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Preparation method of triketone dehydrogenation product

A triketone and dehydrogenation technology, applied in the field of preparation of triketone dehydrogenates, can solve the problems of poor substrate solubility, low conversion rate, long conversion period and the like

Pending Publication Date: 2022-04-12
HUNAN NORCHEM PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The dehydrogenation of triketones is currently mainly accomplished by biotransformation, but due to the poor solubility of the substrate and the difficulty in improving the activity of microorganisms (enzymes), the conversion period is long, the conversion rate is low, separation and purification are difficult, and the yield is difficult to increase.

Method used

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  • Preparation method of triketone dehydrogenation product
  • Preparation method of triketone dehydrogenation product
  • Preparation method of triketone dehydrogenation product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Simply Pylori Seed Culture

[0028] Slap medium: 2% glucose, 2% yeast paste 2%, pH 7.5, agar 2%, 20 min sterilized for 30 min, and inoculate simple node bacteria (ARTHROBACTER SIMPLEX CPCC 140451). Culture Conditions: 25-32 ° C, 1-6 days.

[0029] The first-class seed medium: 2% glucose, 2%, protein 胨 1%, yeast paste 0.5%, potassium phosphate 0.5%, pH 7.0-8.0.121 ° C, sterilization for 30 min. Simply node bacillus under the sterile conditions, culture conditions: 500 ml shake bottled 100ml seed medium, 30-32 ° C, speed of 200 rpm, and culture time 48 h.

[0030] Secondary seed medium: 2% glucose, 2%, protein 胨 1%, yeast paste 0.5%, 5% phosphoric acid dihydrogen, pH 7.0-8.0.121 ° C, sterilized for 30 min. Under aseptic conditions, a seed seed was allowed to inoculate 20%, cultural conditions: 500 ml shake bottled 100ml seed medium, 30 ° C, speed of 200 rpm, and culture time 48 h.

[0031] Fermented tank seed medium: 2% glucose, 2%, protein 胨 1%, yeast paste 0.5%, 0...

Embodiment 2

[0059] According to Example 1, a shake bottle seed culture was carried out, and the primary seed liquid was allowed to enter 100 mL of secondary seed medium for secondary seed culture. After 48 h, 3 g of triketone was added to the cultured secondary seed, 0.05 g Tween-80, 0.5 g Polyether Following agent PPE, 0.5GPMS, conversion conditions: 30 ° C, 200 rpm, conversion time 64 h, after conversion, the translation of HPLC, triketone dehydrate conversion of 88.39%, trinone transformation 9.22%.

Embodiment 3

[0061] According to Example 1, a shake bottle seed culture was carried out, and the primary seed liquid was allowed to enter 100 mL of secondary seed medium for secondary seed culture. After 48 h, 3 g of triketone was added to the cultured secondary seed, 0.05 g Tween-80, 0.5GPMS, 1G polyether ablying agent PPE, conversion conditions: 30 ° C, 200 rpm, conversion time 64h, after conversion, the translation of HPLC, triketone dehydrate conversion is 89.70%, triketone conversion 7.965%.

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Abstract

The invention belongs to the technical field of steroid hormone preparation, and particularly relates to a preparation method of a triketone dehydrogenation product, which comprises the following steps: mixing a microorganism containing 3-ketosteroid-1-dehydrogenase with a raw material triketone, adding a surfactant, a polyether defoamer PPE and an exogenous electron acceptor at the same time, not adding an organic solvent, carrying out microbial fermentation, and after the fermentation is completed, carrying out solid-liquid separation to obtain the 3-ketosteroid-1-dehydrogenase. The reaction route is as follows: the conversion rate of the product is effectively improved, and the yield is improved.

Description

Technical field [0001] The present invention belongs to the technical field of steroid hormone preparation, and is, in particular, to the preparation method of a triketone dehydrot. Background technique [0002] At present, the common method of steroidal drug production is the use of chemical synthesis and microbial transformation, and the key steps are generally achieved by microbial conversion, including A ring C1,2-bit dehydrogenation. The transformation of microbes on steroids is based on a microbial body, and an enzyme that acts on steroid C1, 2-bit dehydrogenation is called 3-steroid-1-dehydrogenase (KSDD) Also known as steroid C1, 2-bit dehydrogenase. The enzyme is usually present in Nocardia Sp.), Rhococcoccus Sp.), Bacillus Sp.), Mycobacterum SP.) And Pylori (Arthrobacter SP) .) Other microorganisms. Factors affecting steroidal biological dehydrogenation conversion are in addition to the solubility of the substrate, the maximum influencing factor is the activity of the t...

Claims

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Application Information

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IPC IPC(8): C12P33/02C12N1/20C12N1/38C12R1/06
Inventor 赵小娟刘喜荣曾春玲孟浩王雅茹叶豪宇
Owner HUNAN NORCHEM PHARMACEUTICAL CO LTD
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