Fermentation method for increasing spore rate of bacillus licheniformis

A technology of Bacillus licheniformis and fermentation method, which is applied in the fermentation of spore rate and the field of fermentation of Bacillus licheniformis, can solve the problems such as unsatisfactory spore rate, and achieve the effect of high spore rate and high number of viable bacteria

Active Publication Date: 2017-12-29
ZHEJIANG JINGXIN PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Bacterial fermentation is affected by many factors such as medium formula, moisture, temperature, etc., but the current literature research on the fermentation of Bacillus licheniformis is only limited to the regulation of fermentation temperature, dissolved oxygen and other influencing conditions (see CN103205388A), and the spore rate is not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1) Seed activation culture: after dissolving the Bacillus licheniformis CGMCC NO.0183 strain preserved in the ampoule tube with sterile saline, inoculate it in the nutrient broth medium (peptone 10g / L, beef extract powder 3g / L, chlorinated Sodium 5g / L, pH7.2), 150rpm, 37 ℃ shaker culture 6h, obtain seed activation culture solution.

[0040] 2) Seed culture: the seed activation culture solution obtained in step 1) is inoculated with 1% inoculum size into 1000mL nutrient broth medium (peptone 10g / L, beef extract powder 3g / L, sodium chloride 5g / L , pH7.2) in shake flasks, 37 ° C, 150 rpm for 6 hours to obtain seed culture solution.

[0041] 3) Fermentation culture: the seed culture solution that step 2) obtains is inoculated in the 15L fermenter with the inoculation amount of 20%, and 10L nutrient broth medium (peptone 10g / L, beef extract powder 3g / L is housed in the fermenter) , sodium chloride 5g / L, pH7.2), cultured at 37°C. The initial ventilation ratio is 1:1, increase...

Embodiment 2

[0044] Step 2) in the above-mentioned embodiment 1) the seed culture solution that obtains is inoculated in the 15L fermentor with 10% inoculum size, and 10L nutrient broth culture medium (10g / L of peptone, 3g / L of beef extract powder is housed in the fermenter) , sodium chloride 5g / L, pH7.0), cultured at 37°C. Initial ventilation ratio 1:0.5, increase ventilation ratio when dissolved oxygen is lower than 20%, decrease ventilation ratio when dissolved oxygen is higher than 50%; speed 200-600rpm, adjust speed according to dissolved oxygen value, decrease when dissolved oxygen rises RPM, increase RPM when dissolved oxygen decreases. Cultivate for 32h. Among them, the pH was adjusted to maintain 7.0 with phosphoric acid during the 0-24h fermentation; the pH was adjusted to maintain 8.5 until 32h at 24h.

[0045] After the fermentation, the fermented liquid was centrifuged at 10000rpm for 10min to obtain 5.1g / L of wet thallus, which was examined under an oil microscope after sta...

Embodiment 3

[0047] Step 2) in the above-mentioned embodiment 1) the seed culture liquid that obtains is inoculated in the 15L fermenter with the inoculum size of 30%, is housed in the fermentor , sodium chloride 5g / L, pH7.5), cultured at 37°C. The initial ventilation ratio is 1:1, increase the ventilation ratio when the dissolved oxygen is lower than 20%, reduce the ventilation ratio when the dissolved oxygen is higher than 50%; the speed is 200-600rpm, adjust the speed according to the dissolved oxygen value, and reduce it when the dissolved oxygen rises RPM, increase RPM when dissolved oxygen decreases. Cultivate for 32h. Among them, the pH was adjusted to maintain 7.5 with phosphoric acid during 0-24h of fermentation; the pH was adjusted to maintain 8 to 32h at 24h.

[0048] After the fermentation, the fermented liquid was centrifuged at 10000rpm for 10min to obtain 5.3g / L of wet thallus, which was examined under an oil microscope after staining with methylene blue, and the spore rat...

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Abstract

The invention discloses a fermentation method for increasing the spore rate of bacillus licheniformis. The fermentation method comprises the two following stages: inoculating the bacillus licheniformis in a culture medium with the pH of 7.0-7.5, and carrying out fermentation culture for 20-26 hours at the temperature of 36-38 DEG C; then regulating and controlling the pH of fermentation liquid to 8.0-8.5, continuing carrying out fermentation culture at the temperature of 36-38 DEG C, stopping fermentation at the moment of 30-36 hours since inoculation, and harvesting thalli with the spore rate of more than 85%.

Description

technical field [0001] The invention belongs to the field of microbial fermentation engineering, relates to a fermentation method of bacillus licheniformis, in particular to a fermentation method for increasing the spore rate of bacillus licheniformis. Background technique [0002] Bacillus licheniformis (Bacillus licheniformis) is one of the most commonly used strains of live bacterial preparations. It is a kind of Bacillus, a single cell, 0.6-0.8×1.5-3 microns, and the cell shape and arrangement are rod-shaped, single Healthy, can produce nearly mesogenic ellipsoid spores, cysts slightly enlarged. Bacillus licheniformis can promote the body to produce antibacterial substances and kill pathogenic bacteria. It can not only produce antibacterial substances, but also has a unique biological deoxygenation mechanism, which can inhibit the growth and reproduction of pathogenic bacteria (Escherichia coli, Clostridium perfringens) , Effectively regulate gastrointestinal dysfunctio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/742A61P1/00C12R1/10
CPCA61K35/742C12N1/20
Inventor 高兴强王晓飞张敏洁
Owner ZHEJIANG JINGXIN PHARMA
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