acid passage times, long production cycle, large demand for butyric acid seed solution, etc. , to achieve the effects of reducing the risk of strain variation, reducing the passage of strains, and shortening the fermentation cycle

Pending Publication Date: 2020-11-06
HENAN JINBAIHE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] The purpose of the present invention is to provide a large-scale production method of Clostridium butyricum, to solve the problems of large demand for butyric acid seed solution, many times of butyric acid passage and long production cycle in the existing method

Method used

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  • Large-scale production method for clostridium butyricum
  • Large-scale production method for clostridium butyricum
  • Large-scale production method for clostridium butyricum

Examples

Experimental program
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Effect test

experiment example 1

[0052] According to the method of above comparative example and embodiment, the total number of live bacteria contained in the bacterium suspension of embodiment is adjusted to 3.5L Clostridium butyricum bacteria liquid (butyric acid seed solution) in the comparative example is the same Order of magnitude, measure the number of 0h viable bacteria inoculated into the seed tank and the bacteria content in the Clostridium butyricum fermenter, the results are shown in Table 1-Table 3.

[0053] Table 1 Comparison of fermentation effects when the number of live bacteria in 0h is 9-28CFU / ml

[0054]

[0055]

[0056] Table 2 The number of live bacteria in 0h is 10 2 Comparison of fermentation effects at CFU / ml level

[0057]

[0058] Table 3 0h live bacteria count is 10 3 Comparison of fermentation effects at CFU / ml level

[0059]

[0060]

[0061] From the above data, it can be seen that when the number of viable bacteria at 0h is basically the same, the 3.5L Clost...

experiment example 2

[0063] Select the total number of viable bacteria as the total number of viable bacteria in the bacterial suspension to be 10 5 、10 6 、10 7 、10 8 、10 9 、10 10 The Clostridium butyricum suspension of CFU was respectively put into 500L first-class seed tanks of Clostridium butyricum. Under the same conditions of other processes, the fermentation effects are shown in Table 4 and Table 5.

[0064] Table 4 total number of different live bacteria (10 5 、10 9 、10 10 CFU) Clostridium butyricum suspension fermentation effect

[0065]

[0066] Table 5 total number of different live bacteria (10 6 、10 7 、10 8 CFU) Clostridium butyricum suspension fermentation effect

[0067]

[0068] Can know by above-mentioned experiment: in clostridium butyricum seed tank 0h live bacteria number is in 10 2 ~10 4 CFU / ml (the fourth group-the sixth group), that is, the total number of live bacteria of Clostridium butyricum inserted is at 10 6 ~10 9 CFU, the time required for Clostrid...

experiment example 3

[0073] In the manner of the reference example, the Clostridium butyricum suspension was inoculated into 50L, 500L, and 1000L seed tanks respectively for anaerobic culture, and then the fermented strains obtained in the seed tank were inoculated into the Clostridium butyricum fermentation Fermentation culture was carried out in the tank, and the results are shown in Table 6.

[0074] The fermentation effect of the method of the embodiment of table 6 in different volume seed tanks

[0075]

[0076] From the experimental results in Table 6, it can be seen that by adjusting the total number of viable bacteria at the time of access, the number of viable bacteria at 0h is guaranteed to reach 10 2 -10 4 CFU / ml, the number of live bacteria can be obtained by large-scale fermentation at 10 8 Clostridium butyricum products above CFU / ml.

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Abstract

The invention relates to a large-scale production method for clostridium butyricum. The method comprises the following steps: (1) preparing a clostridium butyricum suspension with clostridium butyricum colonies of slant culture; (2) inoculating the clostridium butyricum suspension into fermentation broth in a seed tank, and performing anaerobic culture to a logarithmic growth phase to obtain fermentation strains; and (3) inoculating fermentation strains into the fermentation broth in a fermentation tank, and performing large-scale anaerobic fermentation production to obtain a clostridium butyricum product with a living bacteria number greater than 10<8>CFU/ml. The method prepares the clostridium butyricum suspension with a clostridium butyricum lawn on a slant culture medium, and then inoculates the clostridium butyricum suspension into the seed tank for anaerobic fermentation, and experimental results show that the same fermentation effect as a clostridium butyricum fermentation seedsolution can be achieved; the method does not require secondary expanding culture, so that the fermentation cycle is shortened by at least one primary seed fermentation cycle in the entire process flow; and meanwhile, at least one strain passage is also reduced, thus reducing the risk of strain variation caused by excessive strain passage.

Description

technical field [0001] The invention belongs to the field of cultivation of Clostridium butyricum, and in particular relates to a large-scale production method of Clostridium butyricum. Background technique [0002] Clostridium butyricum is an obligate anaerobic Gram-positive bacillus, which has extremely high requirements for anaerobic environment during the growth process. It is cultivated by general liquid submerged fermentation method. The concentration of the bacteria is low, and the cultivation is difficult. Production The high cost restricts its industrial production. [0003] The Chinese invention patent application with application publication number CN110846258A discloses a typical Clostridium butyricum fermentation process, including primary seed preparation, secondary seed preparation and fermentation inoculation. At present, the large-scale production of Clostridium butyricum mostly adopts the following two-stage fermentation process: [0004] (1) Inoculate th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/145
CPCC12N1/20
Inventor 韩娅楠李克克李保红谢璐璐冯利菲李光华
Owner HENAN JINBAIHE BIOTECH CO LTD
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