Method for synthesizing 7alpha-hydroxy-androstenone by efficient utilization of colletotrichum lini

A technology of Trichosporium flax, -OH-AD, applied in the biological field, can solve the problems of low substrate conversion rate, single product, long conversion cycle, etc.

Active Publication Date: 2015-05-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the main species that have the ability to transform AD are mycobacteria ( Mycobacterium ), which not only has a long conversion cycle, low substrate conversion rate, but also easily forms other by-products
In addition, the hydroxylation products of AD are single, only the hydroxylation of 9α, 15α, and 14α has been reported, and there is no research on the hydroxylation of 7α

Method used

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  • Method for synthesizing 7alpha-hydroxy-androstenone by efficient utilization of colletotrichum lini

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1 prepares Spirospora flaxensis ( Colletotrichum lini ) Liquid culture of cells from the ST-1 strain

[0015] (1) Pick a loop of the Neptonia flaxensis strain on the solid PDA medium, inoculate it in a 250 mL Erlenmeyer flask containing 30 mL of seed medium, and place it on a shaker at 200 r / h at 30 °C. Min rotation speed culture 20-24 h to the logarithmic phase, that is prepared C. lini Liquid culture of ST-1 cells.

[0016] (2) The composition and ratio of the seed medium are: peanut cake powder 10-20 g / L, starch 15-30 g / L; glucose 20-50 g / L; yeast powder 10-30 g / L; NH 4 NO 3 5-10 g / L; KCl 0.1-1 g / L; K 2 HPO 4 0.1-1.0 g / L; MgSO 4 ·7H 2 O 0.01-0.1 g / L; FeSO 4 7H 2 O 0.01-0.1 g / L g / L; adjust the pH of the medium to 6.0-6.5 before sterilization, and sterilize under high-pressure steam at 121°C for 20 minutes.

Embodiment 2

[0017] Embodiment 2 Spirospora flaxensis ( Colletotrichum lini ) Shake flask fermentation of the ST-1 strain

[0018] (1) Put the above C. lini The cell liquid culture of ST-1 was inoculated in a sterilized 250 mL Erlenmeyer flask containing 30 mL of fermentation medium at an inoculum size of 8-10% (w / w), at 20-40°C placed on a shaker at a rotational speed of 120-200 r / min, fermented for 20-24 h, and the bacteria entered the logarithmic growth phase.

[0019] (2) The composition and ratio of the fermentation medium are: glucose 10-100 g / L; starch 10-30 g / L; yeast powder 5-50 g / L; peptone 5-50 g / L; NaCl 0.2-2 g / L;K 2 HPO 4 0.1-1.0 g / L; KH 2 PO 4 0.1-1.0 g / L; MgSO 4 0.01-0.1 g / L; FeSO 4 7H 2 O 0.01-0.10 g / L; pH 6.5-7.5, sterilized under high-pressure steam at 121°C for 20 min.

[0020] Embodiment 2 Spirospora flaxensis ( Colletotrichum lini ) ST-1 strain transformed into AD

[0021] Accurately weigh an appropriate amount of AD, put it into the bacterial fermenta...

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Abstract

The invention belongs to the biotechnology field, and relates to a method for synthesizing novel 7alpha-hydroxy-androstenone (7alpha-OH-AD) steroid by the efficient utilization of colletotrichum lini ST-1 hydroxylation androstane-4-alkene-3, 17-diketone (AD). The method has the advantages that the conversion rate reaches up to 90% by adopting a two-step conversion method under the condition that the feeding amount of a substrate is 10g / L, and the total yield of the 7alpha-OH-AD product is 79.32%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for efficiently utilizing Spirospora flaxensis ( Colletotrichum lini ) ST-1 Hydroxylation of androst-4-ene-3,17-dione (AD) to 7α-hydroxy-androstenone (7α-OH-AD). Background technique [0002] Androst-4-ene-3,17-dione (AD) is the product of phytosterols after the side chains are excised by microorganisms. Due to the structural characteristics of the AD mother nucleus, it is known as the "mother of hormones and medicinal gold" in the field of steroid drug synthesis, with a global annual output value of nearly 10 billion US dollars. Using biotransformation technology, one or more hydroxyl groups are introduced into different sites of the AD mother nucleus to form a variety of steroidal drug prodrugs, which are widely used in the synthesis of anabolic hormones, androgen, spironolactone, and can also be used Used in the synthesis of more than 100 kinds of drugs such...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/06C12R1/645
Inventor 许正宏吴燕史劲松李会李恒张旦旦许鸿瑜
Owner JIANGNAN UNIV
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