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Bacillus cereus high-density fermentation method and application

The technology of Bacillus cereus and Bacillus cereus is applied in the field of Bacillus cereus high-density fermentation, and can solve the problems that the high-density fermentation of Bacillus cereus cannot be fully satisfied, pH-stat strategy feed control is slow, and the like

Active Publication Date: 2021-08-31
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The basic operation method of pH-stat is similar to that of DO-stat. It is also to set the expected maximum pH value and minimum pH value in advance. When the pH of the fermentation broth exceeds this range, the system will automatically start or limit the feeding of nutrients. It also has its own shortcomings, because when the nutrients in the fermentation broth are exhausted, the change of pH tends to be slower than the change of DO, so compared with DO-stat, the feeding control of pH-stat strategy is relatively slow
Both the DO-stat method and the pH-stat method have their own defects and cannot fully meet the requirements of high-density fermentation of Bacillus cereus.

Method used

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  • Bacillus cereus high-density fermentation method and application

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Embodiment 1

[0037] The impact of the feeding method of embodiment 1 different fed batches on the high-density cultivation of Bacillus cereus

[0038] (1) The Bacillus cereus ATCC 14579 preserved in the laboratory was transferred to NA solid slant medium, cultured in a constant temperature incubator at 30°C for 24 hours, passed twice and stored at 4°C.

[0039] (2) Pick two rings of activated bacterial lawn from the culture medium, inoculate them in a 250mL shake flask filled with 50mL LB liquid medium, and cultivate them at 30°C and 200r / min for 14-16h to obtain first-class seeds liquid.

[0040] (3) The primary seed solution obtained in step (2) was inoculated in a 500 mL Erlenmeyer flask containing 100 mL of seed medium with an inoculum volume ratio of 2% and cultivated for 14-16 hours to obtain a seed solution.

[0041] (4) Inoculate the seed liquid obtained in step (3) with an inoculum size of 10% by volume in a fermenter containing 2.5L of the initial fermentation medium, temperatur...

Embodiment 2

[0053] The impact of embodiment 2 different feeding mediums on the high-density cultivation of bacillus cereus

[0054] (1) The Bacillus cereus ATCC 14579 preserved in the laboratory was transferred to NA solid slant medium, cultured in a constant temperature incubator at 30°C for 24 hours, passed twice and stored at 4°C.

[0055] (2) Pick two rings of activated bacterial lawn from the culture medium, inoculate them in a 250mL shake flask filled with 50mL LB liquid medium, and cultivate them at 30°C and 200r / min for 14-16h to obtain first-class seeds liquid.

[0056] (3) The primary seed solution obtained in step (2) was inoculated in a 500 mL Erlenmeyer flask containing 100 mL of seed medium with an inoculum volume ratio of 2% and cultivated for 14-16 hours to obtain a seed solution.

[0057] (4) Inoculate the seed liquid obtained in step (3) with an inoculum size of 10% by volume in a fermenter containing 2.5L of the initial fermentation medium, temperature (32°C ± 0.5), pH...

Embodiment 3

[0070] Embodiment 3 The influence of the lye that maintains pH differently on the high-density cultivation of Bacillus cereus

[0071] (1) The Bacillus cereus ATCC 14579 preserved in the laboratory was transferred to NA solid slant medium, cultured in a constant temperature incubator at 30°C for 24 hours, passed twice and stored at 4°C.

[0072] (2) Pick two rings of activated bacterial lawn from the culture medium, inoculate them in a 250mL shake flask filled with 50mL LB liquid medium, and cultivate them at 30°C and 200r / min for 14-16h to obtain first-class seeds liquid.

[0073] (3) The primary seed solution obtained in step (2) was inoculated in a 500 mL Erlenmeyer flask containing 100 mL of seed medium with an inoculum volume ratio of 2% and cultivated for 14-16 hours to obtain a seed solution.

[0074] (4) Inoculate the seed liquid obtained in step (3) with an inoculum size of 10% by volume in a fermenter containing 2.5L of the initial fermentation medium, temperature (...

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Abstract

The invention discloses a high-density fermentation method of bacillus cereus and application, and belongs to the technical field of fermentation engineering. According to the high-density fermentation method, high-density fermentation of the bacillus cereus is carried out through a strategy of regularly and instantaneously supplementing nutrient substances required by thallus growth, converting different alkali liquor at a thallus growth stage to maintain pH in combination with a three-stage dissolved oxygen regulation and control mode. After fermentation is finished, the dry weight of the bacillus cereus reaches 40-48 g / L and is increased by 70-135% in comparison with traditional continuous fermentation, and is increased by 328% in comparison with a non-fed-batch fermentation method. The method provided by the invention effectively increases the dry weight of thalli of the bacillus cereus, is simple and easy to implement, and has a wide application prospect.

Description

technical field [0001] The invention relates to a high-density fermentation method and application of Bacillus cereus, belonging to the technical field of fermentation engineering. Background technique [0002] Bacillus cereus (Bacillus cereus) is a common probiotic that can produce spores and has strong stress resistance. It is resistant to acid and alkali, high temperature and some toxic chemical toxins. In nature, it has a close symbiotic relationship with most animals and plants, and can produce a variety of probiotic factors. Bacillus cereus is used in wastewater treatment, soil remediation, production of high-yield enzymes, bacteriocins, as immune active substances, prepared into biological preparations for the treatment of plant diseases, prevention and treatment of human and animal diarrhea, etc. , food, health products, environmental protection and other aspects are commonly used materials. [0003] As a probiotic preparation, the basis for its function in the bod...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/085
CPCC12N1/20
Inventor 詹晓北胥聆铭朱莉张晓凤
Owner JIANGNAN UNIV
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