Method for producing 11 alpha-hydroxycarvenone by efficient conversion of canrenone

A technology of hydroxycanrenone and canrenone, which is applied in the field of microbial preparation, can solve problems such as low conversion rate, low conversion rate, and restrictions on product application and promotion, and achieve reduced production costs, high conversion rate, and shortened conversion time Effect

Inactive Publication Date: 2013-08-21
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In 1996, Ng; John S. etc. used intermittent feeding. Although the amount of feed was greatly increased, the conversion time was prolonged and the conversion rate was not high, which greatly increased the cos

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The production strain is Aspergillus ochratum (AS, TCCC41060), which was cultured on a PDA slant at a constant temperature of 25°C for 6 days to produce yellow spores. Then transfer to YPDA slant, culture at 30°C for 5 days to produce golden yellow spores. Store in refrigerator at 4°C and use after 10 days. Add 10 mL of 2‰ (w / v) Tween 80 solution into the production slope, wash the spores, count and adjust the spore suspension concentration to 4×10 8 pcs / 100mL.

[0025] Seed medium (g / L): Glucose 20, soybean peptone 20, yeast extract 20, adjust the pH to 5.0-6.0, sterilize at 121°C for 20 minutes, add 1mL spore suspension to every 100mL seed medium, and place on a shaker Temperature 30°C, culture at 180r / min for 18h to obtain seed culture solution;

[0026] Fermentation medium (g / L): glucose 15, corn steep liquor 20, yeast extract 20, K 2 HPO 4 2. Adjust the pH value to 5.8, insert the well-grown seed culture solution into a 7L (4.5L liquid loading) fermenter with ...

Embodiment 2

[0028] The production strain is Aspergillus ochratum (AS, TCCC41060), which was cultured on a PDA slant at a constant temperature of 30°C for 3 days to produce yellow spores. Then it was transferred to YPDA slant and cultured at 28°C for 6 days to produce golden yellow spores. Store in refrigerator at 4°C and use after 10 days. Add 10 mL of 2‰ (w / v) Tween 80 solution into the production slope, wash the spores, count and adjust the spore suspension concentration to 6×10 8 pcs / 100mL.

[0029] Seed medium (g / L): Glucose 20, soybean peptone 20, yeast extract 20, adjust the pH to 5.0-6.0, sterilize at 121°C for 20 minutes, add 1mL spore suspension to every 100mL seed medium, and place on a shaker Temperature 28°C, 190r / min cultivation for 20h to obtain the seed culture solution;

[0030] Fermentation medium (g / L): glucose 15, corn steep liquor 20, yeast extract 20, K 2 HPO 4 2. Adjust the pH value to 5.8, and insert the well-grown seed culture solution into a 7L (liquid fillin...

Embodiment 3

[0032] The production strain is Ochra (AS, TCCC41060), which is cultured on a PDA slant at a constant temperature of 28°C for 4 days to produce yellow spores. Then transfer to YPDA slant and culture at 25°C for 5 days to produce golden yellow spores. Store in refrigerator at 4°C and use after 10 days. Add 10 mL of 2‰ (w / v) Tween 80 solution into the production slope, wash the spores, count and adjust the spore suspension concentration to 8×10 8 pcs / 100mL.

[0033] Seed medium (g / L): Glucose 20, soybean peptone 20, yeast extract 20, adjust the pH to 5.0-6.0, sterilize at 121°C for 20 minutes, add 1mL spore suspension to every 100mL seed medium, and place on a shaker Temperature 25°C, 200r / min cultivation for 24h to obtain the seed culture solution;

[0034] Fermentation medium (g / L): glucose 15, corn steep liquor 20, yeast extract 20, K 2 HPO 42. Adjust the pH value to 5.8, insert the well-grown seed culture solution into a 7L (liquid content 4.5L) fermenter with an inocul...

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Abstract

A microbial method by efficient conversion of canrenone belongs to a microbial preparation technology and relates to an efficient-conversion method for producing 11 alpha-hydroxycarvenone by using aspergillus ochraceus as a strain and by high-density culture. By the adoption of the method, problems, such as long production cycle, high cost, low conversion rate and the like, existing in present canrenone microbial transformation technologies are solved. By high-density culture, concentration of thalli in a nutrient solution per unit volume is raised and a high concentration is maintained, high conversion rate is guaranteed, conversion time is shortened, and production costs are greatly reduced. The method provided by the invention lays a foundation for industrial production and is of great significance.

Description

technical field [0001] The invention belongs to the technical field of microorganism preparation, and relates to a method for producing 11α-hydroxycanrenone through microbial transformation, in particular to a method for producing 11α-hydroxycanrenone through high-density cultivation using high-density culture to achieve efficient transformation of ochrax as the strain Methods. Background technique [0002] Canrenone (Canrenone) belongs to a steroid hormone cardiovascular drug, which is mainly used clinically as a non-selective aldosterone receptor antagonist to treat cardiovascular diseases. Cardiovascular diseases are mainly related to the renin-angiotensin-aldosterone system (RAAS), and the use of aldosterone receptor antagonists can block the effect of RAAS, thereby reducing the incidence and mortality of cardiovascular diseases. However, due to its negative interference to the quantitative detection of the cardiotonic agent Digoxin (Digoxin), and the dangerous conseque...

Claims

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Application Information

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IPC IPC(8): C12P33/10C12R1/66
Inventor 别松涛王家明路福平
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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