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Method for preparing hydrocortisone by resting cell multi-round fermentation

A technology for converting hydrocortisone and resting cells, which is applied in the field of microbial steroid pharmacy and medical engineering, can solve the problems of poor specificity of 11β-hydroxylase, limited HC yield, low feeding concentration, etc., to save bacteria The effect of cultivating steps, shortening the cycle, improving conversion rate and specific productivity

Pending Publication Date: 2021-04-06
HUAZHONG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, due to the poor specificity of Absidia coeruleus 11β-hydroxylase, many by-products, and low feed concentration, the yield of HC is limited.

Method used

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  • Method for preparing hydrocortisone by resting cell multi-round fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0037] A method for preparing hydrocortisone by multi-round fermentation of resting cells, the steps are as follows:

[0038] Strain preparation: inoculate the blue columbine mold on the PDA solid slant medium under sterile conditions, control the culture temperature at 28±0.5°C, and cultivate for 7-9 days until the gray spore filaments that fill the test tube grow and grow. spore.

[0039] Fermentation culture: the spores of the above strains are eluted with a certain amount of sterile water under sterile conditions, inoculated into the shake flask medium, and cultivated at a temperature of 28±0.5°C and a rotational speed of 200r / min until the pH is 4.0 , and the culture period was 24h, and the fermented bacterial liquid of Absidia indigo was obtained.

[0040] The shake flask culture medium (by percentage) is: glucose 1.2%, corn steep liquor 1%, yeast extract 0.4%, ammonium sulfate 0.6%, the rest is water, pH=6.3, sterilized at 121° C. for 30 minutes.

[0041] Substrate fe...

Embodiment 2

[0044] A method for preparing hydrocortisone by multi-round fermentation of resting cells, the steps are as follows:

[0045] (1) Strain preparation: Inoculate the blue columbine mold on the PDA solid slant medium under aseptic conditions, control the culture temperature at 28±0.5°C, and cultivate for 7 to 9 days until it grows into a tube full of bacteria. Gray spores and spores.

[0046] (2) Fermentation culture: Then, the spores of the above strains are eluted with a certain amount of sterile water under sterile conditions, inoculated into the shake flask culture medium, and cultivated at a temperature of 28±0.5°C and a rotational speed of 200r / min. Until the pH is 4.0, the culture period is 24 hours, and the fermented bacterial liquid of Absidia indigo can be obtained.

[0047] The shake flask culture medium (by percentage) is: glucose 1.2%, corn steep liquor 1%, yeast extract 0.4%, ammonium sulfate 0.6%, the rest is water, pH=6.2, sterilized at 121° C. for 30 minutes.

...

Embodiment 3

[0051] A method for preparing hydrocortisone by multi-round fermentation of resting cells, the steps are as follows:

[0052] (1) Strain preparation: Inoculate the blue columbine mold on the PDA solid slant medium under aseptic conditions, control the culture temperature at 28±0.5°C, and cultivate for 7 to 9 days until it grows into a tube full of bacteria. Gray spores and spores.

[0053] (2) Fermentation culture: Then, the spores of the above strains are eluted with a certain amount of sterile water under sterile conditions, inoculated into the shake flask culture medium, and cultivated at a temperature of 28±0.5°C and a rotational speed of 200r / min. Until the pH is 4.0, the culture period is 24 hours, and the fermented bacterial liquid of Absidia indigo can be obtained.

[0054] The shake flask culture medium (by percentage) is: glucose 1.2%, corn steep liquor 1%, yeast extract 0.4%, ammonium sulfate 0.6%, the rest is water, pH=6.4, sterilized at 121° C. for 30 minutes.

...

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Abstract

The invention discloses a method for preparing hydrocortisone by resting cell multi-round fermentation. The method comprises the following steps: by taking vitiligo blue as a production strain, carrying out seed amplification culture, adding a substrate RSA into fermentation liquor, and performing converting to generate hydrocortisone; after transformation is finished, collecting mycelia centrifugally, taking water as a fermentation medium to feed a substrate RSA for fermentation transformation, and so on, performing multiple rounds of resting cell transformation. Particularly, in the resting cell multi-round transformation process, transformation medium water only needs to be simply boiled, high temperature, high pressure, any PH adjustment and other aspects like culture medium sterilization are not needed, the strain culture step is omitted, materials and power needed by preparation and sterilization of the culture medium are also saved, and the production cost is reduced; and the conversion rate and the specific productivity of hydrocortisone are improved.

Description

technical field [0001] The invention belongs to the field of microbial steroid pharmacy and medical engineering, and in particular relates to a method for preparing hydrocortisone by multi-round fermentation of resting cells. Background technique [0002] Steroid drugs have important physiological activities and are the second largest class of drugs after antibiotics. Hydrocortisone (HC for short), also known as cortisol, has a chemical name of 11β, 17α, 21-trihydroxypregna-4-ene-3,20-dione, which belongs to the class of adrenocortical hormone drugs and is the current hormone It has anti-inflammatory and anti-allergic effects, and is mainly used in clinical substitution therapy for adrenal insufficiency. At the same time, because it has a certain influence on glucose metabolism, it is also used in the treatment of diseases such as glucose and hyperglycemia, and can also be used as the precursor of many steroidal corticosteroid drugs in production. Since the microbial synth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/08C12R1/65
CPCC12P33/08
Inventor 徐明琴付林冯蕾廖俊田玉林曾建华
Owner HUAZHONG PHARMA
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