Process for producing pyrroloquinoline quinone disodium salt by fermentation method

Abstract

The invention provides a process for producing pyrroloquinoline quinone disodium salt by a fermentation method, and relates to the field of microbial fermentation. The process for producing the pyrroloquinoline quinone disodium salt by the fermentation method comprises the following steps: step 1, proliferation: inoculating an original strain Methylovous sp.MP688 into a bacterial basic culture medium; step 2, culturing: culturing in a bacterial basic culture medium for 4-7 days, adding fermentation liquor, stirring into slurry, putting into a fermentation tank, and adding sufficient fermentation liquor; 3, material exchange: after fermentation for 2-3 days, placing a micro-filtration membrane, with one side of the micro-filtration membrane being original liquid in the fermentation tank and the other side being fresh fermentation liquid; and 4, purification: collecting the fresh fermentation liquid in the fermentation tank, and purifying the pyrroloquinoline quinone disodium salt. Substances are replaced through the micro-filtration membrane, so that acquisition of a target product and nutrition supplement of methylotrophic bacteria are realized, and the fermentation time of the methylotrophic bacteria is prolonged.

Description

technical field

[0001] The invention relates to the technical field of microbial fermentation, in particular to a process for producing pyrroloquinoline quinone disodium salt by fermentation. Background technique

[0002] Pyrroloquinoline quinone (PQQ) is the coenzyme of the third type of oxidoreductase, and is also known as a new vitamin with various physiological functions.

[0003] PQQ is a cofactor involved in redox reactions and acts as a coenzyme for many enzymes, such as dehydrogenases, oxidases, hydratases, and decarboxylases. PQQ contains o-quinone structures that can directly participate in redox reactions, and the above enzyme proteins can catalyze the oxidation reactions of non-phosphorylated substrates, such as alcohols, aldehydes and aldoses.

[0004] It has important medical value in the prevention and treatment of liver injury and liver poisoning, promoting the production of nerve growth factors, regulating the level of free radicals, resisting radiation dam...

Examples

Embodiment 1

[0029] The embodiment of the present invention provides a process for producing pyrroloquinoline quinone disodium salt by fermentation, comprising the following steps:

[0030] Step 1. Proliferation: Inoculate the original bacterial species methylotrophic bacteria Methylovorus sp.MP688 into the bacterial basal medium, add dihydrogen phosphate ion and monohydrogen phosphate ion on the 1b basic machine to adjust the pH of the bacterial basal medium to 6.5 ~7.0, lb medium uses tryptone (Tryptone) 10g / L, yeast extract (Yeast extract) 5g / L, sodium chloride (NaCl) 10g / L, through this medium to expand the population of methylotrophic bacteria MP688 .

[0031] Step 2. Cultivation: After culturing the bacterial basal medium for 4 to 7 days, add the fermentation broth, and then stir it into a slurry to make it easy to mix with the fermentation broth. The methylotroph MP688 is evenly distributed, and then placed in the fermenter 1, 5, and add a sufficient amount of fermentation broth to...

Embodiment 2

[0039] This embodiment is the structure of a fermenting device in Embodiment 1: the fermenter 1 is interspersed with an outer tube 4 , the lower end of the outer tube 4 is fixedly connected with a microfiltration membrane 2 , and an inner tube is arranged inside the outer tube 4 . During the fermentation process, that is, the pyrroloquinoline quinone and metabolites produced in the proliferation and production of methylotrophic bacteria MP688 pass through the microfiltration membrane 2 and enter the interior of the outer tube 4, while the nutrients inside the outer tube 4 enter the fermenter 1 In the fermented stock solution, the inner tube 3 is injected with fresh fermented broth at this time, and material replacement occurs at the microfiltration membrane 2, and the replaced fresh fermented broth is discharged and collected from the outer tube 4 for extracting pyrroloquinoline quinone.

[0040] In addition, there are equipments that can ensure the normal proliferation and gro...

Embodiment 3

[0042]This embodiment is the structure of another fermentation device in Embodiment 1: a liquid storage container 6 is provided inside the fermenter 5, and a microfiltration membrane 7 is provided on the side wall of the liquid storage container 6, and the liquid storage container 6 is equipped with Into fresh fermented liquid, and liquid storage container 6 is placed in fermentor 5, and the pyrroloquinoline quinone and metabolite that produce in the multiplication production of methylotrophic bacteria MP688 pass through microfiltration membrane 7 and enter the inside of outer tube 4, simultaneously Nutrients in the liquid storage container 6 enter the fermentation stock solution in the fermenter 5 . Regularly take out the liquid storage container 6 and replace it with fresh fermented liquid, the original one is used to extract pyrroloquinoline quinone

[0043] In addition, there are equipments that can ensure the normal proliferation and growth of methylotrophic bacteria MP68...

technical field

[0001] The invention relates to the technical field of microbial fermentation, in particular to a process for producing pyrroloquinoline quinone disodium salt by fermentation. Background technique

[0002] Pyrroloquinoline quinone (PQQ) is the coenzyme of the third type of oxidoreductase, and is also known as a new vitamin with various physiological functions.

[0003] PQQ is a cofactor involved in redox reactions and acts as a coenzyme for many enzymes, such as dehydrogenases, oxidases, hydratases, and decarboxylases. PQQ contains o-quinone structures that can directly participate in redox reactions, and the above enzyme proteins can catalyze the oxidation reactions of non-phosphorylated substrates, such as alcohols, aldehydes and aldoses.

[0004] It has important medical value in the prevention and treatment of liver injury and liver poisoning, promoting the production of nerve growth factors, regulating the level of free radicals, resisting radiation dam...

Examples

Embodiment 1

[0029] The embodiment of the present invention provides a process for producing pyrroloquinoline quinone disodium salt by fermentation, comprising the following steps:

[0030] Step 1. Proliferation: Inoculate the original bacterial species methylotrophic bacteria Methylovorus sp.MP688 into the bacterial basal medium, add dihydrogen phosphate ion and monohydrogen phosphate ion on the 1b basic machine to adjust the pH of the bacterial basal medium to 6.5 ~7.0, lb medium uses tryptone (Tryptone) 10g / L, yeast extract (Yeast extract) 5g / L, sodium chloride (NaCl) 10g / L, through this medium to expand the population of methylotrophic bacteria MP688 .

[0031] Step 2. Cultivation: After culturing the bacterial basal medium for 4 to 7 days, add the fermentation broth, and then stir it into a slurry to make it easy to mix with the fermentation broth. The methylotroph MP688 is evenly distributed, and then placed in the fermenter 1, 5, and add a sufficient amount of fermentation broth to...

Embodiment 2

[0039] This embodiment is the structure of a fermenting device in Embodiment 1: the fermenter 1 is interspersed with an outer tube 4 , the lower end of the outer tube 4 is fixedly connected with a microfiltration membrane 2 , and an inner tube is arranged inside the outer tube 4 . During the fermentation process, that is, the pyrroloquinoline quinone and metabolites produced in the proliferation and production of methylotrophic bacteria MP688 pass through the microfiltration membrane 2 and enter the interior of the outer tube 4, while the nutrients inside the outer tube 4 enter the fermenter 1 In the fermented stock solution, the inner tube 3 is injected with fresh fermented broth at this time, and material replacement occurs at the microfiltration membrane 2, and the replaced fresh fermented broth is discharged and collected from the outer tube 4 for extracting pyrroloquinoline quinone.

[0040] In addition, there are equipments that can ensure the normal proliferation and gro...

Embodiment 3

[0042]This embodiment is the structure of another fermentation device in Embodiment 1: a liquid storage container 6 is provided inside the fermenter 5, and a microfiltration membrane 7 is provided on the side wall of the liquid storage container 6, and the liquid storage container 6 is equipped with Into fresh fermented liquid, and liquid storage container 6 is placed in fermentor 5, and the pyrroloquinoline quinone and metabolite that produce in the multiplication production of methylotrophic bacteria MP688 pass through microfiltration membrane 7 and enter the inside of outer tube 4, simultaneously Nutrients in the liquid storage container 6 enter the fermentation stock solution in the fermenter 5 . Regularly take out the liquid storage container 6 and replace it with fresh fermented liquid, the original one is used to extract pyrroloquinoline quinone

[0043] In addition, there are equipments that can ensure the normal proliferation and growth of methylotrophic bacteria MP68...

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