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Method for promoting conversion of plant sterols into 9 alpha-hydroxyandrostenedione by Mycobacterium fortuitum

A phytosterol and hydroxyl technology, applied in the field of ultrasonic vibration, can solve the problems of prolonged transformation time, poor contact between substrate and microbial cells, poor solubility, etc.

Inactive Publication Date: 2016-07-06
TIANJIN JINYAO GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Yang Ying pointed out in the article (research progress of microbial degradation of sterol side chain and conversion of androst-4-ene-3,17-dione, Microbiology Bulletin, 2006 33(6), 142-145) that sterol substrates in aqueous solution The problem of poor solubility in the medium and serious product (substrate) inhibition has plagued the entire steroid industry. Sterols are hydrophobic compounds with extremely low solubility in water, which leads to poor substrate and microbial cells. contact, the conversion rate is low and the conversion time is prolonged
The use of ultrasonic crushing, surfactants and organic solvents to dissolve substrates has achieved certain results, but surfactants and organic solvents have certain inhibitory effects on cell activity, which restricts its application.
[0008] Simultaneously take phytosterol as raw material and utilize Mycobacterium fortuitum (mycobacterium) bacterial strain fermentation to obtain 9α-hydroxy androstenedione, the time is longer, often up to 72 hours, and the problem of long-term fermentation is that at first it is easy to cause excessive fermentation of microorganisms, will get The product is converted into 9α-hydroxytestosterone and even degrades the steroid ring. Secondly, if the reaction time is too long, the bacteria will increase, and the conversion specificity of the culture bacteria will decrease, thereby increasing impurities. Thirdly, the long-term reaction will also cause the production cycle. , manufacturing costs, labor costs, etc. are rising, which is not conducive to industrialization

Method used

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  • Method for promoting conversion of plant sterols into 9 alpha-hydroxyandrostenedione by Mycobacterium fortuitum
  • Method for promoting conversion of plant sterols into 9 alpha-hydroxyandrostenedione by Mycobacterium fortuitum
  • Method for promoting conversion of plant sterols into 9 alpha-hydroxyandrostenedione by Mycobacterium fortuitum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0055] Strain: Mycobacterium fortuitum ATCC35855.

[0056] Take 10 different batches of corn steep liquor and ferment them respectively according to the quantity and process in this example. Except corn steep liquor, other materials are the same batch of materials.

[0057] Seed culture stage:

[0058] Seed medium (per liter):

[0059]

[0060] Medium volume: 200ml / flask

[0061] Inoculum volume: Inoculate 3ml of cell suspension from agar slant per 200ml of new medium

[0062] Container: 1000ml conical flask

[0063] Stirring: Shaker (1”throw) 200rpm

[0064] Temperature: 30°C

[0065] Growth time: 72 hours culture

[0066]Note: A good culture organism is indicated if a yellow cell pellet forms at the bottom

[0067] method:

[0068] Inoculum for biotransformation was prepared in culture medium. Inoculate Erlenmeyer flasks from the agar surface (cultivated in glass test tubes), inoculate under sterile conditions, pipette sterile water (5 mL) onto the agar shaking ...

Embodiment 4

[0089] Method is similar to embodiment 1-3, but throws substrate concentration and becomes 4% (phytosterol 40g), and fermentation culture stage parameter changes as follows:

[0090] Biotransformation experiments were performed in stainless steel fermenters with efficient bottom agitation. The main parameters of the fermenter used are: the total volume of the fermenter is 10 liters, the working volume is 3 liters, the amount of inoculum is 0.4 L, the ventilation rate is 0.5 L / L / min, and the stirring is 400 rpm.

[0091] The experimental results are shown in the table below

[0092]

Embodiment 5

[0094] Method is the same as embodiment 4, but throws substrate concentration and becomes 7% (phytosterol 70g),

[0095]

[0096] Proved by the above examples, the preparation of 9α-hydroxyl-androst-4ene-3,17dione by MycobacteriumfortuitumATCC35855 as substrate by phytosterol composition, in the case of ultrasonic vibration, achieved better results, the conversion rate Significantly improved, thus through conventional technical methods, the yield and quality of the product can be significantly improved. Especially in the case of high-concentration feeding, such as 7%, the conversion rate is obviously better than the method without ultrasonic vibration through 2 times of ultrasonic vibration.

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Abstract

The invention relates to a method for converting plant sterols into 9 alpha-hydroxyandrostenedione by using a Mycobacterium fortuitum ATCC 35855 bacterial strain. Especially the method uses ultrasonic wave vibration.

Description

field of invention [0001] The invention relates to a method for converting phytosterols into 9α-hydroxy androstenedione by using Mycobacterium fortuitum ATCC35855 strain, in particular ultrasonic vibration is used in the method. Background technique: [0002] Androst-4-ene-3, 17-dione (androst-4-ene-3, 17-dione, referred to as androstenedione or AD) is an irreplaceable intermediate of steroid hormone drugs, which plays an important role in the body. very important regulation. It can be said that almost all steroid hormone drugs are produced with AD as the starting material. Such as for the production of sex hormones, progesterone, anabolic hormones and corticosteroids, but also for the synthesis of more than 100 kinds of drugs such as hydrocortisone, prednisone oxide, progesterone, estrenol, dexamethasone, etc. It is an essential raw material for the production of anti-pregnancy drug mifepristone and various family planning drugs. AD is now being used by some developed co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/00C12R1/33
Inventor 孙亮
Owner TIANJIN JINYAO GRP
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