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Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes

An -OH-ADD, mixed bacteria fermentation technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms and other directions, can solve the problems of reducing product yield, increasing post-processing costs and fermentation energy consumption, etc. Improve conversion rate, reduce production cost and complete conversion effect

Active Publication Date: 2012-12-19
HUNAN LONGTENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Preparation of 11α-OH-ADD from 4-AD, the prior art is carried out in two steps, namely C 1,2 dehydrogenation and C 11 Hydroxylation reaction requires post-treatment after the previous step reaction is completed, which increases product post-processing cost and fermentation energy consumption, and reduces product yield

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Slant and seed cultivation methods are as described in the summary of the invention. In 1L of water, the fermentation substrate 4-AD is 20g, the components and weight of the fermentation medium are methanol 20g, TW80 (Tween 80) 2g, yeast extract 10g, and molasses 10g , sodium nitrate 5g, disodium hydrogen phosphate 2g, sodium dihydrogen phosphate 2g, the pH value is 7.0. Ochra Aspergillus ochraceus ATCC 18500 The inoculum size was 20%, Arthrobacter Arthrobacter simplex ATCC 6946 The inoculum amount was 10%. Use a 50-liter tank for conversion, sample volume 70%, tank pressure 0.05 MPa, air flow: 0.5 VVM, temperature: 30°C, stirring: 150 rpm, conversion for 50 hours, sampling TLC analysis, the conversion is basically complete, the sample is washed with ethyl acetate Ester (EA) extraction, the EA layer was sent to HPLC, the content of 11α-OH-ADD was 96.1%, the content of 4-AD was 0.5%, the content of ADD was 0.7%, and the content of 11α-OH-4-AD was 1.2%.

[0032] Afte...

Embodiment 2

[0034] Incline, seed culture method as described in the summary of the invention, in 1L of water, fermentation substrate 4-AD 19g, the components and weight of the fermentation medium are methanol 21g, TW80 2.1g, yeast powder 10.5g, molasses 9.5g, nitric acid Potassium 4.75g, dipotassium hydrogen phosphate 1.9g, potassium dihydrogen phosphate 1.9g; pH value is 6.8. Ochra Aspergillus ochraceus ATCC 18500 The inoculum size was 19.5%, Arthrobacter Arthrobacter simplex ATCC 6946The inoculum amount was 10.5%. Use a 50-liter tank for conversion, sample volume 70%, tank pressure 0.055MPa, air flow: 0.45 VVM, temperature: 30°C, stirring: 150 rpm, conversion for 48 hours, sampling TLC analysis, the conversion is basically complete, and the sample is extracted with EA , send the EA layer to HPLC, the content of 11α-OH-ADD is 95.6%, the content of 4-AD is 0.6%, the content of ADD is 1.0%, and the content of 11α-OH-4-AD is 0.9%.

[0035] After the conversion is complete, inactivate a...

Embodiment 3

[0037] Incline, seed cultivation method as described in the summary of the invention, in 1L of water, the fermentation substrate 4-AD 21g, the components and weight of the fermentation medium are methanol 19g, TW80 1.9g, yeast powder 9.5g, molasses 10.5g, nitric acid Potassium 5.25g, dipotassium hydrogen phosphate 2.1g, potassium dihydrogen phosphate 2.1g; pH value is 7.2. Ochra Aspergillus ochraceus ATCC 18500 The inoculum size was 19%, Arthrobacter Arthrobacter simplex ATCC 6946 The inoculum amount was 10%. Use a 50-liter tank for conversion, sample volume 70%, tank pressure 0.045 MPa, air flow: 0.55VVM, temperature: 30°C, stirring: 150 rpm, conversion for 48 hours, sampling TLC analysis, the conversion is almost complete, and the sample is extracted with EA , send the EA layer to HPLC, the content of 11α-OH-ADD is 97.11%, the content of 4-AD is 0.23%, the content of ADD is 0.33%, and the content of 11α-OH-4-AD is 1.52%.

[0038] After the conversion is completed, inact...

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Abstract

The invention relates to a production method of steroids medicines, and concretely relates to a method for conversing 4-AD to produce 11 alpha-OH-ADD by mixed fermentation of two microbes. According to the invention, 4-AD is taken as a raw material, and the mixed fermentation is carried out through aspergillus ochraceus ATCC18500 and Arthrobacter simplex ATCC6946. Compared with hydroxylation and dehydrogenation reaction by individually using the two above bacteria, 11 alpha-OH-ADD can be obtained through mixed fermentation of 4-AD, two above bacteria have cooperative effect on the fermentation and conversion, The method has the following advantages: the overall yield is high, the conversion is more complete, a step of post-treatment is reduced, and the energy consumption is reduced. The mixed fermentation is capable of greatly reducing the production cost, increasing the conversion rate, reducing the conversion time, and relatively reducing the post-treatment and the energy consumption.

Description

technical field [0001] The invention relates to a production method of steroid drugs, in particular to a method for preparing 11α-OH-ADD by fermenting 4-AD with mixed bacteria. Background technique [0002] Steroidal drugs have anti-inflammatory, contraceptive, diuretic, anti-toxic, anti-allergic, and anti-shock effects, and are widely used in eczema, asthma, and rheumatic diseases. In recent years, the scope of application of steroidal drugs in the medical field has been continuously expanded, and they are widely used in the treatment of rheumatism, cardiovascular disease, collagenous diseases, lymphatic leukemia, human organ transplantation, anti-tumor, bacterial encephalitis, skin diseases, endocrine disorders, Senile diseases, etc. Steroid drugs also have a certain effect on certain cancers, especially hormone-dependent tumors. Steroid drugs have a wide range of actions and high yields, and are the second largest class of drugs in the pharmaceutical industry after antib...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P33/08C12R1/66C12R1/06
Inventor 刘喜荣孟浩
Owner HUNAN LONGTENG BIOTECH
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