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Method for increasing yield of cyclo-lipopeptide antibiotics of bacillus amyloliquefaciens Q-426

A technology of amylolytic spores and cyclolipopeptides, which is applied in the field of biomedicine and can solve problems such as high difficulty and low content of cyclolipopeptides, and achieve the effect of increasing production

Inactive Publication Date: 2018-10-02
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the presence of various cyclolipopeptides and their homologues in the fermentation broth, and the content of each cyclolipopeptide is low, it is very difficult to obtain high-purity products in large quantities.

Method used

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  • Method for increasing yield of cyclo-lipopeptide antibiotics of bacillus amyloliquefaciens Q-426
  • Method for increasing yield of cyclo-lipopeptide antibiotics of bacillus amyloliquefaciens Q-426
  • Method for increasing yield of cyclo-lipopeptide antibiotics of bacillus amyloliquefaciens Q-426

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Experimental program
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Embodiment 1

[0040] A method for improving the production of bacillus amyloliquefaciens Q-426 cyclolipopeptide antibiotics, comprising the steps of:

[0041] (1) Synthesis of Pre-ComX P by Fmoc solid phase method 10

[0042] Using Fomc-protected tyrosine-substituted 2-chlorotrityl chloride resin as the starting material, using the Fomc solid-phase synthesis method, sequentially linking nine Fomc-protected amino acids, using frozen trifluoroacetic acid, 1,2 - The mixture of ethanedithiol and m-cresol was used as the lysate, and the Pre-ComX P 10Cut off from the 2-CTC resin; the amino acids are Fmoc-Lys(Boc)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Gly-OH, Fmoc- Gly-OH, Fmoc-Phe-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ile-OH and Fmoc-Val-OH; the trifluoroacetic acid, 1,2-ethanedithiol, m-cresol The volume ratio is 95:4:1.

[0043] Purification by reverse semi-preparative chromatography to obtain pure Pre-Com P 10 , the purity reaches 99%. Its liquid chromatogram is shown in figure 2 , molecu...

example 1

[0051] The seed solution was inoculated in 300ml liquid medium with 1% inoculum amount, and the bacterium was cultivated to the logarithmic growth phase on a shaker at 37°C and 180rpm, and 50ul of the reaction solution prepared in step 2 (signal molecule ComX P 10 ), continue to cultivate for 72h.

example 2

[0053] The seed liquid is inoculated in 300ml liquid culture medium with 1% inoculum amount, 37 ℃, 180rpm shaking table is cultivated to the logarithmic growth phase of the bacterium, adds the reaction solution 50ul (buffer solution+ComQ+ GPP, without Pre-ComXP 10 ), continue to cultivate for 72h.

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Abstract

The invention relates to the technical field of biomedicine, and in particular, relates to a method for increasing the yield of cyclo-lipopeptide antibiotics of bacillus amyloliquefaciens Q-426. According to the main technical schemes, the method comprises the following steps: (1) Pre-ComX P10 is synthesized by a Fmoc solid-phase method; (2) signal molecules ComX P10 are prepared; and (3) the bacillus amyloliquefaciens Q-426 is fermented, and the signal molecules ComX P10 are added in the fermentation process, wherein the bacillus amyloliquefaciens is isolated from compost of Dalian City, Liaoning Province and is identified as bacillus amyloliquefaciens by a traditional and molecular biological method. A large number of the Pre-ComX P10 are synthesized by the chemical synthesis method outside a biological system, and are added to the fermentation process of the bacillus amyloliquefaciens, and the yield of the lipopeptide antibiotics of the bacillus amyloliquefaciens is increased by artificial regulation and control.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for increasing the yield of bacillus amyloliquefaciens Q-426 cyclolipopeptide antibiotics. Background technique [0002] Since the discovery and application of penicillin in clinical treatment in 1943, it has played an important role in human treatment and victory over infectious diseases. In recent years, with the abuse of antibiotics in human disease treatment and the excessive addition of antibiotics in animal feed, a variety of drug-resistant superbugs have emerged. Existing antibiotics have been unable to control the infection of drug-resistant superbugs. Human health poses a great risk. According to relevant reports, the "super drug-resistant bacteria" produced by the abuse of antibiotics will threaten the lives of millions of Chinese people every year by 2050, and will also cause losses of 20 trillion US dollars for China. [0003] Cyclic lipopep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C12N1/20C07K7/06C07K1/06C07K1/04C12R1/07
CPCC07K7/06C12P21/02C12N1/205C12R2001/07
Inventor 权春善张丽影吴懿王路路范圣第
Owner DALIAN NATIONALITIES UNIVERSITY
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