A kind of high-density fermentation method of duck Escherichia coli

A high-density fermentation and Escherichia coli technology, which is applied in the field of microbial fermentation, can solve the problems of low requirements for culture conditions and difficulty in achieving high-density fermentation, and achieve the effects of simple preparation methods, increased concentration, and reduced production costs

Active Publication Date: 2021-08-03
GUANGDONG HAID ANIMAL HUSBANDRY & VETERINARY RES INST
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  • Abstract
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Problems solved by technology

[0004] Pathogenic Escherichia coli is a facultative anaerobic bacterium, which does not require high culture conditions. It can grow normally in ordinary LB broth medium, but it is difficult to achieve high-density fermentation. High-density fermentation of pathogenic duck E. coli It is rarely reported

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  • A kind of high-density fermentation method of duck Escherichia coli

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Embodiment 1

[0029] The growth curve and cell morphology of duck Escherichia coli (O78 serotype) were compared under the conditions of four different fermentation methods. The differences between the four fermentation methods are shown in the table below:

[0030] Table 1 Differences between the four fermentation methods

[0031] Example 1 Based on the high-density fermentation medium of the present invention, adding glucose Comparative example 1 Based on the high-density fermentation medium of the present invention, without adding glucose Comparative example 2 Based on LB broth medium, add glucose Comparative example 3 Based on LB broth medium, no glucose

[0032] The specific fermentation method of embodiment 1 is as follows (medium volume is 1L):

[0033] (1) Prepare the preferred duck Escherichia coli high-density fermentation medium and related reagents, and carry out the tank-killing treatment and assembly of the fermenter:

[0034] a. Optimum h...

Embodiment 2

[0062] Embodiment 2 A kind of high-density fermentation method of duck escherichia coli

[0063] According to the inoculum size, inoculate duck Escherichia coli into the high-density fermentation medium for fermentation (add an appropriate amount of sterilized antifoaming agent to the high-density fermentation medium before inoculation), and in the fermentation process, use sulfuric acid solution and ammonia water Adjust the pH of the fermentation broth to 7.0, and the fermentation temperature to 37.5°C; gradually increase the speed setting from 100rpm to 400rpm, and gradually increase the air ventilation rate from 1.5L / min to 5L / min (increase the speed at different time points, and the OD of the fermentation broth 600 It takes about 3 to 4 hours for the nm to reach 5.0 to 8.0. When inoculated, the speed is set to 100rpm, the ventilation rate is 1.5L / min, the 0.5h is set to 150rpm, 2L / min, the 1h is set to 200rpm, 3L / min, and the 2h is set to 300rpm, 4L / min. min, set to 400rpm...

Embodiment 3

[0065] Embodiment 3 A kind of high-density fermentation method of duck escherichia coli

[0066] According to the inoculum size, inoculate duck Escherichia coli into a high-density fermentation medium for fermentation (add an appropriate amount of sterilized antifoaming agent to the high-density fermentation medium before inoculation), and in the fermentation process, use sulfuric acid solution and ammonia water Adjust the pH of the fermentation broth to 7.5, and the fermentation temperature to 36.5°C; gradually increase the speed setting from 100rpm to 400rpm, and gradually increase the air ventilation rate from 1.5L / min to 5L / min (increase the speed at different time points, and the OD of the fermentation broth 600 It takes about 3 to 4 hours for the nm to reach 5.0 to 8.0. When inoculated, the speed is set to 100rpm, the ventilation rate is 1.5L / min, the 0.5h is set to 150rpm, 2L / min, the 1h is set to 200rpm, 3L / min, and the 2h is set to 300rpm, 4L / min. min, set to 400rpm a...

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Abstract

The invention discloses a method for high-density fermentation of duck coliform bacteria. Based on a high-density fermentation medium, a fermenter is used to carry out high-density fermentation of duck coliform bacteria. During the fermentation process, set pH to 7.0 to 7.5, temperature to 36.5 to 37.5°C, rotational speed to 90 to 420 rpm, and air volume to 1.2 to 5.2 L / min. Add an appropriate amount of defoamer before inoculation, after inoculation, wait until the bacteria OD 600 When the nm reaches 4.0-6.0, add 45-55 (w / v)% glucose solution at a flow rate of 10-25mL / h / L, and the amount of glucose added is 50-100g of glucose per increase in the density of the fermentation medium. The method for high-density fermentation of duck Escherichia coli according to the present invention is based on a high-density fermentation medium, which is rich in nutrients, can realize high-density fermentation of duck Escherichia coli by feeding glucose, and can be used for large-scale fermentation and production of duck Escherichia coli vaccines.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a high-density fermentation method of duck Escherichia coli. Background technique [0002] Duck colibacillosis is caused by pathogenic Escherichia coli, and the pathological manifestations mainly include acute sepsis, air sac inflammation, panophthalmitis, synovitis, salpingitis and peritonitis. With the continuous expansion of the breeding scale and the continuous deterioration of the water environment for waterfowl breeding, colibacillosis has become one of the more serious bacterial infectious diseases that endanger waterfowl farming. Since antibiotics are widely used clinically to treat bacterial diseases such as Escherichia coli, a large number of drug-resistant strains have been produced. Therefore, vaccination is an effective preventive approach in the prevention and treatment of Escherichia coli. [0003] For bacterial vaccines, how to increase...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/19
CPCC12N1/20
Inventor 龚凤平袁建丰马海彬罗梦萍
Owner GUANGDONG HAID ANIMAL HUSBANDRY & VETERINARY RES INST
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