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A kind of bacillus and its high-density culture method

A technology for high-density culture and bacillus, applied in the field of bacillus 13002 and its high-density culture, can solve the problems of insufficient optimization conditions, single method selection, limited scope of application, etc., and achieve the effect of overcoming the inhibition of substrates and products

Active Publication Date: 2019-08-20
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Specific methods include adding neutralizers to control pH, adding carbon sources to reduce substrate inhibition, etc., but there are still shortcomings such as limited scope of application, insufficient optimization conditions, and single method selection.

Method used

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  • A kind of bacillus and its high-density culture method
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  • A kind of bacillus and its high-density culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Put the bacterial suspension of Bacillus 13002 in the logarithmic phase growth in the seed culture medium, and carry out 2 to 3 continuous activation cultures to obtain the seed culture liquid; the seed culture medium is: calculated according to the mass ratio, the bacteria 1.0 parts of peptone, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 1.0 parts of NaCl, distilled water to 100 parts, and NaCl 2 HPO 4 -NaH 2 PO 4 Adjust the pH of the buffer to 6.5, and cool it for later use after sterilization;

[0044] (2) Inoculate the seed culture solution in the fermentation medium at a volume ratio of 6:100, and carry out shake flask culture at 45°C until the 24h cultivation is completed to obtain the fermentation medium; the fermentation medium is: according to the quality Ratio calculation, 1.0 parts of bacteriological peptone, 1.0 parts of yeast extract powder, 2.0 parts of glucose, 1.0 parts of NaCl, distilled water to 100 parts, adjusted pH to 6.5, steril...

Embodiment 2

[0076] Preparation of Bacillus 13002 bacterial powder

[0077] (1) Put the bacterial suspension of Bacillus 13002 in the logarithmic phase growth in the seed culture medium, and carry out 2 to 3 continuous activation cultures to obtain the seed culture liquid; the seed culture medium is: calculated according to the mass ratio, the bacterial 1.0 parts of peptone, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 1.0 parts of NaCl, distilled water to 100 parts, and NaCl 2 HPO 4 -NaH 2 PO 4 Adjust the pH of the buffer to 6.5, and cool it for later use after sterilization;

[0078] (2) Cleaning, sterilizing, cooling, and calibrating the pH electrode of the 4L automatic fermenter;

[0079] (3) In an automatic fermenter, insert the seed culture solution into the fermentation medium at a ratio of 6:100 by volume, and cultivate it at 45°C to measure the total number of live bacteria corresponding to 0h and 4h; the fermentation medium The formula is: calculated according to...

Embodiment 3

[0085] (1) Put the bacterial suspension of Bacillus 13002 in the logarithmic phase growth in the seed culture medium, and carry out 2 to 3 continuous activation cultures to obtain the seed culture liquid; the seed culture medium is: calculated according to the mass ratio, the bacterial 1.0 parts of peptone, 0.5 parts of yeast extract powder, 2.0 parts of glucose, 1.0 parts of NaCl, distilled water to 100 parts, and NaCl 2 HPO 4 -NaH 2 PO 4 Adjust the pH of the buffer to 6.5, and cool it for later use after sterilization;

[0086] (2) Inoculate the seed culture solution in the fermentation medium with a volume ratio of 2:100, and carry out shake flask culture at 45°C; the fermentation medium is: calculated according to the mass ratio, 1.5 parts of bacteriological peptone, yeast 2.5 parts of extract powder, 1.5 parts of glucose, 0.5 parts of NaCl, with Na 2 HPO 4 -The volume of the citric acid buffer was adjusted to 100 parts, and the pH was adjusted to 6.5, and cooled for ...

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Abstract

The invention discloses a bacillus and a high-density cultivation method for the same. A strain is Bacillus sp. 13002, and a preservation number is CGMCC No.7431. The high-density cultivation method includes (1), activating and cultivating bacterial suspension of the bacillus 13002 in seed cultivation media to obtain seed cultivation liquid; (2), inoculating the seed cultivation liquid in fermentation cultivation media, fermenting and cultivating the seed cultivation liquid for 16h-36h and collecting fermented cultivation liquid; (3), centrifuging the fermented cultivation liquid, then collecting bacterial mud, cleaning the bacterial mud by the aid of normal saline, adding bacterium protective agents into the bacterial mud, then uniformly mixing the bacterium protective agents and the bacterial mud with one another to obtain mixtures and drying the mixtures to obtain bacterial powder. The bacillus 13002 in the bacterial suspension grows at log phases. The bacillus and high-density cultivation method have the advantages that the total amount of viable bacteria in the freeze-dried bacillus can reach 0.730*10<11>CFU / g at least, accordingly, the problem of bacterial growth inhibition due to acid generated in cultivation procedures can be effectively solved, and the growth rate of the bacillus 13002 can be increased to a great extent.

Description

technical field [0001] The invention relates to the field of microbial fermentation engineering, in particular to a bacillus 13002 and a high-density culture method thereof. Background technique [0002] Bacillus coagulans is a spore-producing, Bacillus-like lactic acid bacterium. Gram-positive bacteria, the metabolic type is facultative anaerobic type, aerobic conditions can produce more biomass than anaerobic conditions. Bacteria are rod-shaped, with terminal spores and no flagella. The optimum growth temperature is 40°C-50°C, the optimum growth pH is generally 6.6-7.0, and fermentation can produce lactic acid. At the same time, Bacillus coagulans is a potential stock in the field of probiotics. It has stress resistance such as high temperature resistance, bile salt resistance, and acid resistance. It also has a certain ability to inhibit some antibiotics and certain harmful bacteria. In recent years, Bacillus coagulans has become one of the research hotspots at home an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/07
Inventor 刘冬梅金迅周全兴陈舒然
Owner SOUTH CHINA UNIV OF TECH
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