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Method for improving conversion efficiency of converting phytosterol into androstenedione

A technology of phytosterols and diketones, applied in the field of preparation of androst-4-ene-3, which can solve the problems of low solubility, increase of miscellaneous bacteria, excessive microbial fermentation, etc.

Inactive Publication Date: 2015-06-10
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
[0006] Utilize mycobacterium to carry out fermentation at the same time, the time is longer, often up to 72 hours, many papers all have been introduced to this, for example CN03804973, and the problem of long-time fermentation is that at first it is easy to cause excessive fermentation of microorganisms, the obtained AD / ADD is converted into 9α-hydroxytestosterone and even the steroid ring is degraded. Secondly, the reaction time is too long, which will increase the number of miscellaneous bacteria, and the conversion specificity of the culture bacteria will decrease, thereby increasing impurities. Thirdly, the long-term reaction will also cause production cycles. , manufacturing costs, labor costs, etc. are rising, which is not conducive to industrialization
[0007] The application of weak electric field in the process of phytosterol fermentation has not been found yet.

Method used

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  • Method for improving conversion efficiency of converting phytosterol into androstenedione
  • Method for improving conversion efficiency of converting phytosterol into androstenedione
  • Method for improving conversion efficiency of converting phytosterol into androstenedione

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1-a

[0060] Species: Mycobacterium: Mycobacterium fortuitum. NRRL B-8153.

[0061] 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.

[0062] Seed culture stage:

[0063] Seed medium (per liter):

[0064] 40ml corn syrup

[0065] 5.2g NaNO 3 ,

[0066] 0.8g NH 4 h 2 PO 4

[0067] 5.3g glucose

[0068] The rest is sterile water

[0069] Medium volume: 200ml / flask

[0070] Inoculum volume: Inoculate 3ml of cell suspension from agar slant per 200ml of new medium

[0071] Container: 1000ml conical flask

[0072] Stirring: Shaker (1”throw) 200rpm

[0073] Temperature: 30°C

[0074] Growth time: 72 hours culture

[0075] Note: A good culture organism is indicated if a yellow cell pellet forms at the bottom

[0076] method:

[0077] Inoculum for biotransformation was prepared in culture medium. Inoculate Erlenmeyer ...

Embodiment 2-1-a

[0097] 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.

[0098] The process of this embodiment is the same as the method of Example 1-1-a, and the detection method is the same, but the strain is changed to mycobacterium: Mycobacterium sp NRRL-B3683, and the culture medium is changed as follows:

[0099] Seed culture stage:

[0100] Seed medium (per liter):

[0101] 10ml corn syrup

[0102] 8.7g NaNO 3 ,

[0103] 0.3g NH 4 h 2 PO 4

[0104] 15g glucose

[0105] Na 2 HPO 4 and NaH 2 PO 4 Adjust the pH of the buffer to 7.0

[0106] The rest is sterile water

[0107] Medium volume: 200ml / flask

[0108] Inoculum volume: Inoculate 3ml of cell suspension from agar slant per 200ml of new medium

[0109] Container: 1000ml conical flask

[0110] Stirring: Shaker (1”throw) 200rpm

[0111] Temperature: 30°C

[01...

Embodiment 3-1-a

[0125] 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.

[0126] The process of this embodiment is the same as the method of Example 1-1-a, and the detection method is the same, but the strain is changed to mycobacterium: Mycobacterium sp NRRL-B3805, and the culture medium is changed as follows:

[0127] Seed culture stage:

[0128] Seed medium (per liter):

[0129] 25ml corn syrup

[0130] 4g NaNO 3 ,

[0131] 1g NH 4 h 2 PO 4

[0132] 1g FeCl 2

[0133] 15g glucose

[0134] Na 2 HPO 4 and NaH 2 PO 4 Adjust the pH of the buffer to 7.0

[0135] The rest is sterile water

[0136] Medium volume: 200ml / flask

[0137] Inoculum volume: Inoculate 3ml of cell suspension from agar slant per 200ml of new medium

[0138] Container: 1000ml conical flask

[0139] Stirring: Shaker (1”throw) 200rpm

[0140] Tempera...

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Abstract

The invention dsiclsoes a method of preparing androstane-4 alkene-3,17 diketone and / or androstane-1,4-diene-3,17-diketone by taking a phytosterol composition as a substrate. The method is characterized by comprising the following steps: adding the phytosterol composition into a reactor which contains nocardia or mycobacterium microorganisms and a culture medium for fermentation, regulating the PH value to be larger than 7, fermenting at 20-37 DEG C, converting the phytosterol composition into androstane-4 alkene-3,17 diketone and / or androstane-1,4-diene-3,17-diketone, wherein the conversion process is carried out in an electric field.

Description

field of invention [0001] The invention relates to a fermentation method for promoting the transformation of phytosterols into androsteroids by a weak electric field, in particular to a method for preparing androst-4-ene-3,17-diketone. 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 devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/02C12R1/365C12R1/32
Inventor 卢彦昌李静赵春霞张杰
Owner TIANJIN JINYAO GRP
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