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Preparation method of theelin by microbial conversion

A technology for microbial transformation and estrone, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of troublesome separation and purification, low feeding concentration and high production cost, and achieves easy separation and purification, feeding concentration. High, raw material cost reduction effect

Inactive Publication Date: 2008-03-26
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to solve the problems existing in the above-mentioned technology, overcome the disadvantages of low feed concentration, troublesome separation and purification, and high production cost for the preparation of estrone by the existing microbial method, and provide a method with simple operation and a production environment that meets environmental protection requirements. At the same time, the process of producing estrone by microbial conversion method with high feeding concentration, high conversion rate, high yield and low cost, and the product is single, easy to separate and purify

Method used

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  • Preparation method of theelin by microbial conversion
  • Preparation method of theelin by microbial conversion
  • Preparation method of theelin by microbial conversion

Examples

Experimental program
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Effect test

Embodiment 1

[0040] The production strain is Arthrobacter simplex AS1.94 * ), the production strain was cultured at a constant temperature of 30°C on an agar slant for 2 days, the thallus had grown completely on the slant, and was light milky yellow in appearance, plump and shiny; most of the thallus were in the shape of Rod-shaped, some of which are angled. Store at 4°C for 5 days and use. Wherein, the liquid used for making the bacterial seed suspension is fresh seed culture medium, and the sterilization condition is: 121° C. for 20 minutes.

[0041] The seed medium consists of carbon source, nitrogen source and inorganic phosphorus, namely:

[0042] Glucose 8g / L

[0043] Corn syrup 10g / L

[0044] K H 2 PO 4 1.5g / L

[0045] pH 7.0

[0046] The prepared seed suspension was added to the above seed medium, and cultured at 32° C. for 18 hours at a shaking table with a rotation speed of 150 r / min to obtain a seed culture suitable for inoculation.

[0047] Fermentation medium comp...

Embodiment 2

[0056] The bacterial strain was cultured on an agar slant for 1 day, and the liquid used for making the bacterial seed suspension was sterile water.

[0057] For primary seed culture, culture at 30°C for 22 hours at a shaker speed of 170r / min to obtain a seed culture suitable for inoculation;

[0058] During secondary seed cultivation, insert 120mL of well-grown seed culture solution into 4L of the above-mentioned fermentation medium; control the dissolved oxygen level in the fermentation solution at 40% during the fermentation process, and keep the pH value at 7.0-7.2 during the fermentation process Cultivate for 20 hours to obtain a bacterial culture solution suitable for feeding.

[0059] Substrate (I, where R 1 =HCO-), that is, 19-formyloxy-3,17-diketone-pregn-4-ene is pulverized, with a particle size of 14-16μ, and is quickly put into the cultured fermentation broth under sterile conditions. It is 72g, and 0.8g polyether defoamer is added. During the transformation pro...

Embodiment 3

[0062] The strain was cultured on the agar slant for 2 days, and the liquid used to make the bacterial seed suspension was sterile water.

[0063] For primary seed culture, culture at 32°C for 20 hours at a shaker speed of 180r / min to obtain a seed culture suitable for inoculation;

[0064] During secondary seed cultivation, insert 200mL of well-grown seed culture solution into 4L of the above-mentioned fermentation medium; control the dissolved oxygen level in the fermentation solution at 40% during the fermentation process, and keep the pH value at 7.0-7.2 during the fermentation process Cultivate for 22 hours to obtain a bacterial culture solution suitable for feeding.

[0065] Substrate (I, where R 1 =H) That is, 19-hydroxyl-3,17-dione-pregn-4-ene is pulverized, with a particle size of 14-16 μ, and is quickly put into the cultured fermentation broth under aseptic conditions, with a feeding amount of 80 g, and added 1.2g polyether defoamer. During the transformation proc...

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Abstract

A process of preparing phenolic ketone by way of micro-conversion. Strain is simple bacillus. Substrate is steroids (1 and 2). The process contains preparation of suspension, culture of seed solution, after-fermentation, substrate conversion and separation of product. Inventory rating is 12-20g / L fermenting solution. Level of O2 is between 30% and 50%. Time is between 30 and 60h. Adopt acetone-water method so that the product agrees USP26.

Description

technical field [0001] The invention belongs to microorganism preparation technology, and relates to a method for preparing estrone by using Arthrobacter simplex as strain and steroid I or II as substrate by microbial transformation method. Background technique [0002] Estrone is an important intermediate in the synthesis of 19-desmethyl steroids and other estrogenic drugs (such as estradiol and estradiol benzoate, norethindrol, estriol, etc.). The key to the preparation of estrone is the 19-normethyl and A-ring aromatic rings. At present, chemical synthesis methods are mostly used at home and abroad, such as using diketene to remove the 19-methyl group at a high temperature of 600 ° C to aromatize the A ring. , the yield is only 20%; when diketene has C 6 When the double bond is used, the yield can be increased to 40%. Then, Tsuda et al. found a new method to aromatize the A ring of diketene, and the reaction conditions are relatively mild and simple, that is, in the pres...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C12N1/20C12R1/06
Inventor 杜连祥路福平王敏别松涛
Owner TIANJIN UNIV OF SCI & TECH
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