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Mycobacterium and application thereof

A technology of mycobacteria and classification and naming, applied in the field of microorganisms, can solve the problems that 9α-hydroxycanrenone has not been found in literature reports, and achieve the effect of good industrialization prospects

Active Publication Date: 2019-12-13
JIANGSU JIAERKE PHARMA GRP CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]However, there is no literature report on the conversion of canrenone into 9α-hydroxycanrenone by microbial transformation technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1）

[0033] This example is the construction of mycobacterium JYC-07 with high production of 3-sterone-9α hydroxylase.

[0034] S1. Screening of Mycobacterium JYC.

[0035] S11. Take 5g of activated sludge from the sewage treatment system of Jiangsu Jiaerke Pharmaceutical Group Co., Ltd., add it to 45mL of sterilized liquid enrichment medium, and then shake it on an air bath shaker at a speed of 180rpm and a temperature of 35°C. The enrichment culture was carried out for 96 hours to obtain the enrichment solution.

[0036] The formula of the above liquid enrichment medium is as follows: ammonium sulfate 0.6g / 100mL, sodium citrate 0.5g / 100mL, magnesium sulfate 0.05g / 100mL, potassium dihydrogen phosphate 0.02g / 100mL, disodium hydrogen phosphate 0.03g / 100mL, Tween-80 0.05g / 100mL, canrenone 0.15g / 100mL, pH 7.0.

[0037] After gradient dilution of the above-mentioned enrichment solution, spread it evenly on a solid enrichment medium plate, culture it in a constant temperature incubato...

Embodiment 1）

[0060] This embodiment is a preparation method of 9α-hydroxycanrenone, which has the following steps:

[0061] ①Seed cultivation.

[0062] The mycobacterium JYC-07 was inoculated from the eggplant bottle into 20 mL of seed medium with an inoculation loop, and the seed culture was carried out at a rotation speed of 180 rpm and a temperature of 30° C. for 36 hours to obtain a seed culture solution.

[0063] The composition of the seed medium in this embodiment is as follows: glucose 1.8g / 100mL, peptone 1.0g / 100mL, corn steep liquor 2.0g / 100mL, the balance is water; initial pH value is 7.2; sterilization.

[0064] ② Fermentation culture.

[0065] Inoculate 5 mL of the seed culture solution obtained in step ① into 95 mL of fermentation medium, that is, the inoculum size is 5% (V / V), and carry out fermentation at a speed of 200 rpm and a temperature of 30°C for 48 hours to obtain a fermentation liquid.

[0066] The fermentation medium composition of the present embodiment is as f...

Embodiment 2）

[0073] This embodiment is a preparation method of 9α-hydroxycanrenone, which has the following steps:

[0074] ①Seed cultivation.

[0075] The mycobacterium JYC-07 was inoculated from the eggplant bottle into 20 mL of seed medium with an inoculation loop, and the seed culture was carried out at a rotation speed of 200 rpm and a temperature of 32° C. for 48 hours to obtain a seed culture solution.

[0076] The composition of the seed medium in this embodiment is as follows: glucose 1.5g / 100mL, yeast extract 1.2g / 100mL, corn steep liquor 2.0g / 100mL, the balance is water; initial pH value is 7.0; sterilization.

[0077] ② Fermentation culture.

[0078] Inoculate 10 mL of the seed culture liquid obtained in step ① into 90 mL of fermentation medium, that is, the inoculation amount is 10% (V / V), and carry out fermentation at a speed of 220 rpm and a temperature of 30°C for 48 hours to obtain a fermentation liquid.

[0079] The fermentation medium composition of the present embodim...

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PUM

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Abstract

The invention discloses a mycobacterium and application thereof. The mycobacterium is classified and named as mycobacterium JYC-07, and is preserved in China Center for Type Culture Collection on June9th, 2017, and the preservation number is CCTCC NO : M 2017325. The strain Mycobacterium sp.JYC capable of converting canrenone into 9alpha-hydroxycarvenone by microorganisms is obtained by screeningand separating from a natural environment, the strain Mycobacterium sp.JYC-07 capable of producing 3-sterone-9alpha hydroxylase at high yield is further obtained through genetic engineering, the strain can efficiently convert canrenone into 9alpha-hydroxycarvenone, the conversion rate is over 90 percent, and the strain has very important significance for expanding the application of steroid raw material drug canrenone and good industrial prospects.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a mycobacterium and its application. Background technique [0002] As the second largest class of drugs in the world after antibiotics, steroid drugs play a very important role in regulating the body. Different steroid drug molecular structures are derived from steroid drug intermediates. 9α-hydroxycanrenone is an important intermediate in the synthesis of steroid drugs and plays an important role in the industrial production of steroids. Using 9α-hydroxycanrenone as a precursor can synthesize hydrocortisone, hydrocortisone acetate, cortisone acetate, cortisone, prednisolone, prednisolone acetate, prednisone, and ipre Ketone, Betamethasone, Dexamethasone, Triamcinolone Acetonide, Triamcinolone Acetate Acetate, Budesonide, Desonide, Ciclesonide and other corticosteroid steroid raw material products with important commercial value . Therefore, it is imperative...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P33/20C12R1/32
CPCC12N1/20C12P33/20C12R2001/32C12N1/205
Inventor 顾向忠蒋澄宇
Owner JIANGSU JIAERKE PHARMA GRP CORP
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