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Novel method for adding decanoic acid in fermentation process of daptomycin

A fermentation process and technology of daptomycin, applied in the field of microbial fermentation, can solve the problems of affecting the effect of fed addition, easy crystallization and precipitation, etc.

Inactive Publication Date: 2012-10-03
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to explore a new method for adding capric acid in the fermentation process of daptomycin, to solve the problem that capric acid is solid at the culture temperature (30° C.) and is easy to crystallize even if it is dissolved in a solvent, thus affecting the feeding effect The problem

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] The invention produces daptomycin by fermenting Streptomyces roseospora NRRL11379.

[0010] Process steps

[0011] 1. Prepare slant medium, the medium composition is: soluble starch 2g / L, KNO 3 0.1g / L, NaCl 0.05g / L, K 2 HPO 4 0.05g / L, MgSO 4 0.05, FeSO 4 0.01g / L, agar powder 2g / L 0; pH 7.5. After subpackaging, sterilize at 121°C for 20 minutes for later use. The preserved strains were transferred to slant medium and cultured at 30°C for 120h.

[0012] 2. Prepare seed medium, the composition of which is: glucose 5g / L, dextrin 15g / L, peptone 10g / L, yeast powder 10g / L; pH 7.0. Sterilize at 121°C for 20min. Fill each 250mL Erlenmeyer flask with 30mL seed medium. Inoculate the fresh slant spores into the seed medium, and then culture them in a shaker at 220r / min at 30°C for 36h.

[0013] 3. Prepare the fermentation medium, the composition of the medium is: dextrin 10.62g / L, yeast extract 1.59g / L, casein hydrolyzate 1.28g / L, L-aspartic acid 1.50g / L, potassium sul...

Embodiment 2

[0016] The invention produces daptomycin by fermenting Streptomyces roseospora NRRL11379.

[0017] Process steps

[0018] 1. Prepare slant medium, the medium composition is: soluble starch 2g / L, KNO 3 0.1g / L, NaCl 0.05g / L, K 2 HPO 4 0.05g / L, MgSO 4 0.05, FeSO 4 0.01g / L, agar powder 2g / L 0; pH 7.5. After subpackaging, sterilize at 121°C for 20 minutes for later use. The preserved strains were transferred to slant medium and cultured at 30°C for 120h.

[0019] 2. Prepare seed medium, the composition of which is: glucose 5g / L, dextrin 15g / L, peptone 10g / L, yeast powder 10g / L; pH 7.0. Sterilize at 121°C for 20min. Fill each 250mL Erlenmeyer flask with 30mL seed medium. Inoculate the fresh slant spores into the seed medium, and then culture them in a shaker at 220r / min at 30°C for 36h.

[0020] 3. Prepare the fermentation medium, the composition of the medium is: dextrin 10.62g / L, yeast extract 1.59g / L, casein hydrolyzate 1.28g / L, L-aspartic acid 1.50g / L, potassium sul...

Embodiment 3

[0023] The invention produces daptomycin by fermenting Streptomyces roseospora NRRL11379.

[0024] Process steps

[0025] 1. Prepare slant medium, the medium composition is: soluble starch 2g / L, KNO 3 0.1g / L, NaCl 0.05g / L, K 2 HPO 4 0.05g / L, MgSO 4 0.05, FeSO 4 0.01g / L, agar powder 2g / L 0; pH 7.5. After subpackaging, sterilize at 121°C for 20 minutes for later use. The preserved strains were transferred to slant medium and cultured at 30°C for 120h.

[0026] 2. Prepare seed medium, the composition of which is: glucose 5g / L, dextrin 15g / L, peptone 10g / L, yeast powder 10g / L; pH 7.0. Sterilize at 121°C for 20min. Fill each 250mL Erlenmeyer flask with 30mL seed medium. Inoculate the fresh slant spores into the seed medium, and then culture them in a shaker at 220r / min at 30°C for 36h.

[0027] 3. Prepare the fermentation medium, the composition of the medium is: dextrin 10.62g / L, yeast extract 1.59g / L, casein hydrolyzate 1.28g / L, L-aspartic acid 1.50g / L, potassium sul...

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Abstract

The invention relates to a novel method for adding decanoic acid in the fermentation process of daptomycin, namly lipase and ethyl decanoate are simultaneously added in the fermentation cultivation process of streptomyces roseosporus, the ethyl decanoate is hydrolyzed by the lipase so as to add the precursor decanoic acid to produce the daptomycin, and the novel method is a method for improving the yield of the daptomycin. The method has the advantages that: the novel method for adding the decanoic acid is explored, the decanoic acid is slowly added by an enzymolysis method of the ethyl decanoate, and the toxicity of the decanoic on cells is reduced, so that the yield of the daptomycin is improved. The method comprises the following steps of: optimizing the addition amount of the ethyl decanoate and the lipase; and optimizing the addition time of the ethyl decanoate and the lipase. The yield of the daptomycin can be improved by when the novel method provided by the invention is used for producing the daptomycin.

Description

technical field [0001] The invention relates to a microbial fermentation method, in particular to a method for producing daptomycin by slowly adding precursor capric acid in an enzymatic way. Background technique [0002] 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 ...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12R1/465
Inventor 吴旻王旻王泽根王航
Owner CHINA PHARM UNIV
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