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Complex extraction method for separation and purification of pyrroloquinoline quinine in fermentation broth

A technology for separation and purification of pyrroloquinoline quinone, which is applied in the field of separation and purification of water-soluble vitamins, can solve the problems of not finding and preparing PQQ, etc., and achieve the effect of facilitating large-scale industrial production, fast process and simple conditions

Active Publication Date: 2015-09-09
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to literature research, no simple and easy method for preparing PQQ from microbial fermentation broth has been found at present, and the industrial scale production of PQQ has been realized.

Method used

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  • Complex extraction method for separation and purification of pyrroloquinoline quinine in fermentation broth
  • Complex extraction method for separation and purification of pyrroloquinoline quinine in fermentation broth
  • Complex extraction method for separation and purification of pyrroloquinoline quinine in fermentation broth

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Effect test

Embodiment 1

[0028] The complex extraction method separates and purifies the pyrroloquinoline quinone in the fermented liquid, and its steps are as follows:

[0029] (1) Fermenting Gluconobacter oxydans DSM 2003 to obtain a fermentation broth;

[0030] (2) After the fermentation broth is centrifuged at high speed, the supernatant is subjected to complex extraction in a two-solute extraction system, in which trioctylamine is used as a complexing agent and n-hexane is used as a diluent to obtain a supernatant rich in pyrroloquinoline quinone. The volume ratio of trioctylamine and n-hexane in the dual-solute extraction system is 1:1, the volume ratio of the extraction system to the fermentation broth is 1:1, and the extraction times are 3 times;

[0031] (3) Back-extracting the upper layer rich in pyrroloquinoline quinone with ammonia water, concentrating the ammonia water layer under reduced pressure, and freeze-drying to obtain the crude product of pyrroloquinoline quinone, the concentratio...

Embodiment 2

[0034] The complex extraction method separates and purifies the pyrroloquinoline quinone in the fermented liquid, and its steps are as follows:

[0035] (1) Fermenting Gluconobacter oxydans DSM 2003 to obtain a fermentation broth;

[0036] (2) After the fermentation broth is centrifuged at high speed, the supernatant is subjected to complex extraction in a two-solute extraction system, in which trioctylamine is used as a complexing agent and n-hexane is used as a diluent to obtain a supernatant rich in pyrroloquinoline quinone. The volume ratio of trioctylamine and n-hexane in the dual-solute extraction system is 3:1, the volume ratio of the extraction system to the fermentation broth is 3:1, and the number of extractions is 5 times;

[0037] (3) Back-extracting the upper layer rich in pyrroloquinoline quinone with ammonia water, concentrating the ammonia water layer under reduced pressure, and freeze-drying to obtain the crude product of pyrroloquinoline quinone, the concentr...

Embodiment 3

[0040]The complex extraction method separates and purifies the pyrroloquinoline quinone in the fermented liquid, and its steps are as follows:

[0041] (1) Fermenting Gluconobacter oxydans DSM 2003 to obtain a fermentation broth;

[0042] (2) After the fermentation broth is centrifuged at high speed, the supernatant is subjected to complex extraction in a two-solute extraction system, in which trioctylamine is used as a complexing agent and n-hexane is used as a diluent to obtain a supernatant rich in pyrroloquinoline quinone. The volume ratio of trioctylamine and n-hexane in the dual-solute extraction system is 2:1, the volume ratio of the extraction system to the fermentation broth is 2:1, and the number of extractions is 4 times;

[0043] (3) Back-extracting the upper layer rich in pyrroloquinoline quinone with ammonia water, concentrating the ammonia water layer under reduced pressure, and freeze-drying to obtain the crude product of pyrroloquinoline quinone, the concentra...

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Abstract

The invention discloses a complex extraction method for separation and purification of pyrroloquinoline quinine in fermentation broth. The method comprises the steps of: (1) fermenting bacteria to obtain a fermentation broth; (2) subjecting the fermentation broth to high-speed centrifugation, subjecting a supernatant liquid to complex extraction in a two-solute extraction system using trioctylamine as a complexing agent and n-hexane as a diluent, to obtain an upper liquid rich in pyrroloquinoline quinine; (3) subjecting the upper liquid rich in pyrroloquinoline quinine to back extraction with ammonia water, concentrating the ammonia water layer under reduced pressure and lyophilizing to obtain a pyrroloquinoline quinine crude product; and (4) dissolving the pyrroloquinoline quinine crude product in ultrapure water, adjusting the pH to 3-4 by HCl, adding ethanol, stirring at 20-25 DEG C for 5-6 h, and standing for 12-24 h to obtain pyrroloquinoline quinine. The above production method has simple condition and quick process, is easy for large-scale industrial production, and has important meaning to promote the industrialization of pyrroloquinoline quinine.

Description

technical field [0001] The invention relates to the separation and purification of water-soluble vitamins, in particular to a post-treatment method for producing pyrroloquinoline quinone by fermentation of Gluconobacter oxydans. Background technique [0002] Pyrroloquinoline quinone (PQQ) is a water-soluble quinone compound widely found in food and animals, and is the coenzyme of glucose dehydrogenase and alcohol dehydrogenase. PQQ has been proven to have very important physiological functions and is considered a new B vitamin. Studies have shown that PQQ can be used to treat neurological and psychiatric disorders, because PQQ can prevent the formation of amyloid and inhibit the cytotoxicity of the C-terminal shortened variant synuclein. Moreover, PQQ can effectively prevent neurodegeneration caused by oxidative stress. In short, PQQ has the following four functions related to the nervous system: (1) anti-oxidation and scavenging free radicals; (2) affecting the function o...

Claims

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Application Information

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IPC IPC(8): C07D471/04
CPCC07D471/04
Inventor 杨雪鹏马科胡仙妹叶建斌邵化
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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