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Method for producing tacrolimus by fermentation

A tacrolimus, fermentation culture technology, applied in the field of fermentation production of tacrolimus, can solve the problems of low yield, high cost, low yield, etc., and achieve the effect of reducing feedback inhibition, increasing production, and increasing production

Active Publication Date: 2014-04-02
CHENGDU YATU BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These patents report the production of tacrolimus with different streptomyces strains, the yield is not very high, the tacrolimus production strains of streptomycessp.MA6858 mentioned in the US5,116,756 patent and streptomyces tsukubaensis mentioned in US5,624,842, The production of tacrolimus is very low, and the highest concentration of tacrolimus in the fermentation broth is only 37.8 mg / L, which leads to high product cost after industrial production and causes a heavy burden to patients

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of slant medium: Weigh 10g of glucose, 20g of malt powder, 5g of yeast powder, 1.5g of sodium chloride, and 15g of agar, dilute to 1L with deionized water, and sterilize at 0.1MPa for 30min.

[0023] Preparation of liquid medium: weigh 10g of glucose, 20g of corn flour, 5g of yeast powder, 20g of soluble starch, 15g of glycerol, and 2g of calcium carbonate, dilute to 1L with deionized water, and sterilize at 0.1MPa for 30min.

[0024] Preparation of fermentation medium: Weigh 1.5g of yeast powder, 40g of soluble starch, 30g of corn steep liquor, 20g of glycerin, 1.5g of dipotassium hydrogen phosphate, 2g of calcium carbonate, dilute to 1L with deionized water, and sterilize at 0.1MPa for 30min , that is.

Embodiment 2

[0026] Utilize the various mediums provided in embodiment 1 to carry out the method for following fermentation production tacrolimus, comprise the following content:

[0027] a. Prepare the seed solution of Streptomyces sp1908 CGMCC No.1288: inoculate the preserved Streptomyces sp1908 CGMCC No.1288 strain on the slant medium, cultivate it at 30°C for 50 hours, and then inoculate it in the liquid medium , incubate in a shaker at 30°C for 50h (the rotation speed of the shaker is 100-300rpm) to obtain the Streptomyces seed solution;

[0028] b. Inoculation: inoculate the seed solution obtained in step a into the fermentation medium, and the inoculum size is 1%;

[0029] c. Fermentation culture: carry out fermentation culture with the culture medium after inoculation obtained in step b, the temperature of fermentation culture is 28~30 ℃, the rotating speed of fermentation culture is 150~200rpm;

[0030] d. Add pipecolic acid: after 120 hours of fermentation, add 1.0% pipecolic ac...

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PUM

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Abstract

The invention provides a method for producing tacrolimus by fermentation, belonging to the field of biomedicines. According to the method of producing tacrolimus by fermentation, parameters such as a strain-activating bevel culture medium component, a liquid culture medium component, seed liquid inoculum size and a fermentation medium component are optimized to maintain the growing activity and the fermenting activity of streptomycete, so that the production of tacrolimus is improved. In the fermenting process, piperidine acid is added into the fermentation medium and the adding manner such as adding time and addition is optimized, so that the feedback inhibition of products is reduced, and meanwhile, the output of the final product tacrolimus is increased, so that the final output of tacrolimus can reach 385 microgram / mL.

Description

[0001] technical field [0002] The invention belongs to the field of biomedicine, and in particular relates to a method for producing tacrolimus by fermentation. Background technique [0003] Over the past ten years, organ transplantation in my country has developed rapidly, ranking first in Asia and second in the world. Immunosuppressant drugs used to reduce rejection after organ transplantation also show rapid growth. The immunosuppressant commonly used to prevent and suppress rejection is cyclosporine, but its use is often accompanied by side effects. The development of new immunosuppressants has become particularly necessary. At present, a new type of immunosuppressant, tacrolimus, is being widely used. [0004] Tacrolimus, also known as FK-506, is a macrolide antibiotic extracted from soil actinomycetes by Fujisawa Pharmaceutical Company of Japan in 1984. It has strong immunosuppressive properties. It is more effective than cyclosporine in preventing the rejection o...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/465
Inventor 朱辉莫洪
Owner CHENGDU YATU BIOLOGICAL TECH
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