Method for fermenting daptomycin by adding caprate

A technology of fed-adding decanoate and daptomycin, applied in the field of microbial fermentation, can solve the problems of cytotoxicity, affecting the effect of fed-adding, easy crystallization and precipitation, etc., and achieves the effects of low toxicity, guaranteed effect and widened path.

Active Publication Date: 2010-11-10
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, methyl oleate, ethanol, etc. are used to dissolve capric acid, but capric acid dissolved in methyl oleate or ethanol is still highly toxic to cells, and it is easy to crystallize and precipitate when the temperature is low, which affects the feeding effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Fermentation of Daptomycin with Feed Potassium Decanoate as Precursor

[0019] (1) shake flask culture

[0020] Table 1 shake flask culture medium formula

[0021] Medium components

[0022] Put 30ml of the culture medium prepared according to the formula in Table 1 in a 250ml Erlenmeyer flask, sterilize it with steam, and inoculate each bottle with 0.5ml of spore suspension when it is cooled to 30°C, the shaker speed is 220rpm, and the culture temperature is 30°C , culture time 24h.

[0023] (2) Seed tank cultivation

[0024] In the 10L seed tank, prepare 7L medium according to the formula in Table 2. After the medium is sterilized, when the temperature drops to 30°C, insert the shaken spore suspension in the shaker bottle, the inoculation amount is about 2%, and the culture temperature is 30°C. ±1°C, tank pressure 0.03-0.05MPa, air flow rate 1:1vvm, stirring speed 300-400rpm, and ensure that the dissolved oxygen is not less than 20% during the peak period...

Embodiment 2

[0033] Fermentation of daptomycin by adding sodium caprate as precursor

[0034] Compared with Example 1, the difference of this example is: during the cultivation process of the fermenter, after about 28 hours of cultivation, feed the fermenter with a flow rate of 0.5-1.5ml / / h in each liter of fermentation broth Add 10.0% sodium caprate solution until the fermentation ends, and generally at about 120 hours, the total sugar reaches below 0.5, the hyphae autolyze, and the pH value of the fermentation broth begins to rise rapidly. At this time, the effect of daptomycin in the fermentation broth The price reaches 1000U / ml.

Embodiment 3

[0036] Fermentation of daptomycin by feeding ammonium caprate as precursor

[0037] Compared with Example 1, the difference of this example is that during the cultivation process of the fermenter, at about 28 hours of cultivation, the fermenter was fed with a flow rate of 1.0-2.0ml / / h per liter of fermentation broth Add 8.0% ammonium caprate solution until the fermentation ends, and generally at about 160 hours, the total sugar reaches below 0.5, the hyphae autolyze, and the pH value of the fermentation broth begins to rise rapidly. At this time, the effect of daptomycin in the fermentation broth The price reaches about 1700U / ml.

[0038] In addition, the present invention is not only suitable for fermenting daptomycin in small tanks, but also suitable for fermenting daptomycin in large tanks.

[0039] In summary, the present invention uses 5.0% potassium caprate solution or 10.0% sodium caprate solution or 8.0% ammonium caprate solution as the precursor for fermenting daptom...

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PUM

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Abstract

The invention relates to a method for fermenting daptomycin by adding caprate. In the fermentation tank culturing process, the daptomycin is produced by adding 5.0 percent potassium decanoate solution, 10.0 percent sodium decanoate solution or 8.0 percent ammonium decanoate solution, namely the decanoates are used as precursors for fermenting the deptomycin. The method has the advantages that: while the daptomycin fermenting path is enlarged, the adding effect can be ensured because the precursors have small toxicity to cells, can exist in a liquid form at normal temperature and cannot easily crystallize due to low temperature.

Description

【Technical field】 [0001] The invention relates to a microbial fermentation method, in particular to a method for fermenting daptomycin by adding capric acid salt. 【Background technique】 [0002] With the widespread use of antibiotics in clinical practice, the problem of bacterial resistance is becoming more and more serious. Methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), penicillin-resistant Streptococcus pneumoniae (PRSP), and Macrolide Gram-positive infections, like other Gram-positive pathogenic infections, are on the alarming rise. Glycopeptide antibiotics represented by vancomycin are effective against almost all Gram-positive (G + ) Bacteria have antibacterial effect and are applied to severe G + Bacteria and drug-resistant bacteria infection, vancomycin is the last line of defense against such refractory diseases. However, with the emergence of VRE in the 1980s, glycopeptide-resistant enterococci continued to increase. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/465
Inventor 郑卫程元荣周剑张引高悟岩刘颖黄建峰江红林如
Owner FUJIAN INST OF MICROBIOLOGY
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