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Bacitracin and preparation method of zinc salt thereof

The technology of bacitracin zinc and bacitracin is applied in the field of preparation of bacitracin and its zinc salt bacitracin zinc, which can solve the problems of low total yield, difficult recovery and the like, and achieves short processing time, simple and fast operation, and simplified extraction. The effect of the purification process

Inactive Publication Date: 2016-04-20
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method needs to use a large amount of inorganic salts, recovery is difficult, and the total yield of this method is also on the low side

Method used

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  • Bacitracin and preparation method of zinc salt thereof

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preparation example Construction

[0030] Preparation of Seed Solution of Bacillus licheniformis Strain

[0031] The slant culture of the Bacillus licheniformis strain was inoculated into the seed medium, and cultured at 30° C. and 200 rpm for 20 hours to obtain a seed solution.

[0032] The seed medium described therein is obtained by the following method: 5.0 grams of cornstarch, 10.0 grams of glucose, 10.0 grams of yeast powder, 2.0 grams of ammonium sulfate, 2.0 grams of sodium chloride, 1.5 grams of magnesium sulfate heptahydrate, add tap water to be constant to 1000mL, pH7.0, sterilized at 121°C for 30 minutes.

[0033] The bacitracin producing bacterium of the present invention is the strain numbered ATCC10716.

[0034] Fermentation medium composition:

[0035] 30.0 grams of corn starch, 20.0 grams of glucose, 15.0 grams of cottonseed protein powder, 15.0 grams of yeast powder, 2.0 grams of ammonium sulfate, 4.0 grams of corn steep liquor, 1.5 grams of potassium dihydrogen phosphate, 2.5 grams of sodiu...

Embodiment 1

[0040] Get 500ml of bacitracin fermentation broth, fermentation unit 765U / ml, add 15 grams of superparamagnetic microspheres with a diameter of 20 μm to the culture broth, stir for half an hour, and magnetically separate the superparamagnetic microspheres in the fermentation broth. Collect the superparamagnetic microspheres and wash them twice with 30ml of deionized water, collect the washed superparamagnetic microspheres, soak them in 25ml of 2% ammonia solution for 20 minutes, separate the superparamagnetic microspheres, and collect the desorption solution once; then Soak in 20ml of 2% ammonia solution for 20 minutes, separate the superparamagnetic microspheres, and collect the secondary desorption liquid; combine the two desorption liquids to 43ml, and the HPLC content is 7828U / ml. Adjust the pH value to 6.0 with 2mol / L hydrochloric acid, extract with 20ml of n-butanol to obtain 24ml of n-butanol extract, add 120ml of acetone to the n-butanol extract, and precipitate bacitra...

Embodiment 2

[0042]2L of bacitracin fermentation broth, fermentation unit 744U / ml, 80 grams of superparamagnetic microspheres with a diameter of 20 μm were added to the culture broth, stirred for half an hour, and the superparamagnetic microspheres in the fermentation broth were separated magnetically. Collect the superparamagnetic microspheres and wash the magnetic field twice with 160ml deionized water, collect the washed superparamagnetic microspheres, soak them in 160ml4% ammonia solution for 20 minutes, separate the superparamagnetic microspheres, and collect the desorption solution once; Soak it in 160ml of 4% ammonia solution for 20 minutes, separate the superparamagnetic microspheres, and collect the secondary desorption solution; combine the two desorption solutions into 300ml, and the HPLC content is 4762U / ml. Adjust the pH value to 8.0 with 1 mol / L sulfuric acid, extract twice with 150ml of n-butanol to obtain 180ml of n-butanol extract, add 1440ml of acetone to the n-butanol ext...

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Abstract

The invention discloses bacitracin isolated and purified from a bacitracin fermentation broth and a method for preparing a zinc salt thereof, that is zinc bacitracin. The method includes: adsorbing bacitracin active components in the fermentation broth with super-magnetic microspheres, and preparing high-activity bacitracin via desorption, extraction and precipitation; back extracting an extract into a water phase, reacting with a zinc salt to obtain the zinc bacitracin. The super-magnetic microspheres are highly selective for the bacitracin active components and have weak ability to adsorb impurity bacitracin F, and the introduction of the super-magnetic microsphere medium gives a great simplification in the purifying process and a great increase in yield; the process is high in efficiency, low in energy consumption and low in solvent usage with bacitracin yield up to greater than 70 %, and both the prepared bacitracin and zinc salt comply with the standards of United States Pharmacopoeia.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a preparation method of bacitracin and zinc halobacitracin zinc. Background technique [0002] Bacitracin (Bacitracin) was originally a strain of Bacillus subtilis isolated by Johnson from patient wound pollutants and cultivated as a polypeptide antibiotic. It can be produced by Bacillus subtilis (bacillus sabtilis) and Bacillus licheniformis (Bacillus licheniformis). Due to its strong antibacterial effect on Gram-positive bacteria, its stability is enhanced after complexing with metal ions, and its antibacterial activity is higher. The local application of the drug has little irritation, rare allergic reactions, and is resistant to enzymes. Bacteria develop resistance to it slowly, and it is extremely rare to obtain drug-resistant strains. Moreover, its activity is not affected by pus, sputum, blood, exudate and necrotic tissue, so it is often use...

Claims

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Application Information

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IPC IPC(8): C07K7/58C07K1/14C07K1/107
CPCC07K7/58
Inventor 张炜张雪霞耿文飞冷凤任风芝
Owner NCPC NEW DRUG RES & DEV
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