Preparation method of 15alpha hydroxylated levorotatory ethyl sterene diketone

A technology of ethyl sterenedione and hydroxylation, which is applied in the field of biocatalysis, can solve the problems of harmful effects, cytotoxicity, pollution, etc., and achieve the effect of sufficient source of raw materials, reduced use of organic solvents, and simple operation process

Inactive Publication Date: 2011-12-21
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that organic solvents can be used in steroid conversion as solubilizers to promote the effective dissolution of substrates and achieve the purpose of increasing conversion rates, but organic solvents are very toxic to cells and will pollute the environment , its harmful effects cannot be underestimated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] (1) Preparation of spore suspension: Inoculate Penicillium raistrickii (ATCC 10490) on the slant culture medium, wash the slant spores with sterile water (a Tween-80 aqueous solution with a content of 1‰) after cultivation , prepare the spore suspension;

[0037] (2) Mycelia culture: insert the prepared spore suspension into the fermentation medium, and the inoculum size is 10 8 ~10 11 each / ml, the culture temperature is 28-32°C, the culture time is 24-48 hours, and the spherical mycelium obtained after the culture is mature is obtained by suction filtration to obtain the fungus cake;

[0038] The biocatalyst used is the spherical mycelium of Penicillium raceae with a diameter of 0.9-3.0 mm for 18-72 hours of cultivation time; 5-8% of the total.

[0039](3) Biotransformation in the two-liquid phase system: put the bacteria cake obtained by suction filtration into the water phase of the ionic liquid / water two-phase system, the mass ratio of the bacteria cake to the wa...

Embodiment 1

[0049] A method for preparing 15α-hydroxylated L-ethylstenedione. In this example, a phosphate buffer solution with a pH of 5.8 is used as the water phase, and the ionic liquid phase is 1-butyl-3-methylimidazolium hexafluorophosphate. , that is [BMIM][PF6], V (ionic liquid phase): V (water) = 0.25: 1, the total liquid volume of the two-liquid phase system is 30ml, and the steps are:

[0050] (1) Preparation of spore suspension: Penicillium raistrickii (ATCC 10490) was inoculated on the slant medium, and the slant spores were inoculated with sterile water (a Tween-80 aqueous solution with a content of 1% by mass) after cultivation. After washing, the spore suspension is prepared;

[0051] (2) Mycelia culture: insert the prepared spore suspension into the fermentation medium, and the inoculum size is 10 9 each / ml, the culture temperature is 28°C, the culture time is 36h, and the spherical mycelium obtained after the culture is mature is obtained by suction filtration to obtain ...

Embodiment 2

[0058] A preparation method of 15α-hydroxylated L-ethylstenedione. In this embodiment, a phosphate buffer solution with a pH of 5.8 is used as the water phase, and the ionic liquid phase is 1-hexyl-3-methylimidazolium hexafluorophosphate , that is [HMIM][PF6], V (ionic liquid phase): V (water) = 0.5: 1, the total liquid volume of the two-liquid phase system is 30ml, and the steps are:

[0059] (1) Preparation of spore suspension: inoculate Penicillium raistrickii (ATCC 10490) on the slant culture medium, and use sterile water (a Tween-80 aqueous solution with a content of 1‰ (m / m)) after cultivation Wash the slant spores to prepare a spore suspension;

[0060] (2) Mycelia culture: insert the prepared spore suspension into the fermentation medium, and the inoculum size is 10 9 each / ml, the culture temperature is 28°C, the culture time is 36h, and the spherical mycelium obtained after the culture is mature is obtained by suction filtration to obtain the fungus cake;

[0061] (...

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PUM

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Abstract

The invention relates to a preparation method of 15alpha hydroxylated levorotatory ethyl sterene diketone. The preparation method comprises the following steps of: performing penicillium raistrickii (ATCC 10490) fermentation; and adding a substrate levorotatory ethyl sterene diketone to perform biotransformation. The preparation method is characterized in that: a biotransformation system consistsof an ionic liquid phase and a buffer liquid phase; and the volume ratio of the ionic liquid phase to the buffer liquid phase is (0.25-1.25):1. In an ionic liquid / water double-liquid phase system, hydroxylase contained in the penicillium raistrickii (ATCC 10490) thalli is used for catalyzing the 15alpha position of the levorotatory ethyl sterene diketone to generate hydroxylation reaction to generate the 15alpha hydroxyl levorotatory ethyl sterene diketone, wherein an organic solvent is not required to be added in the preparation process, so use of the organic solvent with severe damage effect on the environment is reduced, the reaction condition is mild, a transformation device is simple, sources of the raw materials are sufficient, the preparation process is simple, production cost is low, damage of environmental pollution is small, and production cost can be reduced.

Description

technical field [0001] The invention belongs to the field of biocatalysis, in particular to a preparation method of 15α hydroxylated L-ethylstenedione. Background technique [0002] As the population grows faster and faster, the emergence of steroidal contraceptives has brought hope for the effective control of excessive population growth. Gestodene is one of the main components of the new generation of contraceptives. It is based on the prototype and progesterone Receptor binding, with high affinity and almost no androgen and estrogen activity, rapid and complete oral absorption, is one of the most ideal oral contraceptives at present, the 15α hydroxylated L-ethyl sterene bis Ketone is a key intermediate in the synthesis of "gestodene". [0003] Generally, steroids have strong hydrophobicity (solubility range is 10 -5 ~10 -4 mol / L), while steroidal invertase is an intracellular enzyme, and the substrate can only contact the enzyme to carry out the conversion reaction by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/06C12R1/80
Inventor 毛淑红路福平胡晓杰刘莎李玉杜连祥
Owner TIANJIN UNIV OF SCI & TECH
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