Method for efficiently preparing pneumocandin B0

A concentrated solution, separation and acquisition technology, applied in the direction of peptides, etc., can solve the problems of low separation process efficiency, difficult environmental control, low process efficiency, etc., and achieve the effect of improving purity and yield, high recovery rate, and high separation efficiency.

Inactive Publication Date: 2015-12-02
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method uses a dynamic axial compression column to replace the traditional silica gel column, the solid-liquid separation in the process still uses traditional centrifugal and rotary evaporation. The separation process has low efficiency, high energy consumption, and large losses.
[0006] In the prior art, centrifugation or evaporation is most

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1

[0034] Take 50L of the fermentation broth of Numocantine B0, and filter the fermentation broth with a tubular membrane with a membrane pore size of 0.2 μm under an operating pressure of 0.5 MPa. 21kg of bacteria and 23L of filtrate were obtained.

[0035] Add 25L of ethanol to the mycelium for extraction, stir well for 30 minutes and filter to obtain 26L of ethanol extract. The bacteria were leached for the second time with 25L of ethanol, and a total of 52L of extract was obtained after filtration.

[0036] Under the operating pressure of 2.5 MPa, the leaching solution is concentrated with a tubular membrane with a molecular weight cut-off of 600 to obtain a concentrated solution a2L. At an operating pressure of 2.0 MPa, the filtrate was concentrated with a tubular membrane with a molecular weight cut off of 2000 to obtain a concentrated solution b8L.

[0037] Mix concentrated liquid a and concentrated liquid b, adjust the pH to 4.0, and stir for a few minutes, ...

Example Embodiment

[0050] Example 2:

[0051] Take 50L of the fermentation broth of Numocantine B0, and filter the fermentation broth with a tubular membrane with a membrane pore size of 0.3um under the operating pressure of 0.5MPa. Obtain 20kg of bacteria and 24L of filtrate.

[0052] Add 30L of ethanol to the mycelium for extraction, fully stir for 30 minutes, and filter to obtain 32L of ethanol extract. The bacteria were leached for the second time with 30L of ethanol, and a total of 64L of extract was obtained after filtration.

[0053] At an operating pressure of 2.5 MPa, the filtrate is concentrated with a tubular membrane with a molecular weight cut-off of 600 to obtain a concentrated solution a2L. At an operating pressure of 2.0 MPa, the filtrate is concentrated with a tubular membrane with a molecular weight cutoff of 3000 to obtain a concentrated solution b10L.

[0054] Mix concentrated liquid a and concentrated liquid b, adjust the pH to 3.5, and stir for a few minutes, and there will be a...

Example Embodiment

[0060] Example 3:

[0061] Take 50L of the fermentation broth of Numocantine B0, and filter the fermentation broth with a tubular membrane with a membrane pore size of 0.2 μm under an operating pressure of 0.5 MPa. 21kg of bacteria and 23L of filtrate were obtained.

[0062] Add 50L of ethanol to the mycelium for extraction, stir well for 30 minutes and filter to obtain 53L of ethanol extract.

[0063] At an operating pressure of 3.0 MPa, the filtrate was concentrated with a tubular membrane with a molecular weight cut off of 300 to obtain a concentrated solution a2L. Under the operating pressure of 2.5MPa, the filtrate was concentrated with a tubular membrane with a molecular weight cutoff of 3000 to obtain a concentrated solution b10L.

[0064] Mix concentrated liquid a and concentrated liquid b, adjust the pH to 4.5, and stir for a few minutes, and there will be a large amount of precipitates to obtain a suspension. After a few minutes of free sedimentation, after testing, it wa...

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PUM

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Abstract

The invention discloses a method for efficiently preparing pneumocandin B0 by separating pneumocandin B0 from fermentation broth. As a membrane separation technique is adopted to prepare pneumocandin B0, the method is relatively applicable to industrial continuous operation. Due to the adoption of an acid precipitation method, a great deal of pigment can be reduced, and B0 loss can be reduced; through silicagel column purification, an isomeride C0 can be effectively removed, and the extraction purity can be up to 99%. The whole process is simple, convenient and feasible, low in power consumption, small in reagent toxicity, and beneficial for industrial application.

Description

technical field [0001] The invention relates to the field of medicinal chemistry, in particular to a method for efficiently preparing pneumocidine B0. Background technique [0002] Neomercontin B 0 (pneumocandinB 0 )By Glarealozoyensis produced secondary metabolites, Glarealozoyensis In addition to producing pneumocantine B during fermentation 0 In addition to the structural analogue A 0 and isomer C 0 . As the precursor compound of the antifungal drug caspofungin, the preparation of high-purity pneumocontin B 0 It is essential for the refined purification of caspofungin. [0003] Chinese invention patent 200910133118.0 discloses the preparation of pneumocandine B 0 The method, the main steps are: a) centrifugal nemocontin B 0 Fermentation broth, take the mycelia, and extract neomocontin B with methanol 0 ; b) Evaporate the methanol extract to dryness, and then use n-butanol to extract Neomocontin B 0 ; c) the n-butanol extract is evaporated to dryness, and then ...

Claims

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Application Information

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IPC IPC(8): C07K7/56C07K1/36C07K1/34C07K1/16
Inventor 黄和冯昆达宋萍秦婷婷纪晓俊沈文和
Owner NANJING UNIV OF TECH
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