Method of adopting semi-continuous fermentation technology to produce pyrroloquinoline quinone

A technology of pyrroloquinoline quinone and fermentation technology, which is applied in the field of fermentation engineering, can solve the problems of short maintenance time, limit the fermentation output of PQQ, and spend a lot of time, and achieve the effect of maintaining volume

Active Publication Date: 2019-04-16
NEW FOUNDER HLDG DEV LLC +2
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

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 of adopting semi-continuous fermentation technology to produce pyrroloquinoline quinone
  • Method of adopting semi-continuous fermentation technology to produce pyrroloquinoline quinone
  • Method of adopting semi-continuous fermentation technology to produce pyrroloquinoline quinone

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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 solution OD 600 The value is 0.95.

[0045] Fermentation tank fed-batch...

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Abstract

The invention provides a method of adopting a semi-continuous fermentation technology to produce pyrroloquinoline quinone. The method comprises the steps that on the basis of a conventional feeding fermentation technology, steps are conducted for multiple circulations that discharging is conducted partly and fresh fermentation mediums are replenished, and then fermentation continues; the OD600 value of fermentation liquid is controlled within a certain range, so that during a semi-continuous fermentation circulation period, the fermentation liquid is always in a high-speed synthesis stage of the pyrroloquinoline quinone, and thus the production efficiency is improved. Compared with the conventional feeding batch fermentation technology, by the adoption of the method, the production efficiency is improved by 25-50%, the unit consumption of main fermentation raw materials is lowered by 12-20%, the production continuity is improved, and the method 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 Applications(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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