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Method for separating and purifying pyrroloquinoline quinone by using reverse micelle surface modified magnetic nano solid-phase extraction material

A solid-phase extraction material and a technology for pyrroloquinoline quinone, which are applied in the field of separation and purification of pyrroloquinoline quinone by magnetic nano solid-phase extraction materials, can solve the problems of low concentration of fermentation products, increased difficulty in separation and purification of PQQ, etc., and achieve mass transfer efficiency. The effect of high, high production efficiency and simple equipment

Inactive Publication Date: 2022-03-08
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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  • Claims
  • Application Information

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Problems solved by technology

However, the yield of PQQ produced by the fermentation method of Mitsubishi Gas Company in Japan is only 10 grams per liter. Compared with other fermented fine chemical products, the concentration of PQQ fermentation products is low
In addition, due to the characteristics of PQQ’s strong polarity, water solubility, and complex components of fermentation broth, conventional natural product separation and purification methods are only suitable for the separation of medium-polarity fat-soluble compounds. Therefore, it is more difficult to separate and purify PQQ from fermentation broth. The development of PQQ must solve the technical problems of separation and purification

Method used

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  • Method for separating and purifying pyrroloquinoline quinone by using reverse micelle surface modified magnetic nano solid-phase extraction material
  • Method for separating and purifying pyrroloquinoline quinone by using reverse micelle surface modified magnetic nano solid-phase extraction material
  • Method for separating and purifying pyrroloquinoline quinone by using reverse micelle surface modified magnetic nano solid-phase extraction material

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Embodiment 1

[0036] The preparation of the magnetic nano solid phase extraction material of embodiment 1 reverse micelle surface modification

[0037] A preparation method of magnetic nano solid phase extraction material, comprising the following steps:

[0038] 1) Preparation of Fe by silicon embedding 3 o 4 @SiO 2 ; put 10g Fe 3 o 4 Dissolve in 800mL ethanol solution (consisting of 200mL deionized water and 600mL absolute ethanol), ultrasonically disperse for 30min; then add 10mL 10% (v / v) ammonia water dropwise while stirring, after stirring for 30min, add normal Tetraethyl silicate (TEOS) 60mL, stirred at room temperature for 24h; after the reaction, washed with ethanol for 3 to 5 times to remove the remaining reactants, and then washed with deionized water for 3 to 5 times, and the sample was placed at 60°C Vacuum dry at high temperature for 12h, set aside.

[0039] 2) Fe 3 o 4 @SiO 2 silanization; Take 5mL n-dodecyltrimethoxysilane or n-octadecyltrimethoxysilane and stir at ...

Embodiment 2

[0042] The separation and purification of embodiment 2 pyrroloquinoline quinone

[0043] A method for separating and purifying pyrroloquinoline quinone using a magnetic nanometer solid-phase extraction material modified on the surface of reverse micelles, comprising the following steps:

[0044] (1) Preparation of fermented liquid: Utilize Gluconobacter oxidans to prepare fermented liquid containing pyrroloquinoline quinone, the specific method is:

[0045] Each liter of medium contains 40g sorbitol, 20g yeast extract, 10g (NH 4 ) 2 SO 4 , 4g KH 2 PO 4 , 10gMgSO 4 ·H 2 O.

[0046] Gluconobacter oxydans was inoculated in the fermentation medium according to the inoculum amount of 5%, and cultured with shaking at 28°C for 3 days to obtain the seed liquid; then the seed liquid was inoculated into the enrichment medium according to the inoculum amount of 10%, Shake culture at 28°C for 5 days, centrifuge the culture solution at 5°C and 9000r / min for 15min, collect the super...

Embodiment 3

[0050] The separation and purification of embodiment 3 pyrroloquinoline quinone

[0051] A method for separating and purifying pyrroloquinoline quinone using a magnetic nanometer solid-phase extraction material modified on the surface of reverse micelles, comprising the following steps:

[0052] (1) Preparation of fermented liquid: Utilize Gluconobacter oxidans to prepare fermented liquid containing pyrroloquinoline quinone, the specific method is:

[0053] Each liter of medium contains 40g sorbitol, 20g yeast extract, 10g (NH 4 ) 2 SO 4 , 4g KH 2 PO 4 , 10gMgSO 4 ·H 2 O.

[0054] Gluconobacter oxydans was inoculated in the fermentation medium according to the inoculum amount of 5%, and cultured with shaking at 28°C for 3 days to obtain the seed liquid; then the seed liquid was inoculated into the enrichment medium according to the inoculum amount of 10%, Shake culture at 28°C for 5 days, centrifuge the culture solution at 5°C and 9000r / min for 15min, collect the super...

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Abstract

The invention provides a method for separating and purifying pyrroloquinoline quinone by using a reverse micelle surface modified magnetic nano solid-phase extraction material. The method comprises the following steps: (1) preparing fermentation liquor; (2) preparation of a magnetic nano solid-phase extraction material: preparing Fe3O4 (at) SiO2 by silicon embedding, silanizing the Fe3O4 (at) SiO2, and carrying out self-assembly modification on the surface of the magnetic nano solid-phase extraction material by using a supramolecular solvent; (3) adsorption; (4) eluting; and (5) crystallizing and purifying. The method provided by the invention effectively overcomes the problem that the traditional organic solvent physical extraction method has low and insufficient extraction rate on the water-soluble component PQQ. According to the method, a high-purity PQQ product is prepared by extracting and enriching PQQ from fermentation liquor by using a reverse micelle surface modified magnetic nano solid-phase extraction material. Compared with liquid-liquid extraction, equipment such as an extraction tower or a centrifugal extractor is not needed for liquid-liquid two-phase separation, and the equipment is simple and convenient to operate.

Description

technical field [0001] The invention relates to a method for separating and purifying pyrroloquinoline quinone using a magnetic nanometer solid-phase extraction material modified on the surface of reverse micelles, belonging to the field of biochemical industry. Background technique [0002] Pyrroloquinoline quinone (PQQ) can reversibly transfer electrons, and is used as an oxidoreductase cofactor in living systems. It is the third type of coenzyme after riboflavin and nicotinamide, and has important physiological activities for living organisms. [0003] At present, the preparation methods of PQQ mainly include microbial fermentation and chemical synthesis. The existing synthesis methods of PQQ include chemical synthesis and microbial fermentation. The chemical total synthesis method needs more than ten steps of reaction to obtain, and the yield is very low; and it needs to use a large amount of toxic reagents and heavy metal catalysts, which is relatively harmful to the e...

Claims

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Application Information

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IPC IPC(8): B01J20/10B01J20/28B01D15/10B01D15/42B01J20/30C12P17/18C12R1/01
CPCB01J20/103B01J20/06B01J20/28009C12P17/182B01D15/424B01D15/10
Inventor 马科杨雪鹏钟桂芳王光路王冰洋胡仙妹程源航苏泽宇
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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