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Amino acid adding method for raising sirolimus fermentation yield

A technology for sirolimus and amino acid is applied in the field of improving the fermentation yield of sirolimus, which can solve the problem of low fermentation level of sirolimus strain, complex metabolic pathway of sirolimus, and regulation of rapamycin biosynthesis. The mechanism is not clear and other problems, to achieve the effect of clear purpose, low fermentation cost, and improved fermentation level

Inactive Publication Date: 2011-09-28
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the fermentation level of sirolimus strains is still low, and the biosynthetic metabolic pathway of sirolimus is relatively complex. So far, the regulatory mechanism of rapamycin biosynthesis is still unclear, and it is also one of the current research hotspots.

Method used

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  • Amino acid adding method for raising sirolimus fermentation yield

Examples

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Effect test

Embodiment 1

[0011] Example 1: The components of the fermentation medium are glucose 40g / L, glycerol 50g / L, cottonseed powder 20g / L, lysine salt 10 g / L, NaCl 0.5 g / L, K 2 HPO 4 0.1 g / L.

[0012] The initial pH of the medium was controlled at 6.5, the sterilization temperature was 121°C, and the sterilization time was 30 min; 0.5% proline was added to the initial fermentation medium. The fermentation process adopts the shake flask fermentation process, the liquid volume of 500mL shake bottle is 50 mL, the shaker speed is 220 r / min, and Streptomyces hygroscopicus S. hygroscopicus ATCC 29253 culture seed solution, the inoculum size was 10%, the culture temperature was 30 ℃, the sirolimus fermentation yield increased by 12.39% compared with the control group.

Embodiment 2

[0013] Example 2: The components of the fermentation medium are glucose 40g / L, glycerin 50g / L, cottonseed powder 20g / L, lysine salt 10 g / L, NaCl 0.5 g / L, K 2 HPO 4 0.1 g / L.

[0014] The initial pH of the medium was controlled at 6.5, the sterilization temperature was 121°C, and the sterilization time was 30 min. A mass concentration of 0.5% phenylalanine was added to the initial fermentation medium. The fermentation process adopts the shake flask fermentation process, the liquid volume of 500mL shake bottle is 50 mL, the shaker speed is 220 r / min, and Streptomyces hygroscopicus S. hygroscopicus NRRL5491 was cultured with seed solution, the inoculum size was 10%, and the culture temperature was 30°C. The fermented sirolimus yield increased by 6.8% compared with the control group.

Embodiment 3

[0015] Example 3: The components of the fermentation medium are glucose 40g / L, glycerol 50g / L, cottonseed powder 20g / L, lysine salt 10 g / L, NaCl 0.5 g / L, K 2 HPO 4 0.1 g / L.

[0016] The initial pH of the medium was controlled at 6.5, the sterilization temperature was 121°C, and the sterilization time was 30 min. Glutamic acid and 0.5% glutamine were added to the initial fermentation medium at a mass concentration of 0.5%. The fermentation process adopts the shake flask fermentation process, the liquid volume of 500mL shake bottle is 50 mL, the shaker speed is 220 r / min, and Streptomyces hygroscopicus S. hygroscopicus ATCC 29253 cultured seed liquid, the inoculum amount was 10%, and the culture temperature was 30°C. The fermentation yield of sirolimus glutamic acid was increased by 21.63% compared with the control group; the fermentation yield of sirolimus glutamine was increased compared with the control group 39.74%.

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Abstract

The invention brings forward a method for raising sirolimus fermentation yield by adding amino acid into a fermentation medium. During fermentation, the initial pH value of the medium is controlled within the range from 5.5 to 7.0, the disinfection temperature ranges from 115 DEG C to 121 DEG C and the disinfection process lasts for 30 min; simultaneously, amino acid at mass concentration of 0.2-1% is added into the initial fermentation medium, wherein the amino acid is one or more combinations selected from the group consisting of proline, phenylalanine, glutamic acid and glutamine, valine, alanine, and leucine or threonine. According to the invention, with the combination of sirolimus biosynthesis characteristics, amino acid which is related with the supply of sirolimus synthesis precursor is obtained by screening, and the addition of a certain concentration of amino acid into the initial medium can greatly raise the sirolimus fermentation level. The method provided by the invention has advantages of clear purpose and low fermentation cost and the like, can be used to conduct the metabolic regulation of the sirolimus fermentation by adding amino acid and applied for industrial production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for increasing the fermented yield of sirolimus. Background technique [0002] Sirolimus is a macrocyclic triene lactone compound with a molecular structure as (I), produced by fermentation of Streptomyces hygroscopicus, and is a new type of potent immunosuppressant with antifungal, antiproliferative and antitumor activities. [0003] [0004] Due to its specific binding and inhibition of mammalian target of rapamycin (mTOR), sirolimus is currently being developed as an anti-rejection drug for organ transplantation and a drug for the treatment of autoimmune diseases; at the same time Derivatives developed on the basis of sirolimus such as Temsirolimus (temsirolimus), Everolimus (everolimus), Deforolimus (AP23573, MK-8669) have been used as targeted, low-toxic anti-tumor cancer drugs into the clinic. [0005] At present, the fermentation level of sirolimus st...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/55
Inventor 邹祥李进
Owner SOUTHWEST UNIVERSITY
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