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The method and application of producing pyrroloquinoline quinone by using Gluconobacter oxydans

A technology for oxidizing glucose and pyrroloquinoline quinone is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., which can solve the problem of not finding PQQ and the like, and achieves convenient large-scale industrial production, fast process, and simple conditions. Effect

Active Publication Date: 2019-06-28
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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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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  • The method and application of producing pyrroloquinoline quinone by using Gluconobacter oxydans
  • The method and application of producing pyrroloquinoline quinone by using Gluconobacter oxydans
  • The method and application of 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 bacterial classification is inserted into the fermentation medium according to the ratio of 5% (every liter contains 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 f...

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Abstract

The invention discloses a method for producing pyrroloquinoline quinone by utilizing Gluconobacter oxydans. The steps are as follows: (1) Fermenting Gluconobacter oxydans DSM 2003 to obtain a fermentation liquid; (2) High-speed centrifugation of the fermentation liquid to obtain the supernatant Liquid, enriched by HP‑20 type macroporous resin chromatographic column to obtain the elution fraction rich in pyrroloquinoline quinone; (3) concentrating the elution fraction rich in pyrroloquinoline quinone under reduced pressure, Purified by amide column chromatography to obtain pure pyrroloquinoline quinone. The Gluconobacter oxidans strain provided by the present invention has a PQQ output of 125 mg / L under conventional culture medium and culture conditions, and the above-mentioned production and preparation method has simple conditions, quick process, and is convenient for large-scale industrial production, which is helpful for promoting the industrialization of PQQ have important meaning.

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 Patents(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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