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Method for extracting daptomycin from fermentation broth

A technology for daptomycin and fermentation broth, applied in the field of fermentation broth, can solve the problems of acid hydrolysis of daptomycin, large pollution, increase the cost of separation and purification, etc., and achieves reduction of acid hydrolysis, low cost, significant economic and social value Effect

Inactive Publication Date: 2012-06-13
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Some of the purification technologies mentioned in these patents use acetonitrile as the eluent, which is more polluting to the environment; Advanced ion exchange packing and reverse phase column chromatography packing greatly increase the cost of separation and purification

Method used

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  • Method for extracting daptomycin from fermentation broth
  • Method for extracting daptomycin from fermentation broth
  • Method for extracting daptomycin from fermentation broth

Examples

Experimental program
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Effect test

Embodiment 1

[0025] 1) The daptomycin fermentation broth was centrifuged at 4800rpm for 15min to remove the mycelia to obtain the fermentation supernatant (see figure 1 ). Adjust the pH of the supernatant to 5.0, add 1 / 4 volume of n-butanol, extract 3 times, 95% of the daptomycin in the supernatant enters the organic phase, 3 volumes of pH 7.0 sodium acetate-acetate buffer Daptomycin was back extracted into the aqueous phase.

[0026] 2) The pH of the stripping solution was adjusted to 5.0, and the sample was loaded on a chromatographic column filled with HP20 macroporous adsorption resin. The flow rate was 1.0mL / min. -Sodium acetate-acetate buffer gradient elution, collect macroporous adsorption eluate.

[0027] 3) The macroporous adsorption eluent was loaded onto a DEAE Sepharose FF ion-exchange chromatography column that was pre-balanced with sodium acetate-acetic acid buffer at a flow rate of 2mL / min. The buffer was fully washed, and 50mM pH 7.0 sodium acetate-acetic acid buffer was...

Embodiment 2

[0029] 1) The daptomycin fermentation broth was centrifuged at 4800 rpm for 15 minutes to remove the mycelia to obtain the fermentation supernatant. Adjust the pH of the supernatant to 4.5, add 1 / 4 volume of n-pentanol, extract 4 times, and back-extract the daptomycin into the water phase with 3 volumes of pH 7.5 sodium acetate-acetate buffer.

[0030] 2) The pH of the stripping solution was adjusted to 4.5, and the sample was loaded on a chromatographic column filled with HZ816 macroporous adsorption resin, the aspect ratio was 20, and the flow rate was 1.0mL / min. %, 40% isopropanol-sodium acetate-acetate buffer gradient elution, and the macroporous adsorption eluate was collected.

[0031] 3) The macroporous adsorption eluent is loaded on the DEAE52 ion exchange chromatography column which has been pre-balanced with sodium acetate-acetate buffer at a flow rate of 2mL / min. Wash thoroughly, use 50mM pH7.0 sodium acetate-acetic acid buffer as the base solution, add NaCl for li...

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Abstract

A method for extracting daptomycin from fermentation broth relates to fermentation broth. Supernatant liquor is obtained after the fermentation broth is centrifuged, the daptomycin enters an organic phase after the supernatant liquor is extracted by an organic solvent, sodium acetate-acetic acid buffer solution is used to perform re-extraction to reach a water phase so as to obtain strip-extraction liquid, the strip-extraction liquid is purified through hydrophobic interaction chromatography, isopropanol-sodium acetate-acetic acid buffer solution with constant intensity is used for elution to obtain hydrophobic interaction chromatography eluent, the eluent is separated and purified through ion-exchange column chromatography to obtain ion-exchange eluent, the sodium acetate-acetic acid buffer solution is used as basic liquid, and neutral salt is added for elution so as to complete extraction of the daptomycin from the fermentation broth. Organic solvent extraction and resin chromatography technology is applied comprehensively, and the daptomycin with purity more than 85% can be separated and purified from the fermentation broth by using the method. Compared with other separating and purifying technology, the method is moderate in operating condition, low in cost and capable of effectively reducing acidolysis of the daptomycin. The daptomycin serving as novel lipopeptide antibiotics has good medical market prospect.

Description

technical field [0001] The invention relates to a fermented liquid, in particular to a method for extracting daptomycin from the fermented liquid. Background technique [0002] Antimicrobial resistance is one of the serious challenges facing the world today. The way to overcome bacterial resistance is to prevent the rapid increase and flood of drug-resistant bacteria caused by the abuse of antibiotics; the second is to continuously research and develop new antibacterial drugs that can inhibit bacteria according to the mechanism of action of bacterial drug resistance; It is the research and development of new drugs that will not make bacteria resistant. Only in this way can we effectively control the increasingly serious problem of drug-resistant bacterial infections. [0003] Daptomycin is a new type of lipopeptide antibiotic obtained by fermentation and extraction of Streptomyces roseospora. Its chemical structure and mechanism of action are different from all existing an...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/20C07K1/18C07K1/14
Inventor 敬科举张萍卢英华吴意珣凌雪萍
Owner XIAMEN UNIV
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