Mycobacterium neoaurum and application thereof in preparation of pregn-4-en-3-one,9,21-dihydroxy-20methyl

A new mycobacterium aureus and hydroxymethyl technology, applied in the field of bioengineering, can solve problems such as inability to produce quickly and efficiently, and achieve the effects of short production cycle, improved product purity, and increased content

Active Publication Date: 2020-06-30
广东本科生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is to overcome the defect that 9OA-BA in the prior art cannot be produced quickly and efficiently, thereby providing a new mycobacterium aureus and its preparation of 9α-hydroxyl-20α-hydroxymethyl- Application of Pregn-4-en-3-one

Method used

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  • Mycobacterium neoaurum and application thereof in preparation of pregn-4-en-3-one,9,21-dihydroxy-20methyl

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preparation example Construction

[0031] The present invention also provides a preparation method of 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3-one, the preparation method comprising converting sterols into The step for 9α-hydroxy-20α-hydroxymethyl-pregn-4-en-3-one.

[0032] Among them, the English name of 9α-hydroxy-20α-hydroxymethyl-pregn-4-en-3-one is: pregn-4-en-3-one, 9,21-dihydroxy-20methyl (referred to as 9OH-BA, CAS number: 69155-84-6), the chemical structure is as follows:

[0033]

[0034] Wherein, the sterols include at least one of phytosterol, animal sterol, wood sterol and cholesterol.

[0035] The conditions of fermenting and cultivating the seeds of Mycobacterium neogolden in the transformation medium containing sterols are: the seed inoculum size is 5-15% (V / V), the culture temperature is 30-34°C, and the pH of the transformation medium is The value is 6.5-8.5, the ventilation rate is 0.05-1.5vvm, and the stirring rate is 200-400r / min. The fermentation time is preferably 100-150h.

[00...

Embodiment 1

[0052] This embodiment relates to a production method of 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3one, comprising the following steps:

[0053] (1) Seed cultivation

[0054] Inoculate Mycobacterium neoaurum BK076 from the solid slant of the eggplant bottle into the seed medium, and cultivate at 200r / min at 32 for 40h to obtain seeds suitable for inoculation.

[0055] The seed medium includes the following components (g / L): yeast powder 10, glucose 10, diammonium hydrogen phosphate 5, magnesium sulfate heptahydrate 3, the balance is water, and the pH is adjusted to 7.5.

[0056] (2) Microbial transformation into 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3one

[0057] The seed solution obtained in step (1) is inoculated in a 10L fermenter for transformation culture, the seed inoculum size is 10% (V / V), the pH value of the transformation medium is 6.7, the transformation temperature is 32 ° C, and the ventilation rate is 0.5 vvm, the stirring speed is 250r / min, and the fermenta...

Embodiment 2

[0064] This embodiment relates to a production method of 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3one, comprising the following steps:

[0065] (1) Seed cultivation

[0066] Inoculate Mycobacterium neoaurum BK076 from the solid slant of the eggplant bottle into the seed medium, and culture at 32°C for 40 hours at 200r / min to obtain seeds suitable for inoculation.

[0067] The seed culture medium includes the following components (g / L): 10 peptones, 10 glucose, 5 glycerol, 5 diammonium hydrogen phosphate, 3 magnesium sulfate heptahydrate, the balance is water, and the pH is adjusted to 7.5.

[0068] (2) Microbial transformation into 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3one

[0069] The seed solution obtained in step (1) is inoculated in a 10L fermenter for transformation culture, the seed inoculum size is 10% (V / V), the pH value of the transformation medium is 6.7, the transformation temperature is 32 ° C, and the ventilation rate is 0.8 vvm, the stirring speed is 250r / ...

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Abstract

The invention relates to the field of bioengineering, in particular to Mycobacterium neoaurum and application thereof in preparation of pregn-4-en-3-one,9,21-dihydroxy-20methyl. The Mycobacterium neoaurum is classified and named as Mycobacterium neoaurum BK076, the preservation institution is China Center for Type Culture Collection, and the preservation number is CCTCCM20191115. The bacterial strain is a new bacterial strain obtained by mutagenesis screening, and can efficiently convert sterol substances to produce the pregn-4-en-3-one,9,21-dihydroxy-20methyl. The sterol conversion rate of substrate plants reaches 90% or above, the concentration of the pregn-4-en-3-one,9,21-dihydroxy-20methyl in fermentation liquor can reach 36 g/L or above, and the conversion rate reaches up to 85% or above.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a new mycobacterium aureus and its application in the preparation of 9α-hydroxyl-20α-hydroxymethyl-pregn-4-en-3-one. Background technique [0002] Steroid drugs are clinically essential drugs, which refer to steroid substances, usually including adrenaline, androgen, estrogen, etc., which can play a very important role in regulating the body, such as the adrenal gland Corticosteroids have anti-inflammatory, anti-allergic, anti-shock, anti-allergic effects, and are widely used to treat skin diseases such as rheumatoid arthritis, bronchial asthma, and eczema, and can treat and relieve collagen diseases, anaphylactic shock, etc. It is also an indispensable drug for the treatment of endocrine diseases such as Addison's. Generally speaking, steroid hormone drugs have strong anti-infection, anti-allergy, anti-virus and anti-shock and other important functions, so they are often...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P33/00C12R1/32
CPCC12P33/00C12N1/20C12R2001/32C12N1/205
Inventor 林钦鹏林汉标林智德陈斯涛陈峰吴婷
Owner 广东本科生物工程股份有限公司
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