Method for producing pyrroloquinoline quinone by semi-continuous fermentation process

A technology of pyrroloquinoline quinone and fermentation process, which is applied in the field of fermentation engineering, can solve the problems of limiting PQQ fermentation output, maintaining a short time, spending a lot of time, etc., so as to improve equipment utilization and production efficiency, improve production efficiency, and save energy. consumption effect

Active Publication Date: 2022-05-10
NEW FOUNDER HLDG DEV LLC +2
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  • Application Information

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Problems solved by technology

[0004] At present, the traditional batch fermentation production process of PQQ has two outstanding problems. One is that each batch of fermentation takes a lot of time to cultivate and proliferate bacteria. The other is that due to the continuous accumulation of bacteria and metabolites, the maintenance time of PQQ high-speed synthesis Very short, limiting the further improvement of PQQ fermentation yield

Method used

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  • Method for producing pyrroloquinoline quinone by semi-continuous fermentation process
  • Method for producing pyrroloquinoline quinone by semi-continuous fermentation process
  • Method for producing pyrroloquinoline quinone by semi-continuous fermentation process

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Shake flask seed culture of embodiment 1 PQQ

[0036] Prepare the seed bottle medium according to the basic medium formula, the seed bottle volume is 30ml / 250ml, after sterilization, add 0.3ml of methanol to each bottle before inoculation.

[0037] From the inoculation of the slant strain to the seed bottle, the seed bottle cultivation process: cultivation temperature 30°C, shaker speed 250rpm, cultivation time 48h, cultured shake flask seed solution OD 600 The value is 0.92.

Embodiment 2

[0038] Shake flask fermentation of embodiment 2 PQQ

[0039] Prepare fermentation bottle culture medium according to the formula of the basic medium, and the volume of the fermentation bottle is 30ml / 250ml. After sterilization and before inoculation, add 0.6ml of methanol to each bottle.

[0040] The shake flask seed liquid cultivated according to the method in Example 1 was transferred to the fermentation bottle, and the fermentation bottle cultivation process: the transfer amount was 10% v / v, the cultivation temperature was 30° C., the shaking table speed was 250 rpm, and the cultivation time was 5 days. The fermentation units of the three fermentation bottles were 9.2mg / L, 11.7mg / L, and 13.6mg / L, respectively, with an average of 11.5mg / L.

[0041] During the cultivation process of PQQ's production strain, Phytophagy microbes, the pH of the fermentation broth drops rapidly, and gradually deviates from the optimum growth pH 6.90. Because it is inconvenient to supplement ammon...

Embodiment 3

[0042] The fed-batch fermentation experiment of embodiment 3 PQQ

[0043] The fermentation equipment used in this example is a set of dual fermentation small test tank equipment, the volume of the seed tank is 5L, and the volume of the fermenter is 50L.

[0044] Seed tank seed culture: the formula of the seed tank culture medium is the same as that of the basic medium formula, and the seed tank filling capacity is 3L / 5L. After sterilization, add 60ml of methanol before inoculation, and then shake the seeds cultivated according to the method in Example 1 The solution was inoculated into the seed tank according to the inoculation amount of 1.0% (30ml). The seed tank cultivation process: the cultivation temperature was 30°C, the ventilation rate was 0.8vvm, the stirring speed was 100-300rpm, the stirring speed was related to the dissolved oxygen (DO), and the control DO≥30 %, culture time 48h, cultured seed liquid OD 600 The value is 0.95.

[0045] Fermentation tank fed-batch f...

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Abstract

The invention provides a method for producing pyrroloquinoline quinone by a semi-continuous fermentation process. The method is based on a conventional feed-fed fermentation process, and continues to ferment after performing partial discharge and replenishing fresh fermentation medium through multiple cycles. OD of the fermentation broth 600 The value is controlled within a certain range, so that during the cycle of semi-continuous fermentation, the fermentation broth is in the high-speed synthesis period of pyrroloquinoline quinone, thereby improving the production efficiency. Compared with the traditional fed-batch fermentation process, the invention improves the production efficiency by 25-50%, reduces the unit consumption of main fermentation raw materials by 12-20%, improves the production continuity, and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, in particular to a method for producing pyrroloquinoline quinone by a semi-continuous fermentation process. Background technique [0002] Pyrroloquinoline quinine (Pyrroloquinoline quinine, PQQ) is a water-soluble quinone compound, which was first discovered in microorganisms and acts as a redox coenzyme of membrane-bound dehydrogenase in bacterial cells. Subsequent studies have shown that it is also widely present in In animals and plants, it is the coenzyme of glucose dehydrogenase and alcohol dehydrogenase. Japanese scholars believe that PQQ fully meets the characteristics of vitamins (Nature 2003; 422:832), and advocates listing it as the 14th vitamin. Current studies have found that PQQ can regulate the level of free radicals in the body, stimulate the growth of the body, promote the synthesis of nerve growth factors, protect the heart, prevent liver damage, reduce inflammat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/66C12R1/01
CPCC12P7/66
Inventor 赵子刚张葵
Owner NEW FOUNDER HLDG DEV LLC
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