Process for fermenting rapamycin with high yield

A technology of rapamycin and fermentation technology, which is applied in the field of high-yield rapamycin fermentation technology, can solve the problems of low cell density, unreasonable feeding, high viscosity, etc., to reduce metabolic intermediate products, increase content, and increase production Effect

Inactive Publication Date: 2014-02-05
HEILONGJIANG WEIKAIER BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] From the above patent documents, it can be obtained that the fermentation unit of rapamycin reaches more than 600mg/L, but there are the following problems: the culture medium is unreasonable and the viscosity is high, resulting in low dissolved oxygen content in the large-scale fermentation process, which is not conducive to bacterial fermentation. During the fermentation process, there is no feeding, and the cell densit

Method used

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  • Process for fermenting rapamycin with high yield
  • Process for fermenting rapamycin with high yield
  • Process for fermenting rapamycin with high yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Evenly spread the rapamycin spore suspension diluted to a certain concentration on a plate filled with 30ml slant medium, the medium is soybean powder 1g / L, soluble starch 10g / L, K 2 HPO 4 0.5g / L, MgSO 4· 7H 2 O 1g / L, agar 20g / L, cultivated in a biochemical incubator at 28°C for 11 days, picked a single colony and cultivated it on the slant of the test tube.

[0037] Use an inoculation spatula to take a ring from the slant of the cultured rapamycin-producing bacteria and inoculate it into a 250ml Erlenmeyer flask containing 30ml of seed medium. The shaker speed is 200r / m, the temperature is 30°C, and cultured for 52h.

[0038] Inoculate the seed culture liquid into a 400L primary seed tank with 200L seed culture liquid according to the inoculation amount of 1% by volume, and the air volume is 2.0m 3 / h, the tank pressure is 0.02MPa, the stirring speed is properly adjusted so that the dissolved oxygen is not lower than 10%, the culture temperature is 30°C, and cultur...

Embodiment 2

[0043] Inoculate the prepared seeds with 3% inoculum by volume and inoculate the seed culture solution into a 400L primary seed tank with 250L seed culture solution, with an air volume of 2.0m 3 / h, the tank pressure is 0.02MPa, the stirring speed is properly adjusted so that the dissolved oxygen is not lower than 10%, the culture temperature is 30°C, and cultured for 48h. Seed medium comprises the following components and content: peptone 10g / L, yeast extract 6g / L, glucose 20g / L, L-lysine 5g / L, NaH 2 PO 4 0.5g / L, adjust the pH to 6.8 before sterilization, and sterilize at 121°C for 45min.

[0044] Inoculate the seed culture solution into a 2200L secondary seed tank with 1500L seed medium according to the volume ratio of 10% inoculum, and the air volume is 2.2m 3 / h, the tank pressure is 0.02MPa, the stirring speed is properly adjusted so that the dissolved oxygen is not lower than 10%, the culture temperature is 30°C, and cultured for 48h.

[0045] Put the seed culture so...

Embodiment 3

[0047] Inoculate the prepared seeds with 3% inoculum by volume and inoculate the seed culture solution into a 400L primary seed tank with 250L seed culture solution, with an air volume of 2.0m 3 / h, the tank pressure is 0.02MPa, the stirring speed is properly adjusted so that the dissolved oxygen is not lower than 10%, the culture temperature is 30°C, and cultured for 48h. Seed medium includes the following components and contents: peptone 10g / L, yeast extract 6g / L, glucose 20g / L, L-lysine 5g / L, NaH 2 PO 4 0.5g / L, adjust the pH to 6.8 before sterilization, and sterilize at 121°C for 45min.

[0048] Inoculate the seed culture solution into a 2200L secondary seed tank with 1500L seed medium according to the volume ratio of 10% inoculum, and the air volume is 2.2m 3 / h, the tank pressure is 0.02MPa, the stirring speed is properly adjusted so that the dissolved oxygen is not lower than 10%, the culture temperature is 30°C, and cultured for 48h.

[0049] Put the seed culture...

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Abstract

The invention provides a process for fermenting rapamycin with high yield, and belongs to a method for preparing antibiotics by utilizing fermentation of microorganism in the technical field of biology. According to the method, enzymolytic cotton seed powder, dextrin, glycerinum, yeast extract, L-lysine, KH2PO4, K2HPO4, (NH4)2SO4 and trace elements as culture media, wherein the trace elements contain Co<2+>, glycerinum and phosphate buffer are fed in the early phase of the fermenting culture process, and L-pipecolic acid and shikimic acid are fed in the synthesizing phase. By adopting the method, the methyl derivative can be reduced in the growing phase, and the content of rapamycin is increased; intermediate products can be reduced, and rapamycin can generate key enzyme activity; the beneficial effect of high yield of rapamycin can be achieved.

Description

technical field [0001] The invention belongs to a method for producing antibiotics by microbial fermentation in the field of biotechnology, in particular to a high-yield rapamycin fermentation process. Background technique [0002] Rapamycin (Rapamycin, RAPA), also known as Sirolimus (Sirolimus), molecular structure such as figure 1 The molecular formula of RAPA is C 51 h 79 NO 13 , molecular weight 99lKD, white solid crystal, melting point 183-185 ℃, specific rotation [ɑ] 25 D =153 0 (Methanol), lipophilic, soluble in methanol, ethanol, acetone, chloroform and other organic solvents, very slightly soluble in water, almost insoluble in ether, low and neutral pH in plasma, very fast degradation at 37°C, in plasma The half-life is 1~5h. Rapamycin is a new type of immunosuppressant of nitrogen-containing triene macrolide composed of 36-membered rings. It has anti-fungal, anti-proliferation and anti-tumor effects. In recent years, rapamycin has been successfully used cli...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/55
Inventor 姜浩魏玮刘云娜李岩姜巍
Owner HEILONGJIANG WEIKAIER BIOTECH CO LTD
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