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Method for purification of ascomycin from streptomyces fermentation liquid

A technology of ascomycin and streptomyces, applied in the field of bioengineering, can solve the problems of market occupation, affecting product purity and product form, and high separation cost

Active Publication Date: 2013-12-25
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problems in the separation and extraction of ascomycin at this stage are the low yield of ascomycin and the high cost of separation.
In addition, due to the isomerism of ascomycin in the water phase, the current extraction and separation methods mostly contain water phase, and the product contains ascomycin structural analogues, which affect the purity and shape of the product
However, there is no report on the industrial production of ascomycin in my country, and its market is basically occupied by foreign companies such as Japan and the United States.

Method used

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  • Method for purification of ascomycin from streptomyces fermentation liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1) Filtrate the Streptomyces CGMCC No.2519 fermentation broth, and collect the filtrate and mycelia respectively. The mycelium was extracted 3 times by ultrasound, the organic solvent used in the ultrasound-assisted extraction was methanol, the mass volume ratio (g / ml) of mycelia to methanol was 1:15, the ultrasonic frequency was 45KHz, the power was 120W, the For 30 minutes, the pH of the system was 6.5, the extract was collected and combined with the filtrate to obtain the combined solution;

[0044] 2) After the combined liquid obtained in step 1) is filtered through the microfiltration membrane, the permeated liquid of the microfiltration enters the ultrafiltration membrane for filtration to obtain the filtrate, which is transferred to the bed layer equipped with SP207 macroporous resin for adsorption; Wash with 60% ethanol of the column bed volume of the macroporous resin, desorb with ethyl acetate of 1 times the column bed volume of the macroporous resin, and coll...

Embodiment 2

[0050] 1) Filtrate the Streptomyces CGMCC No.2519 fermentation broth, and collect the filtrate and mycelia respectively. The mycelium was extracted 4 times with ultrasonic assistance, the organic solvent used in the ultrasonic-assisted extraction was ethyl acetate, the mass-volume ratio (g / ml) of mycelium to ethyl acetate was 1:10, the ultrasonic frequency was 35KHz, and the power was 180W , the ultrasonic extraction time is 15 minutes, the pH of the system is 7, the extract is collected, combined with the filtrate to obtain the combined solution;

[0051] 2) After the combined solution obtained in step 1) is filtered through the microfiltration membrane, the microfiltration permeate enters the ultrafiltration membrane for filtration to obtain the filtrate, which is transferred to the bed layer equipped with SP825 macroporous resin for adsorption; Wash with 55% methanol of the column bed volume of the macroporous resin, desorb with acetone twice the volume of the column bed of...

Embodiment 3

[0057] 1) Filtrate the Streptomyces CGMCC No.2519 fermentation broth, and collect the filtrate and mycelia respectively. The mycelium is extracted 5 times by ultrasonic wave. The organic solvent used in the ultrasonic wave-assisted extraction is ethanol. For 45 minutes, the pH of the system was 7.5, the extract was collected and combined with the filtrate to obtain the combined solution;

[0058] 2) The combined solution obtained in step 1) is filtered through the microfiltration membrane, and the permeate of the microfiltration enters the ultrafiltration membrane for filtration to obtain the filtrate, which is transferred to the bed layer equipped with D101 macroporous resin for adsorption; Wash with 50% ethanol of the column bed volume of the porous resin, desorb with methanol of 3 times the volume of the column bed of the macroporous resin, and collect the desorption liquid at the temperature of the column bed at 30°C;

[0059] 3) The desorption solution collected in step ...

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Abstract

Belonging to the field of bioengineering, the invention discloses a method for purification of ascomycin from a streptomyces fermentation liquid. The method adopts the techniques of fermentation liquid filtration, mycelium ultrasonication, macroporous resin adsorption, silicagel column chromatography and crystallization so as to obtain pure ascomycin. By employing ultrasonication, the method of the invention has advantages of substantially improved extraction efficiency, convenient operation, simple process, low cost, and high efficiency, thus being suitable for industrial production.

Description

technical field [0001] The invention relates to a process for purifying ascomycin from fermented liquid, belonging to the field of bioengineering, in particular to biopharmaceuticals, methods for extracting, separating and purifying fermentation products, and in particular to a method for purifying ascomycin from streptomyces fermented liquid Methods. Background technique [0002] Ascomycin (Ascomycin, Immunomycin, FK-520) is a macrocyclic lactone compound with three carbon atoms, and it is an ethyl analog of the immunosuppressant FK-506 (tacrolimus, tacrolimus). Ze Pharmaceutical Company isolated from soil containing Streptomyces hygroscopicus. Since the discovery of the immunosuppressive activity of its structural analog tacrolimus, the research on the immunosuppressive activity of ascomycin and its derivatives has been rapidly promoted. Studies have found that ascomycin has a strong immunosuppressant effect, and currently ascomycin has been used as an intermediate in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D498/18
Inventor 杨文革胡永红王瑶函
Owner NANJING TECH UNIV
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