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Method for separating and purifying pyrroloquinoline quinone in fermentation liquid by molecularly imprinted solid-phase extraction method

A pyrroloquinoline quinone, solid-phase extraction technology, applied in chemical instruments and methods, organic chemistry, other chemical processes and other directions, to achieve the effect of promoting industrialization development, simple process operation, and convenient large-scale industrial production

Active Publication Date: 2016-12-07
郑州尼采生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there have been related studies on the production of PQQ by microbial fermentation. The original strain of PQQ used is 0.07-7mg / L, and the fermentation time is about 2-5 days. However, these studies have not provided simple and feasible methods. Separating and purifying PQQ from microbial fermentation broth, making the preparation of PQQ realize the method of industrial scale production

Method used

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  • Method for separating and purifying pyrroloquinoline quinone in fermentation liquid by molecularly imprinted solid-phase extraction method
  • Method for separating and purifying pyrroloquinoline quinone in fermentation liquid by molecularly imprinted solid-phase extraction method
  • Method for separating and purifying pyrroloquinoline quinone in fermentation liquid by molecularly imprinted solid-phase extraction method

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Effect test

Embodiment 1

[0040] A method for separating and purifying pyrroloquinoline quinone in fermentation broth by molecular imprinted solid phase extraction, comprising the following steps:

[0041](1) Preparation: Utilize pyrroloquinoline quinone producing bacteria to prepare a fermentation broth containing pyrroloquinoline quinone. The specific method is: inoculate Gluconobacter oxydans in the fermentation medium according to the inoculation amount of 5%, and inoculate the fermentation medium at 28° C. Shake culture for 3 days to obtain seed liquid; then inoculate the seed liquid into enriched medium according to the inoculum amount of 10%, shake culture at 28°C for 5 days, centrifuge the culture liquid at 5°C and 9000r / min for 15min, and collect the supernatant to obtain a fermented liquid rich in pyrroloquinoline quinone whose pyrroloquinoline quinone concentration is 100-200mg / L;

[0042] (2) Extraction and separation: the fermentation broth containing pyrroloquinoline quinone is extracted ...

Embodiment 2

[0048] A method for separating and purifying pyrroloquinoline quinone in fermentation broth by molecular imprinted solid phase extraction, comprising the following steps:

[0049] (1) Preparation: Utilize pyrroloquinoline quinone producing bacteria to prepare a fermentation broth containing pyrroloquinoline quinone. The specific method is: inoculate Gluconobacter oxydans in the fermentation medium according to the inoculation amount of 5%, and inoculate the fermentation medium at 28° C. Shake culture for 3 days to obtain seed liquid; then inoculate the seed liquid into enriched medium according to the inoculum amount of 10%, shake culture at 28°C for 5 days, centrifuge the culture liquid at 5°C and 9000r / min for 15min, and collect the supernatant to obtain a fermented liquid rich in pyrroloquinoline quinone whose pyrroloquinoline quinone concentration is 100-200mg / L;

[0050] (2) Extraction and separation: the fermentation broth containing pyrroloquinoline quinone is extracted...

Embodiment 3

[0056] A method for separating and purifying pyrroloquinoline quinone in fermentation broth by molecular imprinted solid phase extraction, comprising the following steps:

[0057] (1) Preparation: Utilize pyrroloquinoline quinone producing bacteria to prepare a fermentation broth containing pyrroloquinoline quinone. The specific method is: inoculate Gluconobacter oxydans in the fermentation medium according to the inoculation amount of 5%, and inoculate the fermentation medium at 28° C. Shake culture for 3 days to obtain seed liquid; then inoculate the seed liquid into enriched medium according to the inoculum amount of 10%, shake culture at 28°C for 5 days, centrifuge the culture liquid at 5°C and 9000r / min for 15min, and collect the supernatant to obtain a fermented liquid rich in pyrroloquinoline quinone whose pyrroloquinoline quinone concentration is 100-200mg / L;

[0058] (2) Extraction and separation: the fermentation broth containing pyrroloquinoline quinone is extracted...

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Abstract

The invention discloses a method for separating and purifying pyrroloquinoline quinone in fermentation liquid by a molecularly imprinted solid-phase extraction method. According to the method, the pyrroloquinoline quinone is obtained by performing separation and purification on the fermentation liquid rich in the pyrroloquinoline quinone through a pyrroloquinoline quinone molecularly imprinted polymer. According to the method, silicon dioxide is used as a base material, the pyrroloquinoline quinone is used as a template molecule, methacrylic acid is used as a functional monomer, ethylene glycol dimethacrylate is used as a cross-linking agent, and the pyrroloquinoline quinone molecularly imprinted polymer is obtained through a thermal polymerization technique. Adsorptivity experiments prove that the pyrroloquinoline quinone molecularly imprinted polymer prepared by the method disclosed by the invention has good specific selective properties of adsorbing target objects namely the pyrroloquinoline quinone, and the largest adsorption volume can reach 10mg / mg (molecularly imprinted polymers). Solid-phase extraction is performed on the target object namely the pyrroloquinoline quinone by using the polymer as a solid-phase extraction filling, so that the purpose of efficiently separating and purifying the pyrroloquinoline quinone from the fermentation liquid is realized.

Description

technical field [0001] The invention relates to a method for separating and purifying pyrroloquinoline quinone in a fermentation broth by using a molecular imprinted solid-phase extraction method, belonging to the technical field of biochemical industry. 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. Since PQQ has very important physiological functions, PQQ is also considered as a new B vitamin. Studies have shown that PQQ can prevent the formation of amyloid and inhibit the cytotoxicity of the mutant synuclein with shortened C-terminus, so PQQ can be used for the treatment of neurological and psychiatric disorders. In addition, PQQ can also effectively prevent neurodegeneration caused by oxidative stress. In simple terms, PQQ has the following four functions related to the nervous system: (1) anti-oxidation, scavenging...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D471/04B01J20/26B01J20/30
CPCB01J20/268C07D471/04
Inventor 马科杨雪鹏钟桂芳叶建斌崔君竹闫记
Owner 郑州尼采生物科技有限公司
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