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Method for producing Daptomycin by fed-batch methyl oleate fermentation

A technology of daptomycin and oleic acid is applied in the field of daptomycin production by fermenting methyl oleate to achieve the effect of guaranteeing effect

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] So far, there have been no reports of increased daptomycin production with the addition of methyl oleate alone during feeding

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0015] A method for producing daptomycin by fermenting methyl oleate in stream, comprising the following contents:

[0016] Cultivate the seed liquid of Streptomyces roseospora; inoculate the seed liquid into the fermentation medium; then cultivate and ferment, the fermentation temperature is 30-32°C; after 20 hours of fermentation, add oil with a mass concentration of 2.0% to the fermentation liquid Acetate methyl ester solution (the solvent is a mixture of ethanol and water at a volume ratio of 1:20), the feeding speed is: feed the fermenter at a flow rate of 1.5-3.0ml / h per liter of fermentation broth After 48-144 hours of fermentation and cultivation, the fermentation is terminated, and the obtained fermentation broth is extracted and separated to obtain the daptomycin.

Embodiment 2

[0018] A method for producing daptomycin by fermenting methyl oleate in stream, comprising the following contents:

[0019] 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 roseospora seed solution.

[0020] 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;

[0021] 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 ...

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Abstract

The invention provides a method for producing Daptomycin by fed-batch methyl oleate fermentation. A methyl oleate solution is supplemented into a fermentation tank in a Streptomyces roseosporus fermentation culture process to produce the Daptomycin. Thus, the method provided by the invention can enhance the yield of the Daptomycin. The method provided by the invention widens the path for producing Daptomycin by a fermentation process; and the methyl oleate solution can exist in the form of liquid at normal temperature, and thus, can not crystallize due to low temperature, thereby ensuring the fed-batch effect. Besides, the method is suitable for laboratory small-tank fermentation of Daptomycin, and also suitable for scale-up production in industrialized big tanks.

Description

[0001] technical field [0002] The invention belongs to the production field of daptomycin, and in particular relates to a method for producing daptomycin by fermenting methyl oleate in stream. 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 bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/465
Inventor 朱辉朱春燕林亿均
Owner CHENGDU YATU BIOLOGICAL TECH
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