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Method and application for producing pyrroloquinoline quinone by using gluconobacter oxydans

A technology for oxidizing glucose and pyrroloquinoline quinone, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of not finding and obtaining PQQ, etc., and achieve the convenience of large-scale industrial production, simple conditions, and fast process Effect

Active Publication Date: 2015-02-04
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

According to literature research, there is no simple and easy way to obtain PQQ from microbial fermentation broth, and realize the industrial scale production of PQQ

Method used

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  • Method and application for producing pyrroloquinoline quinone by using gluconobacter oxydans
  • Method and application for producing pyrroloquinoline quinone by using gluconobacter oxydans
  • Method and application for producing pyrroloquinoline quinone by using gluconobacter oxydans

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Embodiment Construction

[0019] Utilize the method for producing pyrroloquinoline quinone by Gluconobacter oxydans, its steps are as follows:

[0020] (1) Fermentation culture of glucose oxidizing bacteria

[0021] The strains were inserted into the fermentation medium at a ratio of 5% (each liter contained 40 g sorbitol, 20 g yeast extract, 5 g (NH 4 ) 2 SO 4 , 2 g KH 2 PO 4 , 5 g MgSO 4 ·H 2 O), cultured with shaking at 28°C for 3 days. When cultivating in the fermenter, inoculate at 10%, and the concentration of the medium is 2 times of normal. The strains were inoculated into the enrichment medium, cultured with shaking at 28° C. for 5 days, and the culture was centrifuged at 9000 r / min at 5° C. for 15 minutes to obtain a supernatant containing PQQ.

[0022] (2) Enrichment and purification of PQQ

[0023] The supernatant was applied to HP-20 type macroporous resin chromatography column, eluted with methanol-water gradient (0%, 10%, 70%, v / v), 10 column volumes for each gradient elution, a...

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Abstract

The invention discloses a method for producing pyrroloquinoline quinone by using gluconobacter oxydans. The method comprises the following steps: (1) fermenting the gluconobacter oxydans DSM2003 to obtain fermentation liquor; (2) carrying out high-speed centrifugation on the fermentation liquor, taking supernate and enriching by using an HP-20 type macroporous resin chromatographic column to obtain an eluted fraction which is rich in pyrroloquinoline quinone; and (3) concentrating the eluted fraction which is rich in pyrroloquinoline quinone under reduced pressure and purifying by using polyamide column chromatography to obtain a pyrroloquinoline quinone pure product. According to the method, under a common culture medium and common culture conditions, the PQQ (pyrroloquinoline quinone) yield of the gluconobacter oxydans strain can reach 125mg / L. The production and preparation method is simple in condition, fast in process and convenient for industrial production in a large scale and has important meaning for industrialization of PQQ.

Description

technical field [0001] The invention relates to the application of a microbial strain, in particular to a process for producing and preparing pyrroloquinoline quinone by using Gluconobacter oxidans DSM 2003. Background technique [0002] Pyrroloquinoline quinine (PQQ) is a water-soluble quinone compound, a coenzyme of glucose dehydrogenase and alcohol dehydrogenase, and is considered a new B vitamin (Nature 2003; 422:832). The molecular weight of PQQ is 330. It was first discovered in microorganisms, and subsequent studies have shown that it also exists in animals and plants. As the prosthetic group of many important enzymes, PQQ participates in the electron transfer in the respiratory chain and can regulate the level of free radicals in the body. It has important physiological functions. It is known that PQQ has the following four functions related to the nervous system: (1) anti-oxidation and scavenging free radicals; (2) affecting the function of the respiratory chain an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/01
Inventor 杨雪鹏马科胡仙妹钟桂芳叶建斌刘寅马歌丽
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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