Polymyxin B production fermentation culture medium, preparation method and uses thereof

A fermentation medium and polymyxin technology, applied in the biological field, can solve the problem of low fermentation yield

Active Publication Date: 2015-07-15
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The relevant medium formulations involved in existing reports have the problem of low fermentation yield

Method used

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  • Polymyxin B production fermentation culture medium, preparation method and uses thereof

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Experimental program
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Effect test

preparation example Construction

[0037] Preparation method of fermentation medium

[0038] The culture medium provided by the invention is treated with amylase before sterilization.

[0039] Specifically, the preparation method of the fermentation medium provided by the invention comprises the following steps:

[0040] In the first step, the organic nitrogen source, whole wheat flour, inorganic salt and water are mixed according to the above formula to obtain a mixture 1;

[0041] In the second step, amylase is applied to the gelatinized or ungelatinized mixture 1 to obtain the fermentation medium provided by the present invention.

[0042] The amylase involved in the above method is selected from α-amylase, β-amylase, glucoamylase or isoamylase; preferred α-amylase; by the weight of the whole wheat flour dropped in the first step, the amylase The dosage is 2-1500U / g; preferably 2-1000U / g, more preferably 100-1000U / g; the action temperature of the amylase is 50-80°C, and the action time is 10-60 minutes; pr...

Embodiment 1

[0063] Culture strain: Paenibacillus polymyxa ATCC10401

[0064] Seed medium / %: corn steep liquor 2.0; glucose 5.0; ammonium sulfate 0.4; sodium chloride 0.1; calcium carbonate 0.3; pH6.8~7.0.

[0065] Fermentation medium / %: whole wheat flour 8.0; corn steep liquor powder 0.4; ammonium sulfate 0.9; potassium dihydrogen phosphate 0.03; calcium carbonate 0.03.

[0066] The fermentation medium was gelatinized, and BF7658α-amylase (Xingtai Wanda Bioengineering Co., Ltd.) was added. The amount of amylase was 100U / g, and the reaction was performed at 50°C-80°C for 30 minutes; after that, it was sterilized at 121°C for 20 minutes.

[0067] Cultivation method: inoculate the strains preserved on the slant into the seed medium, and cultivate at 28°C for 18 hours. Add 4% inoculum to 18L of the above-mentioned fermentation medium (the total volume of the fermenter is 35L), stir at 400r / min, under 50kPa tank pressure, ventilation at 1.2vvm, and culture at 30°C for 48h.

[0068] Determina...

Embodiment 2

[0071] Other operations are the same as in Example 1, except that the starting strain is NRRL B-719. After testing, the yield of polymyxin B obtained by cultivation was 3.14g / L.

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Abstract

The present invention discloses a polymyxin B production fermentation culture medium, a preparation method and uses thereof, wherein the culture medium comprises, by weight, 0.1-1.5 w/w% of an organic nitrogen source, 5.0-15.0 w/w% of whole wheat powder, and 0.3-1.2 w/w% of an inorganic salt.

Description

technical field [0001] The invention relates to the field of biotechnology; in particular, it relates to a fermentation medium and its preparation method and application. Background technique [0002] Polymyxin B (Polymyxin B, PMB) is a basic cyclic peptide antibiotic produced by Bacillus polymyxa or Paenibacillus polymyxa, which has good activity against Gram-negative bacteria. Colistin B drugs include polymyxin B sulfate injection and compound topical preparations of polymyxin B sulfate and other antibiotics, which are used to treat skin wounds, peritonitis, urinary system and respiratory system caused by Gram-negative bacteria Infection at other sites; PMB can also significantly inhibit the acute phase response induced by endotoxin, so it can also be used for sepsis. [0003] At present, the preparation of polymyxin B adopts the fermentation method. After the seeds of Bacillus polymyxa or Paenibacillus polymyxa are cultured, they are inoculated in a fermenter and cultiva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/12C12R1/01
Inventor 马川川那可赵文杰魏晓东刘永双张守雨
Owner SHANGHAI INST OF PHARMA IND CO LTD
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