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Double liquid phase fermentation method for degrading plant sterol and preparing androstenedione

A technology of phytosterol and androstenedione, which is applied in the biological field, can solve the problems of insoluble raw materials, etc., and achieve the effects of increasing fermentation concentration, reducing use, and increasing feeding amount

Inactive Publication Date: 2013-08-21
北京明新高科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is to overcome the problem of insoluble raw materials in the fermentation process, and design a method to improve the utilization rate of strains, increase the conversion rate of phytosterols, and improve the extraction rate of products

Method used

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  • Double liquid phase fermentation method for degrading plant sterol and preparing androstenedione

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Strain: Mycobacterium sp.DE6823

[0023] Strain intake: 10%

[0024] Fermentation medium: corn starch 10%, NH 4 NO 3 0.1%, K 2 HPO 3 0.1%, aviation kerosene 0.2%, defoamer 0.002%, the concentration of sunflower oil is 20g / L, and the concentration of phytosterol is 22g / L.

[0025] Fermentation tank volume: 10L, added according to 50% fermentation broth.

[0026] Stirring speed: 160r / min

[0027] Fermentation time: 8d

[0028] pH range: 8.2-8.4

[0029] After the fermentation is completed, filter and remove the solids in the fermentation broth, add 5g / L of K 2 CO 3 , stirring, standing still, the two phases are separated, take the oil layer, add ethanol, stir fully, pass through the membrane device with polypropylene membrane, the ethanol phase will flow out quickly, and the oil phase will be intercepted by the membrane due to its large viscosity and molecular weight. live. Concentrate the ethanol solution containing AD at a volume ratio of 10:1, add 5% activ...

Embodiment 2

[0031] Bacterial strain: with embodiment 1

[0032] Strain intake: 8%

[0033] Fermentation medium: corn starch 8%, NH 4 NO 3 0.2%, K 2 HPO 3 0.1%, aviation kerosene 0.3%, defoamer 0.005%, the concentration of soybean oil is 20g / L, and the concentration of phytosterol is 22g / L.

[0034] Fermentation tank volume: 100L, added according to 50% fermentation liquid.

[0035] Stirring speed: 300r / min

[0036] Fermentation time: 9d

[0037] pH range: 8.2~8.4

[0038] After the fermentation is completed, filter and remove the solid in the fermented liquid, add 8g / L Na 2 CO 3, stirring, standing still, the two phases are separated, take the oil layer, add acetone, stir well, pass through the membrane device with polypropylene membrane, the acetone phase will flow out quickly, and the oil phase will be intercepted by the membrane due to its large viscosity and molecules live. Concentrate the acetone solution containing AD at a volume ratio of 10:1, add 5% activated carbon t...

Embodiment 3

[0040] Bacterial strain: with embodiment 1

[0041] Strain intake: 12%

[0042] Fermentation Medium: Corn Starch 12%, NH 4 NO 3 0.1%, K 2 HPO 3 0.2%, aviation kerosene 0.5%, defoamer 0.002%, the concentration of rice bran oil is 26g / L, and the concentration of phytosterol is 22g / L. Fermentation tank volume: 100L, added according to 50% fermentation broth.

[0043] Stirring speed: 250r / min

[0044] Fermentation time: 9d

[0045] pH range: 8.2~8.4

[0046] After the fermentation is completed, filter and remove the solid in the fermented liquid, add 8g / L of K 2 CO 3 , stirring, standing still, two-phase separation, take the oil layer, add isobutanol, stir well, pass through the membrane device with polypropylene membrane, the isobutanol phase will flow out quickly, and the oil phase will flow out due to the viscosity and larger molecules , will be intercepted by the membrane. Concentrate the isobutanol solution containing AD at a volume ratio of 10:1, add 5% activate...

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Abstract

The invention provides a method for preparing androstenedione using a double liquid phase fermentation system by fermenting Mycobacterium sp.DE6823, and aseptic distilled water is selected as a fermentation medium water phase, and rice oil, sunflower oil, soybean oil and peanut oil can be selected as an oil phase, after fermentation conversion, a solution containing androstenedione is obtained, and after condensation, decolouring, filtering, condensation, recrystallization and vacuum drying and other operations, and androstenedione whose purity is above 99% is obtained. Different oil phases can be selected, and the conversion rate of the plant sterol is in the range of 59%-85%. The invention solves the problems that the raw materials of the present androstenedione production technology in our country are limited by season and region, thereby effectively reducing destroy to ecological environment; the invention has the characteristics of simple operation, small environmental pollution, high conversion rate, low production cost, etc., and is suitable for industrialized production.

Description

1. Technical field [0001] The invention discloses a method for preparing androstenedione by degrading phytosterols by a fermentation method, which belongs to the field of biotechnology and relates to fermentation and degradation by using a two-liquid phase fermentation system. 2. Background technology [0002] The chemical structural formula of androstenedione is as follows: [0003] [0004] 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 a very important role in the body. It can be said that almost all steroid hormone drugs are produced with AD as the starting material, which can be used to produce sex hormones, progestins, anabolic hormones and corticosteroids, and can be used to synthesize hydrocortisone, Prednisone oxide, progesterone, estrenol, dexamethasone and more than 100 kinds of drugs can also be directly used in the production of anti-early pregna...

Claims

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Application Information

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IPC IPC(8): C12P33/02C12R1/32
Inventor 杨旭锦石慧
Owner 北京明新高科技发展有限公司
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