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Method of preparing 4-androstenedione by fermentatively degrading phytosterol by virtue of single water phase system

A technology of phytosterols and androstenedione, which is applied in the field of microbial transformation of steroidal drugs, can solve problems such as environmental pollution and product extraction difficulties, and achieve the effects of less environmental pollution, fewer types of solvents, and lower difficulty

Inactive Publication Date: 2014-07-16
HEBEI ZHONGSHENG BIOTECH
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  • Claims
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Problems solved by technology

[0005] The present invention proposes a method for preparing 4-androstenedione by fermenting and degrading phytosterols in a single-water phase system, which is used to solve the problems of difficult product extraction and environmental pollution in the existing method for preparing 4-androstenedione from microbial conversion of phytosterols serious problem

Method used

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  • Method of preparing 4-androstenedione by fermentatively degrading phytosterol by virtue of single water phase system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A method for preparing 4-androstenedione by fermenting and degrading phytosterols in a single aqueous phase system, comprising the steps of:

[0038] The first step, strain propagation: the concentration of 10 7 The Mycobacterium smegmatis bacterial classification of each / ml inserts and cultivates in the culture jar that seed medium is housed with 10% inoculation amount, comprises the component of following concentration in the seed medium: glucose 10g / L, K 2 HPO 4 0.6g / L, MgSO 4 ·7H 2 O0.5g / L, (NH 4 ) 2 Fe(SO 4 ) 2 ·6H 2 O0.05g / L, CaCO 3 5.0g / L and citric acid 2.5g / L, the pH of the seed medium is 6.0;

[0039] The second step, sterol emulsification:

[0040] The substrate phytosterol is emulsified under the action of emulsifier Tween 80, and the emulsification temperature is 60°C;

[0041] The third step, fermentation culture:

[0042] The OD of the culture medium in the first step 600 At 20 o'clock, with 50% inoculum size, it is inserted in the fermenter ...

Embodiment 2

[0049] Change the emulsification temperature in Example 1 to 50°C, and the rest of the operations are the same as in Example 2.

[0050] Test results: the conversion rate of phytosterol was 89.6%, the final potency was 13.44g / L, and the purity of the obtained 4-androstenedione was greater than 99.0%.

Embodiment 3

[0052] Change the emulsification temperature in Example 1 to 70°C, and the rest of the operations are the same as in Example 2.

[0053] Test results: the conversion rate of phytosterol was 84.3%, the final titer was 12.65g / L, and the purity of the obtained 4-androstenedione was greater than 99.0%. Comprehensive analysis of Examples 1-3 shows that the most preferred emulsification temperature is 60°C.

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Abstract

The invention provides a method of preparing 4-androstenedione by fermentatively degrading phytosterol by virtue of a single water phase system. The method comprises the following step: 1, strain cultivation, namely inoculating a mycobacterium smegmatis stain into a seed medium and cultivating; 2, sterol emulsification, namely emulsifying the phytosterol as a substrate under the action of a surfactant; 3, fermentation cultivation, namely inoculating the mycobacterium smegmatis stain obtained in the first step in a fermentation medium, adding the phytosterol obtained in the second step and carrying out fermentation cultivation; 4, finished product extraction. According to the method of preparing the 4-androstenedione by fermentatively degrading the phytosterol by virtue of the single water phase system, an emulsifier is added, so that the yield of 4-androstenedione is increased, and the extraction technology of the 4-androstenedione is also simplified; the cost is low, the pollution is little, and the environmental protection pressure is weak.

Description

technical field [0001] The invention relates to the technical field of microbial transformation of steroid medicines, in particular to a method for preparing 4-androstenedione by fermenting and degrading phytosterols in a single aqueous phase system. Background technique [0002] Androst-4-ene-3,17-dione, English name: androst-4-ene-3,17-dione, referred to as 4-androstenedione, is an irreplaceable intermediate of steroid hormone drugs, At present, most steroid hormone products are produced with 4-androstenedione as raw material. [0003] There are two main production processes for 4-androstenedione at present: one is to extract diosgenin from wild Chinese herbal medicines such as Diandrah and turmeric, and then prepare 4-androstenedione through chemical synthesis; the other is It uses microorganisms to convert phytosterols to produce 4-androstenedione. The former has high process cost, heavy pollution, and long production cycle; the latter is simple to operate and short in...

Claims

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Application Information

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IPC IPC(8): C12P33/02C12R1/34
Inventor 宋浩雷张敏然刘倩
Owner HEBEI ZHONGSHENG BIOTECH
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