A biotransformation method for the 19-position hydroxylation of a series of steroid compounds

A biotransformation and compound technology, which is applied in the fields of chemistry and biology, can solve the problems of poor selectivity and low yield, and achieve the effects of mild fermentation reaction conditions, high yield, and simple preparation process

Active Publication Date: 2020-12-01
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of the present invention can be used for efficient hydroxylation of 19-position sterol, can efficiently obtain a series of sterol hydroxylation products, solves the problems of low yield and poor selectivity of steroidal compounds at the 19-position hydroxylation product, and has high industrial application value

Method used

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  • A biotransformation method for the 19-position hydroxylation of a series of steroid compounds
  • A biotransformation method for the 19-position hydroxylation of a series of steroid compounds
  • A biotransformation method for the 19-position hydroxylation of a series of steroid compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022]

[0023] Culture condition 1 (solid culture): Inoculate the strain Thanatephorus cucumeris NBRC 6298 (T. cucumeris NBRC6298) on PDA medium (potato 200g / L, glucose 20g / L and agar 15g / L) and activate it in a constant temperature incubator at 30°C to cultivate. After 6-10 days, inoculate the well-growth strains on the plate into a 250mL Erlenmeyer flask containing 100mL yeast liquid medium (glucose 25g / L, yeast extract 20g / L), and grow for about 2-3 days at 30°C and 200rpm 25mg Cortexolone (Cortexolone) was dropped into the thallus culture medium, continued to cultivate and transformed for about 3-4 days until the conversion rate reached the highest, and obtained the compound 19-OH-cortexolone 19-OH-Cortexolone (transformation rate 27%).

[0024] Culture condition 2 (liquid culture): the strain T. cucumeris NBRC 6298 was inoculated on PDA medium for activation culture in a constant temperature incubator at 30°C. After 6-10 days, inoculate the well-growth strains on th...

Embodiment 2

[0027]

[0028]The strain T.cucumeris NBRC 6298 was inoculated on PDA medium for activation culture in a constant temperature incubator at 30°C. After 6-10 days, inoculate the well-growth strains on the plate into a 250mL Erlenmeyer flask containing 100mL yeast liquid medium (glucose 25g / L, yeast extract 20g / L), and grow for about 2-3 days at 30°C and 200rpm , get 5 mL of well-grown bacterium solution and transfer it in a 250 mL Erlenmeyer flask containing 100 mL of fresh yeast liquid medium, continue to cultivate under the same conditions for 2-3 days, and 25 mg of 17-acetyl-cortodopine (17- acetyl-Cortexolone) and ferrous ammonium sulfate were dropped into the thallus culture medium (iron ion final concentration 1.5mmol / L), continued to cultivate and transformed for about 3-4 days until the conversion rate reached the highest, and obtained compound 19-OH-Cortexolone ( 81% conversion).

Embodiment 3

[0030]

[0031] The strain T.cucumeris NBRC 6298 was inoculated on PDA medium for activation culture in a constant temperature incubator at 30°C. After 6-10 days, inoculate the well-growth strains on the plate into a 250mL Erlenmeyer flask containing 100mL yeast liquid medium (glucose 25g / L, yeast extract 20g / L), and grow for about 2-3 days at 30°C and 200rpm , get 5 mL of well-grown bacterium solution and transfer it to a 250 mL Erlenmeyer flask containing 100 mL of fresh yeast liquid medium, continue to cultivate under the same conditions for 2-3 days, and 25 mg of 21-acetyl-cortodopine (21- acetyl-Cortexolone) and ferrous ammonium sulfate were dropped into the thallus culture medium (iron ion final concentration 1.5mmol / L), continued to cultivate and transformed for about 3-4 days until the conversion rate reached the highest, and obtained compound 19-OH-Cortexolone ( Conversion rate 30%).

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Abstract

The invention discloses a bioconversion method for series steroid compound 19-position hydroxylation and belongs to the field of chemistry and biology. The method comprises the steps as follows: a steroid compound and iron salt are added to a liquid culture medium containing bioconversion strains for culture, wherein the bioconversion strains contain a strain with a steroid compound 19-position hydroxylation function and a heterologous expression strain containing a steroid compound 19-position hydroxylation gene; and the steroid compound contains cortodoxone, 17-position and / or 21-position hydroxyl protected cortodoxone as well as other modification of steroid frameworks on the basis of the defined compounds. The method can be used for efficient steroid 19-position hydroxylation to obtainseries steroid hydroxylated products. The method solves the problems of low yield and poor selectivity of 19-posiiton hydroxylated products of the steroid compounds, used raw materials are cheap andavailable, the reaction condition is mild, the substrate universality is good, the yield is high, the preparation process is simple, and the method has very high industrial application values.

Description

technical field [0001] The invention relates to the fields of chemistry and biology, and more specifically relates to a biotransformation method for the 19-position hydroxylation of a series of steroid compounds. Background technique [0002] Steroids are also known as steroids. Although there are many types of these compounds, they occupy an important position clinically ([1]Fernandes P., Cruz A., Angelova B., Pinheiro H.M., Cabral J.M.S. Enzyme MicrobTechnol.2003,32,688-705 .[2]Tong W.Y., Dong X.Recent Pat.Biotechnol.,2009,3,141-153.[3]Donova M.V.,Egorova O.V.Appl.Microbiol.Biotechnol.,2012,94,1423-1447.[4]Bhatti H.N. ,Khera R.A.Steroids,2012,77,1267-1290), according to their functions can be roughly divided into sex hormones, corticosteroids and anabolic hormones. At present, there are more than 300 kinds of steroid drugs in clinical use, accounting for about 15% of all clinically used drugs. [0003] The basic skeleton of a steroid consists of four fused carbocycles (A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/12C12R1/645
Inventor 瞿旭东周强辉张亚楠王军林
Owner WUHAN UNIV
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