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Method for producing daptomycin by fermentation

A technology of daptomycin and fermentation liquid, which is applied in the field of fermentative production of daptomycin, can solve the problem of low daptomycin concentration, and achieve the effect of increasing synthetic yield and increasing inhibitory concentration

Inactive 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

[0007] When daptomycin is synthesized by fermentation, due to the feedback inhibition in the microbial metabolism process, when the daptomycin produced by fermentation reaches a certain concentration, it will inhibit the further synthesis of daptomycin, making the daptomycin produced by fermentation The concentration of mycotoxin is not high

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0022] A method for producing daptomycin by fed-batch decanal fermentation, comprising the following:

[0023] a. Cultivate Streptomyces roseospora seed liquid;

[0024] b. Inoculation: inoculate the seed solution obtained in step a into the fermentation medium, and the inoculum size is 10%;

[0025] c. Fermentation culture: the inoculated medium obtained in step b is cultivated and fermented, and the fermentation temperature is 30-32°C;

[0026] d. Adding decanal solution: After 18 hours of fermentation, add decanal solution with a mass concentration of 6.0% (solvent is methyl oleate) to the fermentation broth at a rate of 1.0 to 1.0% per liter of fermentation broth. Feed the fermenter at a flow rate of 2.0ml / h until the end of fermentation;

[0027] e. Finishing of fermentation and separation: After 48-168 hours of fermentation, the fermentation is ended, and the obtained fermentation broth is extracted and separated to obtain the daptomycin.

Embodiment 2

[0029] A method for producing daptomycin by fed-batch decanal fermentation, comprising the following:

[0030] a. Cultivate Streptomyces roseospora seed solution: Transfer the preserved Streptomyces roseospora strain to the slant medium, cultivate at 30°C for 120h, then inoculate in liquid medium, and cultivate in a shaker at 30°C for 36h, that is Obtain the described Streptomyces roseospore seed liquid;

[0031] Wherein, the preparation method of the slant medium is as follows: add 20 g of soluble starch, 0.5 g of dipotassium hydrogen phosphate, 0.5 g of magnesium sulfate heptahydrate, 0.01 g of iron sulfate heptahydrate, and 1 g of potassium nitrate into 1 L of deionized water to adjust the pH =7.2, then add 20g of agar, steam sterilize, and then pour the slant plate to get the slant medium;

[0032] b. Inoculation: inoculate the seed liquid obtained in step a into the fermentation medium (pH of the fermentation medium = 6.5~7.0), the inoculation amount is 10%;

[0033...

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Abstract

The invention provides a method for producing daptomycin by fermentation, and belongs to the field of production of daptomycin. In the method for synthesis of daptomycin by fermentation, the parameters such as the concentration and the inoculum size of a culture seed solution and pH of fermentation medium are optimized, the growth activity and the fermentation activity of streptomyces roseosporus are maintained, the production of the daptomycin is improved; in the fermentation process, a decanal solution is fed to the fermentation medium, and the adding time and the adding amount are optimized, so that the inhibition concentration of the daptomycin can be improved, and the synthetic production of the daptomycin is further improved.

Description

[0001] technical field [0002] The invention belongs to the production field of daptomycin, in particular to a method for fermenting and producing daptomycin. Background technique [0003] In the past 10 years, the trend of Gram-positive drug-resistant bacterial infections has been on the rise. At present, the difficulties in the clinical treatment of bacterial infections mainly focus on methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), and glycopeptide-sensitive Staphylococcus aureus (GISA). Pathogens with a high transmission rate and high fatality rate in hospitals, although vancomycin and other glycopeptide antibiotics have achieved certain curative effects in the treatment of such infections, but in recent years, vancomycin-resistant bacteria have been found clinically, and this The trend is gradually increasing. Therefore, research on new antibiotics against drug-resistant bacteria has become a hot topic again. Daptomycin ...

Claims

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Application Information

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IPC IPC(8): C12P21/04C12R1/465
Inventor 朱辉邹敬源
Owner CHENGDU YATU BIOLOGICAL TECH
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