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Method for separating and purifying pyrroloquinoline quinone by using supramolecular solvent extraction method

A technology for separation and purification of pyrroloquinoline quinone, which is applied in the field of biochemical industry, can solve the problems of complex components of fermentation broth and low concentration of PQQ fermentation products, and achieve the effects of convenient rapid continuous operation, low cost and high separation efficiency

Active Publication Date: 2021-02-12
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, pyrroloquinoline quinone is a product of polypeptide modification after gene transcription and translation. Due to factors such as gene transcription control and modification reaction speed limit, compared with other fermented fine chemical products, the concentration of PQQ fermentation products is still low.
In addition, due to the characteristics of PQQ’s strong polarity, water solubility, and complex fermentation broth components, conventional natural product separation and purification methods are only suitable for the separation of medium-polarity fat-soluble compounds.

Method used

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  • Method for separating and purifying pyrroloquinoline quinone by using supramolecular solvent extraction method
  • Method for separating and purifying pyrroloquinoline quinone by using supramolecular solvent extraction method
  • Method for separating and purifying pyrroloquinoline quinone by using supramolecular solvent extraction method

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Experimental program
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Effect test

Embodiment 1

[0034] Utilize the method for separating and purifying pyrroloquinoline quinone by supramolecular solvent extraction, comprising the steps:

[0035] (1) Preparation of fermented liquid: Utilize Gluconobacter oxidans to prepare fermented liquid containing pyrroloquinoline quinone, the specific method is:

[0036] Inoculate the Gluconobacter oxydans in the culture medium according to the inoculum amount of 5%, and culture it with shaking at 28°C for 3 days to obtain the seed liquid; Shaking culture under the conditions for 5 days, the culture solution was centrifuged at 5°C and 9000r / min for 15 minutes, and the supernatant was collected to obtain a pyrroloquinoline quinone-rich fermentation broth with a pyrroloquinoline quinone concentration of 100-200 mg / L.

[0037] 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.

[0038] (2) Preparation of supramolecular solvent: Weigh 1g of dimethyl benzyl dodecyl am...

Embodiment 2

[0043] Utilize the method for separating and purifying pyrroloquinoline quinone by supramolecular solvent extraction, 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] Inoculate the Gluconobacter oxydans in the culture medium according to the inoculum amount of 5%, and culture it with shaking at 28°C for 3 days to obtain the seed liquid; Under the condition of shaking culture for 5 days, the culture solution was centrifuged at 5°C and 9000r / min for 15 minutes, and the supernatant was collected to obtain a pyrroloquinoline quinone-rich fermentation broth with a pyrroloquinoline quinone concentration of 100-200 mg / L.

[0046] 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.

[0047] (2) Preparation of supramolecular solvent: Weigh 1g of dimethyl benzyl ...

Embodiment 3

[0052] Utilize the method for separating and purifying pyrroloquinoline quinone by supramolecular solvent extraction, comprising the steps:

[0053] (1) Preparation of fermented liquid: Utilize Methylobacter extorquens to prepare fermented liquid containing pyrroloquinoline quinone, the specific method is:

[0054] Methylobacterium extorquens was inoculated in the culture medium according to the inoculum amount of 5%, and shake cultured at 30°C for 4 days to obtain the seed liquid; then the seed liquid was inoculated into the medium according to the inoculum amount of 10%, and the Shake culture at ℃ for 4 days, centrifuge the culture solution at 5℃ and 9000r / min for 15min, collect the supernatant, and obtain a pyrroloquinoline quinone-rich fermentation broth with a pyrroloquinoline quinone concentration of 100-200mg / L .

[0055] Each liter of culture medium contains 10mL methanol, 3g (NH 4 ) 2 SO 4 , 1.5g KH 2 PO 4 , 0.2g MgSO 4 ·7H 2 O, 0.03gCaCl 2 ·7H 2 O, 0.005gZn...

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Abstract

The invention relates to a method for separating and purifying pyrroloquinoline quinone by utilizing a supramolecular solvent extraction method. The method comprises the following steps: (1) preparinga fermentation liquor containing pyrroloquinoline quinone by utilizing pyrroloquinoline quinone producing bacteria; (2) dissolving a quaternary ammonium salt surfactant into alkane, slowly adding alkanol into the solution while stirring, and stirring until the solution is clear and transparent to obtain a supramolecular solvent; (3) extracting the fermentation liquor containing pyrroloquinoline quinone by using the supramolecular solvent to obtain a supramolecular solvent phase containing pyrroloquinoline quinone; (4) performing reverse extraction on the supermolecular solvent phase containing pyrroloquinoline quinone by using a sodium chloride solution to obtain a water phase containing pyrroloquinoline quinone; and (5) standing the aqueous phase containing pyrroloquinoline quinone at 4DEG C for 24 hours, and filtering when red crystals appear to obtain pyrroloquinoline quinone. According to the invention, the method is simple to operate, low in cost, less in pollution, high in separation efficiency and convenient for rapid and continuous operation, and can be applied to industrial production of PQQ.

Description

technical field [0001] The invention relates to a method for separating and purifying pyrroloquinoline quinone by a supramolecular solvent extraction method, belonging to the technical field of biochemical industry. Background technique [0002] Pyrroloquinoline quinone (PQQ) is a water-soluble quinone compound that can reversibly transfer electrons. It acts as an oxidoreductase cofactor in living systems and is the third coenzyme discovered after riboflavin and nicotinamide. As a prosthetic group of various enzymes, it has important physiological activities for living organisms. Studies have found that mice lacking PQQ grow slowly, have fragile skin, osteoporosis, abnormal bones, low immune response, poor reproductive ability, and are prone to joint inflammation. It is worth noting that PQQ cannot be synthesized in mammals and must be ingested from the outside to meet the needs of life activities. Therefore, PQQ is considered as an essential vitamin and nutrient for the h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D471/04
CPCC07D471/04
Inventor 马科杨雪鹏史立军秦天苍钟桂芳徐琳苏筱渲王光路代军帅周颖吴贞贞蒋登科程源航
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY