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A kind of fed-batch fed-batch fermentation method of bacillus

A technology of bacillus and fermentation method, which is applied in the field of fed-feed fermentation of bacillus, can solve problems such as the inability to meet the degradation characteristics of Bacillus subtilis vomitoxin, and achieves the improvement of unit output, the increase of the number of viable fermentation bacteria, and the conversion rate of spores. improved effect

Active Publication Date: 2021-03-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The invention patent with application publication number CN104232526A discloses a preparation process of live Bacillus subtilis preparations. The preparation process of live Bacillus subtilis ANSB471 needs to take into account both the number of live bacteria and the deoxygenation activity of vomitoxin. The fermentation method provided in CN104232526A cannot meet the requirements. Bacillus subtilis ANSB01G exerts the best vomitoxin degradation properties

Method used

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Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: the fermentation of Bacillus subtilis ANSB471 that the present invention relates to

[0033] The fermentation method of bacillus subtilis ANSB471, comprises the steps:

[0034] (1) Primary seed preparation: 250mL triangular flask is filled with 100mL primary seed medium, sterilized by moist heat at 115°C for 20min, and inoculated into the primary seed medium with 1mL ANSB471 live bacteria freeze-dried tube, temperature 37°C, Cultivate for 16 hours under the condition of 200r / min.

[0035] The primary seed medium formula (g / L) is: tryptone 8g, yeast extract 1.5g, glucose 1.5g, beef extract 2g, sodium chloride 3g, disodium hydrogen phosphate 2.5g, magnesium sulfate heptahydrate 0.5g, Add distilled water to 1000mL, the pH value is 7.0.

[0036] (2) Secondary seed preparation: 200L secondary seed medium was installed in a 500L fermenter, sterilized by moist heat at 115°C for 20min, inoculated primary seed into secondary seed medium at a ratio of 1:100, and c...

Embodiment 2

[0043] Embodiment 2: the fermentation of bacillus subtilis ANSB471 of the present invention

[0044] The fermentation method of bacillus subtilis ANSB471, comprises the steps:

[0045] (1) Primary seed preparation: 250mL triangular flask is filled with 100mL primary seed medium, sterilized by damp heat at 121°C for 15min, and inoculated into the primary seed medium with 1mL ANSB471 live bacteria freeze-dried tube strain, temperature 40°C, rotation speed Cultivate for 20h under the condition of 180r / min.

[0046] The formula (g / L) of primary seed medium is: tryptone 12g, yeast extract 3g, glucose 4g, beef extract 5g, sodium chloride 6g, disodium hydrogen phosphate 4g, magnesium sulfate heptahydrate 1.5g, add distilled water to 1000mL, pH 7.0.

[0047] (2) Secondary seed preparation: 300L secondary seed medium was installed in a 500L fermenter, sterilized by moist heat at 121°C for 15 minutes, inoculated primary seed into secondary seed medium at a ratio of 1:100, and cultivat...

Embodiment 3

[0054] Embodiment 3: the preparation of bacillus subtilis ANSB471 bacterium powder

[0055] The preparation method of Bacillus subtilis ANSB471 bacterium powder, concrete operation is: take the fermented liquid of embodiment 1 and 2 Bacillus subtilis respectively and heat to 55 ℃, enter concentrator, be to concentrate under vacuum degree of 85kpa, concentrate until the solid content reaches When the concentration is 15%-30%, the material starts to be discharged; the concentrated fermented liquid is dried and granulated by multi-stage low-temperature spray fluidized drying equipment. ×10 11 indivual. The inlet air temperature is 170°C, and the outlet temperature is 60-80°C.

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Abstract

The invention discloses a fed-batch fed-bacillus fermentation method belonging to the technical field of microbial fermentation. The bacillus is cultured in the fermentation medium after primary and secondary seed expansion. During this period, the carbon source and the relative lack of glycine, methionine, tryptophan, threonine, and proline in the basal medium are added through feeding. The mixture obtained by spray-drying the CFU per gram of Bacillus powder is not less than 1.0×10 11 One, significantly increased unit output and reduced unit cost. Bacillus subtilis ANSB471 was used to degrade vomitoxin, and after 48 hours of reaction, the degradation rate of vomitoxin reached 95%.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a fed-batch fed-bacillus fermentation method. Background technique [0002] Deoxynivalenol (DON), also known as deoxynivalenol or deoxynivalenol or vomitoxin, is the most polluting fusarium toxin to grains, seriously threatening the health of humans and livestock. Wheat, corn and other grains and their by-products are susceptible to vomitoxin, and the incidence is relatively high in Canada, the Nordic region, the United States and other foreign regions, and it can occur from south to north in my country. Studies have shown that the contamination rate and content of vomitoxin in the diet of residents in areas with high incidence of esophageal cancer and gastric cancer in my country are quite high. In 1998, the International Agency for Research on Cancer (IARC) listed vomitoxin as one of the three types of carcinogens in its evaluation report. Only by s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/38C12N1/20C12N3/00A23L5/20C12R1/125
CPCA23L5/28C12N1/20C12N1/38C12N3/00
Inventor 马秋刚赵丽红计成郭永鹏郑文革
Owner CHINA AGRI UNIV
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