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Method for producing pyrroloquinoline quinone by regulation and control of methylotrophic bacteria through redox potential

A technology of methylotrophic bacteria and pyrroloquinoline quinone, which is applied in the field of fermentation engineering, can solve the problems of low unit cell yield, low carbon source conversion rate, influence on bacterial growth, etc., and achieves shortening fermentation time, efficient accumulation, and improved Yield and yield effects

Pending Publication Date: 2021-11-26
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are three outstanding problems in the above-mentioned fermentation process: 1. Low dissolved oxygen in the early stage affects the growth of bacteria, resulting in a long fermentation cycle; Autolysis and chelation of PQQ, resulting in low yield per unit cell

Method used

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  • Method for producing pyrroloquinoline quinone by regulation and control of methylotrophic bacteria through redox potential
  • Method for producing pyrroloquinoline quinone by regulation and control of methylotrophic bacteria through redox potential

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Embodiment 1

[0025] A method for regulating the production of pyrroloquinoline quinone by methylotrophic bacteria using oxidation-reduction potential, introducing an oxidation-reduction potential electrode into the system for producing pyrroloquinoline quinone by fermentation of methylotrophic bacteria, and adjusting the aeration ratio and stirring speed in the system Oxidation-reduction potential, realize pyrroloquinoline quinone three-stage control fermentation in fermentation production, and then guarantee the accumulation of thalline growth and pyrroloquinoline quinone; In this embodiment, methylotrophic bacteria adopt denitrification hyphomicrobium denitrificans DSM 1869, specifically including the following steps:

[0026] S1. Strain activation: use a solid medium with a pH of 6.5. During the activation culture, take 1 ring of denitrifying Phytophagic microbe DSM1869, streak it on a plate in the solid medium, and culture it in a constant temperature incubator at 30°C for 4 days; Wher...

Embodiment 2

[0031] A method utilizing oxidation-reduction potential to regulate and control methylotrophic bacteria to produce pyrroloquinoline quinone, in the present embodiment, methylotrophic bacteria adopt denitrifying hyphophilic bacteria DSM 1869, specifically comprising the following steps:

[0032] S1. Strain activation: use a solid medium with a pH of 7.5. During the activation culture, take 1 ring of denitrifying Rhizoctonia DSM1869, streak the plate in the solid medium, and culture it in a constant temperature incubator at 25°C for 4 days;

[0033] S2, seed culture: adopt liquid culture medium, its pH is 7.5, pick 2 loops of activated bacteria after strain activation to inoculate in the Erlenmeyer flask that liquid culture medium is housed during cultivation, culture temperature is 25 ℃, shaker The rotation speed is 200rpm, the culture time is 30-36h, OD 650 1.2~1.5;

[0034] S3, fermentation production: adopt liquid culture medium, inoculate the seed after cultivating in the ...

Embodiment 3

[0040] A method utilizing oxidation-reduction potential regulation and control methylotrophic bacteria to produce pyrroloquinoline quinone, in the present embodiment methylotrophic bacteria adopts Methylobacter contortus ATCC 8457, specifically comprises the following steps:

[0041] S1. Strain activation: use a solid medium with a pH of 7.0. During the activation culture, take 1 ring of Methylobacterium contortus ATCC8457, streak it on a solid medium, and culture it in a constant temperature incubator at 28°C for 3 days;

[0042] S2, seed culture: adopt liquid culture medium, its pH is 7.0, pick 2 loops of activated bacteria after strain activation to inoculate in the Erlenmeyer flask that liquid culture medium is housed during culture, culture temperature is 28 ℃, shaker The rotation speed is 150rpm, the culture time is 24~30h, OD 650 1.6~2.0;

[0043] S3, fermentation production: adopt liquid culture medium, initial pH is 7.5, inoculate the seed after cultivating in the li...

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Abstract

The invention discloses a method for producing pyrroloquinoline quinone by regulation and control of methylotrophic bacteria through redox potential. The method comprises the steps of strain activation, seed culture and fermentation production. A redox potential electrode is introduced into a system for producing the pyrroloquinoline quinine through fermentation of the methylotrophic bacteria, and the change of the redox potential in a high-potential oxidation stage and a low-potential reduction stage in a fermentation production stage is adjusted by controlling a ventilation capacity and a stirring speed in the system, and therefore, growth of bacteria and accumulation of the pyrroloquinoline quinine are ensured. According to the method, fermentation time can be shortened, and meanwhile, the yield and the productive rate of the pyrroloquinoline quinone and the stability of a fermentation system are improved.

Description

technical field [0001] The present invention relates to a kind of method that utilizes oxidation-reduction potential (ORP) to regulate and control methylotrophic bacteria to produce pyrrole quinoline quinone, and belongs to the technical field of fermentation engineering. Background technique [0002] Pyrroloquinoline quinone (PQQ) is the next NAD + / NADP + The coenzyme of the third oxidoreductase found after FMN / FAD, naturally exists in nature, and is the only biological factor found so far that exists in all animal and plant tissues. As a unique physiologically active substance, PQQ has great potential in anti-aging, anti-fatigue, improving immunity, and treatment or adjuvant treatment of diseases such as inflammation, liver disease and osteoporosis, and has good medical and health care development prospects . PQQ cannot be synthesized by mammals themselves, and can only be ingested from food. Its content is relatively high in celery, spinach, green pepper, kiwi fruit a...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12N13/00C12Q3/00C12R1/01
CPCC12P17/182C12N13/00C12Q3/00
Inventor 柯崇榕杨欣伟黄建忠丁灵涛任洋刘孟粟陶勇
Owner FUJIAN NORMAL UNIV
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