Purification method of pyrroloquinoline quinone

Abstract

The invention provides a purification method of pyrroloquinoline quinone. The method comprises the following steps: pretreating a fermentation liquid containing pyrroloquinoline quinone to obtain a filtrate; passing the obtained filtrate through a polyamide resin, then performing rinsing with an acidic aqueous solution, and discarding a leakage liquid; performing desorption with a mixed aqueous solution of an acid and a salt, and collecting a desorption liquid with a chromatographic purity of 90% or more; adjusting pH of the desorption liquid, first performing ultrafiltration, and then performing nanofiltration and concentration; and finally performing crystallization and drying. The method uses the polyamide resin to separate the pyrroloquinoline quinone fermentation liquid, and then thepyrroloquinoline quinone crystal with HPLC purity >= 98% and content >= 96% can be obtained by combination with crystallization. The used polyamide resin is cheap and easy to obtain, the purificationprocess is simple, and the cost is effectively reduced. In addition, resin regeneration of the polyamide resin used in the invention can be completed with lye, and an organic solvent used in the entire process is greatly reduced, so that the wastewater COD is greatly reduced, and the explosion-proof requirement of plant facilities is also reduced.

Description

technical field [0001] The invention relates to the field of microbial fermentation pharmaceuticals, more specifically, to a method for purifying pyrroloquinoline quinone. Background technique [0002] Pyrroloquinoline quinone (Pyrroloquinoline quinone, PQQ), chemical name is 4,5-dihydro-4,5-dioxide-1-hydropyrrole (2,3f) quinone-2,7,9-tricarboxylic acid, alias Methaxatin, is a new coenzyme. In 1979, Salisbury et al. first isolated it from methylotrophic bacteria and identified it as a redox cofactor of membrane-bound dehydrogenase in bacterial cells. It transports electrons with the respiratory chain and is also a new B vitamin. . Some recent studies have found that PQQ has an abnormally high redox cycle ability, and has great application potential in anti-aging of nerve cells and anti-cancer. Meanwhile, in 2008, the U.S. Food and Drug Administration (FDA) approved cognitive function food with PQQ as the main ingredient. Therefore, PQQ has a good application prospect as ...

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Problems solved by technology

[0005] The focus at this stage is to purify PQQ from the fermentation broth. It has been reported that Zhao Yongfang from Wuhan University and others used DEAE-glucan gel for adsorption and desorption, and the obtained eluate was concentrated to obtain the crude product. Adsorption and desorption are carried out on the column, and the purified product is obtained by concentrating the desorption solution, but the DEAE-glucan gel and Seppak C18 used in this process are relatively expensive, and the cost of large-scale production is very high

There is a U.S. patent using Mitsubishi’s HP20 resin instead of the more expensive DEAE-glucan gel to purify the PQQ fermentation filtrate, but it only compares the recovery of PQQ between the two fillers, and does not give a separation After the purity of PQQ, there is no mention of the subsequent purification

There are also complex extraction separation and purification methods, ion-pair two-phase extraction separation combined with anion exchange resin adsorption purification methods, etc., but the above methods need to use a large amount of organic solvents such as n-hexane, methanol or ethanol, and the industrial application costs are high and pollute the environment.

In the Chinese patent CN107056782A, non-polar macroporous resin is used for adsorption, and then hydrophilic silica gel-based C18 is used to prepare packing for separation. Although only buffer and water are used in the separation process, there will be obvious stains on the resin after use. Dead adsorption, each use must be regenerated with high-concentration organic solvents (such as acetone, ethanol with a concentration above 95%) to ensure the next separation performance

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