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A kind of method utilizing supramolecular solvent extraction method to separate and purify pyrroloquinoline quinone

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: 2021-07-23
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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  • A kind of method utilizing supramolecular solvent extraction method to separate and purify pyrroloquinoline quinone
  • A kind of method utilizing supramolecular solvent extraction method to separate and purify pyrroloquinoline quinone
  • A kind of method utilizing supramolecular solvent extraction method to separate and purify pyrroloquinoline quinone

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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 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 dodecyl am...

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 a supramolecular solvent extraction method, comprising (1) using a pyrroloquinoline quinone producing bacterium to prepare a fermentation broth containing pyrroloquinoline quinone; (2) surface-activating a quaternary ammonium salt Dissolve the solvent in alkanes, slowly add alkanol to the solution while stirring, and stir until the solution is clear and transparent, which is the supramolecular solvent; (3) extract the fermentation broth containing pyrroloquinoline quinone with the supramolecular solvent to obtain The supramolecular solvent phase containing pyrroloquinoline quinone; (4) back-extracting the supramolecular solvent phase containing pyrroloquinoline quinone with sodium chloride solution to obtain an aqueous phase containing pyrroloquinoline quinone; (5) at 4° C. Under certain conditions, the aqueous phase containing pyrroloquinoline quinone was allowed to stand for 24 hours, and filtered when red crystals appeared to obtain pyrroloquinoline quinone. The invention has the advantages of simple operation, low cost, less pollution, high separation efficiency, convenient rapid and continuous operation, and can be applied to the industrialized 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 Patents(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