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Method for preparing bacitracin and method for preparing zinc salt of bacitracin

The technology of bacitracin and acidifying agent 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 simple and easy process, improved purity, and convenient industrial production. Effect

Inactive Publication Date: 2016-05-11
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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  • Method for preparing bacitracin and method for preparing zinc salt of bacitracin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Bacitracin fermentation broth is 15L, and the fermentation unit is 840U / ml. Add glacial acetic acid to adjust the pH value to 4.0, add 800 grams of perlite filter aid, and filter to obtain bacitracin filtrate. Add 3% sodium hydroxide solution to the filtrate to adjust the pH to 6.0, and then pass it into the macroporous cation exchange resin column HD8 for adsorption. Desorb with 2% ammonia water at a flow rate of 0.5L / h. When bacitracin flows out of the desorption outlet, connect the 001X8 resin column and the D315 resin column in series for desalting and decolorization. Collect the destained solution containing bacitracin.

[0029] The above-mentioned decolorization solution was adjusted to a pH value of 6.0 with 3% sodium hydroxide solution, extracted twice with 800ml of n-butanol to obtain a n-butanol extract, and the n-butanol extract was vacuum-concentrated at 50 degrees Celsius, and the concentrated solution was 200ml. Add 1600ml of acetone, and the bacitracin ...

Embodiment 2

[0031] Bacitracin fermentation broth 8L, fermentation unit 893U / ml. Add oxalic acid to adjust the pH value to 3.0, add 400 grams of perlite filter aid, and filter to obtain bacitracin filtrate. The filtrate was adjusted to pH 6.5 and passed into a macroporous cation exchange resin D001 column for adsorption. The resin loading capacity was 0.8 L and the flow rate was 1.5 L / hr. Desorb with 2.5% ammonia water at a flow rate of 0.4L / hr. When bacitracin flows out of the desorption outlet, connect the 001X7 resin column and the D304 resin column in series for desalting and decolorizing. The volume of the desalting and decolorizing resins are both 250ml. Collect 0.98 L of decolorization solution containing bacitracin.

[0032] Adjust the pH value of the above decolorization solution to 6.5 with 5% sodium hydroxide solution, extract twice with n-butanol, concentrate the n-butanol extract under vacuum at 50 degrees Celsius to obtain 150ml of concentrated solution, add 750ml of acetone...

Embodiment 3

[0034] Bacitracin fermentation liquid is 7.2L, and the fermentation unit is 851U / ml. Add 2 mol / l sulfuric acid aqueous solution to adjust the pH value to 3.5, add 300 grams of perlite filter aid, and filter to obtain bacitracin filtrate. The filtrate was adjusted to pH 9.0 with 1% sodium hydroxide solution and passed into a macroporous cation exchange resin HD8 column for adsorption. The resin loading capacity was 0.55 L and the flow rate was 1.0 L / hr. Desorb with 4% ammonia water at a flow rate of 0.25L / hr. When bacitracin flows out from the desorption outlet, connect the 001X2 resin column and the D301 resin column in series for desalting and decolorizing. The volume of the desalting and decolorizing resins is both 250ml. Collect the destained solution containing bacitracin.

[0035]Adjust the pH value of the above decolorization solution to 8.0 with 5% sodium hydroxide, extract twice with n-butanol, add acetic acid aqueous solution to the n-butanol extract to adjust the pH...

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Abstract

The invention discloses a method for separating bacitracin from bacitracin fermentation liquid and purifying the bacitracin and a method for preparing zinc salt bacitracin zinc of the bacitracin. The method includes the steps that the fermentation liquid is acidized and filtered, and the filtered liquid is adsorbed and desorbed through macroporous cation exchange resin; after the desorbed liquid is desalted, decolorized, extracted and precipitated, the high-activity bacitracin is prepared. The extracted liquid is subjected to back extraction and enters an aqueous phase to be reacted with zinc salt, and the bacitracin zinc is obtained. The whole technology is simple and easy to operate, and the activity of the product is retained to the maximum degree; the technology is efficient, low in energy consumption and suitable for industrial production, the yield of the bacitracin is 60% or above, and the prepared bacitracin and the prepared zinc salt both meet the United States pharmacopeia standard.

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): C07K7/58C07K1/18C07K1/14
CPCC07K7/58
Inventor 张炜冷凤耿文飞张雪霞任风芝
Owner NCPC NEW DRUG RES & DEV
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